PROGRAMME & ABSTRACT BOOK

PLANT BIOLOGY SCANDINAVIA 2015 26TH CONGRESS OF THE SCANDINAVIAN PLANT PHYSIOLOGY SOCIETY 9

WWW.SPPS2015.ORG

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WELCOME TO PLANT BIOLOGY SCANDINAVIA 2015 – the 26th Congress of the SPPS It is a pleasure to welcome you to Stockholm and to PLANT BIOLOGY SCANDINAVIA 2015, the 26th Congress of the Scandinavian Plant Physiology Society (SPPS 2015). The Scandinavian Plant Physiology Society has since 1947 organised congresses in Scandinavia to promote the interaction between plant experimental biologists. For a number of years mainly Scandinavian scientists and a small number of international invited keynote speakers attended these congresses. In more recent years, there is a tendency that SPPS congresses attract scientists from all over the world. This is certainly true for the SPPS 26th Congress, where participants come from all continents and from more than 30 countries. The 26th SPPS Congress aims to create a forum for the exchange of results and discussion of new questions in the broad field of experimental plant biology. Under headings such as: Development, Epigenetics and gene regulations, High throughput biology, Photobiology, Abiotic stress and Biotic interactions, there will be talks from international and Scandinavian keynote speakers and shorter presentations selected from abstract submissions. Talks by SPPS Early Career award winners and Physiologia Plantarum award winner also fit into these topics as do a high number of poster presentations. In addition to scientific news, the 26th SPPS congress aims to be a forum for broader discussions related to plant science: science policy, how to reach out with scientific knowledge to the general public, how to foster an interest in plant science in the young generation and how to transfer new knowledge into practical applications. Presentations regarding such matters will be given by SPPS award winner, SPPS popularization prize winner and invited keynote speakers in the session Education and outreach and the opening session From the lab. to the field. The venue of the congress is the magnificent Aula Magna, located at Stockholm University in the beautiful city of Stockholm. So in addition to the congress, you as a participant get the opportunity to experience the capital of Sweden with its refreshing, unpretentious and dynamic atmosphere. We hope you will enjoy the congress and your stay in Stockholm!

Kind regards On behalf of the Organising Committee Lisbeth Jonsson, Congress Chair

The congress is organised with economic support from the Swedish Research Council Formas (www.formas.se)

Societas Physiologiae Plantarum Scandinavica Scandinavian Plant Physiology Society Scandinavian Plant Physiology Society (SPPS) is a scientific society that promotes all aspects of experimental plant biology from molecular cell biology and biochemistry to ecophysiology. Currently, a significant part of the members come from countries outside Scandinavia and membership is open to everybody interested in plant biology. Activities The main activities of SPPS are SPPS congresses and publishing of the journal Physiologia Plantarum. The Society publishes a Newsletter and may financially support the organisation of other conferences and initiatives in the field of plant science in Scandinavia. SPPS also provides travel grants for junior members to visit other lab’s in Scandinavia, to participate in conferences in Scandinavia which are organised or supported by SPPS and to FESPB Congresses. SPPS awards are given to persons with merits in the field of plant science. The society also promotes teaching of plant biology and has established an Education Committee for that purpose. SPPS is affiliated to the Federation of European Societies for Plant Biology (FESPB) and has signed a memorandum of understanding with the European Plant Science Organization (EPSO). SPPS is a funding member of the Global Plant Council (GPC). Organisation and administration The General assembly of SPPS is held in connection with the SPPS congress. The General Assembly elects a Council and other officials to govern the Society between the assemblies. The location of assemblies alternates between the Nordic countries Denmark, Finland, Norway and Sweden which are all represented in the Council. The office of SPPS is located in the country of the President and other key officials. More information www.spps.fi

SPPS 2015 local organising committee Lisbeth Jonsson, Stockholm University, Stockholm, Sweden (Chair) Lars Hennig, SLU and Linnean Center for Plant Biology, Uppsala, Sweden Anna Ohlsson, KTH – Royal Institute of Technology, Stockholm, Sweden Katharina Pawlowski, Stockholm University, Sweden Jens Sundström, SLU and Linnean Center for Plant Biology, Uppsala, Sweden Session organisers Ove Nilsson, SLU, Umeå, Sweden Claudia Köhler, SLU Uppsala, Sweden Stefan Jansson, Umeå University, Sweden Dario Leister, University of Copenhagen, Denmark Jaakko Kangasjärvi, University of Helsinki, Finland Jens Stougaard, Aarhus University, Denmark

SPPS council (2013-2015) Jaakko Kangasjärvi, President, Finland Tom Hamborg Nielsen, Vice President, Denmark Anna Kärkönen, Secretary General, Finland Kurt Fagerstedt, Treasurer, Finland Lisbeth Jonsson, Journal responsible, Sweden Elina Oksanen, Member, Finland Jorunn Elisabeth Olsen, Member, Norway Deputy members

Cornelia Spetea-Wiklund, Sweden Pai Pedas, Denmark Sissel Torre, Norway Eevi Rintamäki, Finland

Programme for Sunday 9 August and Monday 10 August

Programme for Tuesday 11 August

Sunday 09 August 2015

Tuesday 11 August 2015

09.00 - 17.30

Optional excursions

09.00 - 09.45

SPPS Early Career Awardees

09.00 - 15.00

I. Uppsala, incl. Linnaeus Gardens

09.05 - 09.25

09.30 -17.30

II. Boat Trip Birka lake Mälaren

Understanding growth dynamics in the Arabidopsis thaliana root cambium

Ari Pekka Mähönen, University of Helsinki, Finland

16.00 - 19.15

Registration

09.25 - 09.45

Nathaniel Street, Umeå University, Sweden

17.00 - 17.10

Welcome Words

Lisbeth Jonsson, Congress Chair

Applying next generation sequencing to genomics studies of aspen species and Norway spruce

17.10 - 18.00

Opening session: From the Lab to the Field: Improving Crop Yield and Nutritional Quality

Wilhelm Gruissem, ETH Zürich, Switzerland

09.45 - 10.15

High throughput biology

Chair: Stefan Jansson, Umeå University, Sweden

09.45 - 10.15

Systems biology of plant senescence and death

Keynote Speaker: Hong Gil Nam, IBS & DGIST, South Korea

10.15 - 10.45

Coffee/tea

10.45 - 12.00

High throughput biology, continued

10.45 - 11.15

Natural variation in aspen

Stefan Jansson, Umeå University, Sweden

11.15 - 11.30

Whole-genome sequencing of medicinal plant, Senna tora

Sang-Ho Kang, National Academy of Agricultural Science, Jeonju, South Korea

11.30 - 11.45

Gene family evolution in Fagales and Betulaceae – molecular evidence from the Silver Birch (Betula pendula) genome

Sitaram Rajaraman, University of Helsinki, Finland

11.45 - 12.00

Effects of apoplastic H2O2 on phenolic metabolism and gene expression in a lignin-forming cell culture of Norway spruce

Anna Kärkönen, University of Helsinki, Finland

18.00 - ca 19.30 Get together reception Monday 10 August 2015 09.00 - 09.10

09.15 - 09.45

09.45 - 10.15

Welcome Words

Lisbeth Jonsson, Congress Chair, Astrid Söderbergh Widding, Vice-Chancellor of Stockhom University

SPPS Awardee My view on how to foster more of transformative research

Gunnar Öquist, Umeå University Sweden

Development

Chair: Ove Nilsson, SLU, Umeå, Sweden

Photoperiodic regulation of tree growth and development

Speaker: George Coupland, Max Planck Institute for Plant Breeding Research, Cologne, Germany

10.15 - 10.45

Coffee/tea

12.00 - 13.00

Lunch

10.45 - 12.00

Development, continued

13.00 - 14.30

Poster Session 2 (Posters with even numbers)

10.45 - 11.15

Photoperiodic regulation of tree growth and development

Keynote Speaker: Ove Nilsson, SLU, Umeå, Sweden

14.30 - 15.30

Photobiology

Chair: Dario Leister, University of Copenhagen, Denmark

11.15 - 11.30

The role of floral meristem genes during patterning of the Asteraceae capitulum

Paula Elomaa, University of Helsinki, Finland

14.30 - 15.00

How chloroplasts talk to the nucleus: news from GUN1

Dario Leister, University of Copenhagen, Denmark

11.30 - 11.45

HY5 integrates the effect of light and temperature in thermoperiodic control of shoot elongation

Jorunn Elisabeth Olsen, Norwegian University of Life Sciences, Norway

15.00 - 15.30

Chloroplast retrograde signaling

Keynote Speaker: Lixin Zhang, Chinese Academy of Sciences, Beijing, China

11.45 - 12.00

Identification of two microproteins involved in the regulation of flower initiation

Moritz Graeff, University of Copenhagen, Denmark

15.30 - 16.00

Coffee/tea

16.00 - 17.00

Photobiology, continued

16.00 - 16.15

Modulation of the thylakoid membrane ultrastructure and its impact on photosynthetic performance

Mathias Pribil, University of Copenhagen, Denmark

12.00 - 13.00

Lunch

13.00 - 14.30

Poster Session 1 (Posters with uneven numbers)

14.30 - 15.30

Epigenetics and gene regulations

Chair: Claudia Köhler, SLU, Uppsala, Sweden

16.15 - 16.30

Rose bengal as singlet oxygen phtosensitizer in leaves

Éva Hideg, University of Pécs, Hungary

14.30 - 15.00

Mechanisms of transcriptional gene silencing in Arabidopsis thaliana

Keynote Speaker: Thierry Lagrange, Université de Perpignan, France

16.30 - 16.45

Improving the activation of Rubisco in wheat

Alejandro Perdomo López, Rothamsted Research, UK

15.00 - 15.30

Epigenetic mechanisms driving plant speciation

Claudia Köhler, SLU, Uppsala, Sweden

16.45 - 17.00

15.30 - 16.00

Coffee/tea

Light-driven biosynthesis: Using photosynthetic electron transport in synthetic biology

Poul Eric Jensen, University of Copenhagen, Denmark

16.00 - 17.00

Epigenetics and gene regulations, continued

16.00 - 16.15

Functional characterization of RNA binding proteins co-purified with HUB1 in Arabidopsis thaliana

Sabine Le Gall, Ghent University, Belgium

17.30 - latest 01.00

Visit to the Bergius Botanic Garden and buffet at the Old Orangery Restaurant

16.15 - 16.30

Temperature - dependent formation of epigenetic memory in Norway spruce

Carl Gunnar Fossdal, Norwegian Forest and Landscape Institute, Norway

16.30 - 16.45

Molecular role of TSN in cytoplasmic messenger ribonucleoprotein complexes during stress

Emilio Gutierrez Beltran, SLU, Uppsala, Sweden

16.45 - 17.00

Plant sex chromosomes: structure and function

Boris Vyskot, Czech Academy of Sciences, Brno, Czech Republic

19.00 - 20.00

Welcome reception, City Town Hall offered by the City of Stockholm

Programme for Wednesday 12 August

Programme for Thursday 13 August

Wednesday 12 August 2015

Thursday 13 August 2015 09.00 - 09.15

Poster prize winner announcements

Torgny Näsholm, SLU, Umeå, Sweden

09.15 - 10.15

Biotic interactions

Chair: Jens Stougaard, Aarhus University, Denmark

Abiotic stress

Chair: Jaakko Kangasjärvi, University of Helsinki, Finland

09.15 - 09.45

Functional overlap and niche specialization of the Arabidopsis leaf and root microbiotas

DREB regulatory networks for abiotic stress response in plants

Keynote Speaker: Kazuko YamaguchiShinozaki, The University of Tokyo, Japan

Keynote Speaker: Paul Schulze-Lefert, Max Planck Institute of Plant Breeding Research, Cologne, Germany

09.45 - 10.15

Signals and receptors involved in endosymbiosis

Reactive oxygen species in plant signaling: Subcellular compartments and specificity

Jaakko Kangasjärvi, University of Helsinki, Finland

Jens Stougaard, Aarhus University, Denmark

10.15 - 10.45

Coffee/tea

09.00 - 09.35

Physiologia Plantarum Awardee

09.00 - 09.35

Plant Nitrogen Nutrition – the simple and the complex version

09.35 - 10.35 09.35 - 10.05 10.05 - 10.35 10.35 - 11.00

Coffee/tea

10.45 - 12.00

Biotic interactions, continued

11.00 - 12.00

Abiotic stress, continued

10.45 - 11.00

PP2A phosphatase as a regulator of ROS signaling in plants

11.00 - 11.15

Molecular basis of variation in stomatal responsiveness to elevated CO2

Saijaliisa Kangasjärvi, University of Turku, Finland

11.00 - 11.15

Richard Hilleary, University of Wisconsin, US

11.15 - 11.30

NAC transcription factor RAF controls senescence and Bernd Müller Röber, plant development under abiotic stress conditions in Arabi- University of Potsdam, Germany dopsis thaliana

Glutamate-like receptor channels GLR3.3 and GLR3.4 modulate MAMP-dependent root-to-shoot calcium waves

11.15 - 11.30

Transcript patterns in infected vs. uninfected cells of Datisca glomerata nodules

Marco Salgado, Stockholm University, Sweden

11.30 - 11.45

Functional significance of oxylipin induced heat shock protein accumulation in Arabidopsis thaliana

Miriam Münch, University of Würzburg, Germany

11.30 - 11.45

The expanded family of IDA- and PIP/PIP-like peptides: peptide ligand motifs, evolution and stress induction

Atle Magnar Bones, Norwegian University of Science and Technology, Norway

11.45 - 12.00

Chloroplast antioxidant dynamic control ROS-signaling upon repetitive cold stress

Margarete Baier, FU, Berlin, Germany

11.45 - 12.00

Closing words

12.00 - 13.00

Lunch

Afternoon

Optional guided boat tour in Stockholm

Karin S. L. Johansson, University of Gothenburg, Sweden

12.00 - 13.00

Lunch

13.00 - 14.00

Poster Session 3 (All posters)

14.00 - 15.00

SPPS General Assembly

15.00 - 15.30

Coffee/tea

15.30 - 17.00

Education and outreach

Chair Jodi Maple-Grödem, University of Stavanger, Norway

15.30 - 16.00

Who represents plant scientists at European level, how can you have a say and get involved?

Keynote Speaker: Karin Metzlaff, EPSO, Brussels, Belgium

16.00 - 16.30

Plant, politics and public perception

SPPS Popularisation Prize Winner: Stefan Jansson, Umeå University, Sweden

16.30 - 17.00

The Gatsby Plant Science Summer School

Keynote Speaker: Celia Knight, Leeds University, UK

19.00 - latest 01.00

Congress dinner and party, The Vasa Museum

SPPS Awards

SPPS 2015 Poster Prizes

SPPS awards have been established to recognise the work and achievements done in advancement of plant physiology and plant science in general. In the general assembly of 2011, it was decided to establish three new awards in addition to the older Popularisation Prize and there are currently four awards given out by SPPS. The awards are presented during the biannual SPPS Congress and membership of the Society is not a requirement for receiving an award. A call for nominations is opened by the Council prior to the SPPS Congress. The awards are not, however, necessarily presented at every Congress, but depend on the nominations received and the decision of the Council.

The poster prizes are designed to reward the considerable work that goes into preparing a poster for presentation at the Congress and as an encouragement especially to PhD-students and young postdoctors.

SPPS Popularisation Prize SPPS Popularisation Prize is a monetary award of honor to encourage plant biologists to bring research results to the public. The prize can be given to a person who has clearly popularised plant biology in newspapers, journal articles, books, television, radio or other public forums. This year’s winner of the SPPS Popularisation Award is Stefan Jansson (Plants, politics and public perception, Wednesday, 16.00 - 16.30)

SPPS Early Career Award SPPS Early Career Award is a monetary award to an early career scientist based in Scandinavia. The award is granted to a young, highly talented scientist, who has shown good progress and made significant, independent contributions to Scandinavian plant biology. All young plant biologists based in Scandinavia are eligible for the award if they have received their doctoral degree less than 10 years before the SPPS Congress. This year, there are two SPPS Early Career Award winners: Ari Pekka Mähönen (Understanding growth dynamics in Arabidopsis thaliana) and Nathaniel Street (Applying next generation sequencing to genomics studies of aspen species and Norway spruce), Tuesday 09.00 - 09.40.

SPPS Award SPPS Award is a monetary award given to a scientist based in Scandinavia in recognition of his/her outstanding, merited contribution to the science of plant biology in Scandinavia. This year’s winner of the SPPS Award is Gunnar Öquist (SPPS Awardee: My view on how to foster more of transformative research, Monday 10 August, 09:10-09:40)

Physiologia  Plantarum  Award Physiologia Plantarum Award is a monetary award to a scientist that has made outstanding contributions to plant science in the areas that are covered by Physiologia Plantarum.  This could be either lifetime contributions or recent breakthrough-type of contributions. This year’s winner of the Physiologia Plantarum Award is Torgny Näsholm (Plant nitrogen nutrition – the simple and the complex version, Wednesday 12 August, 09.00 - 09.30)

A panel of judges will attend the poster sessions to evaluate the posters and ask the presenting authors questions about their work. The panel will select the three best posters for the following prizes: 1st prize

2000 SEK

2nd prize

1000 SEK

3rd prize

1000 SEK

The poster prizes will be announced in the morning on Thursday August 13th.

General information

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We do not allow the use of personal laptops for presentations. Please bring your presentation on a CD, DVD or memory stick. At the end of the conference, all presentations will be deleted so no copyright issues will arise.

Contact information If you have any questions or need help, please go to the registration desk when at the conference venue, otherwise please contact: [email protected] ti Fresca

Information for speakers Please bring your presentation to the technician in the auditorium (at least 1 hour before your presentation). The technician will transfer the presentation to the congress server and make sure your presentation runs smoothly.

About Stockholm Please visit the official tourist information www.visitstockholm.com

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Venue Aula Magna, Stockholm University Frescativägen 6, SE-106 91 Stockholm Sweden

Stockholm University Campus Map

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Stockholm Metro Map SPPS 2015 EXHIBITION 6 Bergmann Labora 7 Ninolab 8 SPPS/Physiologia Plantarum 9 LICOR

Social Programme

www.regentinstruments.com [email protected]

Image Analysis for Plant Science since 1991

Sunday 9 August Pre-congress tour A day with Linneaus in Uppsala Travelling from Stockholm, we will meet at the Central Station on the platform for the train from Stockholm to Uppsala leaving at 09.11. You need to buy your own ticket before entering the train. If you are joining in Uppsala, we will meet at the platform for the train arriving from Stockholm at 09.49. Pre-congress tour Boat trip to Birka in lake Mälaren We meet at 9.30 at Stadshusbron, Klara Mälarstrand 2, where the boat starts. This is situated close to the City Hall (Stadshuset) but on the other side of the bridge (see map). Departure is at 10.00 and arrival at Birka is 11.45. The boat leaves Birka at 15.30 and reaches Stockholm at 17.30 www.stromma.se/stockholm/utflykter/dagsutflykter/birka-vikingastaden/ Get together in the evening at the congress venue – Aula Magna (included in registration fee)

Monday 10 August Welcome reception at Stockholm City Hall offered by the City of Stockholm (incl. in registration fee) Address: Hantverkargatan 1, Stockholm. For those who wish guidance to the City Hall, we will meet outside of the Central station at 18.30 and walk together to the City Hall.

Tuesday 11 August Tour of Bergius Botanic Garden and buffet dinner at the Old Orangery Restaurant in the garden. After the closing of the session on Tuesday afternoon, we will meet at the entrance of Aula Magna and walk together to the meeting place for the guided tours. To those not attending any guided tour, please refer to the map at http://www.bergianska.se/english/visit-us to find the Old Orangery Restaurant.

Wednesday 12 August Congress dinner & party in Skeppshallen at the Vasa Museum. The dinner starts at 19.00 at the Vasa Museum. Address: Galärvarvsvägen 14, 11521 Stockholm More information at www.vasamuseetsrestaurang.se

Thursday 13 August Optional guided boat tour of Stockholm Further information will be given at the conference secretariat, where you will be able to register for this tour during the congress.

WinSCANOPY™

• Canopy structure and Solar radiation • Plant canopy health/stress (NDVI) • Camera, calibrated fisheye lens, self-leveling mount, electronic compass,...

WinCELL™

• Wood-cell structure parameters over annual rings • Analysis of one or more rings per image • Data computed on yearly basis in a format compatible with WinDENDRO™

WinDENDRO™

• Tree rings from disks, cores, X-ray films and digital X-ray systems • Cross-dating graphic, correlation functions, detrending,... • Wood density and earlywood/latewood boundary

WinFOLIA™

• Leaf morphology • Leaf shape (Fractals), aspect ratio and form coefficient • Healthy, diseased and pest damage areas

WinSEEDLE™

• Seed and needle morphology • Counts and classification of seeds and needles • Healthy, diseased and pest damage areas

WinRHIZO™

• Washed root morphology, topology, link, and architecture • Automatic analysis of Arabidopsis seedlings • Healthy, diseased and pest damage areas

WinRHIZO™ Tron

• Morphology, architecture and topology of roots growing in soil • Roots must be traced manually with a mouse or by touching the screen of a tablet computer

Need More Than One Software Program? Take Advantage of Our New Suites. Image acquisition hardware sold separately.

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ORAL PRESENTATIONS

[1] FROM THE LAB TO THE FIELD: IMPROVING CROP YIELD AND NUTRITIONAL QUALITY

[2] MY VIEW ON HOW TO FOSTER MORE OF TRANSFORMATIVE RESEARCH

Wilhelm Gruissem ETH Zurich, Department of Biology, Switzerland

Gunnar Öquist,1 1 Department of Plant Physiology, Umeå Plant Science Centre, Umeå University, Sweden

Newest projections expect that by 2050 the world population will grow to more than 9 Billion people. To provide sufficient food for these people requires an increase in crop production by at least 50%. Usable agricultural production areas are already at their limits, and therefore we have to significantly improve the yield of our major crop plants. Major crops such as maize, wheat, rice, potato and cassava are rich in starch and together they represent more than 85% of the carbohydrate calories consumed worldwide. People for whom these crops are the primary staple food often suffer from malnutrition because the seeds, tubers and roots of these plants do not contain enough of the necessary vitamins and minerals such as iron for a healthy diet. For example, 1.6 billion people worldwide suffer reduced productive capacity due to iron-deficiency anemia. To meet the challenges of population growth and public health we also have to radically change the way of breeding our crop plants. Utilization of the genetic potential of crop diversity available in seed banks around the world can make an important contribution to reach this goal. In addition, gene technology strategies are required to accelerate crop breeding and to increase the iron and vitamin content of major staple crops such as rice, cassava and wheat. But it also needs better consumer education and acceptance of modern breeding methods.

I am not going to give a scientific talk since I closed my laboratory 10 years ago. I am going to talk about the nature of scientific research, and I will reflect on how we in the Nordic countries could take steps to foster more of ground-breaking or transformative research, research that opens up for new perspectives and new, often unexpected opportunities for mankind. Certain criteria need to be in place for increasing the probability for transformative research to develop. These criteria are: recognising and nurturing talents, trust and freedom, long-term perspectives, creative and dynamic research environments, and finally, beyond and across disciplines. These five criteria have a clear focus on individuals with novel ideas and they were formulated in the “Aarhus Declaration” of 2012 (www.excellence2012.dk). I will refer to the Academy Report “Fostering breakthrough research: A comparative study”, which was made by my selves and Professor Mats Benner, Lund University, in 2012. I will emphasise the role of academic leadership, in contrast to a today prevailing managerial leadership, and I will stress the importance of recruitment and career opportunities given to young people in order to foster renewal in science. I will discuss the complementary roles of universities and granting agencies in promoting excellence in research. If time allows, I may also exemplify with some experience from my own research, experiences that in different ways have formed my way through academia.

[3] SEASONAL FLOWERING IN ANNUAL AND PERENNIAL PLANTS

[4] PHOTOPERIODIC REGULATION OF TREE GROWTH AND DEVELOPMENT

George Coupland,1

Ove Nilsson,1

1

Max Planck Institute for Plant Breeding, Germany

We study the mechanisms by which the life cycles of annual and perennial plants are synchronised to the changing seasons. Arabidopsis thaliana is a model annual species and we have shown how circadian-clock regulation of transcription of specific regulatory genes and photoreceptor signaling combine to promote flowering of this species in response to long summer days. This process activates expression in the leaf of a protein that moves systemically to the shoot meristem and reprogrammes transcription leading to flower development. Perennial relatives of A. thaliana show interesting differences in their responses to seasonal cues, and we have developed Arabis alpina as a model perennial. This species flowers predominately in response to winter cold, and shows characteristic features of perennials such as only becoming sensitive to environmental cues after it reaches a certain age and cycling between episodes of flowering and vegetative development within its life cycle. Using a combination of forward genetics, genomics and comparative analyses in a phylogenetic context we have defined some of the mechanisms by which flowering regulation differs between annuals and perennials, contributing to the divergence of these life histories during evolution.

Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences (Slu), Sweden

1

Day length controls flowering time in many plants. The day-length signal is perceived in the leaf, and this signal is transduced to the shoot apex where floral initiation occurs. In Arabidopsis, the day-length response depends on the induction of the FLOWERING LOCUS T (FT) gene by the gene CONSTANS (CO), but also partially on a CO-independent mechanism involving the gene GIGANTEA (GI). In contrast to annual plants like Arabidopsis, forest trees display a perennial growth behaviour characterized by a very extended juvenile phase before flowering, and, in temperate regions of the world, an annual cycling between growth and dormancy. We have shown that the CO/FT regulatory module is functionally conserved in the aspen tree and that at least the FT homologs control the timing of flowering. However, unexpectedly, the CO/FT module also controls the short-day induced growth cessation and bud set normally occuring in the fall. We suggested that the differences in critical daylength for growth cessation can be explained by a difference in the phase of expression of the aspen ortholog of the gene CO leading to induction of FT at different critical daylengths. However, ectopic expression of CO is not sufficient to prevent growth cessation, suggesting that also other factors are needed together with CO. We have found that one such factor is likely to be the Populus homolog of GI which seems to have a much more important role in the day-length perception in Populus than in Arabidopsis. We are also investigating how the genes involved in the control of growth cessation and bud set are affecting bud burst in the spring and how some of these genes might be linked not only to the regulation off flowering, but also to other aspects of the juvenility-to-maturity transition in trees, including vegetative phase change. Our ultimate goal is to understand how the same regulatory pathways that control flowering in Arabidopsis are playing key roles for local adaptation of trees by controlling seasonal patterns of growth and dormancy.

[5] THE ROLE OF FLORAL MERISTEM GENES DURING PATTERNING OF THE ASTERACEAE CAPITULUM

[6] HY5 INTEGRATES THE EFFECT OF LIGHT AND TEMPERATURE IN THERMOPERIODIC CONTROL OF SHOOT ELONGATION

Yafei Zhao,1, Teng Zhang,1, Suvi K. Broholm,1, Katriina Mouhu,1, Sari Tähtiharju,1, Teemu H. Teeri,1, Paula Elomaa,1

Jorunn Elisabeth Olsen,1, Micael Wendell,1, Amsalu Gobena Roro,1, Ivana Todorcevic,1, Deepak Mahat,1, Jon Anders Stavang,2, Sissel Torre,1

1

Department of Agricultural Sciences, University of Helsinki, Finland

Department of Plant Sciences, Norwegian University of Life Sciences, Norway, 2 Institutt for Biologi, Universitetet I Bergen, Norway 1

A key question in biology is to understand how organismal form is generated. In plants, an important aspect of the form is the architecture of inflorescences that may vary from a single flower to large flower clusters, and is thus one of the major determinants of crop yield and reproductive success of plants. Still, the development and evolution of distinct inflorescence forms remain poorly understood. Most of our knowledge on the molecular underpinnings of inflorescence architecture is based on studies in the classical model plants; Arabidopsis with racemes and Solanaceous species petunia and tomato with cymes. In the large sunflower (Asteraceae) plant family, the inflorescence (capitulum) resembles a solitary flower but is a highly compressed structure consisting of different types of flowers. In the model plant Gerbera hybrida, capitulum development involves rapid expansion of the meristem, initiation of hundreds of individual flowers in a spiral manner, and differential development of distinct flower types. Morphological characterization of early stages inflorescence meristem patterning and flower primordia initiation in combination with functional analyses of key floral meristem identity genes (LEAFY/UNUSUAL FLORAL ORGANS) brings out novel information of the development and evolutionary origin of the capitulum architecture.

Temperature is among the most important determinants of plant growth and development, and accordingly plants have developed adaptive mechanisms to cope with seasonal and diurnal changes in temperature. Plants are able to discriminate between day and night temperature, a phenomenon referred to as thermoperiodism. Thermoperiodic control of shoot elongation is reported for several plant species with larger inhibition of shoot elongation in response to decreased temperature in the early morning compared to at night. However, information on the mechanisms underlying thermoperiodism is limited. Using pea and Arabidopsis thaliana as model systems, we aimed to shed light on the signalling associated with thermoperiodism. We show here that thermoperiodic control of shoot elongation depends on the photomorphogenesis-related transcription factor HY5, which is regulated in a diurnal manner with highest and lowest levels during day and night, respectively. Consistent with this, the sensitivity to temperature alteration was highest and lowest in the middle of the day (not in the morning) and night, respectively. Plants mutated in HY5 did not exhibit thermoperiodic control of hypocotyl or stem elongation. Also, cop1 mutants unable to regulate their level of HY5 in a diurnal manner, responded equally to temperature alteration during day and night. The role of HY5 in inhibition of shoot elongation upon lowered day temperature was associated with HY5-mediated up-regulation of specific GA2oxidases acting in gibberellin inactivation, and down-regulation of specific auxin and brassinosteroid biosynthesis genes. These studies demonstrate that HY5 integrates the effect of light and temperature in thermoperiodic control of shoot elongation.

[7] IDENTIFICATION OF TWO MICROPROTEINS INVOLVED IN THE REGULATION OF FLOWER INITIATION Moritz Graeff, , Daniel Straub, , Tenai Eguen, , Stephan Wenkel, 1

1

1

1

1

Institut for Plant and Environmental Sciences; Copenhagen Plant Science Center, Denmark

MicroProteins (miPs) are small, single domain proteins, which can function as negative regulators of protein complex formation and complex activity by directly interacting with larger proteins, sequestering them in a non-functional sate. Many examples for this kind of protein species in plants and animals have been described. Often miPs can be found to act in developmental processes where they modulate the activity of genetic key components in these pathways. One of the most important and complex transitions in plants is the initiation of flower formation. In the last years many genetic key components of this complex regulatory network have been identified and described but still it remains often unclear how these factors function and are regulated. We performed a screen for potential miPs in the proteome of Arabidopsis thaliana and several other representative plant species and identified two small proteins, conserved among dicotyledonous plants, that affect the process of flower initiation in Arabidopsis. The presented data will show our characterization of the identified miPs and elucidate their role in the complex process of flowering time regulation.

[8] MECHANISMS OF TRANSCRIPTIONAL GENE SILENCING IN ARABIDOPSIS THALIANA Thierry Lagrange,1, Sylvie Lahmy,1, Natacha Bies-Etheve,1, Dominique Pontier,1, Suhua Feng,2, Michèle Laudié,1, Mohamed-Ali Hakimi,3, Richard Cooke,1, Steven E. Jacobsen,2 Laboratoire Génome Et Développement de Plantes, Umr5096 Cnrs/Université de Perpignan Via Domitia, France, 2 Department of Molecular, Cell and Developmental Biology, Hhmi, University of California Los Angeles, United States 3 Laboratoire Adaptation Et Pathogénie des Microorganismes, Umr5163 Cnrs/Université Joseph Fourier, France 1

RNA-directed DNA methylation (RdDM) is the major small RNA-mediated epigenetic pathway in plants. Two RNA polymerase II (PolII)-related enzymes, known as RNA polymerase IV and V (PolIV and PolV), have been functionally linked to the production and/or action of TE-derived sRNAs in RdDM. In the current model for RdDM, PolIV transcribes TE loci and produces 24-nucleotide (nt) siRNAs that are loaded into AGO4-related proteins leading to the assembly of AGO-siRNA effector complexes that guide DOMAINS REARRANGEDMETHYLTRANSFERASE 2 (DRM2) to methylate DNA. Targeting of DRM2 also involves an second type of long-noncoding RNAs (lncRNAs), produced by PolV, that are proposed to act as template to recruit the AGO4-siRNA effector complex via base-pairing interactions. In addition to these core components, several other proteins have been shown to interact either with PolIV/ PolV/AGO4 and assist in their function at various stages of RdDM action. Our current work is concerning the mechanism of action of these proteins in RdDM and gene silencing.

[9] EPIGENETIC MECHANISMS DRIVING PLANT SPECIATION Claudia Köhler,1, David Kradolfer,1, Philip Wolff,1, Jordi Moreno-Romero,1, Nicole Schatlowski,1, Hua Jiang,1, Carolin Rebernig,1, Clément Lafon-Placette,1 1

Department of Plant Biology, Uppsala Biocenter, Swedish University of Agricultural Sciences, Sweden

[10] FUNCTIONAL CHARACTERIZATION OF RNA BINDING PROTEINS CO-PURIFIED WITH HUB1 IN ARABIDOPSIS THALIANA Sabine Le Gall,1, Magdalena Woloszynska,1, Tommaso Matteo Boccardi,1, Stjin Aesaert,1, Stijn Dhondt,1, Leonardo Bruno,2, Bram Slabbinck,1, Frederik Coppens,1, Geert De Jaeger,1, Kristiina Himanen,3, Mieke Van Lijsebettens,1 Department of Plant Systems Biology, Vlaams Instituut Voor Biotechnologie (Vib); Department of Plant Biotechnology and Bioinformatics, Ghent University, Belgium, 2 Dipartimento DI Ecologia, Università Della Calabria, 3 Department of Agricultural Sciences, Finland 1

Polyploidization is a widespread phenomenon among plants and is considered a major speciation mechanism. Polyploid plants have a high degree of immediate post-zygotic reproductive isolation from their progenitors, as backcrossing to either parent will produce mainly nonviable progeny. This reproductive barrier is called triploid block and it is caused by malfunction of the endosperm. Our work has revealed that deregulated parent-of-origin specific genes (imprinted genes) are causal for the response to interploidy hybridizations, revealing an epigenetic basis of this phenomenon. I will discuss epigenetic changes in response to interploidy hybridizations and their consequences for endosperm development. I will furthermore discuss an epigenetic method for the generation of viable triploids, providing an impressive example for the potential of epigenome manipulations for plant breeding. Lastly, I will discuss a recently evolved interspecies hybridization barrier in the genus Capsella that reveals striking similarities to interploidy hybridization barriers, suggesting a common mechanistic basis.

H2Bub is an activating histone mark found in gene bodies and absent from promoter regions. In plants, Histone monoubiquitination1 (HUB1) is a conserved E3 ubiquitin ligase that monoubiquitinates histone H2B in a heterodimeric complex with its homolog HUB2 with the help of the UBC2 conjugating enzyme. H2Bub has been linked to transcriptional activation of genes and pathways related to flowering, cell cycle, dormancy, photomorphogenesis, cuticle and wax composition, circadian clock and pathogen defense. The mechanism by which HUB1/HUB2 targets genes is not known therefore to further elucidate HUB1 regulation and function in plants we investigated HUB1 interacting proteins. A Tandem Affinity Purification on Arabidopsis cell suspension cultures using HUB1 as bait co-purified two proteins containing RNA binding domains: RBP1 and RBP2. RBP1 and RBP2 were functionally characterized in plant growth and development by using mutant alleles. We investigated several aspects of vegetative growth and flowering on macro and cellular level. The role of RBP1 and RBP2 in H2Bub and/or RNA related biology is being examined by transcriptome analysis and ChIP. This research is funded by the European Commission Marie Curie Research Training Network ‘Chromatin in Plants – European Training and Mobility’ (CHIP-ET, FP7-PEOPLE-2013-ITN607880).

