International Seed Testing Association Secretariat, Zürichstrasse 50, CH-8303 Bassersdorf, Switzerland Phone: +41 44 838 60 00 Fax: +41 44 838 60 01 Email: [email protected] - http://www.seedtest.org Document OGM13-05

Rules Proposals for the International Rules for Seed Testing 2014 Edition This document was prepared by the Technical Committees and the Rules Committee of the Association and has been endorsed by the ISTA Executive Committee. The proposals are submitted to the ISTA Ordinary Meeting 2013 for voting by the nominated ISTA Designated Members on behalf of their respective Governments. It is submitted to all ISTA Designated Authorities, ISTA Members and ISTA Observer Organizations for information two months prior to the ISTA Ordinary Meeting 2013. It contains proposed amendments and changes for the ISTA International Rules for Seed Testing and will be discussed and voted on at the Ordinary Meeting 2013 to be held on Tuesday, June 18, 2013 in Antalya, Turkey under Agenda point 11. Consideration and Adoption of the Proposed Rules Changes.

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Introduction to the ISTA Rules Proposals to become effective 1 January 2014 The current version of the ISTA International Rules for Seed Testing is the 2013 edition. Single copies of replacement pages and front covers for the 2013 edition have been sent free to all ISTA Member Laboratories. Extra copies are available for purchase from the ISTA Publications section. As the Rules are an evolving document, it is worth remembering that pages can be headed with different ‘effective from’ dates. The Preface for each edition includes details of changes and when replacement pages were issued. Previous Prefaces as a ‘history of changes’ are available on the ISTA website. The ISTA Rules are the result of the work of the various ISTA Technical Committees with input from many different sources. Thanks go to all the Technical Committee members and the ISTA Secretariat for their help with this year’s proposals. The following Rules Proposals will be discussed at the ISTA Ordinary Meeting in Antalya, Turkey in June 2013 and may be amended during the meeting. If the proposals are accepted by the membership, Amendments will be issued, and they will become the 2014 edition of the ISTA Rules. Please let me know about any problems with these proposals. Many thanks. Steve Jones Chair of Rules Committee Contact details: Dr Steve Jones Canadian Food Inspection Agency Seed Science and Technology Section 301–421 Downey Road Saskatoon, SK, S7N 4L8 Canada Phone: +1 306 975 6505 Fax: +1 306 975 6450 E-mail: [email protected]

Key to text changes : Deleted text New text New text in large blocks, not underlined for ease of reading Any changes made after proposals published to the membership

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Contents

PART A. INTRODUCTION OF EDITORIAL CHANGES

5

A.1. Editorial corrections

5

PART B. NEW SPECIES AND CHANGES TO SPECIES NAMES

6

B.1. Addition of new species to Table 2A.

6

B.2. Changes to the ISTA Stabilized list affecting names used in the ISTA Rules 6 1. Changes to species names

6

2. Changes to assignments of genera to families

9

PART C. RULES CHANGES AND NEW METHODS REQUIRING A VOTE 11 Chapter 1: Certificates

11

C.1.1. Changes to reporting for the tetrazolium test results for coated seed, seed tapes and mats.

11

C.1.2. Inclusion of reporting requirements for tetrazolium testing of seed mixtures Chapter 18. 11 C.1.3. Inclusion of reporting requirements for new Chapter 19.

11

Chapter 2: Sampling

12

C.2.1. Storage of samples after testing

12

C.2.2. Changes to the minimum submitted sample sizes of coated seeds

13

C.2.3. Consequential changes affecting Chapter 11: Testing of Coated Seeds

14

C.2.4. Consequential changes affecting Chapter 16: Rules for size and grading of seeds 15

Chapter 3: The Purity Analysis

16

C.3.1. Adding the use of an anemometer for the uniform blowing method

16

Chapter 5: The Germination Test

21

C.5.1. Harmonisation on seedling evaluation in respect to the evaluation of the cotyledons (50% rule)

21

C.5.2. Growing media for germination test.

23

C.5.3. List of seedling abnormalities

24

C.5.4. Change required due to moving genus Arachis from PSD 11 to PSD 21

25

C.5.5. Duration of germination test for certain grass species

26

C.5.6. Modification to 5.6.4 to add clarity

27

Chapter 6: Biochemical Test for Viability. The Topographical Tetrazolium Test. 28 C.6.1. Amended explanation for testing Helianthus and Bracharia species .

28

Annex to Chapter 7: Seed Health Testing Methods

29

C.7.1. Changes to existing seed health methods to provide a uniform approach

29

C.7.2. Modification to existing seed health method

34

7-028: Detection of infectious tobamoviruses on Lycopersicon esculentum (tomato) by the local lesion assay (indexing) on Nicotiana tabacum plants 34 C.7.3. Modification to existing seed health method OGM13-05 Proposed Changes to the ISTA Rules Edition 2014 Approved by ECOM and RUL on 3 April 2013

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7-019b: Detection of Xanthomonas campestris pv. campestris on Brassica spp. disinfested/disinfected seed

37

C.7.4. Modification to existing seed health method

49

7-021: Detection of Xanthomonas axonopodis pv. phaseoli and Xanthomonas axonopodis pv. phaseoli var. fuscans on Phaseolus vulgaris (Bean) seed

49

C.7.5. New seed health method

56

7-029: Detection of Pseudomonas syringae pv. pisi on Pisum sativum (Pea) seed 56 C.7.6. New seed health method

65

7-007: Detection of Alternaria linicola, Botrytis cinerea and Colletotrichum lini on Linum usitatissimum (Flax) seed 65

Chapter 8: Species and Variety Testing

74

C.8.1. Editorial and Committee review of the whole of Chapter 8.