[11] TEMPERATURE-DEPENDENT FORMATION OF EPIGENETIC MEMORY IN NORWAY SPRUCE

[12] MOLECULAR ROLE OF TSN IN CYTOPLASMIC MESSENGER RIBONUCLEOPROTEIN COMPLEXES DURING STRESS

Carl Gunnar Fossdal,1, Igor Yakovlev,1, Jorunn Elisabeth Olsen,2, Elena Carneros,1

Emilio Gutierrez Beltran,1, Panagiotis N. Moschou,1, Andrei P. Smertenko,2, Peter V. Bozhkov,1

1

Norwegian Forest and Landscape Institute, Norway2 NMBU, Norway

Epigenetic memory formed in the Norway spruce embryos permanently affect the timing of bud burst and bud set in the progenies, vitally important adaptive traits in this long-lived forest species. Epigenetic memory marks are established in response to the temperature conditions prevailing during embryogenesis; the epitype is fixed by the time the embryo is fully developed and is mitotically propagated throughout the tree’s life span. Somatic embryogenesis closely mimics the natural zygotic embryo formation and results in epigenetically different plants in a predictable temperature-dependent manner with respect to altered phenology. Using RNAseq transcriptome analysis of mRNA and noncodingRNA (ncRNA) changes were monitored in somatic embryos under different temperatures. We found distinct differences in mRNA and ncRNA transcriptomes between the genetically identical embryogenic tissues grown under the epitype-inducing temperatures suggesting temperature-dependent canalizing of gene expression during embryo formation, putatively based on chromatin modifications.

Department of Plant Biology, Uppsala Biocenter, Swedish University of Agricultural Sciences and Linnean Center for Plant Biology, Sweden 2 Institute of Biological Chemistry, Washington State University, United States

1

Tudor staphylococcal nuclease (TSN) is an evolutionarily conserved protein involved in transcriptional and post-transcriptional regulation of gene expression in animals [1, 2]. Although TSN was found to be indispensable for normal plant development and stress tolerance, the molecular mechanisms underlying these functions remain elusive. We have found that Arabidopsis thaliana TSN is essential for the integrity and function of cytoplasmic messenger ribonucleoprotein (mRNP) complexes called stress granules (SG) and processing bodies (PB), sites of post-transcriptional gene regulation during stress [3]. We have further revealed that TSN as a key enzymatic component of catabolic machinery responsible for the processing of mRNAs in the mRNP complexes. Notably, TSN is stably associated with both SG and PB, suggesting that it may serve scaffolding role to recruit other proteins to the mRNP complexes. As the first step to address this point and to advance our understanding of molecular role of TSN in SG and PB, we have characterized TSN interactome using tandem affinity purification (TAP) combined with bimolecular fluorescence complementation (BiFC).

1.

Paukku, K., J. Yang, and O. Silvennoinen. Mol Endocrinol, 2003. 17(9): p. 1805-14.

2.

Scadden, A.D. Nat Struct Mol Biol, 2005. 12(6): p. 489-96.

3.

Gutierrez-Beltran, E., et al. Plant Cell, 2015.

[13] PLANT SEX CHROMOSOMES: STRUCTURE AND FUNCTION

[14] UNDERSTANDING GROWTH DYNAMICS IN THE ARABIDOPSIS THALIANA ROOT CAMBIUM

Boris Vyskot,1, Vojtech Hudzieczek,1, Roman Hobza1

Ari Pekka Mähönen1

1

Institute of Biophysics, The Czech Academy of Sciences, Czech Republic

Sex determination in dioecious plants is mostly based on genetics, but morphologically distinct sex chromosomes have only evolved in a few species. Of these, heteromorphic sex chromosomes have been most clearly described in two model species – Silene latifolia and Rumex acetosa. In both species, the sex chromosomes are the largest chromosomes in the genome. They are hence easily distinguished, can be physically separated and analyzed. Here we review some recent experimental data on selected model dioecious species, with a focus on S. latifolia. Phylogenetic analyses show that dioecy in plants originated independently and repeatedly even within individual genera. A cogent question is whether there is genetic degeneration of the non-recombining part of the plant Y chromosome, as in mammals, and, if so, whether reduced levels of gene expression in the heterogametic sex are equalized by dosage compensation. Our recent data on S. latifolia show that both the X and Y chromosomes harbor active genes, but that they are divergent due to genetic degeneration of alleles in the non-recombining region of the Y. Most important achievements include genetic mapping of model dioecious plants, discovery of phylogenetic relationships in sex chromosome evolution, identification of new sex-linked genes, and deciphering a role of repetitive DNA in sex chromosome evolution. Special attention is taken to develop original methods for targeted modification of plant genes and genomes.  Supported by the Czech Science Foundation (P501/12/G090).

1

Institute of Biotechnology/Department of Biosciences; University of Helsinki, Finland

Despite the importance of the vascular cambium in plant biology and in wood production, the molecular and cellular mechanisms underlying cambial activity remain largely unknown. Particularly, it is unknown where the cambial stem cells are located, and how the stem cell niche is organized to drive cambial growth. In animal stem cell studies lineage tracing has been the method for locating the stem cells. In this method single cell clones marked with reporter expression are generated within a population of dividing cells. The clones are transmitted to all daughter cells of the initial cell, resulting in a marked sector within the tissue. By analysing the size and distribution of the sectors the position and the mitotic activity of dividing cells and stem cells can be deduced. In order to understand the cell lineage relations in the Arabidopsis root cambium, we generated GUS/ GFP sectors by using two step CRE-lox based clonal activation system. To understand which cells in the primary tissue contribute to the cambium formation we induced single cell clones during the primary development of the root and analysed the growing sectors during the secondary development. We also generated marked sectors in active cambium to map the position of stem cells and to understand the growth dynamics of the cambial cells. In my presentation I will explain how we are utilizing the lineage tracing data to provide mechanistic understanding of cambium regulation.

[15] APPLYING NEXT GENERATION SEQUENCING TO GENOMICS STUDIES OF ASPEN SPECIES AND NORWAY SPRUCE

[16] SYSTEMS BIOLOGY OF PLANT SENESCENCE AND DEATH Hong Gil Nam,1, Pyung Ok Lim,2, Hye Ryun Woo,2, Daehee Hwang1

Yao-Cheng Lin, , Nicolas Delhomme, , Jing Wang, , Bastian Schiffthaler, , Manfred Grabherr, , Neda Zamani, , Marc P Höpnner,5, Andrea Zuccolo,6, Chanaka Mannapperuma,2, Niklas Mähler,7, Katja Gorenc,2, Kerstin Richau,2, David Sundell,2, Barbara Terebieneic,2, Yves Van de Peer1,1, Eung-Jun Park,8, Torgeir R Hvidsten,7, Stefan Jansson,2, Pär K Ingvarsson,3, Nathaniel Street2 1

2

3

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4

5

1

IBS & DGIST, Rep. of South Korea, 2 Department of New Biology, DGIST, Rep. of South Korea

Department of Plant Systems Biology (Vib) ; Department of Plant Biotechnology and Bioinformatics (Ghent University), Belgium, 2 Umeå Plant Science Centre; Department of Plant Physiology; Umeå University, Sweden, 3 Umeå Plant Science Centre; Department of Ecology and Environmental Science; Umeå University, Sweden, 4 Science for Life Laboratory; Department of Medical Biochemistry and Microbiology; Uppsala University, Sweden, 5 Department of Medical Biochemistry and Microbiology; Uppsala University, Sweden, 6 Institute of Life Sciences; Scuola Superiore Sant’anna, Italy, 7 Department of Chemistry; Biotechnology and Food Science; Norwegian University of Life Sciences, Norway, 8 Department of Forest Genetic Resources Development; Korea Forest Research Institute, Rep. of South Korea 1

We have sequenced the genomes of two Swedish forest trees: The 20 Gbp genome of Norway spruce (Picea abies) is both gigantic and repetitive while the 500 Mbp genome of European aspen (Populus tremula) is highly heterozygous. Both cases challenge current assembly approaches using short read sequencing technology. Useful assemblies of both genomes have been produced, allowing us to identify and explore ‘known unknowns’ in each genome: In Norway spruce we have shown that the large genome resulted from the slow and steady accumulation of a diverse set of LTR TEs, while in aspen we have shown rapid divergence from the genome of Populus trichocarpa. The availability of the genomes also enables discovery of previously ‘unknown unknowns’: In Norway spruce, 24 nt sRNAs, which are known to silence TEs via methylation, were found tobe highly tissue-specific and present at lower abundance than in other plants; In both aspen and Norway spruce, RNA-Seq transcript profiling revealed an extensive set of long intergenic non-coding RNAs (lincRNA) that are under active biological regulation and that have been widely conserved. Armed with these genomes we are employing a systems genetics approach to explore the ‘known unknown’ of how natural variation of complex phenotypic traits is controlled. Additionally we are developing metatranscriptomics studies to explore the diversity and dynamics of the holobiont system in both species. To ensure maximal benefit to the wider community, these resources have been integrated within the PlantGenIE.org web resource, enabling exploration of these genomes and associated genomics data.

Aging, a process of age-dependent functional switching of cells, organs, and organisms, is a life history strategy, eventually leading to senescence and death. Our main question is how plants know when to die and how to die. We previously identified the trifurcate feed forward pathway for age-dependent cell death involving EIN2, ORE1/ANAC092, and miR164 (Science, 2009) in Arabidopsis, which revealed fundamental characteristics of network-level control of plant aging and senescence. We further constructed gene regulatory networks for leaf-expressed NAC genes along aging, and found novel network properties and regulatory genes. To extend the age-associated molecular network, we employed systems-level analyses in Arabidopsis. In-depth analyses of the transcriptomes revealed transition of biological processes toward senescence and death are actively coordinated through differential utilization of various regulatory programs involving transcription factors, splicing variants, antisense transcripts, and small non-coding RNAs. Senescence is one of well-known biological processes to increase plant fitness in given environments. Genome-wide association between SNPs and senescence phenotypes revealed novel regulatory genes for senescence programs which could explain potential roles of senescence programs in their fitness. Senescence and death is in part systemically controlled at the organismal level. We are employing rice as a model system to understand the mechanism underlying the systemic senescence especially during grain filling to improve grain yield and quality. This systemic regulation may contribute to increase grain yield by properly supplying two major nutrients, carbon and nitrogen. Furthermore, the roles of nitrogen remobilizing enzymes in systemic senescence are being elucidated in connection with grain development. In this talk, I will deliver our research efforts and insights toward aging and death.

[17] NATURAL VARIATION IN ASPEN

[18] WHOLE-GENOME SEQUENCING OF MEDICINAL PLANT, SENNA TORA

Stefan Jansson1

Sang-Ho Kang1, Beom-Soon Choi2, Chang-Muk Lee1, Joon-Soo Sim1, Jin-Tae Jeong3, Jae Kyung Sohng4, Tae-Jin Oh4, Hyunhee Kim5, Seong-Han Sohn1, So Youn Won1, and Jung Sun Kim1

1

Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, Sweden

Genomics Division, National Academy of Agricultural Science, RDA, Jeonju, Rep. of South Korea, 2Phyzen Genomics Institute, Rep. of South Korea; 3Department of Herbal Crop Research, NIHHS, RDA, Rep. of South Korea, 4Department of Pharmaceutical Engineering, Institute of Biomolecule Reconstruction, SunMoon University, Rep. of South Korea, 5Department of Life Science, SahmYook University, Rep. of South Korea 1

As high-throughput techniques – in particular DNA sequencing technology – has become cheaper, high throughput studies can now be performed also on other plants than traditional model species like Arabidopsis and rice. This extends the range of scientific questions that can be raised and increases the possibilities to generalize findings from Arabidopsis, but it also poses new challenges as many plant species differ much from Arabidopsis in terms of life history, physiology, genetics and genomics. Trees species are obviously quite different from Arabidopsis, and the genus Populus have been adopted by the scientific community as the – at least so far – best developed model system for trees. We have spent much effort to develop the necessary genetics and genomics tools enabling high-throughput biology in Populus, focusing on one of the most widespread and ecologically important plant species on earth, aspen (Populus tremula). The hurdles for generating a high-quality reference genome sequence for aspen have been massive, as the genome is one of the most heterozygous and variable studied to date, but we have finally reached the point where we can use resequencing data from a natural population of aspens, where we have generated vast amounts of phenotypic data, to perform association mapping. This talk will describe challenges and opportunities, and how these data could tell us more about the local adaptation of aspen, focusing on phenology traits, but also how it is possible to use field experiments with transgenic trees to test predictions.

Senna tora (L.) Roxb. (Cassia tora), a member of Leguminosae (subfamily Caesalpinoideae), is a semiwild annual herb widely grown in different places of south-east Asia. Seeds are extensively used for medicinal applications in gastrointestinal disorders, treatment of skin, and ailments ranging from simple cough, hypertension to diabetes. Despite of its useful applications, there has been little report of molecular and genome study of S. tora. We have recently sequenced the genome of S. tora, CTS-89-2 (2n = 2x = 26), with an estimated haploid genome size around 650 Mb. We assembled into scaffolds representing 98.7% (641 Mb) of the 650 Mb S. tora genome. Genome and transcriptome analysis predicted 45,428 genes for S. tora. On the other hand, we produced 40,000 BAC libraries and generated PacBio single molecule sequences to enlarge the sequence scaffolds of the genome. With the sequenced genome, we are trying to identify the biosynthetic pathway of anthraquinones by transcriptome analysis and metabolic profiling from S. tora. S. tora draft genome sequence, transcriptome, and metabolome will aid the development of medicinally useful species with biologically active metabolites, elucidate a natural pathway of the anthraquinones synthesis, and provide a basis for the breeding of foetid senna with improved agronomic characteristics.

[19] GENE FAMILY EVOLUTION IN FAGALES AND BETULACEAE – MOLECULAR EVIDENCE FROM THE SILVER BIRCH (BETULA PENDULA) GENOME

[20] EFFECTS OF APOPLASTIC H2O2 ON PHENOLIC METABOLISM AND GENE EXPRESSION IN A LIGNIN-FORMING CELL CULTURE OF NORWAY SPRUCE

Jarkko Salojärvi,1, Sitaram Rajaraman,1, Olli-Pekka Smolander,1, Omid Mohammadi,1, Pezhman Safdari,1, Ali Amiryousefi,1, Airi Lamminmäki,1, Juha Immanen,1, Kaisa Nieminen,2, Lars Paulin,1, Petri Auvinen,1, Yrjö Helariutta,1, Jaakko Kangasjärvi1

Teresa Laitinen,1, Kris Morreel,2, Nicolas Delhomme,3, Kaloian Nickolov,4, Adrien Gauthier,1, Gunter Brader,5, Kean-Jin Lim,1, Nathaniel Street,3, Wout Boerjan,2, Teemu H. Teeri,1, Anna Kärkönen1

1

University of Helsinki, Finland, University of Helsinki, Finland; Natural Resources Institute Finland (Luke), Finland 2

Univ. of Helsinki, Dept of Agricultural Sciences, Finland, 2 Vib Department of Plant Systems Biology; Ghent University, Belgium, 3 Umeå Plant Science Centre; Umeå University, Sweden, 4 Univ. of Helsinki, Dept of Agricultural Sciences, Finland; Univ. of Oulu, Dept of Biology, Finland, 5 Univ. of Helsinki, Dept of Biosciences, Finland; Austrian Institute of Technology GmbH, Bioresources, Health & Environment Department, Austria

1

The Silver Birch (Betula pendula) is a commercially and ecologically important deciduous tree which is prevalent across Europe and Asia. Unlike other trees having lengthy generation times ranging from a few years to decades, birch can be made to flower within one year*. It has a small genome size of 440 Mbp and can be inbred and crossed with related birch species. Through Natural Resources Institute of Finland (Luke), we have access to a large number of natural variants and trees with varying wood quality. Birch grows well under laboratory conditions and genetic transformation techniques through Agrobacterium have been developed**. All these properties make birch an ideal model species for tree genetics.

H2O2 is required for extracellular lignin production in a cell suspension culture of Norway spruce (Picea abies) as its scavenging by potassium iodide (KI) represses the formation of extracellular lignin. This observation suggests that in this cell culture peroxidases activate monolignols for lignin polymerization. Phenolic analysis showed that instead of monolignols, KI-treated cultures accumulated di- and oligolignols. Some dimeric structures found in the culture medium can be formed only by intracellular enzymes suggesting that spruce cells can transport diphenols into the apoplast. Interestingly, removal of KI restored extracellular lignin formation, enabling us to study the regulation of lignin, and di- and oligolignol formation.

Our current genome, version 1.1, was obtained by sequencing a fourth generation inbred individual and carrying out a hybrid assembly using 454, Illumina, SOLID and PacBio sequencing. We have predicted approximately 28,000 genes using an Evidence Modeler-based*** genome annotation pipeline trained with gene models from RNA sequencing, ESTs, manually annotated genes and conserved core eukaryotic genes. Present study focuses on understanding gene family evolution within the order Fagales and especially within the family Betulaceae to understand the selection pressure contributing to variation and adaptation to the environment. We have sequenced 12 members of Betulaceae family and are carrying out evolutionary analysis of the genomes using the sequenced Betula pendula genome as reference to understand their divergence. References * K. A. Longman & P. F. Wareing, Early Induction of Flowering in Birch Seedlings, Nature 184, 2037-2038 (1959); doi:10.1038/1842037b0

In the lignin-forming and non-lignin-forming conditions ca. 6400 genes were differentially expressed. Several signal transduction-related genes were strongly affected. Gene expression patterns of antioxidant enzyme and related genes suggests that cultured spruce cells suffer from an oxidative stress in conditions where they produce extracellular lignin, and scavenging of apoplastic H2O2 alleviates this stress. Our data show that removal of apoplastic H2O2 leads to a reduced carbon flux into the shikimate pathway, into synthesis of phenylalanine/tyrosine, and into the phenylpropanoid and monolignol biosynthesis pathways, suggesting that a feedback mechanism exists when there is no strong sink for monolignols for subsequent polymerization. Several peroxidase and laccase genes were either up- or down-regulated when lignin was synthesized. Our focus is to identify candidate proteins for monolignol oxidation and transport, as well as for apoplastic H2O2 production together with putative transcription factors that regulate lignin formation in Norway spruce.

** Lemmetyinen J et al. Activity of the CaMV 35S promoter in various parts of transgenic early flowering birch clones. Plant Cell Rep. 1998;18:243–248 *** Haas et al. Automated eukaryotic gene structure annotation using EVidenceModeler and the Program to Assemble Spliced Alignments. Genome Biology 2008, 9:R7doi:10.1186/gb-2008-9-1-r7

[21] HOW CHLOROPLASTS TALK TO THE NUCLEUS

[22] CHLOROPLAST RETROGRADE SIGNALING

Dario Leister1 1

Copenhagen Plant Science Center, Denmark

Peiqiang Feng,1, Hailong Guo,1, Xiumei Xu,1, Wei Chi,1, Xuwu Sun,1, Rongcheng Lin,1, Congming Lu,1, Lixin Zhang1 Photosynthesis Research Center, Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, China

1

Developmental or metabolic changes in chloroplasts can have profound effects on the rest of the plant cell. Such intracellular responses are associated with signals that originate in chloroplasts and convey information on their physiological status to the nucleus, which leads to large-scale changes in gene expression (retrograde signalling). A screen designed to identify components of retrograde signalling resulted in the discovery of the so-called genomes uncoupled (gun) mutants. Genetic evidence suggests that the chloroplast protein GUN1 integrates signals derived from perturbations in plastid redox state, plastid gene expression and tetrapyrrole biosynthesis in seedlings of Arabidopsis, exerting biogenic control of chloroplast functions. However, the molecular mechanism by which GUN1 integrates retrograde signalling in the chloroplast is unclear. In this contribution, we present a model of how GUN1 controls retrograde signalling on the basis of genetic and biochemical evidence.

Chloroplasts are responsible not only for photosynthesis, but also for the biosynthesis of many essential compounds, such as fatty acids, vitamins, tetrapyrroles and amino acids. Thus, chloroplasts are crucial for establishment and maintenance of the photoautotrophic lifestyle of plants. Chloroplasts are originated from the endosymbiosis which refers to the engulfment of the photoautotrophic free-living cyanobacteria by a eukaryotic cell ancestor, accompanied by a massive transfer of eubacteria genes to the nucleus. This heterogenic arrangement necessitates the coordination of nuclear and plastid gene expression by establishing a bilateral communication. Therefore, chloroplast not only allows the nuclear control of its development by anterograde signals, it also can feedback to the nucleus to regulate gene expression using retrograde signaling. Chloroplast retrograde signaling has emerged to be essential for plant development and adaptive responsiveness to environmental perturbations in order to coordinate development and stress-related gene expressions. We will report the recent progress in the molecular mechanisms underlying signal transduction and the role of retrograde signal in plant growth and development.

[23] MODULATION OF THE THYLAKOID MEMBRANE ULTRASTRUCTURE AND ITS IMPACT ON PHOTOSYNTHETIC PERFORMANCE

[24] ROSE BENGAL AS SINGLET OXYGEN PHOTOSENSITIZERS IN LEAVES Eva Hideg,1, Gyula Czégény,2, Ferhan Ayaydin,3, László Kovács4

Mathias Pribil, , Mathias Labs, , Wenteng Xu, , Dario Leister 1

2

2

1

University of Copenhagen; Department of Plant and Environmental Sciences; Copenhagen Plant Science Center (Cpsc), Denmark, 2 Ludwig-Maximilians-University Munich, Germany

1

Thylakoids of land plant chloroplasts are composed of grana stacks and stroma lamellae, substructures that confer a characteristic three dimensional ultrastructure to the membrane system and support a lateral heterogeneity with respect to the distribution of photosynthetic complexes. Upon changes in environmental conditions thylakoids can undergo various structural rearrangements which account for the high plasticity of the membrane system. The relevance of the dynamic changes in these substructures in terms of regulating photosynthetic processes is poorly understood and little is known about the mechanisms underlying their formation. In Arabidopsis thaliana, the CURT1 protein family was shown to be involved in grana formation by facilitating membrane bending in the grana margins. The degree of membrane bending is thereby correlated with the amount of CURT1 being present in the thylakoid membrane. While a lack of CURT1 proteins (curt1abcd) leads to a general loss in thylakoid membrane curvature the accumulation of CURT1A (oeCURT1A) causes a hyper-membrane bending phenotype. Here, the effects of altered CURT1 levels on photosynthetic performance and growth rate under variable light conditions will be addressed and changes in the assembly and stability of the CURT1 complexes will be discussed in the light of reversible protein phosphorylation. Further, approaches to monitor and dissect CURT1 complex composition and its assembly dynamics will be presented.

Department of Plant Biology; University of Pécs, Hungary, 2 Institute of Biology, University of Pécs, Hungary, 3 Cellular Imaging Laboratory; Biological Research Centre, Hungary, 4 Institute of Plant Biology; Biological Research Centre, Hungary 1

In photosynthetic organisms, chlorophyll molecules may promote the photo-production of singlet oxygen (1O2). Due to the dual role of this ROS (damaging agent or signaling molecule), studies of stress-inducible, triplet chlorophyll mediated 1O2 photo-production are often complemented with experiments utilizing artificial photosensitizers. Effects of various exogeneous photosyntesizers depend on several factors, such as cellular localization, 1O2 quantum yield and additional (other than 1O2-mediated) metabolic interactions. We studied four photosensitizers, Rose Bengal (RB), Methylene Violet, Neutral Red and Indigo Carmine, and found that 1O2 from these dyes affected Photosystem (PS) II to different extents, depending on the microlocalization of the elicitor. Confocal laser scanning microscopy of RB and chlorophyll fluorescence showed that simple infiltration (forcing RB solution into tobacco leaf tissue using a syringe) delivered RB into chloroplasts. Selective excitation of RB with green light lead to oxidative damage of D1, a PS II core protein via 1O2 (Kovács et al. 2014, Photochem. Photobiol. 90:129-136). Recent work was focussed on RB, and explored 1O2-unrelated effects of this dye on photosynthetic electron transport. Experiments with RB-infiltrated leaves or isolated thylakoid membranes in the presence of RB suggested the possibility of PS II → RB electron transfer and chlorophyll → RB energy transfer. Chlorophyll fluorescence, electron transport and spin trapping EPR measurements showed that the effect of this popular photosynthesizer is more complex than prevously thought, and suggest caution when using RB as photosensitizer. Supported by the Hungarian Scientific Research Fund (OTKA NN-85349).

[25] IMPROVING THE ACTIVATION OF RUBISCO IN WHEAT

[26] LIGHT-DRIVEN BIOSYNTHESIS: USING PHOTOSYNTHETIC ELECTRON TRANSPORT IN SYNTHETIC BIOLOGY

Alejandro Perdomo López,1, Elizabete Carmo-Silva1 1

Plant Biology and Crop Science, Rothamsted Research, United Kingdom

Poul Erik Jensen1 1

Wheat is a major world crop and provides 20% of global daily protein and calories. Currently, wheat yields are increasing too slowly to meet the projected double in food demand by 2050. CO2 assimilation during photosynthesis is the primary determinant of plant biomass production. Rubisco plays a key role in CO2 assimilation, but is not optimally poised for crop productivity in current and projected climates. Rubisco is prone to inhibition by unproductive binding of sugar-phosphates that lock active sites in a closed conformation. Re-activation of Rubisco in vivo depends on the interaction with Rubisco activase (Rca), which facilitates the release of sugar phosphate inhibitors from catalytic sites. Rca activity and its consequent effect on Rubisco activation and photosynthesis are modulated by the redox status and ADP/ATP ratio of the chloroplast. In most plant species, Rca is composed of two isoforms: a shorter redox-insensitive β-isoform and a longer redox-sensitive α-isoform. In Arabidopsis thaliana, the two Rca isoforms differ in their ADP/ATP response. In the wheat genome, two Rca genes are present; expression of TaRca1 produces a β-isoform only, whereas alternative splicing of TaRca2 results in either a longer α-isoform or a shorter β-isoform. The three wheat Rca isoforms were purified after expression in E. coli. The regulatory properties of each individual isoform and of Rca in wheat leaf extracts will be characterised. The goal is to identify variation in the properties of Rca isoforms that can be exploited to improve wheat photosynthesis and crop yield.

Copenhagen Plant Science Centre; Dept. Plant and Environmental Sciences; University of Copenhagen, Denmark

The aim of synthetic biology is to engineer biological systems by designing and constructing novel modules to perform new functions for useful purposes. In this context the photosynthetic complexes (photosystem I and II) in the thylakoids of chloroplasts or cyanobacteria can be used to deliver energy and reducing power to novel biosynthetic pathways. Diterpenoids is one group of bioactive compounds with potential uses as pharmaceuticals or other high-value compounds. Key enzymes catalyzing their biosynthesis in medicinal plants are the chloroplast located diterpene synthases and the ER located cytochrome P450s. Through a synthetic biology approach, genes encoding specific diterpene synthases and cytochrome P450s have been engineered into chloroplasts and cyanobacteria and coupled directly to the photosynthetic energy output by relocating the enzymes to the thylakoids. Photo-reduced ferredoxin was shown to be able to deliver electrons directly to the cytochrome P450s without the need for specific reductases thereby avoiding generation of NADPH. Likewise the C20 precursor of diterpenoids, geranylgeranyldiphosphate, was efficiently cyclized into the expected diterpene before being hydroxylated by the cytochrome P450s to yield the diterpenoid. Approaches to optimize electron transfer towards the P450s by constructing fusion proteins between cytochrome P450s and ferredoxin and organization of the new biosynthetic enzymes in thylakoid bound metabolons will be presented. 

[27] PLANT NITROGEN NUTRITION – THE SIMPLE AND THE COMPLEX VERSION

[28] DREB REGULATORY NETWORKS FOR ABIOTIC STRESS RESPONSE IN PLANTS

Torgny Näsholm1

Kazuko Yamaguchi-Shinozaki1

1

Department of Forest Ecology and Management, Swedish University of Agricultural Sciences, Sweden

Terrestrial ecosystems are commonly nitrogen (N) limited and plants and other organisms in these systems therefore strive to maximize their N uptake. Soil solutions of various ecosystems have been shown to contain a wide range of different N compounds. Not surprisingly, therefore, plant roots have been shown capable of absorbing both inorganic and organic N forms, from the well studied absorption of ammonium and nitrate to the less studied uptake of simple organic N forms such as amino acids. Recently, even complex N forms such as peptides and protein have been shown to be part of the plant N diet. The molecular mechanisms enabling for plants to acquire this broad range of N compounds have at least partly been described. Focussing on amino acids, several transporters mediating root acquisition of these compounds have been identified, providing tangible evidence for the claims that such compounds may contribute to plant N nutrition. Three questions follow from these findings: What is the composition of the N arriving through diffusion and mass flow at the surface of plant roots? To what extent are plant transporters active and functional in amino acid acquisition from soil? What are the broader implications of amino acid nutrition for a growing plant? Results from a range of studies, encompassing field investigations of soil N, studies of Arabidopsis thaliana growing under controlled conditions and model calculations of N assimilation consequences will be presented. The multifaceted picture of plant N nutrition emerging from the results of these studies will be discussed.

Laboratory of Plant Molecular Physiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Japan

1

Drought, heat, and freezing are environmental conditions that cause adverse effects on the plant growth and the productivity of crops. Expression of a variety of genes is induced by these stresses in a variety of plants. A cis-acting element DRE/CRT plays an important role in regulating gene expression under cold and drought stress conditions. Arabidopsis DRE-binding proteins, DREB1/CBF and DREB2 function as transcriptional activators in cold- and drought-responsive gene expression, respectively. Overexpression of DREB1A in Arabidopsis activated expression of many stress-inducible genes and resulted in improved tolerance to drought and freezing. Recently, we analysed soybean DREB1-type transcription factors (GmDREB1s) and found that the expression of most GmDREB1 genes in soybean was strongly induced by a variety of abiotic stresses and GmDREB1 proteins activate the expression of numerous soybean-specific stress-responsive genes under diverse abiotic stress conditions. DREB2A induces the expression of dehydration- and heat stress-inducible genes under the corresponding stress conditions. Target gene selectivity is assumed to require stress-specific posttranslational regulation, but the mechanisms of this process are not yet understood. We identified Dpb3-1 as a novel DREB2A interactor. The overexpression of Dpb3-1 in Arabidopsis enhanced the expression of a subset of heat stress-inducible DREB2A target genes, whereas dehydration-inducible genes were not affected. Interaction and expression analyses suggested the existence of a trimer composed of Dpb31 and NF-Y subunits that could synergistically activate the heat stress-inducible gene with DREB2A, which suggests that the identified trimer contributes to the target gene selectivity of DREB2A under heat stress conditions.

[29] REACTIVE OXYGEN SPECIES IN PLANT SIGNALING: SUBCELLULAR COMPARTMENTS AND SPECIFICITY Jaakko Kangasjärvi

1

[30] MOLECULAR BASIS OF VARIATION IN STOMATAL RESPONSIVENESS TO ELEVATED CO2 Karin S. L. Johansson,1, Mohamed El-Soda,2, Mats X. Andersson,1, Anders K. Nilsson,1, Hoda Sadraei,1, Johan Uddling Fredin1 Department of Biological and Environmental Sciences; University of Gothenburg, Sweden, 2 Department of Genetics; Faculty of Agriculture; Cairo University, Cairo 1

1

Division of Plant Biology, Department of Biosciences, University of Helsinki, Finland

Effective responses to both external and internal stimuli will ensure optimal plant growth and survival in an environment where productivity and product quality are adversely affected by biotic and abiotic stresses. Plants must have effective means of defending themselves against invading pathogens and adapting to changes in their environment. The main features of such defense measures involve early recognition and perception of the developing stress, and subsequent activation of induced adaptive and defensive responses leading to both local and systemic resistance. Reactive oxygen species (ROS) that are formed in plant cells as a response to several stresses in several subcellular compartments may be one of the factors that contribute to, and regulate plant stress sensitivity/tolerance and adaptation and acclimation to it through the regulation of nuclear gene expression. Strong evidence has accumulated that ROS play an important role and have specificity for both the species of reactive oxygen and the subcellular localization of the production. The recognition of a stress is followed by involvement of a small number of signal transduction pathways. They also seem to control and potentiate each other’s activities, indicating that cross talk between these pathways may be very common in gene regulation for acclimation and defense. Furthermore, interaction between different subcellular compartments, e.g., apoplast and chloroplasts in ROS-dependent regulation of gene expression is evident; therefore, it seems obvious that combinations of signal molecules and interaction of ROS signaling between different subcellular compartments might direct the specificity in activation of nuclear genes involved in acclimation and defense responses. I will discuss the role of ROS in intra and intercellular signaling involving apoplast-to-chloroplast-to-nucleus signals and especially tne role of the nuclear RCD1 protein on these processes and in nucleus-chloroplast interactions and communication.

Stomata balance the plant’s uptake of CO2 against the loss of water vapour. As drought stress is an increasing problem for agriculture in many parts of the world, an improved understanding of stomatal regulation may have important implications for securing food production in a future climate. Stomata of most plant species close partially in response to elevated CO2 concentrations, thereby reducing the water consumption of the plant and improving its water use efficiency. However, there is a large variation in the magnitude of this response among species and ecotypes. The genetic basis for this variation in stomatal responsiveness is poorly understood, as are the mechanisms for sensing and responding to CO2. In the current study, we have measured the short-term stomatal response to elevated CO2 in a population of Arabidopsis thaliana recombinant inbred lines (RILs) originating from a cross between two parental genotypes showing a large difference in stomatal responsiveness. Hence, this RIL population showed a broad range in stomatal responsiveness. Using quantitative trait locus (QTL) mapping we have identified a genetic locus with a strong effect on stomatal responsiveness to elevated CO2. Candidate genes regulating the stomatal CO2 response will be discussed.

[31] NAC TRANSCRIPTION FACTOR RAF CONTROLS SENESCENCE AND PLANT DEVELOPMENT UNDER ABIOTIC STRESS CONDITIONS IN ARABIDOPSIS THALIANA

[32] FUNCTIONAL SIGNIFICANCE OF OXYLIPIN INDUCED HEAT SHOCK PROTEIN ACCUMULATION IN ARABIDOPSIS THALIANA

Bernd Müller-Roeber,1, Iman Kamranfar,1, Gang-Ping Xue,2, Salma Balazadeh1

Miriam Münch,1, Hsin Chih-Hsuan,1, Stingl-Sinn Nadja,1, Martin J. Müller1

1

University of Potsdam, Institute of Biochemistry and Biology; Max-Planck Institute of Molecular Plant Physiology, Germany, 2 3CSIRO Plant Industry, Australia

1

Adverse environmental conditions such as salinity stress, high temperature and drought limit plant growth and development and typically lead to precocious tissue degeneration and leaf senescence, a process through which nutrients from photosynthetic organs are recycled for the formation of flowers and seeds to secure reaching the next generation under such harmful conditions. Although insight into age-dependent plant aging processes have been unravelled in recent years, the molecular pathways through which abiotic stress affects senescence-triggering gene networks are currently only vaguely defined. We discovered a NAC transcription factor (TF), tentatively called RAF, which plays a central role in abiotic (drought, salinity) stress-triggered senescence and the control of developmental adaptations to stressful environments. RAF is an ABA-responsive TF; RAF overexpressors are hypersensitive to ABA and exhibit precocious senescence while knock-out mutants show delayed senescence. To explore the RAF gene regulatory network, we determined its preferred binding sites by binding site selection assay (BSSA), and performed microarray-based expression profiling using inducible RAF overexpression lines and chromatin immunoprecipitation (ChIP)-PCR. Our studies identified several direct target genes, including several known senescence-associated genes (SAGs), genes linking RAF to salinity and ABA signaling, and genes affecting leaf and root development. Based on our results we conclude that RAF functions as a central transcriptional regulator that coordinates developmental programs with stress-related inputs from the environment. To explore the potential agricultural applications of our findings, we are currently extending our studies towards crop species.

Literature evidence suggests that oxylipins confer basal thermotolerance in Arabidopsis. Elevated temperatures (37°C) have been shown to induce an accumulation of the jasmonates 12-oxo-phytodienoic acid (OPDA), jasmonic acid (JA) and jasmonic acid isoleucine (Ja-Ile). In addition, the Ja-Ile insensitive mutant coi1-1 has been reported to be compromised in basal thermotolerance. Previously, we have shown that exogenously administered OPDA but not JA triggers heat shock protein (HSP) accumulation in the absence of heat suggesting OPDA rather than JA might be a signal involved in thermoregulation.

University of Wuerzburg, Germany

OPDA, but not JA, can be categorized as an “Reactive Electrophile Species” (RES)-oxylipin that induces detoxification and general stress genes, many of which (40 %) are dependent on TGA2,5,6-transcription-factors. In contrast, OPDA- and heat-responsive HSFs and HSP genes are TGA-independent. We show that OPDA induced up-regulation of HSPs are mediated through the master heat shock transcription factor genes HSFA1 and partly through HSFA2. However, short or long term elevated temperatures (37°C) did not trigger jasmonate accumulation. Moreover, a mutant, dde2, deficient in jasmonate biosynthesis, displayed normal heat-induced HSP accumulation and a wild-type like phenotype in different thermotolerance assays. Hence, jasmonates appear not to be involved in thermoregulation. Notably, a variety of abiotic and biotic stresses are associated with dramatically increased OPDA levels and HSP accumulation potentially increasing general stress resistance. Our results indicate that OPDA is as efficient as classical chemical HSP90 inhibitors in inducing HSPs. In the future, we aim to further clarify the OPDA signal transduction mechanisms and the functional significance of the HSP-stress-response.

[33] CHLOROPLAST ANTIOXIDANT DYNAMIC CONTROL ROS-SIGNALING UPON REPETITIVE COLD STRESS

[34] Who represents plant scientists at European level, how can you have a say and get involved?

Joern van Buer,1, Jelena Cvetkovic,1, Ilona Juszczak,2, Ellen Zuther,3, Dirk Hincha,3, Margarete Baier4

Karin Metzlaff, EPSO, Belgium

FU Berlin, Institute of Biology, DCPS, Plant Physiology, Germany, 2 Institute of Molecular Physiology and Biotechnology of Plants; University of Bonn, Germany, 3 Mpi of Molecular Plant Physiology, Germany, 4 Plant Physiology; FU Berlin; DCPS, Germany 1

Temperature variation causes transient photostatic imbalances and generation of reactive oxygen species (ROS) in chloroplasts due to stronger temperature effects on the assimilation reactions than on the photosynthetic electron transport processes. As part of the acclimatization process, the chloroplast antioxidant capacity adjusts within a few days via regulation of nuclear gene expression. In response to mild cold (10 °C) and mild heat stress (30 °C), expression of 2-Cys peroxiredoxin A (2CPA), glutathione peroxiredoxin 7 (GPx7), stroma ascorbate peroxidase (sAPx) and dehydroascrobate reductase (DHAR) increased, while the expression of thylakoid-bound ascorbate peroxidase (tAPx) decreased in Arabidopsis thaliana during acclimatization. Expression of the other components was gradually regulated. While the acclimatization response controls ROS levels during the cold, the cold-induced shift from thylakoid to stroma protection results in a stronger release of superoxide release and weaker H2O2 accumulation when the plants are transferred back to normal temperatures (20 °C). The ROS signature and the expression intensity of most genes for antioxidant enzymes are re-stabilized within 3 days. The priming effect of the first stress stimulus impacts on activation of ROS signaling cascades much longer. Analysis of cold-treated plants demonstrated that a 24 h cold stimulus is sufficient to prepare plants for several days for additional stresses. Priming limits costs into stress signaling and improves plant photosynthetic performance upon light intensity fluctuations.

EPSO, the European Plant Science Organisation, ensures a future for plant research in Europe. Evidence will be presented on the achievements of EPSO in the European Framework Programmes 6, 7 and Horizon 2020 – you know the actions – do you know who invented them? How can you get more involved? EPSO activities in the coming years most relevant for Scandinavian scientists include examples such as: •

F oster inclusion of basic and applied plant research in the Horizon 2020 Societal Challenge programmes / projects

•

E ngage in the EPSO Working Groups to further your science and support the respective policy at European as well as national level

•

 ouble the number of supporting scientists and include mid-career scientists in there to increase D interaction among EPSO members

•

J oin the Fascination of Plants Day – over 960 events were organised in 2015 in 56 countries across the world

•

J oin education proposals based on the Plants for the Future ETP’s Education Action Plan, published 24.3.2015

These and more issues around the future for plant science in Europe will be discussed. Websites: www.epsoweb.org and www.plantday.org

[35] PLANTS, POLITICS AND PUBLIC PERCEPTION Stefan Jansson1 1

Umeå Plant Science Centre, Department of Plant Physiology, Umeå University, Sweden

Publicly funded scientists have got the privilege to use taxpayers’ money to perform what we want; to independently perform research. But taxpayers should, in return, demand that we contribute to make the society better. Compared to scientists involved in applied research, those who work in basic science may find it harder to convincingly show that they work for the public good, our task is to inform public about progress in our field and public events like “Fascination of plants day” and other popular science activities is one important way to inform – perhaps even entertain – society. Public participation in scientific research (PPSR) including “citizen science” is another way of interaction, we have for example engaged schools in monitoring tree phenology. However, we are also obliged to inform decision-makers how they should act to ensure that knowledge is used to the benefit of society. The biggest hurdle in practical application of plant research is the resistance against genetically modified (GM) plants. Science has provided compelling evidence the GM plants are as safe as others and unless they are allowed to be use it will be harder to meet two of the eight Millennium Development Goals; “Eradicate extreme poverty and hunger” and “Ensure environmental sustainability”. Many politicians are aware of this but cannot, in a democracy, really change policy unless public perception changes. Therefore it is our responsibility to inform the public about the pros and cons of GM plants and I will describe how this may be done.

[36] THE GATSBY PLANT SCIENCE SUMMER SCHOOL Celia Knight1 1

The Sainsbury Laboratory; University of Cambridge, United Kingdom

In the UK, there have been growing concerns that a future workforce will be insufficiently trained in plant science to address global issues such as food and energy security in the face of climate change 1. In 2004, The Gatsby Charitable Foundation began funding an annual summer school to enthuse first year undergraduate students about plant science and its career opportunities 2. The school was established at The University of Leeds and is now part of the Gatsby Plant Science Education programme based at The University of Cambridge. An analysis of the impact of the school was published in 2012 3, showing that in its first 4 years students who attended the summer school were almost 4 times more likely to enter plant science-related PhDs than their peers who did not. The Gatsby summer school model shows that a well-considered intervention at the start of an undergraduate study programme can change student attitudes to plant science. This talk will present the elements of the summer school that contribute to its success and invite discussion as to whether components of this model would be useful if applied elsewhere. 1 UK Plant Science Federation (2014): UK Plant Science: Current status and future challenges. [WWW document] URL https://www.societyofbiology.org/policy/groups-and-committees/ukpsf/aboutukpsf/uk-plant-science-status-report [accessed 24th March, 2015] 2 http://www.gatsby.org.uk/plant-science/programmes/gatsby-plant-science-summer-school 3 Levesley A, Jopson SJ and Knight CD. 2012. The Gatsby Plant Science Summer School: Inspiring the Next Generation of Plant Science Researchers. The Plant Cell April vol. 24 no. 4 1306-1315

[37] FUNCTIONAL OVERLAP AND NICHE SPECIALIZATION OF THE ARABIDOPSIS LEAF AND ROOT MICROBIOTAS

[38] SIGNALS AND RECEPTORS INVOLVED IN ENDOSYMBIOSIS Jens Stougaard1

Paul Schulze-Lefert 1

1

Department of Plant Microbe Interactions, Max Planck Institute for Plant Breeding Research, Germany

We have previously shown that healthy roots of Arabidopsis thaliana, grown in natural soils, are colonized by a bacterial consortium with well-defined taxonomic structure. Members of this root microbiota belong mainly to the phyla Actinobacteria, Bacteroidetes, and Proteobacteria*. A comparison of the bacterial root microbiota of A. thaliana with A. thaliana relatives, grown under controlled environmental conditions or collected from natural habitats, demonstrated a largely conserved microbiota structure with quantitative, rather than qualitative, species-specific footprints. Unlike this, the root microbiota of monocotyledonous barley and dicotyledonous A. thaliana, grown in the same soil type, revealed a similar overall structure, but with several bacterial taxa uniquely enriched in the Brassicaceae*** This suggests bacterial root microbiota structure is an ancient plant trait that was already present in the last common ancestor of monocotyledonous and dicotyledonous plants. We have isolated ~60% of the A. thaliana root-enriched microbiota members as pure bacterial cultures and have generated whole-genome sequence drafts for all microbiota members, enabling systematic analysis of root microbiota functions under laboratory conditions. Using gnotobiotic plant growth systems we show that single microbiota members or synthetic bacterial communities protect the plants against several tested soil-borne fungal pathogens, suggesting that indirect pathogen protection is a physiological function of the bacterial root microbiota. I will also illustrate how we utilize synthetic communities and gnotobiotic plant systems to better understand principles underlying root microbiota establishment.

* Bulgarelli et al., Nature 2012 ** Schläppi et al., PNAS 2014 ***Bulgarelli et al., Cell Host&Microbe, 2015

Centre for Carbohydrate Recognition and Signalling, Department of Molecular Biology and Genetics, Aarhus University, Denmark

1

In the model legume Lotus japonicus there are seventeen LysM receptor kinases that could be involved in perception of microbial signal molecules including lipochito-oligosaccharides and chitin derived signal molecules. The possible role of these LysM type serine/threonine receptor kinases in plant-microbe interaction will be discussed. Rhizobial lipochito-oligosaccharides and surface polysaccharides are involved or required for establishing functional infected root nodules. Genetic and biochemical approaches for investigating the function of LysM receptors in perception of microbial signals will be presented and a model for two-step recognition of rhizobial bacteria highlighted.

[39] PP2A PHOSPHATASE AS A REGULATOR OF ROS SIGNALING IN PLANTS

[40] GLUTAMATE-LIKE RECEPTOR CHANNELS GLR3.3 AND GLR3.4 MODULATE MAMP-DEPENDENT ROOT-TO-SHOOT CALCIUM WAVES

Moona Rahikainen,1, Guido Durian,1, Saijaliisa Kangasjärvi1 1

University of Turku, Finland

Richard Hilleary,1, Won-Gyu Choi,1, Kent Jorgenson,1, Simon Gilroy1 1

Organellar reactive oxygen species (ROS) signalling is a key mechanism that promotes the onset of defensive measures in stress-exposed plants. Previously, a combination of mutant analysis, proteomics and metabolomics revealed that a specific regulatory B’γ subunit of protein phosphatase 2A (PP2AB’γ) controls organellar ROS signaling, metabolic adjustments and biotic stress resistance in Arabidopsis thaliana. Currently, we pursue the mechanisms underlying PP2A-dependent metabolic regulation and ROS signalling in leaves. Visualization of protein interactions, together with selected reaction monitoring (SRM) mass spectrometry showed that one of the targets whose phosphorylation level is controlled by PP2A-B’γ is cytoplasmic ACONITASE 3 (ACO3), a central metabolic enzyme functionally connected with oxidative stress responses and cell death regulation in plants. Elucidation of the physiological significance of ACO3 phosphorylation is underway. Parallel efforts are laid on elucidating a novel regulatory mechanism, where PP2A-B’γ controls the function of a protein kinase to adjust primary metabolism and intracellular ROS homeostasis in plants. Latest developments in these studies will be presented.

Department of Botany, University of Wisconsin, Madison; United States

Plants possess an innate immune system comprised of cell surface receptors that recognize a broad spectrum of pathogen molecules known as microbe associated molecular patterns (MAMPs), including a 22 amino acid epitope of bacterial flagellin, flg22. One of the earliest signaling events following MAMP perception in plants is a transient increase in cytosolic [Ca2+], though little direct molecular genetic evidence has surfaced indicating the identity of a calcium channel involved in this canonical signaling event. Through the use of Arabidopsis thaliana plants expressing the genetically encoded calcium sensor Yellow Cameleon Nano-65, we demonstrate that flg22 not only induces local calcium transients at the root tip, but also initiates a systemic ‘wave’ that travels from the root tip towards the shoot at a rate of 70μm/second. The initiation of this MAMP-dependent calcium wave is severely attenuated in glutamate-like receptor glr3.3 mutants, and the propagation rate of this calcium wave is decreased in both glr3.3 and glr3.4 mutants. Further genetic and pharmacological evidence using glr3.3/glr3.4 mutants and calcium channel blockers suggests this wave relays important systemic calcium signals throughout the plant body to modulate molecular and physiological defense responses, including the induction of defense gene expression and improving resistance to the foliar bacterial pathogen Pseudomonas syringae pv. tomato DC3000. These results present the importance of systemic calcium signaling in plant defense responses and the critical role glutamate-like receptors play in mediating both the local and systemic perception of pathogens in plants.

[41] TRANSCRIPT PATTERNS IN INFECTED VS. UNINFECTED CELLS OF DATISCA GLOMERATA NODULES

[42] THE EXPANDED FAMILY OF IDA- AND PIP/PIP-LIKE PEPTIDES: PEPTIDE LIGAND MOTIFS, EVOLUTION AND STRESS INDUCTION

Salgado M. 1, Demina I.V. 1, Gaude N. 2, Krajinski F. 2, Ziegenhain C. 3, Enard W. 3, Pawlowski K. 1

Ane Kjersti Vie,1, Javad Najafi,1, Bin Liu,1, Per Winge,1, Melinka A. Butenko,2, Karina S. Hornslien,1, Robert Kumpf,2, Reidunn B. Aalen,2, Tore Brembu,1, Atle Magnar Bones1

Department of Ecology, Environment and Plant Sciences, Stockholm University, Sweden, 2 Max Planck Institute for Molecular Plant Physiology, Germany, 3 Department of Biology II, Ludwig Maximilian University Munich, Germany 1

Background: Two types of nitrogen-fixing root nodules are known among angiosperms, legume/rhizobia symbioses and actinorhizal symbioses involving woody shrubs from eight different families and Gram-positive actinobacteria of the genus Frankia. There is only one herbaceous actinorhizal species, Datisca glomerata (Cucurbitales; Datiscaceae). In contrast with the situation in legumes, the signaling process that leads to organogenesis and subsequently to the accommodation of Frankia in cortical cells of the the newly formed root nodules is poorly understood. The morphology of nodules of D. glomerata, where the infected cortical cells form an interrupted domain on one side of the acentric stele, provides an opportunity to separate infected and uninfected cortical cells and compare their respective transcriptomes. Therefore, laser capture-microdissection was applied to obtain samples of mature infected and uninfected cortical cells. Results: For each cell type, two 50 bp lenght Paired-End libraries were prepared in a flow-cell using high-troughput RNA-Seq on a Rapid Mode Illumina HiSeq1500 set up. Reads Demultiplexing were assigned with house scripting and assignRG package. NextGenMap (NGM) was used to map the reads of each demultiplexed library to a PolyA-enriched D. glomerata transcriptome previously assembled. Reads that mapped against the assembly were counted in an in-house R script. Hence, RPKM - Reads per kilobase transcript per million reads - values were calculated. Functional annotation was carried out using the Trinotate pipeline. Conclusion: The comparison between the two transcriptomes allowed us to identify infected cell-specific genes, e.g., nodule-specific defensins (Demina et al. 2013). It also yielded information on the metabolic specialization of infected vs. uninfected cells. Demina IV, Persson T, Santos P, Płaszczyca M, Pawlowski K (2013) Comparison of the nodule vs. root transcriptome of the actinorhizal plant Datisca glomerata: actinorhizal nodules contain a specific class of defensins. PLoS One 8, e72442

CMBG; Department of Biology; Norwegian University of Science and Technology, Norway, 2 Department of Biosciences; University of Oslo, Norway

1

Peptide ligands play crucial roles in the life cycle of plants by modulating the innate immunity against pathogens and regulating growth and developmental processes. One well-studied example is INFLORESCENCE DEFICIENT IN ABSCISSION (IDA), which controls floral organ abscission and lateral root emergence in Arabidopsis thaliana. IDA belongs to a family of five additional IDA-LIKE (IDL) members that have all been suggested to be involved in regulation of Arabidopsis development. Here we present three novel members of the IDL subfamily and show that two of them are strongly and rapidly induced by different biotic and abiotic stresses. Furthermore, we provide data that the recently identified PAMP-INDUCED SECRETED PEPTIDE (PIP) and PIP-LIKE (PIPL) peptides, which show similarity to the IDL and C-TERMINALLY ENCODED PEPTIDE (CEP) peptides, are not only involved in innate immune response in Arabidopsis but are also induced by abiotic stress. Expression patterns of the IDA/IDL and PIP/PIPL genes were analysed using in silico data, qRT-PCR and GUS promoter lines. Transcriptomic responses to PIPL3 peptide treatment suggested a role in regulation of biotic stress responses and cell wall modification.

Key words: Arabidopsis, gene expression, INFLORESCENCE DEFICIENT IN ABSCISSION, peptide ligand, evolution

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[43] TARGETED MUTAGENESIS IN SILENE LATIFOLIA USING THE TALENS AND THE CRISPR/ CAS9 NUCLEASES

[44] ANALYSIS OF THE MOLECULAR MECHANISMS REGULATING CELL FATE IN SHAPING PLANT ROOT ARCHITECTURE

Vojtech Hudzieczek,1, Radim Cegan,1, Roman Hobza,1, Boris Vyskot1

Therese Lasses,1, Laszlo Bako1

1

Institute of Biophysics, The Czech Academy of Sciences, Czech Republic

Silene latifolia is a dioecious plant model for studies of sex chromosome evolution, sex determination, sexual dimorfism, sexually transmitted diseases, heavy metal tolerance and invasive species biology. With expansion of massive parallel sequencing during the past decade, a number of candidate genes for important biological traits increased dramatically. However, a functional evaluation of these genes remains complex issue as S. latifolia is considered recalcitrant to Agrobacterium tumefaciens-mediated transformation and to in vitro plant regeneration. Using modern synthetic cytokinin analogs we have established a protocol for in vitro regeneration and selected an ecotype with the highest regeneration rate. Next, we tested several Agrobacterium strains in order to develop an efficient protocol for genetic transformation - the transient and stable transgene expression were assessed by using GUS reporter gene. To study the molecular basis of dioecy in S. latifolia, we designed and assembled the TALENs and the CRISPR/Cas9 nucleases to target coding region of sex-linked genes. To test the efficiency of our constructs, we expressed the nucleases in Silene mesophyll protoplasts and measured the proportion of knock-out alleles by next-generation amplicon sequencing. Our experience with creating targeted mutations using the TALENs and the CRISPR/Cas9 technology in Silene latifolia will be presented.

1

Department of Plant Physiology, Umeå Plant Science Centre, Sweden

In an ever-changing climate, tight control and regulation of development is paramount for plants’ fitness and survival. Shaping of the plant root architecture in response to developmental and environmental signals is crucial for adaptation. Our work aims to better understand the molecular mechanisms underlying cell fate decisions and organogenesis. In this project we focus on a novel chromatin level repressor mechanism that seems to be key for the initiation of lateral root (LR) formation. Our data indicate that repression is established by a complex comprising proteins known to have roles in chromatin remodeling, cell division and differentiation. Using Arabidopsis thaliana as a model system we try to develop our understanding of this repressor-complex’s inherent subunits, targets and regulation. To address the mechanism targeting the repressor complex to gene promoters we performed an Y1H screen for root-specific DNA binding factors potentially acting as part of the complex. The screen identified a set of transcription factors (TFs) and preliminary phenotyping data of TF T-DNA insertion lines indeed show significant changes in LR density that support a potential role in the regulation of LR formation. Next to be assessed are the TFs’ expression profiles, whether they are part of the repressor-complex or not and if they do indeed play a role in regulating LR development. Welcome to my poster where I will present more background and results of the project as well as discuss the future plans we have to try to unravel this interesting story.

[45] JUB1, A CENTRAL REGULATOR OF PLANT GROWTH AND DEVELOPMENT UNDER STRESS CONDITIONS

[46] CHARACTERIZATION OF THE BALANCE BETWEEN ASEXUAL AND SEXUAL REPRODUCTION IN DIPLOID STRAWBERRY

Sara Shahnejat Bushehri,1, Saghar Ebrahimian Motlagh,1, Annapurna Devi Allu,1, Venkatesh Periyakavanam Thirumalai Kumar,1, Bernd Müller-Röber,1, Salma Balazadeh1

Tracey Tenreira,1, Hernould Michel,1, Denoyes Béatrice1 1

UMR1332 Biologie du Fruit et Pathologie; INRA Bordeaux, France

University of Potsdam, Institute of Biochemistry and Biology; Max-Planck Institute of Molecular Plant Physiology, Germany 1

Abiotic stresses cause adverse effects on plant growth and development. Reduction in shoot growth followed by accelerated senescence is a typical plant adaptive response to stress. However, the regulatory mechanisms by which plants integrate stress-derived signals into the growth and developmental program are virtually unknown. Recently, we reported that the Arabidopsis thaliana NAC transcription factor JUNGBRUNNEN1 (JUB1) extends longevity and confers tolerance to various abiotic stresses in part by dampening the cellular H2O2 level. We have shown that JUB1 regulates the cellular H2O2 homeostasis network and primes the plants for upcoming stress through direct regulation of DREB2A, which in a transcription factor regulatory cascade is an upstream regulator of the heat shock factor HSFA2 gene. We now discovered a novel function of JUB1 in balancing growth and stress responses in Arabidopsis. We showed that JUB1 plays an important role in reducing growth hormone (gibberellic acid and brassinosteroid) biosynthesis, which leads to the stabilization of DELLA proteins thus conferring resistance towards various abiotic and biotic stresses. Our results therefore suggest that the JUB1-regulatory network exerts its profound effect on stress tolerance by integrating hormonal and ROS signalling pathways. Furthermore, using phylogenetic footprinting analysis, promoter deletion analysis and yeast-one-hybrid screens we identified several transcription factors as JUB1 upstream regulators that further support JUB1´s role as a central hub for controlling growth under conditions of abiotic stress. Recent data will be presented.

Strawberry (Fragaria sp.) represents an interesting model for studying the relationship between sexual and asexual plant reproduction in polycarpic perennial plants. Strawberry produces both inflorescences and stolons (also called runners), which are lateral stems growing at the soil surface and producing new ramets. Another type of vegetative reproduction occurs through the production of new lateral stems called crowns, which develop in the basal parts of the primary crown and give birth to new crowns with terminal flowering. Flowering behaviour varies from once flowering (OF, flowering once per year during spring) to perpetual flowering (PF, continuous flowering all along the vegetative period) depending on genotypes. To test the hypothesis of a competition between flowering and runnering processes, we have studied a population that segregates for both traits, during the growing season for two years. Firstly, we examined quantitatively the relationship between flowering, runnering and crowns all along the vegetative period, using physiological analyses. Using a clustering approach performed on the 38 perpetual flowering and runnering individuals, we identified different patterns for runnering which depend on the intensity and period of perpetual flowering. Secondly, we examined this relationship using quantitative genetic approach. QTLs linked to the measured traits, emergence of inflorescences, runners and crowns, were identified and interactions were observed. Our results suggest a balance between sexual and asexual plant reproduction.

[47] CHARACTERIZATION OF AN ABA-REGULATED GRAS TRANSCRIPTION FACTOR IN RICE ROOTS

[48] THE ROLE OF EVE1, A NOVEL UBIQUITIN FAMILY PROTEIN, IN SHOOT APICAL MERISTEM DEVELOPMENT OF ARABIDOPSIS

Hay Ju Han,1, Ki Sun Choi,1, Beom Gi Kim,1, Myung Ok Byun,1, In Sun Yoon1

Hoyeun Kim,1

1

Molecular Breeding Division, National Academy of Agricultural Science, RDA, Rep. of South Korea

The growth of young rice root is highly susceptible to environmental stress that involves complex hormonal regulatory networks. ABA and GA antagonistically regulate the growth of rice roots, and GA treatment partially alleviated the growth inhibition of rice roots by ABA or salt stress. By microarray analysis, we have identified ABA-regulated GRAS members in rice roots, the transcriptional regulators of GA signaling pathway. One of the HAM (Hairy Meristem) subfamily members, OsGRAS1, was found to be highly suppressed by ABA and salt stress in rice roots. The OsGRAS1 gene was down-regulated in transgenic rice over-expressing an ABA receptor OsPYL5, indicating that OsGRAS1 is under the control of ABA pathway. The OsGRAS1 protein localized to nucleus and showed transcriptional activation activity. Constitutive expression of OsGRAS1 in transgenic rice increased elongation of primary roots and shoots, suggesting that OsGRAS1 is a positive player of growth control in rice. Supported by grants (SSAC PJ01105103 and PJ01001501) from RDA.

1

Department of Biological Sciences; Seoul National University, Rep. of South Korea

To produce the successful fruit for next generation, shoot growth and development are very important in plants. However, the transition from vegetative to reproductive phase of shoot development is little known. In this study, we report a dominant gain-of-function mutant, eve1-D (eternally vegetative phase1-Dominant), which shows the wavy margin rosette leaves and arrested shoot development. The EVE1 gene encodes a novel ubiquitin family protein containing an ubiquitin domain. EVE1 is expressed very low level in all of the organs. In the eve1-D mutant, the primary shoot apical meristem develops normally during the vegetative stage, however the SAM is not initiated into floral primordial. The eve1-D only shows the axillary SAMs and interrupts the inflorescence development of adult plants, when the wild-type begins to bolt and generates the seeds. Our results explain a role of EVE1 in shoot development for the phase transition from a vegetative to a reproductive stage.

[49] DEVELOPMENT CROSSTALK OF RICE SEEDS IN RESPONSE TO SEED STORAGE PROTEIN STARVATION

[50] IDENTIFICATION AND CHARACTERIZATION OF NEW CO-FACTORS OF THE PLANT MIRNA BIOGENESIS PATHWAY

Young-MI Kim,1, Hye-Jung Lee,1, Yeong-Min Jo,1, Jong-Yeol Lee,1, Sun-Hyung Lim1

Patricia Karlsson,1, Michael D. Christie,2, Danelle Seymour,1, XI Wang,3, Jörg Hagmann,1, Franceli R. Kulcheski,4, Detlef Weigel,1, Pablo A. Manavella5

1

National Academy of Agricultural Science, Rep. of South Korea

Max Planck Institute for Developmental Biology, Germany, 2 Max Planck Institute for Developmental Biology, Germany; Shelston IP, Australia, 3 Max Planck Institute for Developmental Biology, Germany; Bayer Cropscience NV, Innovation Center, Belgium, 4 Max Planck Institute for Developmental Biology, Germany; Centro de Biotecnologia, UFRGS, Brazil, 5 Max Planck Institute for Developmental Biology; Instituto de Agrobiotecnología del Litoral (IAL), Centro Científico Tecnológico Santa Fe (CCT), Argentina

1

Plant storage proteins are exclusively found in seeds, in which they become reserves of amino acids and organic nitrogen for embryonic and developing seedlings. Seed storage protein (SSP) constitutes up to 10~12% of dry weight of seeds in cereal plants, and these proteins are primarily synthesized in aleurone layers surrounding the endosperm of grain. Seed germination is a unique developmental transition from metabolically quiescent seeds to actively growing seedlings. The response of endosperm to germination coordinates nutrient mobilization to support heterotrophic growth until autotrophic photosynthesis is established, which includes hydrolases of the seed reserves, such as nucleic acids, proteins, starch, lipids and mineral complexes. In this study, phenotypic characterization between SSP starvation and seed germination has been investigated particularly in GPGb-RNAi transgenic rice plants, where expressions of SSPs were down-regulated by RNA interference (RNAi) approach. The SSP product was finally accumulated less in dry seeds of GPGb-RNAi transformants than those of wild type. Morphological pattern of starch granules were also altered on dry seeds of transformants. Interestingly, a delayed germination rate was observed in GPGb-RNAi seeds compared with wild type. Gibberellin (GA) known to play as an important hormone in germination process did not improve germination rate of transformants seeds. High concentration of glucose (Gc) is known to inhibit seed germination due to accumulation of abscisic acid, but the suppression induced by exogenous supplement of Gc did not affect germination pattern of transformants. GA- and Gc-insensitive response found in GPGb-RNAi seeds suggests a role of SSP in hormone crosstalk and signal network in regulating nutrient mobilization upon complete of germination in seeds. These data give new aspect of storage protein in the initiation of germination and further eventual productivity of rice.

Plant miRNA biogenesis is a complex process, involving several processing steps that take place in the nucleus, producing a mature miRNA that ultimately gets integrated into the RISC. Numerous proteins involved in miRNA processing and function have been identified. Mutations in the genes encoding these proteins cause a whole range of effects on the plant, from embryonic lethality to minor morphological defects. Recently, a new high-throughput screen for the identification of unknown plant miRNA biogenesis cofactors was developed in our lab. The combination of a luciferase-based reporter system and state-of-the-art sequencing technologies enables the rapid identification of mutant genes in the miRNA pathway. Among the isolated mutants we successfully mapped, using SHORE mapping analysis, the causal mutation of one of the alleles to a gene still uncharacterized in the miRNA context. The identified gene named MSS8 (miRNA-mediated silencing suppressor 8) encodes a KH-containing protein that is predicted to be RNA-binding. Confocal microscopy experiments indicated that MSS8 co-localizes with the miRNA biogenesis machinery while yeast-2-hybrid assays showed that the protein is able to interact with known miRNA-processing factors. The mutant plants showed weak alterations of the levels of pre-miRNAs, mature miRNAs and miRNA targets in mature tissues whereas a very pronounced alteration in the miRNA and target levels was observed in shoot apical meristem enriched samples. Small RNA sequencing and in situ hybridization confirmed MSS8 as a tissue-specific co-factor of the pathway.

[51] THE VERNALIZATION GENE FRIGIDA IN THE FAMILY BRASSICACEAE

[52] PINOID AND LEAF DEVELOPMENT, AN INTIMATE RELATIONSHIP

Oksana Fadina,1, Emil Khavkin1

Kumud Saini,1, Marios Nektarios Markakis,1, Els Prinsen,1, Gerrit T.S. Beemster,2, Kris Vissenberg1

1

Institute of Agricultural Biotechnology, Russia

In the Brassicaceae, FRIGIDA up-regulates expression of the floral repressor FLOWERING LOCUS C. Allelic polymorphisms of these genes contribute to plant adaptation to climate changes; however, opinions vary concerning FRIGIDA polymorphisms in QTLs for flowering time, FRIGIDA latitudinal clines, etc. The structural analysis of FRIGIDA polymorphisms may help solve this problem and elucidate FRIGIDA role in the Quaternary evolution of vegetation. We cloned FRIGIDA from six Brassica species comprising the triangle of U and found that gene structure was orthologous to that in Arabidopsis thaliana. Compared with A. thaliana, FRIGIDA protein in Brassica species lacked the coiled-coil domain in the N-terminal region and differed in the number and structure of specific repeats in C-terminus. Unlike A. thaliana, FRIGIDA in Brassica species was represented by two loci, FRI.a and FRI.b (81% homology). All FRIGIDA sequences from diploid genomes were highly conserved in the corresponding subgenomes of tetraploid species. Analysis of nucleotide diversity of FRIGIDA indicated that one part of the gene was under positive selection, whereas another was under purifying selection. In the context of genome evolution in the Brassicaceae, phylogenetic analysis of all annotated and nonannotated FRIGIDA sequences produced two clusters corresponding to the lineage I and lineage II. However, presently two FRIGIDA loci were found only in lineage II (Brassica and presumably Raphanus and Sinapis), whereas in the genera Arabidopsis, Camelina and Capsella (lineage I) and Eutrema (lineage II), one of two FRIGIDA loci was apparently lost. Three FRIGIDA loci in Camelina sativa suggest retention of the triplicated gene.

Univ. Antwerpen; Lab of Plant Growth and Development, Belgium, 2 Univ. Antwerpen; Lab of Molecular Plant Physiology and Biotechnology, Belgium

1

For over a century auxin has attracted attention of researchers because it is involved in almost all biological processes in plants. To govern and control responses mediated by auxin, its signaling cascade and polar transport are of particular significance. Auxin transport depends on diffusion or AUX1-mediated uptake into cells and efflux by specialised efflux-proteins, PINs and PGPs. PINOID (PID) protein is a major player in the polar recruitment of PINs on the membranes, thus greatly impacting on auxin transport. We try to answer how different expression levels of PID acclimate polar auxin transport and how this affects leaf growth and development in Arabidopsis thaliana. To approach this several T-DNA insertion mutants were studied next to two overexpression lines (Ox) of PID, aiming to extend the existing knowledge of leaf development by focusing on two important growth processes, cell proliferation and expansion. A complementary set of experiments included a detailed kinematic growth analysis, detection of changes in endoreduplication by flowcytometry, free and conjugated indole acetic acid (IAA) measurements, DR5-visualized auxin response localisation and RNA sequencing on PID-Ox lines. Several leaf-related phenotypes were detected in both knock-out and overexpression lines, and they occurred in all three growth dimensions, suggesting that they are due to alterations in auxin transport in the different lines. This was backed up by distinct changes in free and conjugated-auxin levels and increased auxin responses. Next Generation Sequencing pointed to converging pathways induced by stress conditions like drought and to hormonal cross-talk with ethylene, jasmonic acid and salicylic acid.

[53] RESPONSE MODULES IN THE PLASMA MEMBRANE REGULATING ARABIDOPSIS ELONGATION GROWTH

[54] EFFECT OF COLD PRETREATMENT ON MICROSPORE EMBRYOGENESIS IN TOBACCO (NICOTIANA TABACUM L.)