74

C.8.2. New improved A-PAGE method for the verification of Triticum

85

C.8.3. New SDS-PAGE method for the verification of Triticum and xTriticosecale varieties 90

Chapter 11: Testing Coated Seeds

93

C.11.1 Testing methods and reporting for the tetrazolium test for coated seeds

93

Chapter 18: Seed Mixtures

95

C.18.1. Testing methods and reporting for the tetrazolium test for seed mixtures

95

Chapter 19: Testing for Seeds of Genetically Modified Organisms 96 C.19.1. New Chapter for the ISTA Rules

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PART A. INTRODUCTION OF EDITORIAL CHANGES A.1. Editorial corrections General editorial corrections -

Since the 2014 ISTA Rules will be completely reissued, the current “effective from” dates will all be re-set to 1 January 2014.

-

All marks which indicate changes from the previous edition will be removed, except for the latest changes made for the 2014 edition.

-

The English version of the 2014 ISTA Rules will be in A4 format.

-

Wherever ‘shall’ is used it will be replaced with ‘must’, if that is the intent, or where it makes grammatical sense.

-

References to old family names (e.g. Compositae, Gramineae) will be deleted.

-

The Seed Health Methods, currently referred to as “Annexe to Chapter 7”, will now be part of Chapter 7.

-

In all Seed Health Methods, the phrase ‘sponsored by’ will be amended to ‘prepared by’ together with details of the organization which organized the comparative test. Prepared by is also used to list the authors so in this case prepared by will be replaced by Authors. The citation for ISHI-Veg will be updated to International Seed Health Initiative-Vegetables, ISF (ISHI-Veg).

-

In all Seed Health Methods, standardise the way the equation for TSW is presented as: TSW = (weight of seeds / numbers of seeds) x 1000

CURRENT VERSION

PROPOSED VERSION

1.5.2.2. Purity

1.5.2.2. Purity

3.7 Reporting results

3.7 Reporting results

§5… (e.g. Elytrigium repens).

§5… (e.g. Elytrigia repens).

CURRENT VERSION

PROPOSED VERSION

Table 6A Part 1. Agricultural and horticultural seeds Lactuca spp., column 7: ⅓ radicle, measured from radicle tip; ½ of distal end of cotyledons,if superficial; ⅓ at distal end, if pervading

ACCEPTED BY APPLAUSE

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⅓ radicle, measured from radicle tip; ½ of distal end of cotyledons,if superficial; ⅓ at distal end, if pervasive

RESULT

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PART B. NEW SPECIES AND CHANGES TO SPECIES NAMES B.1. Addition of new species to Table 2A. None this year.

B.2. Changes to the ISTA Stabilized list affecting names used in the ISTA Rules The ISTA Stabilized List is updated every 6 years as a result of discussion within the ISTA Nomenclature Committee. The following items have been taken from the document “Proposed Changes to the ISTA List of Stabilized Plant Names” prepared on 1 November 2012. The items of that document have been considered and approved by the ISTA Nomenclature Committee, and the document will be submitted to the ISTA Executive Committee for voting at the ISTA Ordinary Meeting 2013. Only those items from this document that affect the 2014 ISTA Rules are detailed here. The changes below show the changes to species names as they will appear in Table 2A. References to older changes to species names will be removed. Other tables and references to species names will be amended accordingly. Once accepted by vote at the Annual General Meeting, the Stabilised List can come into effect on 1 January 2014, to be consistent with the 2014 ISTA Rules, which also come into effect on 1 January 2014. The following proposal was developed and approved by a vote of the Nomenclature Committee.

1. Changes to species names a. Agricultural and vegetable CURRENT VERSION

PROPOSED VERSION

Andropogon gerardii Vitman Bromus marginatus Nees ex Steud. Cajanus cajan (L.) Millsp. Centrosema pubescens Benth.

Andropogon gerardi Vitman Bromus marginatus Steud. Cajanus cajan (L.) Huth (Centrosema pubescens Benth. see Centrosema molle Mart. ex Benth.) Centrosema molle Mart. ex Benth. (previously Centrosema pubescens Benth.) (Dichondra repens J. R. Forst. & G. Forst. see Dichondra micrantha Urb.) Dichondra micrantha Urb. (previously Dichondra repens J. R. Forst. & G. Forst.) (Lolium ×boucheanum Kunth see Lolium ×hybridum Hausskn.) Lolium ×hybridum Hausskn. (previously Lolium ×boucheanum Kunth) (Lotononis bainesii Baker see Listia bainesii (Baker) B.-E. van Wyk & Boatwr.) Listia bainesii (Baker) B.-E. van Wyk & Boatwr. (previously Lotononis bainesii Baker) (Lycopersicon esculentum Mill. see Solanum lycopersicum L.) Solanum lycopersicum L. (previously Lycopersicon esculentum Mill.)