Klaus Harter,1, Ladwig Friederike,1, Dahlke Renate,2, Stührwohldt Nils,2, Sauter Margret2

Edith Stabentheiner,1, Nadine Neumann,2, Regina Willfurth,2, Petra Bacher,2, Klaus Remele,2, Hartwig Pfeifhofer2

1

Center for Plant Molecular Biology / Plant Physiology; University of Tübingen, Germany, 2 Department of Plant Developmental Biology and Physiology; University of Kiel, Germany

University of Graz, Institute of Plant Sciences, Austria; Nawi Graz, Austria 2 University of Graz, Institute of Plant Sciences, Austria

Several growth modulating factors regulate very early phases of cell elongation independent of gene expression. By applying - amongst other methods - different spectro-microscopical techniques such as in vivo one-chromophor lifetime spectroscopy and high resolution FRET-FLIM, we study growth factor-controlled dynamic responses at the cell wall – plasma membrane – cytoplasm interface. Our subcellular analyses suggest that different response modules exist in the plasma membrane, which consist not only of receptors for growth-modulating factors but also of other proteins such as proton pumps. These functional units appear to directly translate signal perception into defined cellular responses. On the basis of some examples, our findings and conceptual ideas will be discussed.

Male microspores can be reprogrammed to shift from the gametophytic to the embryogenic pathway – a process called “embryogenesis” or “androgenesis” resulting in haploid (doubled haploid) embryos instead of mature pollen. This developmental shift is mediated by stress. Doubled haploid plants are not only important for plant breeding programs, but embryogenic microspores provide a good model system to study basic concepts of developmental biology. However, recalcitrant species, low efficiency, and often bad reproducibility hamper these studies. Cold stress is often used as external trigger for microspore embryogenesis. A better knowledge of the processes going on during cold pretreatment of anthers will help to understanding and controlling this developmental shift. Young flower buds of tobacco (Nicotiana tabacum L.) with microspores in the late uninucleate to early binucleate stage were exposed to low temperatures (main emphasis: 4°C and 8°C) prior to microspore isolation and cultivation. The cold treatment significantly influenced microspore reprogramming, and distinct differences with regard to the temperature could be observed. Whereas microspores from anthers exposed to 4°C showed a higher percentage of pollen maturation (germination) and formation of undifferentiated calli, a treatment of buds at 8°C resulted in a significantly higher rate of embryo formation. Physiological parameters measured mainly in anthers (content of phytohormones, components of the antioxidative system) also revealed differences related to the different pretreatment temperatures and their importance for microspore embryogenesis will be discussed.

1

[55] MEMBRANE PROTEOMICS OF NORWAY SPRUCE RELATED TO LIGNIN FORMATION Enni Väisänen,1, Ogonna Obudulu,2, Joakim Bygdell,3, Junko Takahashi,4, Olga Blokhina,5, Kurt Fagerstedt,5, Gunnar Wingsle,2, Anna Kärkönen6 Department of Biosciences, Division of Plant Biology, University of Helsinki, Finland; Department of Agricultural Sciences, University of Helsinki, Finland, 2 Department of Forest Genetics and Plant Physiology, SLU, Umeå Plant Science Centre (UPSC), Umeå, Sweden, 3 Department of Chemistry, Computational Life Science Cluster (CLIC), Umeå University, Sweden, 4 Department of Biosciences, Division of Plant Biology, University of Helsinki, Finland; Department of Forest Genetics and Plant Physiology, Slu, Umeå Plant Science Centre (UPSC), Umeå, Sweden, 5 Department of Biosciences, Division of Plant Biology, University of Helsinki, Finland, 6 Department of Agricultural Sciences, University of Helsinki, Finland 1

The vessels and tracheids of water-transporting xylem tissue of vascular plants have strong secondary cell walls. Their strength is greatly affected by the macromolecule lignin that is a phenolic polymer of monolignols. The biosynthesis of lignin includes monolignol biosynthesis from phenylalanine via the phenylpropanoid pathway, transport of these precursors to the cell wall, monolignol oxidation, and finally polymerization into lignin. The final steps of lignification, the transport, oxidization, and polymerization of monolignols, occur at the plasma membrane or in the cell wall. Therefore, understanding the roles of plasma membrane-localized proteins involved in this process is of vital importance [1, 2, 3]. We are currently studying the total membrane proteome of developing xylem, phloem, and a lignin-forming tissue culture of Norway spruce (Picea abies) in order to identify candidates for lignification-related membrane proteins such as monolignol transporters.

References: [1] Miao Y, Liu C. 2010. PNAS, USA 107, 22728-22733. [2] Alejandro S, Lee Y, Tohge T, et al. 2012. Current Biology  22, 1207–1212. [3] Lee Y, Rubio MC, Alassimone J, Geldner N. 2013. Cell 153, 402–412.

[56] BIOSYNTHESIS OF CAROTENOIDS DURING BILBERRY (VACCINIUM MYRTILLUS L.) FRUIT DEVELOPMENT Katja Karppinen,1, Laura Zoratti,1, Marian Sarala,1, Elisabete Carvalho,2, Jenni Pukki,1, Helmi Mentula,1, Stefan Martens,2, Hely Häggman,1, Laura Jaakola3 Department of Biology, University of Oulu, Finland 2 Fondazione Edmund Mach, Research and Innovation Center, Italy Climate Lab/Holt, Department of Arctic and Marine Biology, Uit The Artic University of Norway, Norway; Norwegian Institute for Agricultural and Environmental Research, Bioforsk Holt, Norway

1

3

Bilberry (Vaccinium myrtillus L.) fruits are rich with anthocyanin pigments but they also contain other bioactive compounds, such as carotenoids. In the present study, biosynthesis of carotenoids was studied in different stages during bilberry fruit development and ripening and as response to light treatments. Eight key carotenoid biosynthetic genes (PSY, PDS, ZDS, CRTISO, LCYE, LCYB, BCH, ECH) were isolated from bilberry and their expression was analyzed with qRT-PCR. Composition of carotenoids was analyzed from the same samples with HPLC-MS. The most abundant carotenoids in bilberry fruit were lutein and β-carotene accompanied by minor amounts of xanhophylls such as neoxanthin, violaxanthin and zeaxanthin. The expression of the carotenoid biosynthesis genes showed increase in transcript levels of phytoene synthase (VmPSY), phytoene desaturase (VmPDS), carotenoid isomerase (VmCRTISO) and lycopene β-cyclase (VmLCYB) at the onset of fruit ripening. However, the increase in the expression did not lead to the accumulation of carotenoids during ripening in ripe berries and the carotenoid levels decreased during the fruit development. Light conditions during fruit development affected specifically the expression of PSY and LCYB. The results suggest that the carotenoid levels in bilberry fruit are determined by their biosynthesis and degradation of carotenoids to apocarotenoids such as abscisic acid.

[57] GH5 ENZYMES INVOLVED IN PLANT DEVELOPMENT

[58] FVTFL1 INTEGRATES TEMPERATURE AND PHOTOPERIOD SIGNALS TO SUPPRESS FLOWERING OF FRAGARIA VESCA

Yang Wang,1, Shoaib Azhar,2, Rosaria Gandini,3, Christina Divne,3, Ines Ezcurra,1, Henrik Aspeborg1 KTH Royal Institute of Technology, School of Biotechnology, Division of Industrial Biotechnology, Sweden, 2 Wallenberg Wood Science Centre, Department of Fiber and Polymer Technology, KTH Royal Institute of Technology, Sweden, 3 KTH Royal Institute of Technology, School of Biotechnology, Division of Industrial Biotechnology, Sweden; Karolinska Institute, Department of Medical Biochemistry and Biophysics, Sweden 1

In higher plants, enzymes belonging to glycoside hydrolase family 5 (GH5) are active during various developmental processes, but a detailed understanding of their hydrolytic role and participation in cell wall metabolism is lacking. In most plant genomes three different GH5 subfamilies are represented. The most well explored subfamily is GH5_7 containing mannanases, but actually, the catalytic properties of only a few plant mannanases have been revealed. There is only a single characterized enzyme in GH5_14, whereas nothing is known about the enzymes found in subfamily GH5_11. In order to elucidate the function of plant GH5 enzymes, we use a mainly biochemical approach to study the mode of action of these enzymes. Several Arabidopsis GH5_7 mannanases have been generated as recombinant proteins for enzymatic characterization. Distinct differences between the target enzymes were observed when we investigated performance of both hydrolysis and transglycosylation, and our analyses demonstrate diversification of catalytic function among members in the Arabidopsis GH5_7 subfamily. Thus, we begin to understand why Arabidopsis needs several mannanases during its life cycle, and the results also suggest putative roles for these enzymes in Arabidopsis development. Characterizations of plant GH5_14 and GH5_11 enzymes are currently underway.

Marja Rantanen,1, Takeshi Kurokura,2, Panpan Jiang,3, Katriina Mouhu,4, Timo Hytönen4 University of Helsinki; Natural Resources Institute Finland, Finland, 2 Faculty of Agriculture, Utsunomiya University, Japan 3 University of Köln, Germany, 4 University of Helsinki, Finland

1

Photoperiod and temperature interact to control seasonal flowering in perennials, but the mechanisms are largely unknown. Fragaria vesca is a perennial, facultative short day (SD) plant. Flower induction requires SD at intermediate temperatures. At cool temperatures flower induction occurs independently of photoperiod whereas high temperature represses flower induction both in SD and long day (LD)1. Homolog of floral repressor TERMINAL FLOWER1 (FvTFL1) causes seasonal flowering in F.vesca2. FvTFL1 is regulated by F.vesca homolog SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (FvSOC1) that in contrast to Arabidopsis, functions as a floral repressor FvTFL1 dependent manner3. We found that the expression of FvTFL1 correlated with flowering at all temperatures both under SD and LD. Below 13°C FvTFL1 was down-regulated independently of photoperiod, whereas over-expression of FvTFL1 suppressed flowering. At 16°C FvTFL1 was down-regulated only under SD. At non-inductive 23°C FvTFL1 was highly up-regulated. However, silencing of FvTFL1 (RNAi) enabled flower induction at high temperature. The expression of F.vesca homolog of FLOWERING LOCUS T (FvFT1), and FvSOC1 were activated under LD independently of temperature. Critical day length for expression of FvFT1, and flowering was between 14 and 16 hours. In both photoperiods silencing of FvSOC1 resulted in flowering at 16°C, but not at 23°C. Our results indicate that flower induction at intermediate temperatures requires deactivation of FvFT1FvSOC1-FvTFL1 pathway. Cool temperature down-regulates FvTFL1 independently of this pathway. At high temperatures FvTFL1 is activated both in SD and LD by an unidentified factor. We suggest that FvTFL1 integrates photoperiodic and temperature signals to repress flower induction in F.vesca. References: 1Koskela et al. (2012) Plant Physiol. 159, 1043-1054. 3Heide and Sonsteby (2007). Physiol. Plantarum 130, 280-289. 2Mouhu et al. (2013) Plant Cell 25, 3296-3310.

[59] CHROMATIN-LEVEL CO-REGULATION OF CLUSTERED GENES IN STAMEN DEVELOPMENT

[60] ANALYSIS OF ROOT ARCHITECTURE SYSTEM IN POPLAR AND SPRUCE

Johan Reimegård,1, Snehangshu Kundu,2, Ali Pendle,3, Vivian Irish,4, Peter Shaw,3, Naomi Nakayama,5, Olof Emanuelsson,1, Jens Sundström2

Federica Brunoni,1, Irene Perrone,2, Rubén Casanova-Sáez,1, Karin Ljung,1, Catherine Bellini3

KTH-Royal Institute of Technology; Science for Life Laboratory; School of Biotechnology, Division of Gene Technology, Sweden, 2 Department of Plant Biology; Linnean Center for Plant Biology; Swedish University of Agricultural Sciences, Sweden, 3 Department of Cell and Developmental Biology; John Innes Centre, United Kingdom 4 Department of Molecular, Cellular and Developmental Biology; Yale University, United States 5 Institute of Molecular Plant Sciences; University of Edinburgh, United Kingdom 1

Several lines of evidence suggest that co-expression of physically linked genes arranged in operon-like gene clusters occurs much more frequently in eukaryotes than previously anticipated. A bioinformatics pipeline was developed to evaluate large-scale transcriptome datasets for physical clustering tendencies. Independent datasets from male reproductive tissues in Arabidopsis thaliana showed that genes expressed in differentiating stamens tend to form small chromosomal clusters. Clustering may confer a selective advantage as it enables coordinated gene regulation at the chromatin level. Pollen maturation is controlled by MS1 (MALE STERILITY 1), a transcriptional activator that contains a PHD domain; such domains are often linked with changes in chromatin structure. qRT-PCR and mRNA in situ hybridization experiments showed that genes in a sub-set of the clusters become upregulated within 48 hours after MS1 induction. DNA fluorescent in situ hybridization (FISH) combined with structured illumination (SIM) super-resolution microscopy further showed that transcriptional activation of the clustered genes was associated with open chromatin conformation. Stamen development seems to involve transcriptional activation of physically clustered genes through chromatin de-condensation.

Umeå Plant Science Centre, Dept of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, Sweden 2 Istituto Per la Protezione Sostenibile Delle Piante (IPSP-CNR), Italy; Umeå Plant Science Centre, Dept of Plant Physiology, Umeå University, Sweden, 3 Umeå Plant Science Centre, Dept of Plant Physiology, Umeå University, Sweden; Institut Jean-Pierre Bourgin, UMR 1318, Centre de Versailles-Grignon, France

1

Root system architecture (RSA) reflects the spatial configuration of roots of different age and order and is therefore a highly plastic trait that varies both between and within species. RSA can be modulated in several ways: through regulation of primary root or lateral root (LR) growth, through the adventitious root (AR) formation, and through an increase or decrease in root hairs. In Arabidopsis thaliana, it has been showed that similar mechanisms are likely regulating AR and LR development, although the roots originate from different organs. AR initiation is regulated by a transcriptional regulatory module that involves three Auxin Response Factor (ARF) proteins, two of which are also involved in LR initiation. The information gained from the analysis of adventitious root initiation in Arabidopsis prompted us to investigate if similar regulatory mechanisms exist also during adventitious root formation in trees. The role of two transcriptional regulators, ARF8 and ARF17, known in Arabidopsis to act as positive and negative regulators in AR development respectively, was studied in transgenic hybrid aspen (Populus tremula x Populus tremuloides, clone T89). In vitro rooting experiments were performed with transgenic poplars down-regulated for PtARF8 and PtARF17. Furthermore, expression analyses of these genes were carried out on independent poplar transgenic lines in which the PtARF8 and PtARF17 promoters were fused to the GUS reporter gene. We have now started to translate the results in poplar to Norway spruce (Picea abies), which is the economically most important tree species in Scandinavia.

[61] ONTOGENETIC REGULATION IN HORDEUM VULGARE AND ARABIDOPSIS THALIANA MUTANTS LACKING CHLOROPHYLL B

[62] EVOLUTIONARY ASPECTS OF NON-CELL-AUTONOMOUS REGULATION IN ANCIENT TAXA OF VASCULAR PLANTS

Valeriya Dmitrieva,1, Elena Tyutereva,1, Olga Voitsekhovskaja1

Anastasiia Evkaikina,1, Ekaterina Voronina,1, Ksenia Dobryakova,1, Marina A. Romanova,2, Elena Tyutereva,1, Olga Voitsekhovskaja1

1

Komarov Botanical Institute Ras, Russia

1

Recently, it was shown that the stability of pigment-protein complexes of the photosynthetic machinery is an important source of signals for plant cells. These signals carry information not only about environmental conditions, but also about the age of the cell. Over-accumulation of chlorophyll b results in superstabilization of the antenna and in the development of a functional stay-green phenotype based on delayed senescence (Sakuraba et al. 2012). Mutants with reduced levels of chlorophyll b, including mutants which completely lack this pigment, represent a promising but thus far neglected model to study the role of chlorophyll b levels in ontogenetic signaling. We examined the relationship between the levels of chlorophyll b biosynthesis, the stability of antenna complexes and the regulation of flowering and senescence in several Arabidopsis and barley mutants. Financial support of the Russian Scientific Foundation (#14-16-00120) is gratefully acknowledged.

Komarov Botanical Institute Ras, Russia 2 Saint-Petersburg State University, Russia

Plasmodesmata mediate the exchange of information in the form of miRNA, proteins and mRNA between adjacent cells in the course of plant development. This fundamental role of plasmodesmata is well established in angiosperms but has not yet been traced back to the evolutionary ancient plant taxa. Comparative studies on representatives of such taxa including gymnosperms, ferns and lycophytes could shed light on the origin and development on non-cell autonomous regulation of gene expression in plants. The KNOX genes, coding for non-cell-autonoumous homeodomain transcription factors that function as regulators of apical meristems, have been characterized in a large number of angiosperms, and also in some representatives of other taxa. Therefore, the KNOX proteins can be used as models for characterization of cell-to-cell transport. To this end, the patterns of localization of KNOX transcripts, KNOX proteins and of plasmodesmata, respectively, need to be compared in the meristems of the representatives of different taxa. We report on the establishment of the necessary methods for apical meristems of the gymnosperm Picea abies, the fern Ceratopteris richardii, and the lycophytes Huperzia selago and Selaginella kraussiana. Financial support of the Russian Foundation for Basic Research (№13-04-02000) is gratefully acknowledged.

[63] SALT STRESS INDUCES DNA METHYLATION IN ALFALFA (MEDICAGO SATIVA)

[64] EPIGENETIC CONTROL OF BARLEY LEAF SENESCENCE

Abbas Al Lawati,1, Saif Al-Bahry,1, Mahmoud W. Yaish1

Bianka Janack,1, Nicole Ay,1, Andreas Fischer,1, Gunter Reuter,1, Klaus Humbeck1

1

Department of Biology; College of Science; Sultan Qaboos University, Oman

One of the mechanisms that plants can use to adapt to high salinity levels is altering DNA methylation to adjust gene expression at both transcriptional and post-transcriptional levels. Under salt stress, different cultivars can show epigenetic polymorphism at the DNA methylation level and heritable patterns of phenotypic variations that provide raw materials for plant breeding programs aiming to enhance salt tolerance. In this study, Methylation Sensitive Amplified Polymorphism (MSAP) analysis was used to assess the cytosine methylation levels in alfalfa roots exposed to increasing NaCl concentrations (0.0, 8.0, 12.0 and 20.0 ds/m). Eleven indigenous cultivars collected from different geographical locations in Oman were analyzed and a salt-tolerant cultivar was used as a control. The results showed a significant increase in DNA methylation in most genotypes upon exposure to high levels of salinity. Phylogenetic analysis using Amplified Fragment Polymorphism (AFLP) where MspI was used as DNA methylation insensitive endonuclease, and the MSAP where HpaII was used as DNA methylation sensitive endonuclease, showed epigenetic variation within and among alfalfa cultivars when exposed to saline conditions. We found that salinity could prompt global DNA methylation status especially when plants were exposed to a high level of salt (20 ds/m). This increase accounted for 15% (an average across different genotypes) of the total site-specific polymorphic methylated loci in alfalfa as detected by MSAP analysis. Treatment with 5-azacytidine decreased the salinity tolerance in seedlings and the expression of methyl-transferase genes homologues was significantly increased after treatment with salinity. In addition, Identification of polymorphic bands revealed the presence of candidate genes involved in salinity tolerance. Overall, this study showed the importance of DNA methylation in salinity adaptation mechanisms in alfalfa.

1

Martin-Luther-University Halle-Wittenberg; Institute of Biology, Germany

Senescence, the last step of leaf development, involves massive reprogramming of gene expression. Recently, we could show that in Arabidopsis thaliana this step is also controlled by epigenetic mechanisms. We now transferred the methodology to the model crop Hordeum vulgare and analyzed epigenetic control of the gene HvS40, encoding a putative regulator of leaf senescence, which is strongly induced during leaf ageing. We could show that chromatin status at promoter and coding region of this gene changes in a senescence-specific manner. Euchromatic histone modification mark H3K9ac is established at the translational start site and proximal gene body of HvS40 during senescence. In contrast, abundance of the heterochromatic mark H3K9me2 decreases at the same time. In addition to these senescence-specific alterations at associated histones, bisulfite sequencing reveals also a decrease in DNA methylation at one specific CpG motif, 662bp upstream of the translational start site at the HvS40 promoter. While the DNA region upstream of this specific CpG site is heavily methylated in mature and senescent leaves, interestingly no methylation was found at downstream parts of the promoter and gene body at both developmental stages. These results indicate that senescence-specific induction of HvS40 expression involves dynamic mechanisms acting on chromatin status. In addition, alterations in chromatin structure during senescence were analyzed via immunocytology, revealing senescence-specific alterations in spatial distribution of H3K9me2 marks in the nuclei. We now extended these analyses to a genome-wide scale looking at development-specific alterations in the barley epigenome.

[65] CHROMOSOMAL ASSEMBLY OF THE SEQUENCED PHYSCOMITRELLA PATENS GENOME UNCOVERS CONSERVED ANCESTRAL AND DERIVED MADS-BOX GENE SYNTENY

[66] STARCH BIOSYNTHETIC PATHWAY GENES AND THE EVOLUTION OF SORGHUM GRAIN QUALITY

Elizabeth Barker,1, Neil Ashton1

Ian Godwin,1, Bradley Campbell,1, Guoquan Liu,1, Emma Mace,2, Yuri Tusov,1, Edward Gilding,3, Jimmy Botella,1, Bob Gilbert,4, David Jordan4

1

University of Regina, Canada

The University of Queensland; School of Agriculture and Food Sciences, Australia, 2 Qld Department of Agriculture and Fisheries, Australia, 3 The University of Queensland; Institute of Molecular Biosiences, Australia, 4 The University of Queensland; QAAFI, Australia 1

Assembly of the sequenced genome of Physcomitrella patens into 27 mega-scaffolds, presumed to correspond to 27 chromosomes, has confirmed the major predictions of an earlier parsimonious model of expansion of the MADS-box gene family in the Physcomitrella lineage. Additionally, microsynteny has been conserved in the immediate vicinity of some recent duplicates of MADS-box genes. However, comparison of non-syntenic MIKC MADS-box genes and neighbouring genes indicates that chromosomal rearrangements have destroyed shared synteny over longer distances (macrosynteny) around MADS-box genes despite subsets comprising two or three MIKC genes having remained syntenic. In contrast, half of the type I MADS-box genes have been transposed creating new syntenic relations with MIKC genes on extant chromosomes. This implies conserved ancestral synteny of MIKC genes and more recently derived synteny of type I and MIKC genes may be selectively advantageous perhaps enabling shared regulation. Our revised model predicts that the birth rate of MIKC genes in Physcomitrella is higher than that of type I genes. However, this difference is attributable to one early tandem duplication and one early segmental duplication of MIKC genes prior to the two polyploidisations that account for most of the expansion of the MADS-box gene family in Physcomitrella. Furthermore, this early segmental duplication spawned two chromosomal lineages: one with a MIKCC gene, belonging to the PPM2 clade, in close proximity to one or a pair of MIKC* genes and another with a MIKCC gene, belonging to the PpMADS-S clade, characterised by greater separation from syntenic MIKC* genes. Our model has evolutionary implications for the Physcomitrella karyotype.

Sorghum is a major staple cereal with over 500 million people worldwide dependent on it every day in sub-Saharan Africa and India. Through its domestication in Africa, there has been significant narrowing of the diversity of key starch biosynthesis pathway genes. In the western world, sorghum is predominantly utilized as animal feed and bio-industrial end uses, whereas throughout the world it is a major source of alcohol for human consumption in the form of beer and spirits such as the Chinese baijiu. We have found key signatures of directional and balancing selection in major starch biosynthesis genes. Association studies have identified a number of genes that have significant implications for human food uses, in particular the GBSS (waxy) and pullulanase debranching enzymes, which confer enhanced digestibility in monogastric animals. Pullullanase allelic variants encoding two amino acid substitutions have higher enzyme activity which significantly alters starch branching patterns and molecule size in both amylose and amylopectin. We have also expressed some of the key genes ectopically using transgenic approaches. This has lead to altered grain size, digestibility and end-use processing qualities. There are also implications for the growth of plant biomass, where the rates of transient starch turnover have been affected by various starch biosynthesis pathway genes.

[67] STRESS-INDUCED MITOCHONDRIAL PROTEIN DEGRADATION AND PEPTIDE RELEASE Jesper F. Havelund,1, Katarzyna I. Wojdyla,1, Veit Schwämmle,1, Adelina Rogowska-Wrzesinska,1, Allan G. Rasmusson,2, Ian Max Møller3 Department of Biochemistry and Molecular Biology, University of Southern Denmark, Denmark, 2 Department of Biology, Lund University, Sweden, 3 Department of Molecular Biology and Genetics, Aarhus University, Denmark 1

We tested the hypothesis that oxidized peptides released from mitochondria can act as retrograde signals to the nucleus (IM Møller and LJ Sweetlove 2010 Trends Plant Sci 15:370-374). Potato tuber mitochondria were incubated in a medium containing a substrate cocktail and ATP (control), in the same medium either plus FCCP (uncoupled control), or plus methyl viologen and KCN to block electron transport and maximize the production of reactive oxygen species (ROS) (oxidative stress). After 15 min incubation, the mitochondria were pelleted, digested with trypsin, iTRAQ-labelled and the samples pooled. The pooled sample was analyzed by liquid chromatography-mass spectrometry. We found that >50 tryptic peptides decreased in amount relative to other peptides in the same protein after the oxidative stress treatment, but not after FCCP treatment. This indicates that the peptides had been modified, probably by oxidation. The modified proteins represented a wide range of proteins, but the respiratory complexes, especially complex I, the tricarboxylic acid cycle enzymes and ROS-detoxifying enzymes were overrepresented. Many more peptides were released in the presence of the pore-forming peptide alamethicin especially from the matrix and the inner mitochondrial membrane (IMM) consistent with the formation of large pores in the IMM. Twice as many peptides appeared from matrix and IMM proteins during the oxidative stress treatment indicating that ROS reached larger parts of these compartments. We conclude that a detectable protein turnover can be observed in isolated mitochondria respiring in vitro over 15 min and that this protein turnover is accelerated during severe oxidative stress.

[68] THE CYSTEINE-RICH RECEPTOR-LIKE KINASES (CRKS) AS MEDIATORS OF SIGNALING SPECIFICITY WITH A COMPLEX EVOLUTIONARY HISTORY Aleksia Vaattovaara,1, Kerri Hunter,1, Sachie Kimura,1, Chunxiang LI,2, Ari Löytynoja,2, Jaakko Kangasjärvi,1, Jarkko Salojärvi,1, Michael Wrzaczek1 1

University of Helsinki, Department of Biosciences, Finland 2 University of Helsinki, Institute of Biotechnology, Finland

The CRKs are a large group of receptor-like protein kinases (RLKs) in Arabidopsis with 44 members. CRK expression is responsive to stress and our analysis of a crk mutant collection revealed phenotypes related to ROS signaling suggesting an intricate signaling network of CRKs in order to survive in a complex environment. However, in order to extrapolate the phenotypic and functional information from the model species Arabidopsis thaliana understanding of the evolutionary context is required. CRKs contain the plant-specific DUF26 (domain of unknown function 26; PF01657) in their ectodomain. DUF26, hallmarked by a conserved cysteine motif, is also found in the plasmodesmata-located proteins (PDLPs) and cysteine-rich receptor-like secreted proteins (CRSPs). Higher plants encode a large number of DUF26 proteins. Gene family expansion and the impact on the evolution of function are a major challenge in understanding the roles of gene families. In the light of the striking phenotypes observed for crk mutants and to understand why so many DUF26 genes are maintained in plants after duplication events and why expansions are so strikingly diverse in different plant lineages we have identified CRKs, PDLPs and CRSPs from more than 20 plant species covering most plant lineages. DUF26 originates in the first land plants and two DUF26s appear in the lycophytes. In genes with two DUF26, the two domains have differentiated into specific forms with unique sequence context surrounding cysteines. Variation within DUF26 domains between phylogenetic subgroups of the CRKs in Arabidopsis may have functional and structural importance for the extracellular domain.

[69] WHICH ENZYMES PROVIDES SUBSTRATES FOR GLYCOSYLATIONS? USING INHIBITORS TO STUDY NUCLEOTIDE-SUGAR PRODUCING ENZYMES

[70] HIGH THROUGHPUT SCREENING IN PLANTS Stina Lindberg1

Daniel Decker, , Leszek A. Kleczkowski 1

1

1

Umeå Plant Science Center, Sweden

Glycosylations are required for many essential plant processes, such as cell wall formation, and sucrose synthesis, require sugars “activated” with a nucleotide-diphosphate, such as UDP. Two important enzymes involved in this activation are: UDP-glucose pyrophosphorylase (UGPase) and UDP-sugar pyrophosphorylase (USPase), they can catalyze the reversible conversion of UTP and a sugar-1-phosphate to the corresponding UDP-sugar and pyrophosphate (PPi). UGPase can produce UDP-glucose, while USPase also produces several other UDP-sugars (mainly UDP-galactose, UDP-glucoronic-acid and UDP-arabinose UDP-xylose). In order to identify inhibitors for these enzymes, we used a highthrough-put approach, based on the screening of two chemical libraries which contained approximately 18000 compounds. We used an in vitro semi-robotized assay in which PPi was used to couple the activity of UGPase/USPase with luminescence. In this initial screen, 201 putative inhibitors were identified. After removal of false-positives and validation in two additional assay-systems (forward and reverse reaction of the enzymes), 5 compounds remained. The inhibitors showed a dose-dependent response (IC50 between 0.5 and 50 µM) and a subset also appears to inhibit a third pyrophosphorylase (UDP-glcNAC pyrophosphoyrlase). My poster will include details of the inhibitor screening approach, kinetic characterizations of the compounds, attempts at identifying inhibitors that are stronger or more selective against specific pyrophosphorylases (by analyzing compounds that are analogous to the inhibitors), and preliminary in vivo application of UGPase/USPase inhibitors on processes that might require UDP-sugar dependent glycosylation-reactions.

Chemical Biology Consortium Sweden (Cbcs); Laboratories for Chemical Biology Umeå (Lcbu); Umeå University, Sweden 1

In recent years chemical biology has emerged as a powerful technology in areas outside medicine, such as in plant science. Chemical biology represents a powerful tool to study e.g. plant developmental processes. By using high throughput screening to identify small molecules that interfere with a biological process of interest, a large number of compounds can be screened in a short period of time.

Chemical Biology Consortium Sweden (CBCS) is a national network for chemical biology with focus on small molecule screening and compound optimization and profiling. CBCS aims to produce high quality research tools in collaborative academic projects on a national level and does so by providing expertise in assay development, screening, medicinal and computational chemistry and pharmaceutical profiling. CBCS consists of three nodes with complementary capabilities; Laboratories for Chemical Biology Umeå University (LCBU), Laboratories for Chemical Biology Karolinska Institutet (LCBKI), and The Uppsala University Drug Optimization and Pharmaceutical Profiling (UDOPP).

CBCS offers expertise and instrumental assets for conducting chemical biology studies together with national research groups. Projects are based on a collaborative working model in which a majority of downstream in vitro and in vivo model systems/assays are provided by the collaborating group, whereas medicinal and computational chemistry activities as well as pharmaceutical profiling is provided by CBCS.

[71] GRADUAL PHOTOACTIVATION AND ASSEMBLY OF PHOTOSYSTEM II IN DARK-GROWN SPRUCE COTYLEDONS

[72] UV4QUALITY: INCREASING PHYTOCHEMICAL CONTENT AND TOLERANCE TO DESICCATION AND MECHANICAL STRESS OF HORTICULTURAL IMPORTANT PLANTS BY SUPPLEMENTATION WITH UV RADIATION

Andrej Pavlovic,1, Tibor Stolarik,1, Lukas Nosek,1, Roman Kouril,1, Petr Ilik1 1

Centre of the Region Hana for Biotechnological and Agricultural Research, Palacky University, Czech Republic

Irina Kalbina,1, Minjie Qian,1, Marcel A K Jansen,2, Eva Rosenqvist,3, Åke Strid1 School of Science and Technology; Örebro Life Science Center; Örebro University, Sweden, 2 School of Biological, Earth and Environmental Sciences; Enterprise Center, Distillery Field; University College Cork, Ireland, 3 Institute of Plant and Environmental Sciences; Copenhagen University, Denmark 1

Gymnosperms, unlike angiosperms, are able to synthesize chlorophyll and form photosystems in complete darkness. Photosystem I (PSI) formed under such conditions is fully active, but photosystem II (PSII) is present only in the latent form with inactive oxygen evolving complex (OEC). In this work we have studied photoactivation of PSII in dark-grown spruce cotyledons during 24h illumination using chlorophyll a fluorescence rise (FR), thermoluminescence (TL) and protein analysis. In dark-grown cotyledons, clear K and J steps appeared in FR, indicating that alternative reductants act as electron donors to PSII and plastoquinone (PQ) pool. After 5 min illumination, the K step disappeared in FR of cotyledons, which corresponded with measurable oxygen evolution. At the early stage of PSII photoactivation, the H and G steps became the most prominent in FR, reflecting a fast oxidation of electron carriers in PSI with subsequent electron donation to PQ. Prolonged illumination of cotyledons up to 24h resulted in the change of FR into a well-known OJIP curve, which was accompanied by the increasing attachment of PsbO protein to PSII. Interestingly, the 24h illumination was not sufficient for the completion of OEC. We have observed the absence of PsbQ protein in PSII particles, a low maximum yield PSII photochemistry (about 0.7) and a presence of intensive C band and up-shifted Q band in TL curves in cotyledons treated with DCMU. This lack of fully assembled OEC may be related with the absence of PSII supercomplexes detected by native electrophoresis after 24h illumination.

Hypothesis: Ultraviolet (UV) light is a major determinant of quality traits such as the content of phytochemicals (flavonoids, terpenoids) and the resistance to desiccation and mechanical stress in a number of economically important vegetable plants. Background: In Northern Europe, many vegetable crops are produced in greenhouses, where the UV component of sunlight is lacking due to the filtering effect of the cladding material. In certain species this results in stragglier plants with lower nutritional value and higher sensitivity towards desiccation and mechanical damage. Recent research shows that low doses of supplementary UV compensate for this by regulating plant morphogenesis and metabolism. The mechanisms for this remain unclear. We are studying the genetic and physiological consequences of the UV regulation. Results: So far, and by using mass spectrometry analysis, we show that supplementation with UV radiation in greenhouses leads to changes in both the pattern of phytochemical speciation and the quantity of important flavonoids and terpenoids in basil (Ocimum basilicum) and dill (Anethum graveolens). Also, qPCR shows that UV radiation regulates the expression of crucial genes of the phenylpropanoid and terpenoid biosynthesis pathways in basil in a diurnal fashion.

Future perspectives: Our research will lead to the understanding of How UV regulates biosynthesis of phytochemicals How UV regulates stem elongation, branching, leaf thickness and other morphological traits How plants trade growth and biomass accumulation against alteration in metabolism How commercially important plant species compare to model species with respect to UV

This research will also lead to horticultural production of plants with: Increased phytochemical content important for aroma, coloration and human health Increased sturdiness and increased resistance towards desiccation and transport

[73] ORGANIZATION OF PHOTOSYNTHETIC APPARATUS IN THE UNIQUE LIGHT-TOLERANT PHENOTYPE OF THE CHLORINA 3613 BARLEY MUTANT LACKING CHLOROPHYLL B

[74] PROTECTIVE EFFECTS OF VESICULAR ARBUSCULAR MYCORRHIZAL FUNGI ON PISTACHIO (PISTACIA VERA L.) SEEDLING EXPOSED TO DROUGHT STRESS

Elena Tyutereva,1, Wolfram G. Brenner,2, Alexandra Ivanova,3, Katharina Pawlowski,4, Olga Voitsekhovskaja1

Hossein Abbaspour1

1

Komarov Botanical Institute RAS, Russia, 2 Freie Universitaet Berlin, Germany, 3 Komarov Botanical Institute RAS, Russia; Saint-Petersburg State University, Russia, 4 Stockholm University, Sweden

1

The productivity of the barley mutant chlorina 3613 lacking functional chlorophyllide-a-oxygenase (Mueller et al., 2012) and, consequently, the chlorophyll b-containing antenna, can be enhanced considerably by a special cultivation practice (Tyutereva, Voitsekhovskaja, 2011). As a result, chlorina 3613 forms a new, previously unknown phenotype characterized by high tolerance to light and drought stress, and a productivity level comparable to that of the wild type. The light-sensitive phenotype of chlorina 3613 exhibits compromised stomata control and a very low level of photoprotection reflected by high singlet oxygen production, high levels of grana degradation and low NPQ. Contrarily, the light-tolerant phenotype shows the restoration of stomata control and the build-up of efficient photoprotection as judged by wild type levels of singlet oxygen production, restoration of grana, and development of zeaxanthin-independent NPQ. While the light-sensitive phenotype shows delayed flowering and precocious senescence, the transitions between the ontogenetic stages were partially restored in the light-tolerant phenotype. Transcript patterns were compared in both phenotypes vs. the wild type. Minor antenna proteins, especially Lhcb6, accumulated in the light-tolerant chlorina 3613 phenotype. We speculate that this partially restored the arrangement of PSII supercomplexes in the grana thylakoids, improving linear electron transport and repair of PSII, and consequently photosynthesis and productivity. Moreover, as the minor antenna proteins play a role in ABA signaling (Xu et al., 2012), this might be responsible also for the increased drought stress tolerance of the light-tolerant phenotype of chlorina 3613.

Drought stress is considered to be one of the most important abiotic factors limimting plant growths and yield in many areas. Mycorrhizal plants under abiotic stress are well stocked with an array of protective and repair systems that minimize the occurrence of stress damage. This study will sheld light on some different mechanisms that play a role in the protection of pistachio plants colonized by vesicular arbuscular mycorrhizal (VAM) fungi against drought stress. A pot experiment was set up to examine the effects of VAM fungus (Glomus mosseae) and drought on the growth, mineral uptake, soluble sugars, free amino acids, proline, glycine betaine, protein and antioxidant enzyme activity. The results showed that drought stress significantly reduced mycorrhizal colonization and plants growth of VAM-treated was higher than non-VAM treatment in well watered and water stressed plants. The nutrients content, such as P, K, Zn, N and Ca content in M plant were greather than those in NM under well watered and drought stress. The contents of soluble sugars, free amino acids, proline, glycinebetaine and protein increased under drought conditions, these increase were higher in M than NM trated plants. Mycorrhizal colonization altered antioxidant enzyme activity such as peroxidase and superoxide dismutase enzymes and significantly reduced the oxidative damage in plants exposed to drought stress. These results suggested that, VAM fungi may protect pistachio plants against drought stress by alleviating the drought induced oxidative stress.

Financial support of the Russian Scientific Foundation (#14-16-00120) is gratefully acknowledged.

Department of Biology, Faculty of Science, Damghan Branch, Islamic Azad University, Damghan, Iran

Key words: mycorrhiza, drought stress, pistachio, resistance

[75] THE ROLE OF COLONIZATION WITH ARBUSCULAR MYCORRHIZAL FUNGI ON RESISTANCE OF SAFFLOWER (CARTHAMUS TINCTORIUS L.) SEEDLING UNDER SALINITY STRESS

[76] ERF73/HRE1 IS A POSITIVE REGULATOR OF H2O2 PRODUCTION VIA HYPOXIA-INDUCIBLE RBOH GENE EXPRESSION UNDER HYPOXIA SIGNALING

Hossein Abbaspour,1, Roza Quchani,1, Sakineh Saeidi-Sar,1

Chin-Ying Yang1, Yi-Chun Huang1

1

Department of Biology, Faculty of Science, Damghan Branch, Islamic Azad University, Damghan, Iran

Salinity treatment with 3 level (0.5, 6, and 12 dS/m) and mycorrhizal arbuscular inoculation with two level (no inoculation, and inoculation with Glomus intraradices) were applied on cultivar of safflower (Goldasht) in this experiment. Salinity showed great negative effect on all growth and morphological parameters. On the other hand, colonized plant showed higher growth parameters under saline condition in compare with control. Reduced sugar content and pigment content is decreased by salinity, but these traits in colonized plants under saline condition were higher than non-colonized plants. Salinity increased the content of proline but Mycorrhizal plant showed higher content of proline. Mineral elements consist of P, N and Mg was higher in colonized plant, while salinity decreased the absorption of these elements. The content of phosphorous, nitrogen and magnesium were higher in colonized plant than non-colonized ones under stress and non-stress conditions. Considering interaction of salinity and colonization, mycorrhizal inoculated plants had a higher activity related to all measured antioxidant enzyme contain SOD, CAT, POD and APX comparing with non-inoculated ones. Higher activity of the enzymatic antioxidant means higher removal of these compound and higher resistance to stress condition. Overall, it is clear that salinity caused negative effect on cultivar of safflower but this negative effects were lower in inoculated plant, then use of mycorrhizal inoculation is a proper way to control the effect of salinity and maintain plant production to improve the resistance of the plants to salinity stress. Key words: mycorrhiza, salinity stress, Carthamus tinctorius, growth, enzyme antioxidant

1

Department of Agronomy, National Chung Hsing University, Taiwan

Oxygen deficiency (hypoxia) leads to serious physical damage in plant cells, resulting in energy deficit and ultimately cell and tissue death. Under hypoxia, ethylene and H2O2 homeostasis influence the response of AtERF73/HRE1 in hypoxia signaling pathways. However, the molecular functions of AtERF73/HRE1 in H2O2 production are poorly known. To assess the role of AtERF73/HRE1, we used three independent AtERF73/HRE1 knockout lines to detect H2O2 production. Under hypoxic stress, the HRE1-knockout seedlings displayed reduced H2O2 production, indicating that AtERF73/HRE1 might play a negative regulatory role in modulating H2O2 responses. Quantitative reverse-transcription polymerase chain reaction analyses showed that hypoxia-inducible Rboh genes may be differentially affected in HRE1-knockout lines: transcription of Rboh B and D was increased, whereas transcription of Rboh G was reduced. Rboh I was up-regulated at 1 h and down-regulated at 3 h of hypoxia treatment in the HRE1-knockout lines. The transcript levels of antioxidant genes and hypoxia-inducible/ ethylene-responsive genes were affected during hypoxia in the HRE1-knockout lines. Histochemical analysis indicated that AtERF73/HRE1 might be involved in the development of shoots and leaves, and more specifically in guard cells and inflorescence development. Taken together, the results indicate that AtERF73/HRE1 is involved in modulating H2O2 production and acts as a positive feedback regulator of hypoxia signaling pathways.

Keywords: AtERF73/HRE1; hypoxia; hydrogen peroxide; Rboh; antioxidant

[77] MAXIMUM HEAT TOLERANCE OF ALPINE PLANTS: METHODOLOGICAL, (ECO)-PHYSIOLOGICAL, AND ULTRASTRUCTURAL ASPECTS Othmar Buchner, , Andreas Holzinger, , Ilse Kranner, , Gilbert Neuner 1

1

1

1

1

University of Innsbruck; Institute of Botany, Austria

At high elevation sites of the European Alps the increase in atmospheric temperature was found to be twice of that estimated for global warming elsewhere (1890-1998, +1.1°C vs. +0.55°C; Böhm et al. 2001). Particularly in alpine dwarf shrubs already now leaf temperatures can occasionally exceed the critical high temperature thresholds for heat damage to leaves (e.g. Rhododendron ferrugineum: >46°C). For future risk assessment knowledge about the heat vulnerability of alpine plants and of their ability to cope with changed climatic conditions is required. We have developed a novel Heat Tolerance Testing System (HTTS) that allows testing of heat tolerance of whole plants in situ under full natural solar irradiation. Results indicate that heat tolerance of plants is often several degrees (°C) higher than that assessed by conventional methods that use excised plant parts exposing them in darkness. Short-term sub-lethal heat spells (30 min; simulated by the HTTS), led to long-lasting (up to >1 week) reductions of photosynthetic performance. The presence of solar irradiation during short-term heat exposure was additionally shown to have protective effects on photosynthetic functions. For Vaccinium gaultherioides leaves a significant impact of irradiation intensity on the overall heat hardening capacity, xanthophyll cycle pigments and on the pool of antioxidants such as glutathione and ascorbate could be revealed. In Ranunculus glacialis heat and irradiation stress were not causally related to the formation of chloroplast protrusions but rather a significant connection to enhanced catalase-activity (photorespiration) could be evidenced. Our studies demonstrates that, in the European Alps, heat waves can temporarily have a negative impact on photosynthesis and, importantly, that results obtained from experiments performed in darkness and/or on detached plant material may not reliably predict the impact of heat stress under field conditions. Funding: Austrian Science Fund (FWF): Project Nr. P22158-B16 granted to O. Buchner

[78] DROUGHT TOLERANCE AND ITS YIELD PENALTY EFFECT IN POTATO Manuela Haas,1, Heike Sprenger,1, Katharina Rudack,2, Sylvia Seddig,2, Rolf Peters,3, Ellen Zuther,1, Joachim Kopka,1, Dirk Hincha,1, Karin Köhl1 MPI of Molecular Plant Physiology, Germany, 2 Julius-Kühn-Institut, Germany, 3 Versuchsstation Dethlingen, LWK Niedersachsen, Germany 1

Climate models predict an increased likelihood of seasonal droughts for many areas of the world. Potato (Solanum tuberosum L.) produces high calories per unit water invested, but is drought-sensitive. In a marker identification study, we determined drought tolerance of 34 European potato cultivars in pot and field trials. Drought tolerance was quantified as tuber starch yield under drought stress relative to yield under optimal water supply and normalized to the trial median. We found significant tolerance variation within the cultivar panel. This variation was employed to identify and validate leaf metabolite concentration as drought tolerance marker. Correlating drought tolerance with yield under optimal field conditions indicated a yield penalty of tolerance. Distribution of yield data versus tolerance suggested that adjusted breeding strategies can combine tolerance with average yield potential. To that end, sub-populations with increased drought tolerance were selected from a crossing population segregating for drought tolerance. Selection was performed based either on tolerance data from three trials or on a metabolite marker model. Both sub-populations were compared with the remaining population for significant changes in drought tolerance and yield potential. The results will allow assessing the size of the yield penalty and identifying markers to select against yield penalty while increasing drought tolerance.

[79] HEAT STRESS EFFECT ON REMOBILISATION OF NITROGEN AND WATER-SOLUBLE CARBOHYDRATES DURING GRAIN-FILLING IN WHEAT - A FACE STUDY

[80] UV ACCLIMATION – HYDROGEN PEROXIDE IN FOCUS Gyula Czégény,1, Petra Majer,2, Györgyi Sándor,2, András Dér,3, Åke Strid,4, Éva Hideg,1

Allene Macabuhay, , Glenn Fitzgerald, , James Nuttall, , Michael Tausz, , Sabine Tausz-Posch, 1

2

2

3

1

Faculty of Veterinary and Agricultural Sciences; The University of Melbourne, Australia, 2 Department of Environment and Primary Industries, Austria, 3 Department of Forest and Ecosystem Science; The University of Melbourne, Austria 1

Global heating occurs as a consequence of increasing atmospheric CO2 concentrations, and this abiotic mutual occurrence generates interactive effects on plant growth and productivity. This study evaluated the combined effects of elevated [CO2] and short-term heat stress on wheat (Triticum aestivum L.) through physiological traits, biochemical properties and source-sink relationships. More specifically, we investigated whether wheat’s remobilisation of accumulated water-soluble carbohydrates (WSC) and nitrogen under heat stress-induced environment is further aggravated or ameliorated by [CO2] enrichment. Plants were grown in the state-of-the-art Australian Grains Free Air CO2 Enrichment (AGFACE) facility under two [CO2] of ambient (approximately 395 µmol mol-1) and elevated (approximately 550 µmol mol-1). Heat treatments were applied for three days, starting at five days before, and 15 and 30 days after anthesis using purpose-built heat chambers. Significant reduction was observed on dry matter accumulation in grain (19%), primarily with heat applied at 15 days after anthesis. Nitrogen concentration of grain decreased under high temperature by 5% and under elevated [CO2] by 16%; their interaction aggravating the reduction by an average 17%. Concentration of WSC in stem was reduced by 30% under heat stress but elevated [CO2] alleviated this effect with an increase of 23%; and the combined effect generated a 28% increase. This WSC trend, however, didn’t occur in grain as no considerable differences were found. This result confirms the degree of wheat sensitivity to heat during flowering and grain-filling and illustrates the complex interactive effects of combined climate change-driven phenomena on grain quality and quantity of wheat.

Institute of Biology, University of Pécs, Hungary, 2 Institute of Plant Biology, Biological Research Centre, Hugary, 3 Institute of Biophysics, Biological Research Centre, Hungary, 4 Department of Science & Technology, Örebro Life Science Center, Örebro University, Sweden 1

Ultraviolet radiation (280-400 nm) at mid-latitudes of Northern hemisphere is known as a regulation factor of plant development rather than a stressor, although solar UV may become stressful synergistically in response to other abiotic factors (Hideg et al 2013). Lower PAR/UV ratios also can lead to oxidative damage through the shifted balance of pro-oxidants and antioxidants. Hydrogen peroxide (H2O2), the most stable ROS is a well-known signalling molecule produced at several sites in leaves and its levels are well-controlled by scavenging enzymes. Nevertheless it is of special interest, because metabolic H2O2 levels are increasing as an effect of abiotic factors, and it also absorbs solar UV. This study shows that solar UV-B (280-315 nm) has the ability to photoconvert H2O2 to more oxidizing hydroxyl radicals (•OH) (Czégény et al 2014). Thus H2O2 scavenging enzymes have an emphasized role under conditions modifying PAR/UV ratios to avoid the possibile damage by •OH under various light conditions (Majer et al 2014). Supported by grant No. OTKA NN-85349 of the Hungarian Scientific Grant Agency, Grants from the Carl Trygger Foundation, the Knowledge Foundation, and Örebro University’s Faculty for Business, Science and Technology. References Hideg É, Jansen MAK, and Strid Å (2013) UV-B exposure, ROS and stress: inseparable companions or loosely linked associates? Trends Plant Sci. 18, 107-115. Majer, P., Czégény, Gy., Sándor, G., Dix, P. J., Hideg, É. (2014) Antioxidant defence in UV-irradiated tobacco leaves is centred on hydrogen-peroxide neutralization. Plant. Physiol. Biochem. 82, 239-243. Czégény, Gy. Wu, M. Dér, A. Eriksson, L. A. Strid, Å. Hideg, É. (2014) Hydrogen peroxide contributes to the ultraviolet-B (280-315 nm) induced oxidative stress of plant leaves through multiple pathways. FEBS Letters 588, 2255-2261.

[81] ALUMINUM IMPACT AND ACCLIMATION RESPONSE: A COMPARATIVE STUDY OF TWO CONTRASTING WHEAT CULTIVARS

[82] COLD STRESS INCREASES SALT TOLERANCE OF THE EXTREMOPHYTES EUTREMA (THELLUNGIELLA) SALSUGINEA AND EUTREMA (THELLUNGIELLA) BOTSCHANTZEVII

Julietta Moustaka,1, Georgia Ouzounidou,2, Gulriz Baycu,3, Michael Moustakas,4

Anastasia Shamustakimova,1, Tatiyana Leonova,1, Vasiliy Taranov,1, Alexey Babakov,1, Albertus H. de Boer2

1

Univesrity of Creete Voutes Campus, Greece, 2 Institute of Food Technology, Greece, 3 Istanbul University Faculty of Science, Turkey, 4 Aristotle University of Thessaloniki, Greece

1

The presence of phytotoxic aluminum species (Al3+) is considered as the primary factor limiting crop productivity in over 40 % of world’s arable non-irrigated land that is acidic. Al3+ phytotoxicity slows root growth and causes impairment of Photosystem II (PSII) activity. Wheat cultivars that acclimate to Al3+ phytotoxicity and are able to grow and complete their life cycle may be exploited to increase production in those soils and may also be used to understand the mechanisms of Al tolerance. For this purpose, the response of two wheat cultivars (Triticum aestivum L.), with contrasting Al3+ tolerance, cv. Yecora E (tolerant) and cv. Dio (sensitive), exposed to 0, 37, 74 and 148 μM Al for 14 days in hydroponic culture at pH 4.5, was studied. With increasing Al concentration, leaf Ca2+ and Mg2+ content decreased, as well as the quantum efficiency of PSII photochemistry (ФPSII), while a gradual increase in the membrane lipid peroxidation, Al accumulation and PSII excitation pressure occurred. However, the Al-tolerant cultivar retained larger concentrations of Ca2+ and Mg2+ in leaves and kept a larger fraction of the PSII reaction centres in an open configuration, i.e. a higher ratio of oxidized to reduced QA, than the sensitive cultivar. Our results suggest that Al3+ toxicity may be mediated by nutrient deficiency and oxidative stress, and that Al-tolerance of the wheat cultivar Yecora E results, at least partially, from the decreased reactive oxygen species (ROS) formation, that contribute to reduced PSII excitation pressure and a better PSII functioning.

A comparative study was performed to analyze the effect of cold acclimation on improving the resistance of Arabidopsis thaliana, Eutrema (Thellungiella) salsuginea and Eutrema (Thellungiella) botschantzevii plants to salt stress caused by high concentration of NaCl. After low-temperature acclimation of 25-day plants (4°C, 6 days), NaCl was added into the soil at a final concentration of 300 mM for A. thaliana, 400 mM for E. salsuginea and 500 mM for E. botschantzevii. Shoot FW (Fresh weight), sodium and potassium ions accumulation, expression of proton pump genes VAB1, VAB2, VAB3, VP2, HA3 and ion antiporter genes SOS1, HKT1, NHX1, NHX2, NHX5 in plasmalemma and tonoplast were determined after 7 days. Shoot FW of E. salsuginea and E. botschantzevii plants exposed to salt stress after cold acclimation was increased compared to the control plants, in contrast to A. thaliana plants. In the same plants, sodium ion concentration was reduced compared to the control plants, whereas potassium ion concentration change was insignificant. As a result of cold acclimation, the expression of SOS1, VAB1, VAB3, NHX2, and NHX5 genes was significantly enhanced in E. botschntzevii and E. salsuginea plants. In E. salsuginea plants, increased expression of VP2 and HA3 genes was also observed. None of the 10 genes analyzed showed any expression change in A. thaliana plants after cold acclimation. Altogether, the results indicate that cross-adaptation to the consecutive action of low positive temperature and high concentration of NaCl exists in the extremophytes E. salsuginea and E. botschantzevii. This phenomenon may be attributed to the increase of ion transporters gene expression during cold acclimation.

All Russia Institute of Agricultural Biotecnology, Russia, 2 VU University Amsterdam, Netherlands

Key words: Eutrema, Arabidopsis, cold acclimation, salt stress, K+/Na+ accumulation, ion transporter, gene expression.

[83] CO2 AND H2O EXCHANGE OF WOODY SPECIES IN EASTERN FENNOSKANDIA UNDER CLIMATE CHANGE

[84] METABOLISM OF BETULA SPECIES IN THE EASTERN PART OF FENNOSKANDIA UNDER ENVIRONMENT CHANGE

Tatiana Sazonova,1, Vladislava Pridacha,1, Alexander Olchev2

Vladislava Pridacha,1, Elena Novichonok,1, Tatiana Sazonova1

1

Forest Research Institute of Karelian Research Centre of Russian Academy of Sciences, Russia 2 A.N. Severtsov Institute of Ecology and Evolution of Russian Academy of Sciences, Russia

1

The modern climate changes appeared mainly in increase of the air temperature, alterations in the gas composition of the air and land surface moistening conditions may evidently influence the dynamics and rate of biophysical and biochemical processes in plants and soil and, as a consequence, lead to changes in the intensity of CO2 and H2O exchange between plants and ambient air. Measurements of stomatal conductance, rates of photosynthesis and dark respiration and water potential of Scots pine (Pinus sylvestris L.), Norway spruce (Picea abies (L.) Karst.) and silver birch (Betula pendula L.) were provided using the portable photosynthesis system LI-COR 6400XT (Li-Cor Inc., USA) and the pressure chamber Plant Moisture Vessel SKPM 1400 (Skye Instruments Ltd., UK) on the sample plots of FRI KarRC RAS (south Karelia) during the growing seasons in 2009-2014. By studying the patterns in the basic physiological processes (CO2 gas exchange, water exchange, mineral nutrition) against the background of variable hydrometeorological parameters we found the ranges of environmental factors (temperature, relative air humidity, and solar radiation rate) within which the metabolism in pine, spruce and birch is most intensive. Leaf photosynthesis, respiration, stomatal conductance and transpiration data were used in the process-based Mixfor-SVAT model (Olchev et al., 2002, 2008) to derive the possible response of CO2/H2O budgets of forest ecosystems of Eastern Fennoskandia to future changes in the climate and vegetation structure.

Nitrogen is essential for plant metabolism, and his restricted availability often limits plant carbon acquisition and growth. We investigated the impact of nitrogen availability on the carbon and water exchange in the leaf of Betula pendula Roth and B. pubescens Ehrh. in Eastern Fennoskandia (south Karelia). Measurements of stomatal conductance, rates of photosynthesis and transpiration, water potential of B. pendula and B. pubescens were provided using the portable photosynthesis system LICOR 6400XT (Li-Cor Inc., USA) and the pressure chamber Plant Moisture Vessel SKPM 1400 (Skye Instruments Ltd., UK) on the sample plots of FRI KarRC RAS. In response to NH4NO3 treatment, both species demonstrated a rise in stomatal conductance, rates of photosynthesis and transpiration in the leaf. In B. pubescens the rise in the leaf transpiration rate came along with a rise in the water potential of the foliated shoot and a decline in the available water content (WCf ) and saturating water content (WCs) in the leaf. The changes accompanying the rise in leaf transpiration rate in B. pendula were a decline in the water potential of the foliated shoot and stabilization of WCf and WCs of the leaf. This suggests that interspecific distinctions in the dynamics of the indices of water exchange are caused by decrease in hydraulic conductivity of the foliated shoot in B. pendula and variation in cell partial volumes of leaf in B. pubescens.

The study was supported by grants (13-04-00827-а, 14-04-01568-а) of the Russian Foundation of Basic Research (RFBR).

Forest Research Institute of Karelian Research Centre of Russian Academy of Sciences, Russia

The study was supported by grant (13-04-00827-а) of the Russian Foundation of Basic Research (RFBR).

[85] EVALUATION OF BREAD WHEAT CULTIVARS UNDER SALINITY STRESS BASED ON SOME AGRONOMIC TRAITS

[86] SELENIUM SEED PRIMING IN RADISH TO PROMOTE DROUGHT STRESS TOLERANCE Leticia Alves,1, Patricia Cury,1, Priscila Gratão,2, Marina Gavassi,1, Priscila Miranda,1, Lucas Gaion,1, Carolina Monteiro,1

Keyvan Shamsi

1

1

Islamic Azad University Kermanshah Branch Iran, Iran

In order to study the effect of different salinity levels on yield components and yield of three cultivars of bread wheat, a greenhouse experiment; was conducted in Islamic Azad University, kermanshah ,Iran. This factorial experiment was carried out in 3 replications and in the form of Randomize Complete Blocks Design(RCBD). Three cultivars namely Chamran, Marvdasht and Shahryar were placed in 3 salinity levels including 0.6 (control), 8 and 16 dSm-1. The results showed that with increasing salinity, number of grain per spike, number of grain per spikelet, number of fertile spikelet, number of tillers, 1000 grain wheat, plant hight,grain yield, biological yield and harvest index were decreased. The most this traits was observed in Chamran. The negative effect of salinity on plant was due increasing Na+ and decreasing K+ content in the leaves The highest sodium content belonged to Shahryar. Biological yield and harvest index, had the highest correlation with grain yield. The most tolerant and stable cultivar was Chamran which had the highest yield at the salinity level of 16 dSm-1. Keywords: Salinity, Bread Wheat cultivars, Yield,Yield components

Universidade Estadual Paulista “Julio de Mesquita Filho”, Brazil, 2 Laboratório de Fisiologia Vegetal/Fcav/Unesp - Univ. Estadual Paulista/Depto. de Biologia, Brazil

1

Plants in the environment are constantly exposed to stresses during their growth and development, which results in loss crop production. Techniques have been developed to avoid stress in plants and improve the agriculture yield. Seed priming is a recent strategy which can prepare the plant to respond more quickly and efficient to stress. Selenium (Se) is not considered an essential element for higher plants, although in low concentration it can bring benefits in stress tolerance by enhancing their antioxidant system, decreasing lipid peroxidation levels and improving stress tolerance. Brassicaceae species are classified as a primary accumulator and are capable to absorb Se by seeds. The aim of this study was to verify if Selenium seed priming in radish improves germination rate, decrease lipid peroxidation and hydroperoxide content to drought stress. Our results indicated alterations in oxidative stress and germination rate varied according to concentration of Na4Se and time of seed priming, which indicated that Se can improve defense responses to stress condition. These data may be useful to improve the effective use of water in agriculture and decrease the loss production in areas without irrigation. (Financial support by CAPES).

[87] FROST SURVIVAL OF REPRODUCTIVE SHOOTS IS ENSURED BY STRUCTURAL ICE BARRIERS

[88] IDENTIFICATION OF MIRNAS AND THEIR TARGETS EXPRESSED IN LEAVES OF BRACHYPODIUM DISTACHYON DURING DROUGHT STRESS AND SUBSEQUENT RECOVERY

Edith Kuprian,1, Tan Tuong,2, David Livingston,2, Gilbert Neuner,1 1

University of Innsbruck; Institute of Botany, Austria, 2 Usda-Ars; North Carolina State University, United States

Karolina Chwiałkowska,1, Gaurav Sablok,2, Małgorzata Rydzanicz,3, Luis Mur,4, Robert Hasterok,5, Mirosław Kwaśniewski,1 Department of Genetics, Faculty of Biology and Environmental Protection, University of Silesia in Katowice, Poland, 2 Plant Functional Biology and Climate Change Cluster, University of Technology Sydney, Australia, 3 Department of Medical Genetics, Warsaw Medical University, Poland, 4 Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, United Kingdom, 5 Department of Plant Anatomy and Cytology, Faculty of Biology and Environmental Protection, University of Silesia in Katowice, Poland 1

The risk of frost damage to alpine plants increases significantly with increasing elevation [1, 2]. Therefore special protective strategies are necessary not only to sustain freezing stress in order to guarantee vegetative growth, but also to protect the ice-susceptible reproductive organs to ensure a successful sexual reproduction. Woody alpine dwarf shrubs such as Calluna vulgaris protect their reproductive buds, flowers and fruits even during summer by supercooling from frost damage [3]. Ice nucleation starts usually in the apoplast of the vegetative parts and propagates fast throughout the vessels. So an ice barrier is needed that impedes the ice from entering into the reproductive shoots and enables these tissues to supercool. IDTA (infrared differential thermal analysis) recordings revealed a structural barrier at the base of the pedicel, which remains active throughout the whole growing season and all reproductive stages of Calluna vulgaris [3]. The nature of the ice barrier is currently not known. Paraffin embedding and subsequent sectioning allow histological and anatomical characterization of these tissues. First sequences of longitudinal and cross sections of Calluna vulgaris show a special histological composition and cytological peculiarities such as cells in this area are tightly packed, lack intercellulars, and the xylem vessels seem to be pectine-rich in the transition zone between the vegetative part and the reproductive tissue. Structural ice barriers may also play an important role in other species, as they serve as an effective mechanism to avoid freezing and to allow supercooling in extreme down to -40°C in certain plant tissues.

Water deficiency is an adverse environmental stress factor that affects plant growth and development, thus severely limits crop productivity. Plants exposed to drought can employ multiple defense mechanisms, including post-transcriptional gene expression regulation by miRNAs. In this study the results of detailed characterisation of Brachypodium distachyon leaves microtranscriptome and its modulation during drought stress and subsequent recovery are presented. Moreover, putative targets of analyzed miRNAs, predicted based on combined analysis of small RNA and mRNA sequencing are discussed. By using small RNA deep sequencing and stringent bioinformatics analyses we were able to detect 30 conserved miRNA families, comprising 46 MIRNA genes expressed in leaves of B. distachyon seedlings. Among these families, we found 11 mature miRNAs that were differentially expressed in drought-stressed plants. Moreover, 11 novel MIRNA candidates and their putative targets genes were predicted. Consequently, the detailed analysis of microtranscriptome changes induced by water-deficiency stress in B. distachyon leaves, coupled with global transcriptome analysis allowed for identification of the subset of drought-responsive genes, which expression is presumably regulated with the involvement of miRNA pathways. Therefore, our results provide important insight into miRNA-dependent drought stress response networks in plants.

The authors acknowledge financial support from the Polish National Science Centre (grant no. 2012/04/A/NZ3/00572).

[89] CHLOROPLAST ANTIOXIDANT SYSTEM UNDER TEMPERATURE STRESSES AND MEDIUM TERM MEMORY DEVELOPMENT IN ARABIDOPSIS ACCESSIONS

[90] USING OF ETHYLENE MUTANT GRAFTING TOMATO TO STUDY HORMONE SIGNALING BETWEEN ROOTS AND SHOOTS SUBJECT TO WATER DEFICIT

Jelena Cvetkovic,1, Joern van Buer,1, Margarete Baier,2

Lucas Gaion,1, Anieli Baldo,1, Carolina Monteiro,1, Leticia Alves,1, Rogerio Carvalho,1

1

FU Berlin, Institute of Biology, DCPS, Plant Physiology, Germany, 2 Plant Physiology; FU Berlin; DCPS, Germany

Exposing plants to low temperatures are usually resulting in specific physiological, molecular and biochemical changes in plant cells. Process of photosynthesis is necessarily affected by such stress. Photosynthetic capacity is reduced due to unbalanced light energy absorption and consumption by plant metabolic sinks. Reactive oxygen species and the chloroplasts as major sites of their production are important initiators and/or converters of signals to the nucleus under stress conditions. According to many researchers, connection between photosynthesis and other cell processes during cold acclimation is obvious and involves crosstalk among metabolites and photosynthetic redox state. We suggest that the chloroplast antioxidant system behaves as a pro-active priming hub under cold stress. Two weeks under cold conditions are sufficient to develop medium term memory and to be prepared for the next (triggering) challenge. Our focuses are on gene expression and metabolite analysis, especially on genes of ascorbate dependent and independent pathways. Combining these results with observed ecophysiological data provide a picture of signaling and destruction balances caused by high ROS production. Investment into priming costs, but efficient priming is bringing benefits to the plants. Experiments with seven different Arabidopsis thaliana accessions show that the cold priming strategy, costs and benefits have been shaped by adaptation. We acknowledge funding by the German research foundation (SFB 973-C4).

1

Universidade Estadual Paulista “Julio de Mesquita Filho”, Brazil

Drought is a major abiotic stress affecting growth and development plant, estimates that this fact will further worsen in coming years due to climate change. Therefore, the aim of this work is to study the involvement of ethylene in the signaling between root and shoot during drought. Thus, the mutant tomato Never ripe (Nr) that has low sensitivity to the plant hormone ethylene and the cultivar Micro-Tom (MT) was used, the two genotypes were combined by grafting technique for cleft (MT/MT; Nr/Nr; MT/Nr;Nr/MT; MT and Nr) and plants without grafting in both genotypes as control. The plants were transplanted into 350 ml pots filled with commercial substrate and expanded vermiculite 1:1 (v/v) to provide the water deficit stress was made replacement of only 20% of field capacity for seven days and the control received 100% of field capacity throughout the cycle. Were evaluated, dry mass, area, diameter, density and length of roots. Under well hydrated condition the combination Nr/MT shows highest values ​​of area, diameter and dry weight of roots, however plants subjected to water deficit did not change the root development, except grafted plants between Nr and MT (Nr/MT and MT/Nr) which decreased for all parameters. The results show that ethylene is part plant responses to drought and the use of hormonal mutants and grafting technique are important tools for understanding signaling between roots and shoots during stress conditions.

[91] WELWITSCHIA MIRABILIS: EFFICIENT DESIGN AND ADAPTABILITY RELATED TO PHOTOSYNTHETIC GAS EXCHANGE IN THE NAMIB DESERT

[92] RWP-RK PROTEINS : A NEW CLASS OF TRANSCRIPTION FACTORS INVOLVED IN NITROGEN SIGNALING IN PLANTS.

Gert Krüger,1, Anine Jordaan,1, Louwrens Tiedt,1, Reto J Strasser,2, Michele Kilbourn Louw,3, Jacques Berner,1

Camille Chardin,1, Thomas Girin,1, Virginie Gaudon-Brehaut,1, Christian Meyer,1, Anne Krapp,1

1

North-West University, South Africa, 2 University of Geneva, Switzerland, 3 Swakop Uranium, Namibia

The extraordinary survival potential of this endemic plant of the Namib is explained in terms of its special leaf anatomy and resourceful responses to light temperature and drought. Welwitschia is a perennial and therefore cannot escape the severe climatic conditions of the desert. The leaves have a number of xeromorphic features. Modern sample preparation and imaging techniques provided a compelling motivation to re-investigate the anatomy of this fascinating plant from an ecophysiological perspective. SEM and light micrographs revealed new details of leaves, cell wall structures, stomatal anatomy and root hair development of young (potted) and old (in situ) plants. We could demonstrate fast changes in diurnal stomatal conductance measured under controlled and in situ conditions. The fast turgor-driven movements of the armored guard cells are shown to be facilitated by their double hinged cell wall construction (toggle mechanism). Efficient CO2 diffusion into the leaf is allowed by numerous but small intercellular spaces between palisade cells caged in by hypodermal fiber bundles. These act as supportive beams preventing collapse of the mesophyll. Under field conditions chloroplasts seem to assemble at the walls parallel to incoming light. Laboratory experiments proved that the photosynthetic apparatus is stable over the temperature range of 4- 36 °C. Studying the effect of soil volumetric water content on CO2 assimilation proved that CO2 assimilation takes place up to very low soil water content levels. A:Ci response data indicated that although biochemical limitation occurred at severe drought, stomatal limitation was the main constraint under drought stress. Very fast formation of copious new root hairs followed re-watering after severe drought.

1

IJPB INRA Versailles, France

Nitrate, the main nitrogen source in agricultural soils, also acts as a signal that regulates many developmental and metabolic processes, including large changes in gene expression. The Arabidopsis RWP-RK transcription factor NLP7 is a master regulator of this primary nitrate response (Marchive et al., 2013). NLP7 (for NIN-Like Protein 7) is a homolog of the NIN (Nodule INception) protein, which have first been discovered in legumes, allowing nodulation (Schauser and al., 1999). Both NIN and NLP7 proteins possess a highly conserved RWP-RK domain involved in DNA-binding. Proteins containing this specific RWP-RK domain are found in the entire green lineage (Schauser et al., 2005). A phylogenic study of these so-called RWP-RK proteins revealed two major groups: NLP proteins that possess in addition to the RWP-RK domain a PB1 domain, predicted to be a protein-protein interaction domain, and RKD proteins (RWP-RK Domain proteins) which are shorter and contain only the RWP-RK domain. We integrated Brachypodium distachyon in our study since this species is currently used as a model for cereals. The long term goal of our work, as nitrate is one of the most important nutrients for plants, is to exploit new avenues for increasing Nitrogen Use Efficiency (NUE) based on the in-depth knowledge of the nitrate signalling network.

[93] ANTIOXIDANT SYSTEM IN THE RESPONSE OF DROUGHT STRESS IN TROPICAL TREES

[94] AGB, A NOVEL G PROTEIN Β SUBUNIT-LIKE PROTEIN, POSITIVELY REGULATES ABA RESPONSES IN ARABIDOPSIS

Aneth Sarmiento,1, Margarete Baier,1 1

Plant Physiology; Freie Universität Berlin; DCPS, Germany

Minkyun Kim,1, Jaemin Hwang,1 1

Some environmental conditions have forced plants to adapt to survive: e.g. drought stress and hot temperatures. Besides anatomical changes, plants have modified their carbon assimilation strategies. There are three main photosynthetic pathways, C3, C4 and CAM; plants have evolved into one of them, or are even able to switch between them, depending on the environmental conditions. Photosynthesis by itself can lead to production of reactive oxygen species (ROS). This production can get increased by abiotic/ biotic stress. To avoid the negative effects of ROS and to control ROS signaling, a strong and regulated antioxidant system is displayed.

We have evaluated three tropical plants that have different photosynthetic pathways and differ in the tolerance to drought stress in their natural environments. Seeds from Swietenia macrophylla (C3, drough sensitive), Jatropha curcas (C3, probably switching to CAM, drought tolerant) and Clusia rosea (CAM) were imported from Panama and grow in a Greenhouse, under similar conditions as at the natural habitat (28ºC, 12/12h photoperiod, daily watering) until few weeks after germination.

Drought stress experiments were performed to seedlings of the same age. The plant responses were evaluated with respect to ROS levels, photosynthetic activity and water content, at different stress stages. The genes of the antioxidant system are explored. cDNA encoding 2-Cys- peroxiredoxin (2CP) and ascorbate peroxidase (APX) were isolated and gene expression and regulation is under investigation.

We acknowledge funding by IFARHU (Instituto para la Formación y Aprovechamiento de Recursos Humanos), and support by Universidad Tecnológica de Panamá and Freie Universität Berlin.

Seoul National Universty, Rep. of South Korea

The Arabidopsis gene encoding AGB (a putative Arabidopsis G protein beta subunit) was characterized in this study. The predicted structure of AGB has novel features despite the sequence similarity with typical G protein beta subunit proteins: i.e., while typical G protein β subunit proteins consist of an α-helix domain at N-terminus plus seven WD-40 domains making up a hepta-propeller structure, AGB has only six WD-40 domains and extra C-terminal extension that shows no sequence homology to any known functional domains. Transcripts of the nucleus-localized AGB were most abundant in the roots of mature Arabidopsis plants, and induced by ABA, ethephon, mannitol, and NaCl, respectively. AGB-overexpressors exhibited ABA-hypersensitive phenotypes (increased inhibitions of seed germination and seedling growth) and enhanced drought tolerance, while agb knockout mutants showed ABA hyposensitivity. AGB overexpression changed the expression of numerous genes, including those involved in ABA-mediated responses. Taken together, we conclude that AGB positively regulates ABA-mediated development and drought responses in Arabidopsis.

[95] NEVER RIPE TOMATO MUTANT GRAFTING PLANTS SUBMITTED TO CD–STRESSFUL CONDITIONS

[96] EFFECTS OF BORON AND PHOSPHORUS ON BLOSSOM-END ROT (BER) INCIDENCE AND FRUIT QUALITY OF TOMATO

Priscila Gratão,1, Letícia R. Alves,1, Patrícia R. Cury,1, Carolina C. Monteiro,1, Lucas A. Gaion,1, Rogério F. Carvalho,1

Chia-Ying Lee,1, Yi-Ting Lee,1, Shu-I Lin,1

1

Laboratório de Fisiologia Vegetal/Fcav/Unesp - Univ. Estadual Paulista/Depto. de Biologia, Brazil

Environmental pollution with heavy metals as cadmium (Cd) is a global problem, mainly to agricultural soils. Plants exposed to heavy metals increase reactive oxygen species (ROS) production, which can cause serious damages to cellular metabolism and consequently losses in crop production. Malondialdehyde (MDA) is one of several products formed via the decomposition of certain primary and secondary lipid peroxidation products, which can indicate the level of oxidative stress. The aim of this study was to use Never ripe tomato mutant grafting plants with wild-type Micro-Tom to evaluate the crosstalk between antioxidant responses and signaling between plant organs during stress (root-toshoot). With this purpose we used Never ripe tomato mutant plants, which shows a defective gene for ethylene receptor. Our results indicated similar MDA content in roots, being less pronounced in grafted plants. It suggests that the stress induced by Cd can be alleviated by detoxification mechanisms in plants, involving an array of non-enzymatic and enzymatic mechanisms for antioxidants systems. Nonetheless, more evaluations must be done, mainly with antioxidants enzymes. The information available in this work is an important step towards obtaining a better understanding of the physiological changes caused by Cd and its effects on metabolic processes. Financial support by FAPESP

1

Department of Horticulture and Landscape Architecture, National Taiwan University, Taiwan

Blossom-end rot (BER) is a calcium-related physiological disorder commonly encountered during commercial production of tomato (Solanum lycopersicum). It is known that BER results from calcium (Ca) deficiency in parts of fruit tissues and/or abnormal calcium distribution within cells due to misregulation of Ca2+/H+ exchanger (sCAX1) gene, and leads to structure destruction of cell membrane and cell wall. Phosphorus (P) and boron (B) play important roles in maintaining structures of cell membrane and cell wall. Hence, we hypothesized that concentrations of P and B in the supply may affect the incidence and severity level of BER. Accordingly, tomato plants were grown in greenhouse and supplied with various concentrations of P and B using trickle fertigation systems. The observation showed that a higher concentration of P or an intermediate concentration of B in the supply could reduce incidence and/or severity level of BER, but it was cultivar-dependent. In addition, the fruit yield and fruit quality were also varied under different concentrations of P or B.

[97] POST-TRANSLATIONAL MODIFICATIONS OF FERREDOXIN-NADP+ OXIDOREDUCTASE IN RESPONSE TO ENVIRONMENTAL CHANGES

[98] LIPOXYGENASE 6 IS REQUIRED FOR JASMONATE SYNTHESIS IN ROOTS AND PROMOTES STRESS RESISTANCE OF ARABIDOPSIS THALIANA

Magda Grabsztunowicz,1, Nina Lehtimäki,1, Minna M Koskela,1, Käthe M. Dahlström,2, Guy Hanke,3, Anne Rokka,4, Natalia Battchikova,1, Tiina A. Salminen,2, Paula Mulo,1

Ayla Oenel,1, Wiebke Grebner,1, Markus Krischke,1, Agnes Fekete,1, Martin J. Mueller,1, Susanne Berger1 1

University of Wuerzburg, Germany

University of Turku, Department of Biochemistry, Molecular Plant Biology, Finland, Structural Bioinformatics Laboratory, Department of Biosciences, Åbo Akademi University, Finland, 3 Plant Physiology, Faculty of Biology and Chemistry, University of Osnabrück, Germany, 4 Centre for Biotechnology, Finland 1

2

Survival and fitness of plants being constantly challenged by dynamic environment are dependent on rapid adjustment of the metabolism. Changes in enzyme activity are considered as a one of the fastest and most efficient metabolic responses and they can be achieved through post-translational modifications (PTMs), which are known to affect the activity, interactions, as well as localization of a number of proteins. As chloroplast is the central hub of plant metabolism, PTMs of chloroplast proteins are crucial for the effective response of plants to sudden changes in ambient environment. Recent studies have shown that, besides protein phosphorylation, many other types of modifications, including acetylation, methylation and glycosylation, are important in the regulation of plastid metabolism.

In Arabidopisi thaliana ferredoxin-NADP+ oxidoreductase (FNR), a chloroplast protein linking the light reactions of photosynthesis to carbon assimilation, exists as a two isoforms. We have shown that both FNR isoforms are present as two distinct forms with different pI as a result of alternative transit peptide cleavage and partial Nα-acetylation. Additionally, both FNRs contain acetylation of a conserved lysine residue near to the active site but no evidence for in vivo phosphorylation was gained. Structural modeling and experimental evidence suggest that the membrane attachment of FNR or its direct interaction with ferredoxin is not affected by the identified modifications. However, the amounts of differently modified FNR forms change upon transfer from darkness to light, which implies involvement of the modifications in the regulation of FNR function.

Oxylipins are oxygenized polyunsaturated fatty acids, which act as signalling molecules with a variety of different functions. The oxylipin family consists of a wide range of metabolites like the jasmonates 12-oxophytodienoic acid (OPDA), jasmonic acid and jasmonic acid-isoleucin, which are important for developmental processes and play a significant role in responses to abiotic and biotic stresses. 13-Lipoxygenases (13-LOX) catalyze the oxygenation of polyunsaturated fatty acids to 13-hydroperoxyoctadecatrienoic acid which is the first step of the jasmonate biosynthesis and other oxylipin pathways. Recently it has been shown that LOX6 is important for a very fast systemic response in leaves after wounding. In our group the focus has been set on understanding the regulation of the biosynthesis and functions of oxylipins in roots of Arabidopsis. In roots, jasmonate synthesis is dependent on LOX6 that is essential for basal and stress induced jasmonate levels. The lox6 mutant displays a faster wilting phenotype and its roots are eaten faster by detritivors compared to the wild type. Beyond that, LOX6 overexpression lines have been generated to study the formation of LOX6-dependent oxylipins (jasmonates and other products). Comparison of LOX6 overexpressors and the wild type after application of the substrate α-linolenic acid showed that LOX6 expression is not limiting for jasmonate synthesis in roots. In addition, α-linolenic acid treatment caused oxidative stress as indicated by massive non-enzymatic, radical-catalyzed, oxygenation of esterified fatty acids in plant membranes.

[99] PUTATIVE UPSTREAM KINASES OF SOS2

[100] CHLOROPHYLL AND CAROTENOIDS CONTENTS IN NEVER RIPE AND DIAGEOTROPIC TOMATO MUTANTS TO CADMIUM STRESS

Juan de Dios Barajas-López,1, Jian-Kang Zhu,2, Hiroaki Fujii1 1

University of Turku; Department of Biochemistry, Finland 2 Purdue University, United States

Patricia Cury,1, Leticia Alves,1, Priscila Gratão,2, Priscila Miranda,1, Carolina Monteiro,1, Lucas Gaion,1 Universidade Estadual Paulista “Julio de Mesquita Filho”, Brazil, 2 Laboratório de Fisiologia Vegetal/Fcav/Unesp - Univ. Estadual Paulista/Depto. de Biologia, Brazil

1

Reversible protein phosphorylation is one of the most important mechanisms for cell signaling to respond environmental change. The sucrose non-fermenting-1 (Snf1)/AMP-activated protein kinase (AMPK) family is important for metabolic stress responses in eukaryotes. In Arabidopsis, there are 38 protein kinases in the family, designated SnRK. They are classified into three subgroups according to their sequence similarity. SnRK1s play important roles in metabolism, energy balance and growth whereas SnRK2s are important for abscisic acid and osmotic stress signaling. SnRK3 includes SOS2, which is important for salt tolerance. To analyze importance of phosphorylation in the activation loop of SOS2 for salt tolerance, complementation assays with mutated forms of SOS2 were performed. Phosphorylation-mimicking, but not phosphorylation-deficient, form of SOS2 can complement the salt tolerance pathway in the sos2 mutant. Thus, phosphorylation in its activation loop regulates SOS2 activity. We check involvement of GRIK1, which is an upstream kinase of SnRK1 (Plant Physiol 150, 996-1005) in the phosphorylation of SOS2. In our experimental condition with recombinant proteins, GRIK1 can phosphorylate and activate SOS2 in vitro. We also analyze mutant plants lacking GRIK1.

Contamined soils by heavy metals are a limiting factor to agriculture. Cadmium can cause oxidative stress inducing severe physiological disturbances and may reduce the overall photosynthetic capacity of green plants by reduced synthesis of pigments. Therefore, chlorophyll and carotenoids can indicate plant stress level. The aim of this study was to quantify these pigments and indicated plant stress condition in Solanum lycopersicum cv. Micro-tom and their mutants Never ripe (Nr) and diageotropic (dgt), which shows low sensitivity to ethylene and auxin, respectively. Our results indicate that the auxin absence in dgt plants had a favorable effect on photosynthetic pigments biosynthesis. The information available in this work is an important step towards obtaining a better understanding of the physiological changes caused by Cd and its effects on metabolic processes. Financial support by FAPESP

[101] RESPONSES TO DIFFERENT DEGREES OF WATER FLOODING OF SIX WILLOWS FAMILIES

[102] METABOLITE RESPONSES OF SILVER BIRCH TO ELEVATED HUMIDITY

Virginia Luquez,1, Maria Emilia Rodriguez,2, Guillermo Doffo,2, Teresa Cerrillo,3

Jenna Lihavainen,1, Viivi Ahonen,1, Markku Keinänen,1, Sarita Keski-Saari,1, Sari Kontunen-Soppela,1, Anu Sõber,2, Elina Oksanen,1

1

INFIVE; CONICET; UNLP, Argentina, 2 INFIVE, Argentina, 3 Inta Delta, Argentina

University of Eastern Finland UEF, Finland; Department of Biology, Finland, 2 University of Tartu; Institute of Ecology and Earth Sciences, Estonia

1

Morphological and physiological responses of willows to flooding have been characterized, but less is known about what happens in the post-flooding period. Flooding can reduce growth, but after the end of the stress episode, plants could modify some leaf and plants traits to compensate for the biomass losses. The aims of this work were: 1- To analyze morphological and physiological traits related to productivity in willows during the post- flooding period, 2- To analyze if these traits are modified in a different way according to the genotype and the deep of the floodwater. We analyzed three individuals of six willows families, growing in pots in a greenhouse. The plants were subjected to two treatments: 1 – Partial flooding: flooding of the roots only, for 2 months; 2 – Deep flooding: flooding in a pool up to 65 cm of water for 1 month. Both treatments were followed by 1 month of recovery (without flooding). The traits analyzed were growth, leaf size, specific leaf area, root to shoot ratio, leaf nitrogen content, stomatal conductance and electron transport rate. Some traits changed in the same way in both flooding treatments and different genotypes, like shoot to root ratio. But other traits changed in a different manner during the post flooding period, depending on the treatment and genetic background. The results obtained could help to identify traits that minimize biomass losses under flooding in willows, and this would be useful for breeding in flood risk areas.

Climate change scenarios predict that relative humidity (RH) may increase in northern Europe in association with increasing precipitation, cloud cover, atmospheric water vapor and more frequent wet days. RH affects plant growth and nutritional status; under high RH, plants can assimilate carbon with a relatively small cost of water, but at the same time mineral nutrient uptake is impaired due to decreased transpiration rate and mass flow of water. We studied the effects of elevated RH on the leaf metabolites of silver birch (Betula pendula Roth.) in a controlled growth chamber experiment and in a long-term Free Air Humidity Manipulation (FAHM) field experiment. In the chamber experiment, RH was elevated from 60% to 95%, and in the FAHM study RH was elevated by 7-8% over the ambient. Leaves were sampled after 26 days (chamber) and during the 4th growing season (field). Leaf starch content increased under elevated RH in both experiments. Untargeted metabolic profiling with GC-MS revealed that glutamate and aspartate increased, whereas shikimic acid, ribonic acid, glucose, sedoheptulose, glutamine, alanine and valine decreased under elevated RH in both experiments. In addition, elevated RH induced the production of secondary metabolites such as 3-coumaroyl quinic acids and flavonol glycosides in birch leaves in chamber and field experiments.

[103] RCD1 REGULATION AND RECOGNITION OF POLY(ADP-RIBOS)YLATION IN ARABIDOPSIS Julia Vainonen,1, Alexey Shapiguzov,1, Julia Krasensky,1, Natalia Battchikova,2, Kerri Hunter,1, Iulia Danciu,3, Claudia Jonak,3, Michael Wrzaczek,1, Jaakko Kangasjärvi,1

[104] LEAF ANATOMICAL AND BIOCHEMICAL TRAITS OF ANTHYLLIS HERMANNIAE L. IN RESPONSE TO ABIOTIC STRESS Stavroula Mamoucha,1, Nikolaos Chrostodoulakis,1

1

Plant Biology, Department of Biosciences, University of Helsinki, Finland, 2 Molecular Plant Biology, Department of Biochemistry and Food Chemistry, University of Turku, Finland, 3 Gregor Mendel Institute of Molecular Plant Biology, Austria

1

Plants are constantly exposed to a variety of abiotic stresses such as drought, heat and salinity which negatively affect their growth and productivity. Abiotic stress responses are regulated by complex hormonal and redox-dependent signaling pathways leading to transcriptional reprogramming.

Introduction: Plants growing under Mediterranean climatic conditions are exposed to two seasonally separated and totally different in quality environmental stress. This is the reason for their important adaptations in morphological, anatomical, physiological and biochemical features. Seasonal differences in leaf structure seem indispensable for the plant to overcome abiotic conditions of the hot and arid summer and the cold winter. Additionally, Mediterranean plant species are well known for the accumulatation of secondary metabolites (S.M.) which are very important for the survival under abiotic stress.

RADICAL-INDUCED CELL DEATH1 (RCD1) was identified as a key regulator of stress and hormonal signaling pathways in Arabidopsis. RCD1 is a multidomain protein containing the N-terminal WWE domain, inactive PARP-like domain and the C-terminal RST domain. RCD1 and its closest homolog SIMILAR TO RCD1 (SRO1) are the only Arabidopsis proteins containing the WWE domain. Using dot-blot assay and surface plasmon resonance method we show that WWE domain of RCD1 specifically binds poly(ADP-ribose) polymer with high affinity. Poly(ADP-ribosyl)ation is known to be involved in plant stress response, however, no acceptor protein except histones, or proteins involved in recognition of poly(ADP-ribose) chains have been described. We also show that RCD1 is tightly regulated in planta at the protein level. Treatment of Arabidopsis seedling with proteasome inhibitor leads to RCD1 stabilization which is phosphorylation-dependent. We identified four in-vivo phosphorylation sites in RCD1 and show that one of the phosphorylation sites is the target site for ASK protein kinase in-vitro. Currently we address the functional roles of the phosphorylation sites using transgenic Arabidopsis lines and site-directed mutagenesis. The results will provide new information on signal transduction and integration in Arabidopsis.

Faculty of Biology, Greece

Purpose: We investigated anatomical and biochemical profile of the leaf to reveal the plant responses to abiotic stress by employing different strategies. Materials and methods: Summer and winter leaves were detached, fixed, sectioned and investigated using light, transmission (ΤΕΜ) and scanning electron microscopy (SEM) along with histochemical tests. Results: The very xeromorphic summer leaves with large quantities of S.M. are replaced by mesomorphic leaves. The mesophyll is impregnated with huge tannin containing cells which are more developed in winter leaves. Epidermal cells are free of S.M. Conductive tissue is more developed in winter leaves. Histochemical treatments identified many S.M. Conclusions: The differences between leaves are attributed to different abiotic stress Seasonally dimorphism of A. hermanniae L. strongly indicates that climatic conditions correlate with structural differences between the two leaf types.

Acknowledgment: This work was supported by IKY - State Scholarship Foundation, Athens, Greece.

[105] LEAF ANATOMICAL AND BIOCHEMICAL TRAITS OF PHILLYREA LATIFOLIA L. (OLEACEAE) IN RESPONSE TO ABIOTIC STRESS

[106] OZONE: A THREAT TO CROP PRODUCTION Ashutosh Pandey,1, Baisakhi Majumder,2, Sarita Keski-Saari,3, Sari Kontunen-Soppela,3, Vivek Pandey,4, Elina Oksanen,5

Stavroula Mamoucha, , Nikolaos Chrostodoulakis, 1

1

1

Faculty of Biology, Greece

Introduction: Global climate change imposes Mediterranean regions to higher levels of temperatures and aridity. Such abiotic stresses are a complex challenge for the plants of the Mediterranean ecosystems. Consequently, they respond with structural modifications and shifting of their biosynthetic pathways to carbon based secondary compounds which are crucial for their protection under abiotic stress. P. latifolia L. is a sclerophyllous, evergreen tree growing wild in chaparral formations in Greece. It is commonly known as green olive tree or mock privet. The hard leaves are in opposite pairs, small and leathery. The flowers are small and the fruit is a drupe, containing a single seed. Purpose: The main objective of this research was to investigate the anatomical adaptations and the biochemical profile to reveal the plant responses to abiotic stress and the biosynthesis of useful secondary metabolites. Materials and methods: Young and mature leaves were detached, fixed, sectioned and investigated using light, transmission (ΤΕΜ) and scanning electron microscopy (SEM) along with histochemical tests (HI). Results: Leaves are hypostomatic, hairy and compact. The epidermis is unilayered and the palisade tissue is multilayered. Light microscopy revealed numerous huge inosclereids distributed throughout the mesophyll. SEM revealed interesting features of glandular trichomes. HI identified secondary metabolites the subcellular localization of S.M. Conclusions: In order to survive under abiotic factors P. latifolia L. adopts interesting anatomical features and physiological responses. It is very important to have a good knowledge of plants reactions to explain how they encounter stressful environments.

Acknowledgment: This work was supported by IKY - State Scholarship Foundation, Athens, Greece.

University of Eastern Finland; Plant Ecology and Environmental Science, National Botanical Research Institute (CsirNbri), Finland 2 National Botanical Research Institute (Csir-Nbri), Finland 3 University of Eastern Finland, Finland, 4 Plant Ecology and Environmental Science, National Botanical Research Institute (Csir-Nbri), Finland, 5 University of Eastern Finland UEF; Department of Biology, Finland 1

Ozone is considered to be a gaseous pollutant in the lower level of the atmosphere, the troposphere, causing a serious threat to global crop production in major agricultural regions of the world. Various modelling studies predict further increasing ozone levels, in South and East Asia in particular, thereby aggravating the damaging effects of ozone on agriculture. India’s bread basket, the Indo-Gangetic Plains (IGP), have been classified as a ‘hot spot’ for air pollution, owing to the intense agriculture, landuse changes, industrialization, urbanization, population growth and favourable meteorological conditions, causing high emissions of precursors for ozone formation. Field experiments with local crop cultivars of mustard (Brassica campestris L.) and rice (Oryza sativa L.) were conducted in ambient ozone concentrations throughout the growing season. EDU (ethylenediurea) was used as a chemical protectant against the adverse effects of ozone. Both the mustard and the rice cultivars showed sensitivity to prevailing ozone concentrations suffering yield losses, thereby indicating the severity of the ozone-induced risk to agriculture in this region. Mustard seed oil content decreased by 4-5 % in the prevailing ambient ozone concentrations. Out of 18 rice cultivars studied only seven showed adaptability in high-ozone environments in terms of grain yield. EDU-mediated protection against ozone stress was mainly due to the up-regulation of the antioxidative defence system, and its extent and timing varied with the developmental phase of the plant species and/or cultivars. The most responsive parameters in EDU treatments were lipid peroxidation, superoxide dismutase and catalase activities at the vegetative phase, and ascorbate and glutathione content at the flowering phase, under high ambient ozone conditions. These parameters can be used as the most useful indicator parameters, for practical ozone-tolerance screening in mustard and rice cultivars.

[107] ELUCIDATING EARLY STEPS OF SULFATE SENSING MECHANISMS BY BIOSENSORS

[108] LACE PLANT CELL DEATH MORPHOLOGIES INDUCED BY ABIOTIC STRESS

Veli Vural Uslu,1, Guido Grossmann,1, Rüdiger Hell,1

Adrian Dauphinee,1, Trevor Warner,1, Arunika Gunawardena1

1

Center for Organismal Studies, Heidelberg University, Germany

Nutrient uptake from the environment is a common feature of all organisms. Higher organisms need combinations of multiple nutrients for their growth and development. Plants are sessile organisms but they gather the nutrients from the rhizosphere, which is a very unstable and dynamic environment. Therefore, to adapt to the swift changes in the availability of nutrients in the external supply, plants need elaborate sensing and signal transduction systems. Despite extensive studies, nutrient sensing and the signal transduction cascades remained elusive due to technical limitations. Here I present a novel approach to reveal the sequence and the spatiotemporal dynamics of events between external sulfate depletion and sulfate deficiency response, in high resolution. Integration of the new RootChip technology with Biosensors for live-cell imaging enables us to visualize the early events in different environmental conditions in parallel to find out the metabolic parameters of nutrient sensing in unprecedented spatiotynamic resolution. Eventually, this approach makes it possible to build a working model based on these finding to shed a light on the sulfate sensing-signaling mechanisms as well as plant nutrition. Creating such data based models is critical to find out the environmental conditions for optimal and sustainable crop production.

1

Department of Biology, Dalhousie University, Canada

The lace plant (Aponogeton madagascariensis) is an aquatic monocot with a distinct perforated leaf morphology. The holes in lace plant leaves form as part of normal development by means of programmed cell death (PCD). PCD is critical throughout the lives of all multicellular organisms as it allows for the removal of superfluous cells during development and tissue homeostasis. In general, animal cell death is well characterized and better understood compared to plants. The lace plant has emerged as a novel model organism to study cell death in plants primarily due to the predictability of perforation formation and the plant’s nearly transparent leaves, which are ideal for live cell imaging. The morphology of lace plant developmental PCD has been well characterized using long term live cell imaging and other microscopy techniques. In this study, death was induced in lace plant cells that do not undergo PCD during perforation formation using various forms of abiotic stressors (i.e. Heat, NaCl, NaOH and HCL) at different intensities. Live cell imaging was performed and the acquired videos were used to analyze the dynamics, morphologies and time courses of cell death. There were differences and similarities between developmental and environmentally induced cell death, but in all forms of lace plant death the vacuole played a prominent role.

[109] TEMPERATURE AND DAY LENGTH EFFECTS ON EARLINESS IN NORDIC SPRING BARLEY Magnus Göransson,1, Jónatan Hermannsson,2, Áslaug Helgadóttir,2, Åsmund Bjørnstad,3, Morten Lillemo,3, Jón Hallsteinn Hallsson2

[110] INTERACTION BETWEEN AUXIN AND ETHYLENE IN IRON DEFICIENCY RESPONSES IN TOMATO Carolina Monteiro,1, Marina Gavassi,1, Leticia Alves,1, Lucas Gaion,1, Priscila Gratão,2, Renato Prado,1, Rogerio Carvalho1

1

Agricultural University of Iceland, Faculty of Land and Animal Resources, Iceland; Norwegian University of Life Sciences, Department of Plant Sciences, Norway, 2 Agricultural University of Iceland, Faculty of Land and Animal Resources, Iceland, 3 Norwegian University of Life Sciences, Department of Plant Sciences, Norway

1

Range expansion of cereal crops northwards is one way to alleviate predicted negative effects of climate change on today’s global grain baskets. Among economically important cereals, barley is the most tolerant to marginal conditions. Cultivation of spring barley in Iceland has in recent years expanded greatly following breeding efforts and changing weather conditions. The small scale breeding programme has emphasized early flowering and maturity at low temperature. The breeding gene pool is largely based on Nordic cultivars, with additions from the Faroe Islands and Scotland. Field trials have shown that Icelandic breeding lines are earlier than their ancestors, underlining the importance to select advantageous allele combinations in the target environment.

Iron deficiency is the most common micronutrient deficiency on plants in alkaline or calcareous soils and hydroponic cultures. Iron (Fe) is essential nutrient for development of plants, being constituent of several proteins and enzymes. Strategy I in dicots plants can induce development of subapical swelling with abundant root hairs, enhancement of ferric reducing capacity, acidification of the extracellular medium, and increase in the number of Fe2+-transporters (such as IRT1). By this way, auxin and ethylene can regulate these responses, since both of them are enhanced in Fe-deficiency. The aim of this work was to investigate the effect of Fe-deficiency in hormone tomato mutants. For this purpose, we used auxin-insensitive diageotropica (dgt), ethylene-insensitive Never ripe (Nr), ethylene overproduction epinastic (epi) and double mutants - Nr dgt and epi dgt. The mutants and their counterpart Micro-Tom (MT) were grown over 20-day period in the presence of Fe (20 µM Fe-EDDHMA). Afterwards, half part of the plants was grown without Fe during 20-days. Growth analyses, chlorophyll, lipid peroxidation and hydrogen peroxide contents were determined. According to results, we observed reduction in leaf area and chlorophyll in plants treated during 20-days without Fe. Dry mass and length of roots showed a slight increase, while lipid peroxidation and hydrogen peroxide contents increased in mutants when compared to control. Further analysis will be conducted to elucidate the Fe-deficiency responses. Financial support by FAPESP

The local genepool was screened in contrasting day length and temperature in order to gain a better understanding of genotype by environment interaction. We identified one group of day length neutral lines and one group with delayed flowering in short day conditions and high temperature. The collection has been genotyped and genome wide association scans performed to identify genes underlying these traits. Knowledge on the effect of allelic diversity in the breeding material will enable application of marker assisted selection for beneficial allele combinations. This will speed up the barley range expansion which will further facilitate a self-sufficient and diversified local agriculture in the Arctic region.

Universidade Estadual Paulista “Julio de Mesquita Filho”, Brazil, 2 Laboratório de Fisiologia Vegetal/Fcav/Unesp - Univ. Estadual Paulista/Depto. de Biologia, Brazil

[111] CRITICAL ROLES OF FERRITIN-LIKE PROTEINS IN CELLULAR FITNESS OF CYANOBACTERIA

[112] STOMATAL RESPONSES TO AIR HUMIDITY – THE INVOLVEMENT OF ABSCISIC ACID AND ETHYLENE

Karin Stensjö,1 1

Microbial Chemistry, Department of Chemistry-Ångström Laboratory, Uppsala University, Sweden

Louise E. Arve,1, Meseret T. Terfa,1, Jorunn Elisabeth Olsen,2, Sissel Torre1 Norwegian University of Life Sciences; Department of Plant Sciences, Norway, 2 Department of Plant Sciences, Norwegian University of Life Sciences, Norway

1

Maintenance of redox homeostasis, and the ability to control the level of reactive oxygen species is critical for cellular fitness and are crucial factors for efficient production of biofuels and other valuable metabolites. In our research we characterize proteins of importance for cellular adaptation, in the heterocyst-forming, N2-fixing cyanobacterium Nostoc punctiforme, as a strategy to explore and provide metabolic engineering tools to enhance the potential of cyanobacteria for green production. We have recognized the ferritin-like proteins as being of importance for the ability of cyanobacteria to adapt to potentially stressful conditions (1,2,3,4). The most wide-spread ferritin-like proteins in cyanobacteria are the Dps proteins, which corresponding genes, are represented in almost all bacterial genomes. N. punctiforme comprise a larger number of Dps proteins as compared to most other bacteria. The specificity and function of the Dps are only poorly understood. Using a multidimensional strategy: in-vivo characterization by gene-inactivation, overexpression of proteins, cell localization studies as well as in-vitro biochemical characterization of isolated proteins, we try to resolve the functions and roles of the Dps proteins in N. punctiforme. Here we present the roles of Dps-proteins in hydrogen peroxide tolerance, protection of DNA, iron regulation and localization to specific cells, that advise an important role in heterocyst metabolism, N2 fixation and, consequently, bio-H2 production.

[1] Ow SY et al (2008) J Proteome Res 7:1615-1628 [2] Ow SY et al (2009) J. Proteome Res 8 (1): 187–198. [3] Ekman M et al. (2011) J Proteome Res 10:1772-1784 [4] Ekman M et al. (2014) Environ Microbiol 16: 829-44

Stomata play a major role in controlling gaseous exchange of photosynthetic CO2 uptake and transpiration in response to changes in the environment. The regulation of stomatal aperture is important for crop productivity in field and in controlled environment and crucial for plant survival in hash and stressful environment. It has long been apparent that the aerial environment like relative air humidity (RH), CO2 as well as ozone and other gaseous compounds regulate stomatal movement. Plants grown in high RH have a larger stomatal aperture than plants in a drier environment. However, different climate factors modulate the responses and plants produced in high RH are more sensitive to elevated CO2 The mechanisms that determine stomatal responses to high RH are not well understood but is likely to be controlled by the plant hormones like abscisic acid (ABA) and ethylene. However, it appears that different plant species regulate ABA differently in response to high RH. The role of ethylene in stomatal movement is contradictory as ethylene has been shown to induce both stomatal opening and closure. In this work a description of air humidity stomatal responses of different species will be presented and the involvement of ABA and ethylene will be discussed.

[113] UNCOUPLING LOCAL AND SYSTEMIC ABIOTIC STRESS PERCEPTION AND RESPONSES IN ARABIDOPSIS

[114] NUTRIENT REQUIREMENT OF GROWTH IN DIFFERENT THERMAL ENVIRONMENTS Zsofia Stangl,1, Catherine Campbell,2, Vaughan Hurry,2

Melanie Carmody, , Matthew Gordon, , Tiina Blomster, , Barry Pogson, , Jaakko Kangasjärvi, 1

1

2

1

2

1

The University of Helsinki, Finland, 2 The Australian National University, Australia

Long distance stress signals are particularly difficult to study as both local stress perception and signal reception in distal tissues must be taken into account. Spatially, perception of stress by different organs of a plant must be relayed to other parts of the plant in an intracellular, cell-to-cell, and long distance manner; temporally, signals are needed for both sudden protective or gradual developmental adjustments depending on the extremity and longevity of the stress. This transcriptional and physiological study demonstrates the spatial and temporal complexity of systemic abiotic stress responses with the use of high light and ozone treatments in Arabidopsis. Signals produced by the plant from a localised application of stress on a single leaf are monitored in distal leaves using mutant analysis, LUC expression assays, and RT-qPCR of ROS and stress specific marker genes. The requirement of the vasculature in rapid long distance stress signaling is investigated through comparison of systemic signals in orthostichous leaves connected directly through the vasculature compared to unconnected leaves. Finally, an alternative strategy for separating local stress perception and distal signal reception is proposed with the use of a non-invasive and tissue specific inducible system to genetically uncouple an organ for a short amount of time. Work from this study will allow for further understanding of long distance inter-organ communication in plants.

Umeå Plant Science Centre; Dept. of Plant Physiology, Umeå University, Sweden, 2 Umeå Plant Science Centre; Department of Forest Genetics and Plant Physiology; Swedish University of Agricultural Sciences, Sweden

1

We investigated nutrient uptake and growth rates of two birch species (Betula pendula and Betula utilis) with different growth strategies, grown under different temperatures (Tg), in order to assess the interactive effect of temperature and nutrient availability on growth and acclimation. We monitored the nutrient uptake rates and the growth rates at three different Tg, 20°C (control), 10°C and 30°C, under unrestricted nutrient conditions. We found that for fast growing B. pendula the ratio of nutrient uptake to growth was higher under both 30°C and 10°C compared to the control Tg. In contrast, while the slow growing B. utilis responded to non-optimal temperatures with lower growth rates, the ratio of nutrient uptake to growth was not significantly changed. This differential response of the two birch species suggests that fast growing species respond more dynamically to growth temperature than slow growing species. Furthermore our results suggest that increased nutrient uptake supports growth under non-optimal Tg, and therefore nutrient availability might be a restricting factor for maintaining a fast-growth strategy.

[115] GAS EXCHANGE IN THE LEAVES OF LOTUS CORNICULATUS UNDER WATER STRESS Panagiota Kostopoulou,1, Maria Karatassiou,1, Valasia Iakovoglou,2, Paraskevi Exadaktylou,1, Martha Lazaridou -Athanasiadou,2

[116] THE MATRIX METALLOPROTEASE MMP3 IS IMPORTANT FOR RESISTANCE AGAINST FUNGAL DISEASES IN ARABIDOPSIS THALIANA Sungyong Kim,1, Christiane Funk,1, Wolfgang Schröder,1, Katrina Ramonell,2, Walter Gassmann,3

1

Laboratory of Rangeland Ecology, Department of Forestry and Natural Environment, Aristotle University of Thessaloniki, Greece, 2 Department of Forestry, Faculty of Agriculture Technology, Tei of Eastern Macedonia and Thrace, Greece

1

Stomatal conductance is the main factor influencing the basic plant functions of photosynthesis and transpiration through the control of CO2 and water vapour exchange. The aim of this study was to determine the effect of stomatal conductance on gas exchange in leaves of Lotus corniculatus grown under well irrigated and water stressed conditions. Plants from two natural populations of semi-arid areas of Northern Greece (Drama and Kilkis) were selected and transplanted to pots. The second year of plant growth, two irrigation regimes were applied: a) irrigation up to field capacity (IR) and b) interruption of the irrigation so that, a gradually intensive water stress occurred (WS). During late spring of 2014, measurements of leaf water potential (Ψ), assimilation rate (A), transpiration rate (E), stomatal conductance (Gs) and internal CO2 concentration (Ci) were performed on three randomly selected pots, in two day intervals. The ratios A/Ci (apparent carboxylation efficiency) and Ci/Gs (mesophyll efficiency) were also estimated. Water stress significantly reduced all the studied parameters, but had no effect on the apparent carboxylation efficiency (A/Ci). On the contrary, higher mesophyll efficiency was found under water stress conditions. Significant differences between the two regions were found only for the transpiration rate, with the population of Drama showing lower values of E. The results showed linear, positive effect of stomatal conductance on CO2 concentration and transpirational water loss, revealing a higher reduction of transpiration than of internal CO2 concentration (Ci) under water stress conditions.

Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases, containing a conserved catalytic domain (HEXXHXXGXX(H/D)) where zinc binds for activation. MMPs have been characterized in detail in mammals, and they have been shown to play key roles in many physiological and pathological processes, like e.g. morphogenesis or tissue repair. In the annual plant Arabidopsis thaliana five MMPs have been detected, whose functions have not been well characterized so far; At-MMP2 was shown to be involved in early flowering and salt stress. Using Genevestigator At-MMP3 was found to be highly up-regulated after fungi infection, but not after bacterial treatment. Salt and cold treatment also caused up-regulation of its gene expression, while heat treatment had not effect. To study the role of At-MMP3 in resistance to fungal diseases, At-mmp3 KO plants were generated and treated with Erysiphe cichoracearum, which causes powdery mildew disease. The mutant showed severe disease symptoms compared to wild type. At-MMP3 over-expresser lines currently are tested for resistance against fungus disease. Treatment with the bacterium Pseudomonas syringae did not induce any disease symptoms in the At-MMP3 KO mutant. Based on these data, At-MMP3 could function in the plants response to fungal infection.

Acknowledgments: This research has been co-financed by EU and Greek national funds.



Umeå University, Sweden, 2 University of Alabama, United States, 3 University of Missouri-Columbia, United States

[117] ANALYSIS OF BARLEY GENE SEQUENCES BCI-4 AND BCI-7 IN ARABIDOPSIS THALIANA - EFFECTS ON APHID PREFERENCE AND POPULATION GROWTH

[118] DYNAMIC CHANGES IN GLUCOSINOLATES IN MUSTARD (SINAPIS ALBA L.) PLANTS UNDER BACTERIAL STRESS

Aleksandra Losvik,1, Lisa Beste,1, Sara Mehrabi,1, Lisbeth Jonsson,1

Eva Kuchler,1, Hartwig Wilfried Pfeifhofer,1, Maria Müller1

1

Department of Ecology, Environment and Plant Sciences, Stockholm University, Sweden

Two barley genes BCI-4 and BCI-7 with putative defense function against aphids have been studied by expression in Arabidopsis thaliana and evaluation using the generalist peach aphid (Myzus persicae). The BCI-4 gene encodes for a putative calcium binding protein and BCI-7 encodes for a proteinase inhibitor. In barley, the expression of the two gene sequences differs between aphid susceptible and moderately resistant genotypes and was shown to be up-regulated after bird cherry-oat aphid (Rhopalosiphum padi) infestation only in the latter ones. Based on this, the BCI-4 and BCI-7 genes were considered to be potential resistance factors against aphids. The two barley sequences were expressed in A. thaliana using either a constitutive or a phloem-specific promoter. A series of tests were conducted to assess the effects of the transformation on aphid behavior and reproduction. All experiments with aphids were carried out using transgenic lines with confirmed transgene expression and azygous control lines. When compared to control lines, constitutive expression of BCI-7 resulted in altered behavior and reproduction of peach aphid, expressed as reduced settling and lower population growth, respectively. A somewhat reduced population growth was also observed for one of the BCI-7 lines in which transcript was expressed in the phloem. Transformation with the BCI-4 gene sequence caused no changes in aphid behavior or proliferation. Our study shows that expression of aphid-induced barley gene BCI-7 in Arabidopsis provides moderate resistance by affecting the behavior and proliferation of peach aphid.

1

Institute of Plant Sciences, University of Graz, Austria

Brassicaceae are grown around the world for various purposes. A long history of plant breeding has resulted in highly variability in appearance, phytochemistry and use. Several pathogens are particularly damaging Brassicaceae and cause severe diseases, e.g. Alternaria leaf spots (Thomma, 2003) or downy mildew (Jensen et al., 1999; Niu et al., 1983). In our studies we use for the first time as model system mustard and Xanthomonas and we focus the interest on symptom development, as well as on the dynamic specific changes of glucosinolates and antioxidants after infection with black rot pathogen Xanthomonas on mustard plants (Vicente and Holub, 2013). Sinapis alba plants are grown as model organism in climate chambers under controlled conditions for six weeks. After this period the plants are inoculated with Xanthomonas campestris pv. campestris and the symptom development is observed between four hours and five days. At each time-point samples are collected for different morphological and physiological investigations (glucosinolates, glutathione, ascorbic acid). The results show a time-dependent influence on secondary metabolites of mustard plants connected to the action of Xanthomonas. Therefore we conclude that Sinapis alba and Xanthomonas seem to be a perfect model system for studying dynamic changes in quantity and quality of secondary metabolites.

Jensen, B.D., Hockenhull, J., Munk, L., 1999. Plant Pathol. 48, 604–612. Niu, X., Leung, H., Williams, P.H., 1983. J. Am. Soc. Hort. Sci. 108, 775–778. Thomma, B.P.H.J., 2003. Mol. Plant Pathol. 4, 225–236. Vicente, J.G., Holub E.B., 2013. Mol. Plant Pathol. 14, 2-18.

[119] THE INDUCTION OF B-1-3-GLUCANASES BY BIRD CHERRY-OAT APHID IN BARLEY VARIES HIGHLY BETWEEN GENOTYPES WITHOUT ANY CORRELATION WITH APHID SUSCEPTIBILITY OR RESISTANCE

[120] COMPARISON OF BACTERIAL TRANSCRIPTOMES PROVIDES INSIGHTS INTO THE METABOLISM OF CLUSTER II FRANKIA STRAINS IN ROOT NODULES OF CUCURBITALES VS. ROSALES HOST PLANTS

Sara Mehrabi,1, Inger Åhman,2, Lisbeth M.V. Jonsson1

Thanh Van Nguyen,1, Rolf Hilker,2, Jörn Kalinowski,3, Alison Berry,4, Katharina Pawlowski1

1

Department of Ecology, Environment and Plant Sciences, Stockholm University, Sweden, 2 Department of Plant Breeding, Swedish University of Agricultural Sciences, Sweden

Department of Ecology, Environment and Plant Sciences; Stockholm University, Sweden, 2 Department of Bioinformatics and System Biology; Justus Liebig University Giessen, Germany, 3 Center for Biotechnology; University of Bielefeld, Germany, 4 Department of Plant Sciences; University of California, United States

Bird cherry-oat aphid (Rhopalosiphum padi L.) is an important cosmopolitan pest on cereals. We are studying the induced responses to this aphid in barley, aiming to find genes that are adding either to resistance or to susceptibility. Induction of pathogenesis-related (PR) proteins is one of the plant responses upon microbial or insect attack and ß-glucanases constitute one family of PR-proteins. Several b-glucanases are induced in barley by R. padi, but their role in this interaction is not clear. We had found earlier, in a limited number of barley genotypes, that two b-glucanase sequences were expressed in the susceptible but not the moderately resistant lines, suggesting that they were susceptibility factors. Here we have studied the expression of three b-glucanases in a larger selection of 16 barley genotypes, earlier characterized as aphid susceptible or moderately resistant, based on their ability to support nymphal growth. The transcript abundances before and at various time points after aphid infestation were analyzed in relation to aphid resistance. The analysis showed that the pattern of induction or repression of the sequences varied greatly between the different genotypes. However, there was no consistent pattern in relation to aphid growth or aphid settling. The results illustrate that there is variation in the reaction to aphids between genotypes of the same plant species. Furthermore, they did not confirm the idea that one or more of the b-glucanases are related to susceptibility, or to resistance, measured either as antibiosis (growth and reproduction) or antixenosis (behavior) to the aphid.

This study focuses on the nitrogen-fixing symbioses between actinorhizal plants and members of the uncultured Frankia cluster (cluster II) which is important for our understanding of the evolution of nitrogen-fixing root nodule symbioses. Members of the uncultured Frankia cluster nodulate plants from four different families (Datiscaceae, Coriariaceae, Rhamnaceae, Rosaceae) growing on five different continents. Nodule structures of Datiscaceae and Coriariaceae differ from that of Rhamnaceae and Rosaceae. Therefore, we want to compare the bacterial nodule transcriptomes of Datisca glomerata (Datiscaceae) and Ceanothus thyrsiflorus (Rhamnaceae) to investigate the influences of host family on the microsymbiont.

1

Genes encoding proteins that participate in nitrogen fixation are highly expressed in both types of nodules. In Alnus glutinosa, malate is the carbon source supplied to the microsymbiont in the root nodules (Jeong et al., 2004). Our result shows that while the situation is probably similar in D. glomerata, Frankia may receive a different carbon source in C. thyrsiflorus. The expressions of transporters between the two microsymbionts also differ substantially, indicating differences in nutrient exchange in both symbioses. In D. glomerata, Frankia seems to export fixed nitrogen in the form of arginine (Berry et al., 2004). In both transcriptomes, genes encoding enzymes involved in arginine metabolism are expressed at high levels, implying a similar type of nitrogen metabolism in C. thyrsiflorus. There are also significant differences in expression of Frankia genes that encode functions in cell division, sporulation and filamentation in both types of nodule. This might associate with the differences of bacterial morphology in the infected cells.

J. Jeong et al. 2004. Plant Physiol. 134: 969-978. A.M. Berry et al. 2004. Plant Physiol. 135:1849-1862.

[121] DYNAMIC COMPARTMENT SPECIFIC CHANGES IN GLUTATHIONE AND ASCORBATE LEVELS IN PLANTS UNDER DIFFERENT ENVIRONMENTAL CONDITIONS

[122] EARLY SALICYLIC AND JASMONIC ACID RELATED COI1 DEPENDENT DEFENCE RESPONSES IN POTATO TO NECROTROPHIC FUNGI ALTERNARIA SOLANI

Maria Müller,1, Bernd Zechmann2

Dharani Burra,1, Firuz Odilbekov,1, Rosahl Sabine,2, Hedley Pete,3, Liljeroth Erland,1, Erik Andreasson1

1

Institute of Plant Sciences, University of Graz, Austria 2 Center for Microscopy and Imaging, Baylor University, United States

Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Sweden, 2 Department of Stress and Developmental Biology, Leibniz Institute of Plant Biochemistry, Germany, 3 Cell and Molecular Sciences, James Hutton Institute, Invergowrie, United Kingdom

Ascobate and glutathione (and its precursors) are the most important antioxidants in plants. They are involved in the detoxification of reactive oxygen species (ROS), redox signaling, in the modulation of defense gene expression and they are important for the regulation of enzymatic activities. For these reasons levels of glutathione and ascorbate are often used as stress markers in plants.

Early blight caused by Alternaria solani Sorauer causes foliar damage leading to poor quality potato (Solanum tubersoum) tubers resulting in up to 50% losses in production1. Early blight is controlled by the use of fungicides. Worldwide majority of the expenditure on fungicides in early blight control is spent on potato1. Lack of resistant potato varieties warrants the excessive use of fungicides. Numerous reports indicating fungicidal resistance in A. solani populations have been published in the recent past1. Hence there is a need for improved understanding of molecular nature of defence responses to A. solani, results from which can benefit existing breeding programs.

In our studies we focused our interest on the dynamic compartment specific changes of glutathione and precursors as well as ascorbate, to gain thorough knowledge about the subcellular distribution of these antioxidants in plants and on the importance of these antioxidants in certain cell compartments during stress situations. Different agricultural plants (Cucurbita, Nicotiana), as well as Arabidopsis were used as model plants under different environmental conditions in order to detect and quantify subcellular glutathione and ascorbate. For this purpose beside other techniques an immunogold cytohistochemical approach with computer-supported transmission electron microscopy approach was developed and adapted to different plant material. These studies and methods can now be used for the development of new defense strategies for agricultural use in the future, and can protect farmers from possible crop losses induced by environmental stress situations in the future. Acknowledgement: This work was supported by the Austrian Science Fund.

1

We show by using hormone related transgenic potato plants that salicylic acid (SA) restricts pathogen growth and symptom development; while jasmonic acid (JA) related COI1 (coronatine insensitive) is needed for pathogen growth restriction. In addition, time series based microarray analysis revealed that early and rapid defence response to pathogen infection is mediated by SA and COI1 signalling pathways.

Reference 1. Kemmitt, Greg. “Early blight of potato and tomato.” The Plant Health Instructor (2002).

[123] AUTOIMMUNITY TRIGGERED BY ACTIVATION OF PAIRED NLRS Diep Thi-Ngoc Tran,1, Eunyoung Chae,1, Monika Demar,1, Rebecca Schwab,1, Detlef Weigel1 1

Max Planck Institute for Developmental Biology, Germany

[124] INDUCTION OF TRYPTOPHAN-DERIVED AND HYDROXYCINNAMIC ACID AMIDE SECONDARY METABOLITES AND ALTERATION OF THE PRIMARY METABOLISM DURING INFECTION OF WHEAT (TRITICUM AESTIVUM) BY ZYMOSEPTORIA TRITICI Chaouch Sejir,1, Pasquet Jean-Claude,1, Belvert Floriant,2, Lebrun Marc-Henri,3, Dufresne Marie,1, Seng Jean-Marc,1, Gouache David,4, Patrick Saindrenan1 Institute of Plant Sciences of Paris-Saclay, Université Paris Sud, France, 2 Platform Metatoul - EAD6, LISBP - INSA de Toulouse, France, 3 BIOGER, UMR INRA-APT, Campus Agroparistech, France, 4 Arvalis Sciences Du Végétal, France

1

Autoimmunity observed in hybrid plants can result from epistatic interactions between disease resistance (R) genes from separate lineages. R genes often encode nucleotide-binding domain and leucine-rich repeat containing (NLR) proteins, which play key roles in perception of pathogens and activation of immune responses in plants. In the autoimmune hybrid of Arabidopsis thaliana, Uk-3 X Uk-1, which exhibits dwarfism phenotype and spontaneous cell death in leaves, both causal genes encode TIR-NLR proteins - DANGEROUS MIX1 (DM1) and DANGEROUS MIX2d (DM2d). Both genes are located in multi-gene NLR clusters where numerous evolutionary processes contribute to generate diverse NLR members. To investigate how the NLRs as a pair trigger autoimmune responses, we reconstituted the DM1/ DM2d dependent cell death in Nicotiana benthamiana. We found hypersensitive responses (HR) occurred only when both NLRs as full-length proteins were co-expressed, indicating both are required to initiate signaling. Mutations in the P-loop motif in either DM1 or DM2 suppressed HR, suggesting that signaling is dependent on ATP binding and hydrolysis. Co-immunoprecipitation assay revealed DM1/ DM1, DM2d/DM2d homo- and DM1/DM2d heterotypic associations, apparently using a common interaction interface in the N-termini including the TIR domains. Mutations disrupting the heterodimeric association in yeast resulted in the loss of HR in planta. Mutations in the MHD motif that supposedly affect conformation around the ATP binding pocket greatly changed the activity of DM2d but not of DM1. We propose that a highly sensitized and immune competent NLR such as DM2d can activate resting NLR complexes via a heteromeric NLR complex, such as DM1/DM2d.

Septoria tritici blotch (STB) disease of wheat (Triticum aestivum) is caused by the hemibiotroph Zymoseptoria tritici (formerly known as Mycosphaerella graminicola). The asymptomatic and biotrophic phase of Z. tritici lasts approximately 14 days post-inoculation (dpi). Then the fungus brutally accelerates its growth switching its lifestyle to necrotrophy. Here we showed that the tryptophan (Trp) pathway was upregulated in wheat leaves infected with Z. tritici concomitantly with cell death initiation (i.e., 14 dpi). This induction was evidenced by the accumulation of Trp and the enhanced transcript levels of anthranilate synthase (AS.1, AS.2, AS2) and the tryptophan synthase b-subunit (TBS). The tryptophan-derived metabolites tryptamine and serotonin accumulated after cell death initiation concomitantly with a marked increase of the tryptophan decarboxylase (TDC) transcript levels. We also report hydroxycinnamic acid amides accumulation in the infected leaves. Moreover, GC-MS analysis indicated dramatic alteration of the wheat primary metabolism in response to STB with accumulation of sugars, amino-acids and the polyamines putrescine and spermidine. Exogenously supply of diseased leaves with auxin and Trp before cell death initiation significantly reduced pathogen growth. As a whole, our findings indicate that the tryptophan pathway is involved in basal resistance of wheat to STB and its up-regulation is tightly associated with the onset of the first symptoms providing reliable and early markers of the transition to necrotrophy.

[125] MOLECULAR CHARACTERIZATION OF OSWRKY6 THAT BINDS TO THE W-BOX LIKE ELEMENT 1 OF OSPR10A PROMOTER AND CONFERS REDUCED SUSCEPTIBILITY TO PATHOGEN

[126] BIOSYNTHETIC GENE CLUSTERS IN PLANT CHEMICAL DEFENCE AND THEIR DYNAMIC GENOMIC ORGANISATION

Changhyun Choi,1, Seon Hee Hwang,1, Il Lan Fang,1, Soon Il Kwon,1, Sang Ryeol Park,1, Ilpyung Ahn,1, Duk-Ju Hwang,1

Behrooz Darbani,1, Adam Takos,1, Daniela Lai,1, Camilla Knudsen,1, Alexandra Öchsner,1, Fred Rook1

1

NAAS, Rep. of South Korea

WRKY proteins are transcription factors (TFs) that regulate the expression of defense-related genes. Among SA inducible WRKY TFs OsWRKY6 was identified as a positive regulator of OsPR10a by transient expression assays. The physical interaction between the W-box-like element 1 (WLE1), which positively regulates OsPR10a/PBZ1 expression, and OsWRKY6 was verified in vitro. Several PR genes including OsPR10a are constitutively activated; enhanced disease resistance to pathogens was shown in transgenic plants overexpressing (ox) OsWRKY6. By contrast, PR gene induction was compromised in transgenic OsWRKY6-RNAi lines, suggesting that OsWRKY6 is a positive regulator of defense responses. OsWRKY6ox lines displayed lesions on their leaves, and increased OsWRKY6 levels cause cell death. SA levels were higher in OsWRKY6ox lines than that in WT and transcript level of OsICS1, which encodes a major enzyme for SA biosynthesis, was increased in OsWRKY6ox lines compared to WT. OsWRKY6 directly bound to the OsICS1 promoter in vivo. This indicates that OsWRKY6 can directly regulate OsICS1 and then increases SA levels. OsWRKY6 auto-regulates its own expression. OsWRKY6 protein degradation is possibly regulated by ubiquitination. Our results suggest that OsWRKY6 positively regulates defense responses through OsICS1 activation and OsWRKY6 stabilization.

1

University of Copenhagen, Denmark

An enormous diversity of chemical defence compounds are produced by plants. A recent theme in plant biology is that the genes for many of the biosynthetic pathways for such compounds are organised in gene clusters: a functional grouping of non-homologous genes that are localised in close proximity in the genome. Cyanogenic glucosides (α-hydroxynitrile glucosides) are a class of chemical defence compounds that upon tissue damage by herbivores are hydrolysed by specific β-glucosidases, resulting in the release of hydrogen cyanide (HCN). The biosynthesis of cyanogenic glucosides has repeatedly evolved in different plant lineages and independently formed biosynthetic gene clusters for cyanogenic glucosides are present in the genomes of cassava, sorghum, and the model legume Lotus japonicus. The sub-clade within the Lotus genus that contains Lotus japonicus has evolved the ability to additionally produce non-cyanogenic hydroxynitrile glucosides named rhodiocyanosides. A bifurcation of the biosynthetic pathway at the second enzymatic step involves two alternative CYP736 paralogs and an expansion of the gene cluster. In Lotus japonicus, natural variation for the rhodiocyanoside trait is correlated with latitude and caused by repeated loss of the rhodiocyanoside specific CYP736A80 gene. A genomic context of highly repetitive sequences such as retrotransposons likely facilitates these gene cluster dynamics. The self-organisation of biosynthetic pathways into gene clusters likely results from the maintenance, by varying and opposing ecological selection pressures, of alternative allele combinations and chemical defence polymorphisms in natural populations.

[127] THE BIOSYNTHETIC PATHWAY AND ECOLOGICAL ROLE OF HYDROXYNITRILE GLUCOSIDES IN BARLEY-POWDERY MILDEW INTERACTION

[128] ELUCIDATING THE PATHOGENIC EFFECTS OF RAMULARIA COLLO-CYGNI IN BARLEY René Lemcke,1, Michael Lyngkjær,1

Marcus Ehlert, , Birger Lindberg Møller, , Michael Foged Lyngkjær 1

1

1

1

Department of Plant and Environmental Sciences, Denmark

Hydroxynitrile glucosides (HNGs) are widely distributed in the plant kingdom and have diverse plant physiological and ecological functions, e.g. as translocation and storage molecules for nitrogen or as defence compounds against herbivore or pathogen attack. Barley (Hordeum vulgare) contains five leucine-derived hydroxynitrile glucosides: epiheterodendrin, epidermin, sutherlandin, osmaronin and dihydroosmaronin. These compounds are restricted to the leaves, where 99% are stored in the epidermal cell layer and account for 90% of the soluble carbohydrates in this cell type. Interestingly, the obligate biotrophic barley powdery mildew fungus Blumeria graminis f.sp. hordei strictly infects these cells suggesting a crucial role of HNGs in the interaction of barley and powdery mildew. Most likely, the fungus utilizes the HNGs for host recognition and as source for glucose and nitrogen. Little is known about the plant-internal transport of HNGs and the role of HNG-transporters in the interaction. We have found that HNG-biosynthesis genes are clustered within the barley genome. This cluster also harbours genes for a putative transporter of the ABC and MATE family, respectively. Comparing transcript levels in uninfected and infected leaves will provide information about their regulation and role in the interaction of barley and powdery mildew. In addition, immunolocalization of the ABC and MATE transporter will give further information about their putative role in intracellular and intercellular HNG transport, e.g. through the cytoplasm, tonoplast or to the wax layer.

Section for Plant Biochemistry, Department of Plant and Environmental Science, Faculty of Science, University of Copenhagen, Denmark

1

Crop protection receives in times of dramatically increasing global population and, in relation of that rising nutrition requirement, more and more credit in the scientific community by development of resistant varieties against pests and diseases. However, a former hardly recognized pathogen, Ramularia collo-cygni (Rcc), has moved into focus within the last decade because of its increasing infestation in North and Central Europe, New Zealand and South America. As infectious fungus it attacks cereals and develops from an asymptomatic endophyte into a major pathogen that entails yield losses and decreases in grain quality. So far, no cultivars with very high expression of resistance have been identified, and no efficient diagnosis predicting a damaging attack exists. This project concerns the infection biology of Rcc to help avoid Rcc disease outbreaks and protect progress in barley yield increases. Initially, different barley lines will be tested to identify tolerant varieties of spring and winter barley and to investigate the plant-pathogen interaction. The detailed interaction between Rcc and barley will be exploited by genomic and post-genomic tools in a sustainable and efficient manner. The following focus will be set to clear up the basic genetic mechanisms and molecular components underlying Rcc recognition, colonization and disease development and discover genetic and molecular requirements for the endophytic and pathogenic life styles of Rcc.

[129] GLUCOSINOLATE DEGRADATION PRODUCTS: THEIR PHYSIOLOGICAL EFFECTS ON ARABIDOPSIS THALIANA János Urbancsok, , Ane Kjersti Vie, , Ralph Kissen, , Atle Magnar Bones, 1

1

1

2

[130] CYTOKININ MEDIATED RESISTANCE AND BIOCONTROL Dominik Grosskinsky,1, Tafner Richard,2, Maria V. Moreno,3, Sebastian A. Stenglein,3, Inés E. García de Salamone,4, Louise M. Nelson,5, Ondřej Novák,6, Miroslav Strnad,6, Eric van der Graaff,1, Thomas Roitsch,7

1

Norwegian University of Science and Technology, Department of Biology, Norway, 2 CMBG; Department of Biology; Norwegian University of Science and Technology, Norway

University of Copenhagen, Denmark, 2 University of Graz, Austria, 3 Facultad de Agronomía de Azul-Uncpba, Argentina, 4 Universidad de Buenos Aires, Argentina, 5 University of British Columbia, Canada, 6 Palacký University, Czech Republic, 7 University of Copenhagen; Department of Plant and Environmental Sciences; Copenhagen Plant Science Centre, Denmark

Glucosinolates are one of the most important groups of sulfur- and nitrogen-containing secondary plant metabolites, characteristic of the Brassicales. To date, more than 130 different glucosinolates have been identified among plants and this huge diversity is due to a variable side chain of these compounds that can originate from one of 8 amino acids and further undergo secondary modifications (1). Glucosinolates are metabolized by specific enzymes called myrosinases. This enzymic decomposition by myrosinases depends on the presence or absence of special cofactors such as additional proteins, metal ions and pH (2). The substrate and the enzyme are separated from each other but upon tissue disruption they come into contact and the chemical reaction can lead to a variety of biologically important degradation products such as thiocyanates, isothiocyanates, nitriles and epithionitriles. These degradation products play a key role in plant defense responses against pathogens, pests, insects and herbivores, however some of them are also subject of great interest in medical research owing to their dietary cancer preventive effects in humans (3). In order to better understand the biological effects of glucosinolate hydrolysis products we have established and standardized an in vitro system which allows us to evaluate their effects on Arabidopsis thaliana. We exposed several mutant lines to glucosinolate hydrolysis products and investigated their dose-dependent behavior in the presence of chemicals. The remarkable results will be presented and discussed. References

Considering future demands in plant protection and restrictions in the use of classic pesticides, the development of alternative strategies is a major goal in plant sciences.

1.) Agerbirk, N, Olsen, CE (2012) Glucosinolate structures in evolution. Phytochemistry, 77, 16-45. 2.) Bones, AM, Rossiter, JR (2006) The enzymic and chemically induced decomposition of glucosinolates. Phytochemistry, 67, 1053-1067. 3.) Mithen, R, Armah, C, Traka, M (2011) Cruciferous vegetables – and biological activity of isothiocyanates and indoles. In Vegetables, Whole Grains, and Their Derivatives in Cancer Prevention, Mutanen M and Pajari AM (eds), Springer, Dordrecht.

1

Biological control of plant diseases by beneficial microbes offers therein a high potential for integrated plant disease management. Mechanisms contributing to such biocontrol phenomena comprise direct effects on pathogens or virulence factors and strengthening the plant. Cytokinins are phytohormones that are known for long time to be involved in various regulatory processes of plant physiology and development, but have only recently been shown to modulate plant immunity. Complementary experimental approaches, including autoregulated cytokinin synthesis in response to pathogen infection, showed that cytokinins enhance resistance against the virulent hemibiotrophic pathogen Pseudomonas syringae in tobacco. The cytokinin-mediated resistance strongly correlated with an increased level of bactericidal activities and was shown to depend on the two major antimicrobial phytoalexins in tobacco, scopoletin and capsidiol. The specificity of the underlying mechanism is evident from a differential effect of the cis- and trans-isomers of zeatin. The integration into the complex plant defense network is evident from the involvement of salicylic acid and the negative interference of abscisic acid. The mechanism of cytokinin-triggered immunity was also shown to be the basis for the biocontrol activities of a Pseudomonas fluorescence strain that had been identified based on its growth promoting activity. Complementary gain- and loss-of-function approaches with the host plant and the biocontrol strain identified the microbial cytokinin production as a key determinant of the protection of the plant from a bacterial pathogen. The implications of this mechanism for the coevolution of host plants and cytokinin-producing agonists and pathogens and the possible practical application in agriculture to engineer pathogen resistance as well as abiotic and biotic cross tolerance are discussed.

[131] STOMATA SENSITIVITY TO MICROCLIMATIC VARIABLES DECREASES UNDER ELEVATED CO2 - A FACE STUDY

[132] RELATIONSHIP BETWEEN TEA YIELDS AND CLIMATIC FACTORS IN TAIWAN Chin-Ying Yang,1, Mao-Chang Wang,2

Alireza Houshmandfar, , Glenn J. Fitzgerald, , Roger Armstrong, , Allene A. Macabuhay, , Michael Tausz, 1

2

2

1

3

Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Australia, 2 Department of Environment and Primary Industries, Austria, 3 Department of Forest and Ecosystem Science, The University of Melbourne, Australia

1

We employed the widely used boundary line technique to investigate whether relationships of stomatal conductance to environmental factors such as temperature, soil moisture, vapour pressure deficit (VPD), and photosynthetic active radiation (PAR) are similar for ambient and elevated [CO2] conditions. Wheat (Triticum aestivum L.) was grown under ambient [CO2] (approximately 390 µmol mol-1) and elevated [CO2] (approximately 550 µmol mol-1) in the Australian Grains Free Air CO2 Enrichment (AGFACE) facility. Using an open infrared gas analyser, approximately 1500 gas exchange measurements were collected from stem elongation until after completed anthesis on randomly selected healthy and youngest fully grown leaves during 2013 and 2014 wheat seasons. The results suggest that elevated [CO2] does not only decrease stomatal conductance because it increases intercellular [CO2] but also because it decreases sensitivity of stomata to some environmental factors: Vapour pressure deficit induced stomata closure at approximately 2.5 kPa under ambient [CO2] but only at 3 KPa under elevated [CO2]. Optimum temperature for stomatal opening was 2˚C lower under elevated [CO2]. Stomata sensitivity to soil moisture was also significantly lower under elevated than ambient [CO2] conditions. Elevated [CO2] decreased stomatal conductance by on average approximately 30%. Key words: Humidity; Optimum temperature; Soil moisture; Triticum aestivum; Stomatal conductance; Transpiration; Vapour pressure deficit;

National Chung Hsing University; Department of Agronomy, Taiwan, 2 Chinese Culture University; Department of Accounting, Taiwan

1

Climate change has had an impact on global agriculture, and numerous studies have focused on the relationship between agriculture and climatic factors. Tea is a major economic crop of Taiwan, and the beginning of the tea industry dates back 300 years. This study used annual data from the Taiwanese tea industry from between 1953 and 2013 as the sample and investigated the relationships among tea yield value, tea yield quantity, and climatic factors by using regression analysis. Climatic factors included temperature, rainfall, and typhoons. Taiwan has a subtropical climate; during the research period, the mean temperature was 23.45 °C and the mean rainfall was 2179.22 mm. It rains almost every month, and afternoon showers typically adjust high temperatures during the summer. The empirical results suggested that the temperature has a positive relationship with tea yield value and quantity. Typhoons affect Taiwan a mean of 3.39 times per year, and tea farmers typically harvest or moderately prune tea trees before the onset of typhoons; therefore, typhoons have no statistically significant relationship with tea yield quantity. However, typhoons may affect the quality of and market demand for tea. The empirical results suggested that typhoons have a negative relationship with tea yield value. This study uses the Taiwanese tea industry for investigating the relationships among tea yield value, tea yield quantity, and climatic factors by using regression analysis. The results of our study contribute to the literature and provide a valuable reference for practitioners in the tea industry, government, and academia.

Keywords: tea yield value, tea yield quantity, temperature, rainfall, typhoon

[133] THE INVESTIGATION OF THE EFFECT INDOLE ACETIC ACID (IAA) ON LEAF SENESCENCE IN ARABIDOPSIS THALIANA

[134] MORPHOLOGICAL CHARACTERISTICS AND ROOT GROWTH POTENTIAL OF BAREROOT AND CONTAINER TURKEY OAK (QUERCUS CERRIS L.) SEEDLINGS

Nihal Gören Sağlam,1, Vicky Buchanan-Wollaston,2

Esra Bayar,1, Ayse Deligoz,1

1

Istanbul University faculty of Science, Department of Botany, Turkey, 2 School of Life Sciences, University of Warwick, United Kingdom

1

Senescence is a last phase of plant development. It is an important developmental process in plants that eventually leads to whole plant, organ, tissue and cell death through highly regulated, endogenously controlled degenerative processes. Hormones are endogenous components that mediate the effect of environmental factors to regulate senescence. Some hormones such as ethylene, abscisic acid (ABA), jasmonic acid (JA), and salicylic acid (SA) act as inducers of senescence, whereas cytokinins and gibberellins play a role in its suppression. Although, synthetic and natural auxins delay senescence in a major part of the plant tissues, Goren and Cag (2007) showed that the senescence that occurred in the cotyledons of Helianthus annuus L. seedlings was accelerated by the application of IAA. Generally, hormones are not sufficient alone to promote senescence and there is considerable evidence to show that the signalling pathways interconnect to regulate gene expression. The aim of this study is to determine the genes with increased and decreased expression by making microarray in Arabidopsis leaves harvested 4 hours after IAA application at day 27, 31 and 35. Effect of IAA on leaf senescence was investigated by using chlorophyll fluorescence imaging and microarray in the present study. It was seen from the chlorophyll fluorescence imaging studies that applications at different days, especially at day 27 caused a difference in chlorophyll fluorescence values. Microarray results showed that expression levels of the genes playing a role in senescence were significantly increased at day 27 mainly, after IAA application.

In this study, morphology and root growth potential of bare-root and container Turkey oak (Quercus cerris L.) seedlings were compared at the end of the first growth year (February 2014) in the Egirdir Forest Nursery. Container seedlings were significantly larger in growth parameters (root collar diameter, seedling height, shoot and root dry weights, root/shoot ratio, tap root weight) than the bare-root seedlings. The root systems of container seedlings had a larger number of first order lateral than the bare-root seedlings. In addition, bare-root seedlings had poor root growth potential compared to contained seedlings. As a result, container seedlings were taller, greater root collar diameter, heavier, and more first-order lateral roots than bare-root seedlings.

Keywords: Senescence, IAA, chlorophyll fluorescence imaging, microarray, Arabidopsis thaliana

Suleyman Demirel University; Faculty of Forestry; Silviculture Department, Turkey

[135] DNA FINGERPRINTING OF PECAN CULTIVARS

[136] PLANT CELL WALL INTEGRITY IS MAINTAINED THROUGH COOPERATION OF DIFFERENT SENSING MECHANISMS

Anrie Allen,1, Gesine Coetzer,1, Johan Spies,1

Timo Engelsdorf,1, Manikandan Veerabagu,1, Joseph Francis Mckenna,2, Frauke Augstein,1, Dieuwertje Van der Does,3, Håkon Hoel,1, Cyril Zipfel,3, Thorsten Hamann,1

1

University of the Free State, South Africa

Norwegian University of Science and Technology, Norway, 2 Oxford Brookes University, United Kingdom, 3 The Sainsbury Laboratory, United Kingdom

1

Pecan is an important horticultural nut crop originally from North America and is now widely cultivated in South Africa for its economic value. However, a large number of these pecan cultivars are of unknown or questionable pedigree. Molecular techniques must be used to firstly determine the origin of each cultivar and secondly to identify cultivars. The Simple Sequence Repeat (SSR) technique is based on the Polymerase Chain Reaction (PCR) and has been very informative in cultivar identification in many crops. These repeat regions are also known as microsatellite DNA. The ease of use and potential of multiplexing make cpSSRs markers extremely important in genetic diversity studies. The following 7 microsatellites were studied, ntcp40 (mono-nucleotide), wga118 (di-nucleotide), wga242 (di-nucleotide), wga321 (di-nucleotide), ga38 (di-nucleotide), ga39 (di-nucleotide) and cin4 (tri-nucleotide). Each one of the 7 primer sets result in different alleles, enabling the identification of different cultivars and indicating the degree of genetic differences (or heterozygosity) within a cultivar. A total of 150 plants represented 6 different South African cultivars, were used in this study. Although different alleles exist for each primer set, cultivars cannot be identified by using a single primer set. The combination of at least four primer sets are needed for a reliable identification of any specific South African pecan nut cultivar. The results of this study showed that DNA fingerprinting, by means of SSR techniques, were both suitable for genetic diversity analysis and identification of all the South African pecan cultivars.

Plant cell walls fulfill diverse functions during plant development and stress responses. Cell wall integrity (CWI) has to be maintained during these different processes. Although a number of potential CWI maintenance sensors and mediators have been identified, neither their specific functions nor their relationships with each other have been fully resolved. To clarify and standardize the respective functions of candidate proteins, we analyzed responses to CWI impairment in 21 mutant lines of Arabidopsis thaliana. CWI was impaired through cellulose biosynthesis inhibition using isoxaben (ISX) and specificity was ensured by inclusion of the ISX-resistant genotype ixr1-1. The analyses involved quantification of root growth, ISX resistance, ectopic lignin, phytohormone (jasmonic acid and salicylic acid) accumulation and functional assays using osmotica. The resulting datasets were integrated through phenotypic clustering to obtain a global overview of the candidate activities in CWI maintenance. While none of the candidate sensors analyzed mediates the osmosensitivity of ISX-induced responses, the data show that changes in turgor pressure are integral elements of CWI maintenance. The receptor-like kinases (RLKs) THESEUS and FEI2 are required for both ectopic lignin and phytohormone accumulation, whereas RLKs required during plant immunity apparently only modulate responses to CWI impairment. While the putative stretch-activated calcium channel MCA1, the RLK WAK2 and plastid-localized mechanosensitive channels MSL2/3 affect only phytohormone accumulation, the nitrate reductases NIA1 and NIA2 are required for all ISX-induced responses. Our data suggest that CWI in plants is maintained by cooperative function of distinct sensors and sensing mechanisms.

[137] REGULATION OF THE ACTIVITY AND STABILITY OF ACC OXIDASE BY NUCLEOSIDE TRIPHOSPHATE

[138] ANTIOXIDATIVE ACTIVITY OF TOTAL PHENOLIC EXTRACTS FROM OATS (AVENA SATIVA) IN VARAMIN FROM IRAN

Hiroki Iio,1, Takuma Maekawa,1, Tomohide Uno,1, Kengo Kanamaru,1, Hiroshi Yamagata1

Abbas Pazoki1

1

Kobe University; Graduate School of Agricultural Science; Laboratory of Biochemistry, Japan

Ethylene controls plant growth and development as well as responses to stresses. Ethylene biosynthetic pathway involves conversion of S-adenosyl-L-methionine to 1-aminocyclopropane-1-carboxylic acid (ACC). ACC oxidase (ACO) catalyzes the oxidation of ACC to ethylene, the final step of the pathway. ACO belongs to the superfamily of dioxygenases that require Fe2+ as cofactor and bicarbonate as activator. Ascorbic acid has been recently reported to participate in the ring opening of ACC and provide a single-electron to the active site of ACO. However, the detailed action mechanisms of cofactors and other regulatory factors remains to be elucidated. Here we report some new enzymatic characteristics of ACO. Optimum pHs of purified recombinant Arabidopsis thaliana ACO2 (AtACO2) and tomato ACO1 (LeACO1) expressed in E. coli were both 6.9 and reversibly inactivated with a rise of pH. AtACO2 was activated 5-fold by 12 mM NTP but not by NMP. Heat stability of AtACO2 was also remarkably increased by NTP. On the contrary LeACO1 was strongly inhibited by NTP. Although the role of NTP as a modulator of plant enzymes have not been well known, the regulation of ACO by NTP might have some physiological role in certain stages of plant growth. Model structures showed some differences between AtACO2 and LeACO1 in their C-terminal region. A chimera ACO, AtACO21-220 -LeACO1221-318 was not activated by NTP, while LeACO11-217 - AtACO2218-317 was not inhibited but activated by NTP. Taken collectively, a loop and/or helix11 in C-terminal region of ACO are likely to be responsible for the regulation by NTP.

1

Department of Basic Sciences, Islamic Azad University, Varamin Branch, Iran

Seeds of oats was collected from five region of Varamin then were powdered. Total phenolic in the extraction of seeds was determined by spectrometer according to the folin –ciocoltu procedure and calculated as gallic acid equivalents and the antioxidative activity of extracts determinated by autoxidation of linoleic-acid. the highest and lowest plenolic content was seen in Karkhaneghand and Jamalabad region respectively .The maximum of antioxidative activity was recorded in Pishva and the minimum was in Qarchak. The Data of research were analyzed by spss (one-way ANOVA).

[139] CONTROL STRATEGIES AGAINST BOTRYTIS CINEREA ISOLATED FROM STRAWBERRIES

[140] ANALYSIS OF TRANSGENIC BANANA PLANTS WITH ENHANCED RESISTANCE AGAINST BANANA BUNCHY TOP VIRUS

Izumi Sasaki1 1

National Institute of Technology, Oyama College, Japan

Yi-Yin Do,1, Pung-Ling Huang1 1

Botrytis cinerea is a widespread fungus able to infect the aerial parts of many plant species. This study examines the most effective anti-Botrytis strategies leading to reduce pesticides on strawberries. To provide the protection which harmless to humans, higher animals and plants, various approaches were tested. Calcium, yogurt, and 50°C treatments significantly inhibited the spore germination and mycelial growth of B. cinerea. The beta-glucosidase of B. cinerea showed high activities both inside and outside of the fungal cell grown in medium containing calcium. On the other hand, the CMCase and avicelase activities remained low during the incubation. Our findings suggest that the calcium treatments inhibit CMCase production and cause a perturbation of transportations for cell wall-degrading enzymes in B. cinerea.

National Taiwan University, Taiwan

Banana bunchy top disease is a serious disease in tropics caused by Banana bunchy top virus (BBTV). The symptoms include dwarfism in plants and clustering of leaves at the top giving a broom-like appearance on pseudostem. The veins and petioles turn dark green. Infected banana plants often do not bear fruits. In this study, banana plants transformed with antisense BBTV replicase gene were generated and independently transformed plant lines were analyzed for resistance to BBTV. The transgenic banana plants obtained exhibited resistance against BBTV. The consistency of the antisense replicase gene transformation and BBTV resistance indicates that replicase-mediated resistance against BBTV was attained in banana. To exploit the flanking sequences as molecular markers, IPCR (Inverse polymerase chain reaction) and APCR (Anchored polymerase chain reaction) analyze were carried out to determine the border sequences of transgenic lines. To elucidate the mechanism of BBTV resistant transgenic banana lines, we used Northern analysis to determine the accumulation of the small RNA population. The results showed siRNA sequence complementary to transgene accumulated in the transgenic lines. Additionally, infection by BBTV was found to induce the higher siRNA accumulation when compared to un-infected plants. It could be inferred that the resistance mechanism is through post-transcriptional gene silencing of the Rep gene within the BBTV.

[141] CAN ENVIRONMENTALLY INDUCED PHENOTYPIC VARIATION LEAD TO RECURRENT SPECIATION? –INSIGHTS FROM ECOPHYSIOLOGICAL INVESTIGATIONS OF THE MOUNTAIN PLANT HELIOSPERMA PUSILLUM (CARYOPHYLLACEAE)

[142] CHARACTERISATION OF KEY ELEMENTS IN HEXOSE-PHOSPHATE METABOLISM Laura Kathrine Perby,1, Tom Hamborg Nielsen,1 1

Clara Bertel,1, Bozo Frajman,1, Karl Hülber,2, Othmar Buchner,1, Peter Schönswetter,1, Gilbert Neuner,1

Plant and Environmental Sciences, University of Copenhagen; Copenhagen Plant Science Center, Denmark

University of Innsbruck, Institute of Botany, Austria, 2 University of Vienna, Department of Botany and Biodiversity Research, Austria

1

Evolution is strongly influenced by natural selection, favouring individuals adapted in morphology and physiology to the environmental conditions at their growing site. Heliosperma pusillum and H. veselskyi (Caryophyllaceae) from the south-eastern Alps represent an example of morphological and functional adaptation to divergent environmental conditions: H. pusillum has short, glabrous leaves and occurs in open, wet rock habitats in the alpine belt. In contrast, H. veselskyi has elongated leaves and sticky glandular hairs and inhabits shallow caves and cliff overhangs below the timberline. Although phenotypic divergence remains stable across two generations in a common garden, taxa cannot be separated on a genetic basis as shown by AFLP and RADseq data. They rather represent elevation-specific ecotypes; H. veselskyi was shown to have independently evolved multiple times triggered by local environmental conditions. We investigated the functional divergence in relation to microclimatic conditions. Their interplay very likely initiated ecological speciation between the two highly interfertile (as shown by crossing experiments) taxa, which may occur in relatively close proximity. Differentiation in functional traits suggests that different physiological traits are favoured at peculiar growing sites. We further compared field data with data from a common garden experiment in order to estimate to which extent functional traits are driven by short-term-adaptions to the environment. The ecophysiological results will be linked to field studies and phylogenetic investigations, which assessed the underlying molecular mechanisms of differentiation between taxa and the stability of divergence across divergent environments and generations.

Central metabolism of carbohydrates comprises the core reactions that connect all other parts of plant metabolism, such as photosynthesis, respiration, nitrogen-assimilation and formation of structural cell components and secondary metabolites. Consequently, carbohydrate metabolism is essential for plant development and stress tolerance, and it defines the major components of many harvested plant products. Our aim is to characterise key regulatory enzymes of hexose-phosphate metabolism. We will

focus on F2KP (Fru-6-P, 2-kinase/Fru-2,6-bisPase) and PFKs (phosphofructokinases) from A. thaliana. F2KP is a bifunctional enzyme, which phosphorylates and dephosphorylates the C-2 position of Fru6-P. The catalytic regions of F2KP reside at the C-terminal half of the amino acid sequence and is homologous to mammalian enzymes (Nielsen et al., 2004). In addition, F2KP in higher plants comprises a long N-terminus, containing several predicted functional domains. We aim to characterise the molecular function of these motifs. PFKs catalyse the phosphorylation of fructose-6-phosphate to fructose-1,6-bisphosphate. In A. thaliana, the PFKs comprise 5 cytosolic and 2 plastidic isoforms (Mustroph et al., 2007, Nielsen et al 2004). Our studies will aim to characterize the kinetics of selected A. thaliana PFKs and their impact on metabolism as revealed by KO-mutants.

[143] PHENOTYPIC PLASTICITY AND ACCLIMATION CAPACITY UNDER CLIMATE CHANGE: SILVER BIRCH (BETULA PENDULA) PROVENANCES IN A COMMON GARDEN EXPERIMENT

[144] CIRCADIAN PATTERNS OF HYDRAULIC AND XYLEM SAP PROPERTIES: IN SITU STUDY ON HYBRID ASPEN

Sari Kontunen-Soppela,1, Sarita Keski-Saari,1, Antti Tenkanen,1, Matti Rousi,2, Elina Oksanen1

Annika Karusion,1, Arne Sellin,1, Eele Õunapuu-Pikas1

1

University of Eastern Finland UEF; Department of Biology, Finland, 2 Natural Resources Institute Finland, Luke; Vantaa Reseach Unit, Finland

1

Extent of intraspecific variation defines the ability of a species to acclimate to environmental conditions, such as climate change. We have studied geographical variation in acclimation capacity of silver birch (Betula pendula) to warming climate in a long-term multi-site common garden experiment (www.uef.fi/birchadaption). The experimental set-up enables us to evaluate variation due to genotype, provenance and environment. Altogether 26 genotypes, representing 6 provenances (latitudes 60°, 61°, 62°, 65°, 66°, 67°N), grow in three sites in southern (60°N), central (62°N) and northern (67°N) Finland. The plant material is cloned from naturally regenerated stands and the southward latitudinal shift of provenances resembles the future temperature conditions. The results from tree phenology, growth and photosynthetic studies showed large variation and plasticity among the birch provenances, but instead of a clear latitudinal cline, the provenances fall into two groups: the southern and the northern provenances. The height growth of the northern provenances was the smallest in all sites. The growth termination of the northern provenances appears to be more strictly controlled by the photoperiod than in the southern provenances. The northern provenances also showed higher chlorophyll content, total photosynthesis and stomatal conductance than the southern ones.

Physico-chemical properties of xylem sap and tree hydraulic traits are being studied more extensively year by year. Nevertheless, to date complex circadian characterization of these characteristics measured in situ is still lacking. The aim of this study was to characterize circadian rhythms of tree hydraulic traits and physico-chemical properties of xylem sap in hybrid aspen (Populus tremula L. × P. tremuloides Michx.). In addition the study aimed to clarify which environmental drivers govern the daily dynamics of these parameters. All studied hydraulic characteristics including whole-branch hydraulic conductance (Kwb), barebranch hydraulic conductance (KB), leaf blade hydraulic conductance (Klb), petiole hydraulic conductance (KP) and leaf blade relative resistance (Rlb) as well as all xylem sap properties like K+ concentration ([K+]), electrical conductivity (ssap), osmolality (Osm) and pH showed clear daily dynamics. Environmental factors like photosynthetic photon flux density (PPFD), atmospheric water vapor pressure deficit (VPD), air temperature (TA) and relative humidity (RH) had significant impact on both hydraulic and xylem sap properties. Xylem sap pH was negatively correlated with soil water potential (ΨS), which also affected other measured characteristics. Most of the hydraulic traits and xylem sap physico-chemical properties depended on tree size and branch height. We detected strong correlations between [K+] and Kwb, [K+] and Klb, ssap and Kwb, ssap and Klb. However, correlation between [K+] and KB was not seen, as KB was depressed at midday. Other hydraulic properties did not show midday depression. The present study demonstrates an unequivocal coupling between circadian patterns of hydraulic conductance and xylem sap physico-chemical properties. The primary environmental factor responsible for those circadian patterns appears to be photosynthetic photon flux density (PPFD).

Institute of Ecology and Earth Sciences, University of Tartu, Estonia

[145] PHYLOGENETIC ANALYSIS OF SLAC1 PROTEIN FAMILY AND THE ROLE OF SLAC1 MEDIATED FAST STOMATAL CLOSURE IN THE ECOPHYSIOLOGY OF LAND PLANTS Sanna Ehonen,1, Jorma Vahala,1, Jaakko Kangasjärvi,2, Teemu Hölttä,2, Eero Nikinmaa2 1

Department of Biosciences; University of Helsinki, Finland, The University of Helsinki, Finland

[146] ARE LIGNIN MONOMERS SECRETED FROM RAY PARENCHYMAL CELLS IN LIGNIFYING NORWAY SPRUCE XYLEM? K. Fagerstedt,1, A. Kärkönen,2, E. Vaisanen,3, O. Blokhina,1, L. Zhao3

2

The Arabidopsis thaliana S-type anion channel SLAC1 has been shown to be responsible for rapid stomatal closure in response to various environmental factors. The molecular mechanisms involved in SLAC1 regulation have been extensively studied but to our knowledge there has been no attempt to study its role from ecological and ecophysiological perspectives outside laboratory conditions. In order to yield insight into the significance of SLAC1-dependent regulation in the stomatal responses under natural conditions and especially in trees, we have started to elucidate the presence and structure of SLAC1 protein family and related stomatal function in several plant species for which genome sequence information is available. The phylogenetic analysis suggests that Populus species may have lost the functionally important regulatory and structural features of SLAC1 and thus have altered ability to regulate their stomatal aperture, which is evident in measurements of stomatal function. To test the hypothesis further we have prepared transgenic hybrid aspen (Populus tremula x termuloides) lines with potentially altered stomatal regulation. These lines will help us elucidate the role of SLAC1 in the ecophysiology of land plants.

Department of Biosciences, University of Helsinki, Finland, 2 Department of Agricultural Sciences, University of Helsinki, Finland, 3 Department of Agricultural Sciences, University of Helsinki, Finland; Department of Biosciences, University of Helsinki, Finland 1

In Norway spruce (Picea abies L. Karst.) lignin forms a major part of the xylem cell walls and renders the tracheids water resistant while giving support to the cell walls and the whole tree trunk. In Norway spruce lignin is polymerized mainly of coniferyl alcohol but the origins of this monolignol is not known. In the present investigation we examined the possibility whether coniferyl alcohol is produced in the ray parenchymal cells as in angiosperms where neighboring cells have been shown to contribute to biosynthesis of monolignols used for lignin formation in tracheary elements (Pesquet et al. 2013, Smith et al. 2013). Hence, we used Zeiss PALM MicroBeam laser capture microdissection system to isolate ray parenchyma from thin tangential cryomicrotome sections (30 μm) of developing Norway spruce xylem of a mature tree. Total RNA extracted from the collected material was used for low input sequencing on Illumina platform to reveal any transcripts related to monolignol biosynthesis and secretion. As a control material we used xylem tracheid cells isolated from similar cryomicrotome sections. The deep sequencing protocol gives also information on the mRNA frequency in a particular cell type. The results are mapped to the whole genome sequencing data of Norway spruce to identify the key genes that are active in these tissues during xylem development and to characterize cell type-specific monolignol synthesis and transport.

Pesquet E. et al. (2013) Plant Cell 25: 1314-1328. Smith R. et al. (2013) Plant Cell 25: 3988-3999.

[147] TRANSCRIPTIONAL AND METABOLIC PROFILING OF ARBUSCULAR MYCORRHIZAL AND PHOSPHATE-TREATED MEDICAGO TRUNCATULA LEAVES

[148] INSIGHTS FROM HIGH RESOLUTION PHENOTYPING OF ARABIDOPSIS THALIANA ROOTS Klaus Palme,1

Lisa Adolfsson1, Hugues Nziengui1, Azeez Suliman Beebo1, Jila Aboalizadeh1, Benoît Schoefs2, Cornelia Spetea1 1 2

1

University of Freiburg; Institute of Biology, Germany

Department of Biological and Environmental Sciences, University of Gothenburg, Gothenburg, Sweden Mer, Molécules, Santé, MicroMar - EA2160, LUNAM Université, Le Mans, France

Arbuscular mycorrhizas (AM) provide both nutritional (mostly phosphate, Pi) and non-nutritional benefits to plants (including plant defense) and lead to improved growth. However, how AM-regulated signaling pathways in leaves are interconnected has not yet been fully elucidated. Here we used both molecular and metabolic approaches to investigate systemic changes in leaves from control-, AM- and Pi-treated Medicago truncatula plants. Microarray-based genome-wide profiling and quantitative polymerase reaction chain (qPCR) revealed that genes of jasmonate-, flavonoid/anthocyanin- and terpentenoid biosynthesis pathway were activated in AM-leaves and repressed in Pi-leaves, while gene involved in Fe homeostasis were co-repressed in AM- and Pi-leaves. In agreement with the transcriptomics data, we found increased anthocyanin accumulation restricted to AM-leaves, and lower Fe levels in both treatments as compared to control leaves. Based on these data, we propose a conceptual model in which AM-induced jasmonate biosynthesis and signaling plays a central role in regulating plant fitness through the control of secondary metabolite biosynthesis in a Pi-independent manner.

The study of root development relies on techniques for the accurate visualization of tissue and organ structure to understand cell patterning and patterns of gene expression. Currently available techniques for three-dimensional (3D) imaging are limited with respect to the thickness of roots and the resolution that can be achieved. In order to achieve a detailed functional and quantitative understanding of roots, cellular features of roots must be quantified in the three-dimensional context of cells and tissue layers. We therefore aimed, besides genetically, molecularly and functionally characterizing root development, at developing an intrinsic root coordinate system (iRoCS) as a reference model for analysis of the Arabidopsis root apical meristem. iRoCS can be used to rapidly parameterize image data within a single framework in a standardized way. It enables large cohorts of roots to be annotated, making statistical analyses accessible and giving an unbiased evaluation of previously hidden developmental phenotypes. iRoCS was used to study root patterning and shown to even distinguish subtle changes in the distribution of cell division in different cell layers in knock-out mutants.

Physiologia Plantarum

[149] REGULATION OF FLOWERING TIME BY SVP-FLM EXPRESSION LEVEL CONTROL IN ARABIDOPSIS THALIANA

An International Journal for Plant Biology

Jechang Woo, , Kihyun Lim 1

1

1

Department of Biological Science; College of Natural Sciences; Mokpo National University, Rep. of South Korea

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Physiologia Plantarum is an international journal committed to publishing the best full-length original research papers that advance our understanding of the primary physiological, biochemical, molecular and genetic mechanisms governing plant development, growth and productivity; including plant interactions with the biotic and abiotic environment. The journal also welcomes submission of shorter breakthrough manuscripts containing novel, exciting but solidly underpinned research that merits rapid publication. The journal publishes papers on all aspects of all organizational levels of experimental plant biology ranging from molecular and cell biology, biochemistry and biophysics to ecophysiology and global change biology.

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S E P T E M B E R 2ISSN0 0031-9317 14

Aims and scope

Physiologia Plantarum is an international journal committed to publishing the best full-length original research papers that advance our understanding of the primary physiological, biochemical, molecular and genetic mechanisms governing plant development, growth and productivity; including plant interactions with the biotic and abiotic environment. The journal also welcomes submission of shorter breakthrough manuscripts containing novel, exciting but solidly underpinned research that merits rapid publication. The journal publishes papers on all aspects of all organizational levels of experimental plant biology ranging from molecular and cell biology, biochemistry and biophysics to ecophysiology and global change biology. Physiologia Plantarum also publishes Minireviews with the aim of providing a forum for the exchange of information on recent scientific advances and technical developments.

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An International Journal for Plant Biology Regular manuscripts should present original hypothesis-driven research in one of the five subject areas covered

Regular manuscripts should present original hypothesis-driven research in one of the five subject areas covered by the journal (Biochemistry and Metabolism; Ecophysiology, Stress and Adaptation; Uptake, Transport and Assimilation; Development, Growth and Differentiation; Photobiology and Photosynthesis). The experiments described should address questions pertinent to the research topic and the outcomes should advance our understanding of the processes being studied, not simply repeat known results in a different species.

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Pivotal role of the AREB/ABF-SnRK2 pathway in ABRE-mediated transcription in response to osmotic stress in plants

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Minireviews, which are subject to peer review, should cover the background of a topical research problem, emphasizing the state of the art and serving as a focal point for further research. They must be written concisely, with no more than 50 references and a maximum of 8 printed pages (one printed page of full text is equivalent to around 5600 characters, including spaces). Minireviews will be considered in all areas of experimental plant science and it is recommended that the Editor-in-Chief be contacted in advance to confirm the suitability of the chosen topic.

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Minireviews, which are subject to peer review, should cover the background of a topical research problem, emphasizing the state of the art and serving as a focal point for further research. They must be written concisely, with no more than 50 references and a maximum of 8 printed pages (one printed page of full text is equivalent to around 5600 characters, including spaces). Minireviews will be considered in all areas of experimental plant science and it is recommended that the Editor-in-Chief be contacted in advance to confirm the suitability of the chosen topic.

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Physiologia Plantarum is an international journal committed to publishing the best full-length original research papers that advance our understanding of the primary physiological, biochemical, molecular and genetic mechanisms governing plant development, growth and productivity; including plant interactions with the biotic and abiotic environment. The journal also welcomes submission of shorter breakthrough manuscripts containing novel, exciting but solidly underpinned research that merits rapid publication. The journal publishes papers on all aspects of all organizational levels of experimental plant biology ranging from molecular and cell biology, biochemistry and biophysics to ecophysiology and global change biology.

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Printed in Singapore by C. O. S. Printers Pte Ltd

Physiologia Plantarum also publishes Minireviews with the aim of providing a forum for the exchange of information on recent scientific advances and technical developments.

Regular manuscripts

Anpresent International Journal Biology Regular manuscripts should original hypothesis-driven research infor one of Plant the five subject areas covered

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Physiologia Plantarum

PHYSIOLOGIA PLANTARUM

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An International Journal for Plant Biology

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Yasunari Fujita, Takuya Yoshida and Kazuko Yamaguchi-Shinozaki DOI: 10.1111/j.1399-3054.2012.01635.x

APRIL 2015

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by the journal (Biochemistry and Metabolism; Ecophysiology, Stress and Adaptation; Uptake, Transport and Assimilation; Development, Growth and Differentiation; Photobiology and Photosynthesis). The experiments described should address questions pertinent to the research topic and the outcomes should advance our understanding of the processes being studied, not simply repeat known results in a different species.

Regulation of cambial activity in relation to environmental conditions: understanding the role of temperature in wood formation of trees

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Shahanara Begum, Satoshi Nakaba, Yusuke Yamagishi, et al. DOI: 10.1111/j.1399-3054.2012.01663.x

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Minireviews, which are subject to peer review, should cover the background of a topical research problem, emphasizing the state of the art and serving as a focal point for further research. They must be written concisely, with no more than 50 references and a maximum of 8 printed pages (one printed page of full text is equivalent to around 5600 characters, including spaces). Minireviews will be considered in all areas of experimental plant science and it is recommended that the Editor-in-Chief be contacted in advance to confirm the suitability of the chosen topic.

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concisely, with no more than 50 references and a maximum of 8 printed pages (one printed page of full text is equivalent to around 5600 characters, including spaces). Minireviews will be considered in all areas of experimental plant science and it is recommended that the Editor-in-Chief be contacted in advance to confirm the suitability of the chosen topic.

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by the journal (Biochemistry and Metabolism; Ecophysiology, Stress and Adaptation; Uptake, Transport and Assimilation; Development, Growth and Differentiation; Photobiology and Photosynthesis). The experiments described should address questions pertinent to the research topic and the outcomes should advance our understanding of the processes being studied, not simply repeat known results in a different species.

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As mentioned flowering repressor factor, 35S:FLC also showed early flowering time however overexpression of SVP by 35S:FLC is still not clear and relation between SVP-FLM, SVP-FLC on the point of expression level should be investigated further.

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PHYSIOLOGIA PLANTARUM

Flowering is an essential period that life pattern changes for posterity. These changes are regulated by many environmental cues, and we analyzed the function of the SVP and FLM genes known to be regulated by temperature. Those genes were reported that suppress flowering under normal condition but accelerate flowering by degradation of gene product at high temperature condition. When OX(overexpression;35S) plants and knock out mutants(KO) were grown long day cycles at 22℃, flowering time of svp:KO and flm:KO showed 7 days earlier than wild-type, but 35S:SVP showed 12 days earlier than that of wild type. Flowering time of 35S:FLM is also early almost same as 35S:SVP. It means that in the case of the expression level of SVP gene is higher than that of FLM, flowering time was accelerated and those results are inconsistent with other studies. In other words, it may suggest that flowering is suppressed by SVP-FLM linkage such as the expression-level ratio of the SVP and the FLM.

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JANUARY 2015

APRIL 2015

Understanding plant cold hardiness: an opinion Lawrence V. Gusta and Michael Wisniewski DOI: 10.1111/j.1399-3054.2012.01611.x

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Editor-in-Chief: Vaughan Hurry Publish your next paper in Physiologia Plantarum Open Access Option available

Impact Factor: 3.262* Published on behalf of the Scandinavian Plant Physiology Society

Physiologia Plantarum is an international journal committed to publishing the best full-length original research papers that advance our understanding of the primary physiological, molecular and genetic mechanisms governing plant development, growth and productivity; including plant interactions with the biotic and abiotic environment. * Source: Thomson Reuters 2014 Citation Data

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Regulation of potassium transport in plants under hostile conditions: implications for abiotic and biotic stress tolerance Sergey Shabala and Igor Pottosin DOI: 10.1111/ppl.12165

Proteomic analysis of S-nitrosylated proteins in potato plant Hiroaki Kato, Daigo Takemoto and Kazuhito Kawakita DOI: 10.1111/j.1399-3054.2012.01684.x

Downregulation of the lycopene epsilon-cyclase gene increases carotenoid synthesis via the beta-branch-specific pathway andenhances salt-stress tolerance in sweetpotato transgenic calli

Sun Ha Kim, Yun-Hee Kim, Young Ock Ahn, et al. DOI: 10.1111/j.1399-3054.2012.01688.x

Find out more at www.physiologiaplantarum.com

AUTHOR INDEX Abbaspour, Hossein, 74, 75 Adolfsson, Lisa, 147 Al Lawati, Abbas, 63 Allen, Anrie, 135 Alves, Leticia, 86, 90, 110 Ashton, Neil, 65 Aspeborg, Henrik, 57 Baier, Margarete, 33 Balazadeh, Salma, 45 Bayar, Esra, 134 Beltran, Emilio Gutierrez, 12 Bertel, Clara, 141 Bones, Atle Magnar, 42 Brunoni, Federica, 60 Buchner, Othmar, 77 Burra, Dharani, 122 Carmody, Melanie, 113 Chardin, Camille, 92 Chwiałkowska, Karolina, 88 Coupland, George, 3 Cury, Patricia, 100 Cvetkovic, Jelena, 89 Czégény, Gyula, 80 Dauphinee, Adrian, 108 Decker, Daniel, 69 Dmitrieva, Valeriya, 61 Do, Yi-Yin, 140 Ehlert, Marcus, 127 Ehonen, Sanna, 145 Elomaa, Paula, 5 Engelsdorf, Timo, 136 Evkaikina, Anastasiia, 62 Fadina, Oksana, 51 Fagerstedt, K., 146 Fossdal, Carl Gunnar, 11 Fujii, Hiroaki, 99 Fujii, Hiroaki, 99 Godwin, Ian, 66 Grabsztunowicz, Magda, 97 Graeff, Moritz, 7 Gratão, Priscila, 95 Göransson, Magnus, 109 Harter, Klaus, 53 Hideg, Eva, 24 Hilleary, Richard, 40 Houshmandfar, Alireza, 131 Hudzieczek, Vojtech, 43 Humbeck, Klaus, 64 Hwang, Duk-Ju, 125 Ilik, Petr, 71 Jansson, Stefan, 17, 35

NOTES Jensen, Poul Erik, 26 Johansson, Karin S.L., 30 Jaakola, Laura, 56 Kang, Sang-Ho, 18 Kangasjärvi, Saijaliisa, 39 Kangasjärvi, Jaakko, 29 Karlsson, Patricia, 50 Karusion, Annika, 144 Kim, Hoyeun, 48 Kim, Minkyun, 94 Kim, Sungyong, 116 Kim, Young-Mi, 49 Knight, Celia, 36 Kontunen-Soppela, Sari, 143 Krüger, Gert, 91 Kuchler, Eva, 118 Kuprian, Edith, 87 Kärkönen, Anna, 20 Köhl, Karin, 78 Köhler, Claudia, 9 Lagrange, Thierry, 8 Lasses, Therese, 44 Lazaridou-Athanasiadou, Martha, 115 Le Gall, Sabine, 10 Leister, Dario, 21 Lemcke, René, 128 Lihavainen, Jenna, 102 Lin, Shu-I, 96 Lindberg, Stina, 70 López, Alejandro Perdomo, 25 Losvik, Aleksandra, 117 Luquez, Virginia, 101 Macabuhay, Allene, 79 Mamoucha, Stavroula, 104, 105 Mehrabi, Sara,119 Metzlaff , Karin, 34 Müller, Maria, 121 Müller-Röber, Bernd, 31 Münch, Miriam, 32 Mähönen, Ari Pekka, 14 Møller, Ian Max, 67 Nam, Hong Gil, 16 Nilsson, Ove, 4 Näsholm, Torgny, 27 Oenel, Ayla, 98 Olsen, Jorunn Elisabeth, 6 Ouzounidou, Georgia, 81 Palme, Klaus, 148 Pandey, Ashutosh, 106 Pazoki, Abbas, 138

Perby, Laura Kathrine, 142 Pribil, Mathias, 23 Pridacha, Vladislava, 84 Rajaraman, Sitaram, 19 Rantanen, Marja, 58 Roitsch, Thomas, 130 Rook, Fred, 126 Sağlam, Nihal Gören, 133 Saindrenan, Patrick, 124 Saini, Kumud, 52 Salgado, Marco, 41 Sarmiento, Aneth, 93 Sasaki, Izumi, 139 Sazonova, Tatiana, 83 Schulze-Lefert, Paul, 37 Shamsi, Keyvan, 85 Shamustakimova, Anastasia, 82 Stabentheiner, Edith, 54 Stangl, Zsofia, 114 Stensjö, Karin, 111 Stougaard, Jens, 38 Street, Nathaniel, 15 Strid, Åke, 72 Sundström, Jens, 59 Tenreira, Tracey, 46 Torre, Sissel, 112 Tran, Diep Thi-Ngoc, 123 Urbancsok, János, 129 Uslu, Veli Vural, 107 Vainonen, Julia, 103 Van Nguyen, Thanh, 120 Voitsekhovskaja, Olga, 73 Vyskot, Boris, 13 Väisänen, Enni, 55 Woo, Je Chang, 149 Wrzaczek, Michael, 68 Yamagata, Hiroshi, 137 Yamaguchi-Shinozaki, Kazuko, 28 Yang, Chin-Ying, 76, 132 Yoon, In Sun, 47 Zhang, Lixin, 22 Öquist, Gunnar, 2

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