Dichondra repens J. R. Forst & G. Forst.

Lolium ×boucheanum Kunth

Lotononis bainesii Baker

Lycopersicon esculentum Mill.

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CURRENT VERSION

PROPOSED VERSION

Lycopersicon hybrids

(Lycopersicon hybrids see Solanum (sect. Lycopersicon) hybrids) Solanum (sect. Lycopersicon) hybrids (previously Lycopersicon hybrids) (Lycopersicon spp. see Solanum (sect. Lycopersicon) spp.) Solanum (sect. Lycopersicon) spp. (previously Lycopersicon spp.) Pascopyrum smithii (Rydb.) Barkworth & D. R. Dewey (Paspalum wettsteinii Hack. see Paspalum virgatum L.) Paspalum virgatum L. (previously Paspalum wettsteinii Hack.) Petroselinum crispum (Mill.) Fuss

Lycopersicon spp.

Pascopyrum smithii (Rydb.) Á. Löve Paspalum wettsteinii Hack.

Petroselinum crispum (Mill.) Nyman ex A. W. Hill b. Tree and shrub CURRENT VERSION

PROPOSED VERSION

Cupressus macrocarpa Hartw. ex Gordon Eucalyptus citriodora Hook.

Cupressus macrocarpa Hartw.

Eucalyptus ficifolia F. Muell.

Eucalyptus maculata Hook.

Mahonia aquifolium (Pursh) Nutt.

Pinus heldreichii H. Christ Pinus patula Schiede ex Schltdl. & Cham. Pinus ponderosa C. Lawson

(Eucalyptus citriodora Hook. see Corymbia citriodora (Hook.) K. D. Hill & L. A. S. Johnson) Corymbia citriodora (Hook.) K. D. Hill & L. A. S. Johnson (previously Eucalyptus citriodora Hook.) (Eucalyptus ficifolia F. Muell. see Corymbia ficifolia (F. Muell.) K. D. Hill & L. A. S. Johnson) Corymbia ficifolia (F. Muell.) K. D. Hill & L. A. S. Johnson (previously Eucalyptus ficifolia F. Muell.) (Eucalyptus maculata Hook. see Corymbia maculata (Hook.) K. D. Hill & L. A. S. Johnson) Corymbia maculata (Hook.) K. D. Hill & L. A. S. Johnson (previously Eucalyptus maculata Hook.) (Mahonia aquifolium (Pursh) Nutt. see Berberis aquifolium Pursh) Berberis aquifolium Pursh (previously Mahonia aquifolium (Pursh) Nutt.) Pinus heldreichii Christ Pinus patula Schltdl. & Cham. Pinus ponderosa P. Lawson & C. Lawson

c. Flower, spice, herb and medicinal CURRENT VERSION

PROPOSED VERSION

Armeria maritima Willd. Asparagus densiflorus (Kunth) Jessop

Armeria maritima (Mill.) Willd. (Asparagus densiflorus (Kunth) Jessop see Asparagus aethiopicus L.)

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CURRENT VERSION

Asparagus setaceus (Kunth) Jessop

Centaurea americana Nutt.

Centaurea dealbata Willd.

Cnicus benedictus L.

Coleus blumei Benth.

Cymbalaria muralis P. Gaertn. et al. Geranium hybrids* Gerbera jamesonii Bolus ex Hook. f. Helichrysum bracteatum (Vent.) Andrews

Helipterum humboldtianum (Gaudich.) DC.

Helipterum manglesii (Lindl.) F. Muell. ex Benth.

Helipterum roseum (Hook.) Benth.

Kochia scoparia (L.) Schrad.

OGM13-05 Proposed Changes to the ISTA Rules Edition 2014 Approved by ECOM and RUL on 3 April 2013

PROPOSED VERSION Asparagus aethiopicus L. (previously Asparagus densiflorus (Kunth) Jessop) (Asparagus setaceus (Kunth) Jessop see Asparagus plumosus L.) Asparagus plumosus L. (previously Asparagus setaceus (Kunth) Jessop) (Centaurea americana Nutt. see Plectocephalus americana (Nutt.) D. Don) Plectocephalus americana (Nutt.) D. Don (previously Centaurea americana Nutt.) (Centaurea dealbata Willd. see Psephellus dealbatus (Willd.) K. Koch) Psephellus dealbatus (Willd.) K. Koch (previously Centaurea dealbata Willd.) (Cnicus benedictus L. see Centaurea benedicta (L.) L.) Centaurea benedicta (L.) L. (previously Cnicus benedictus L.) (Coleus blumei Benth. see Plectranthus scutellarioides (L.) R. Br.) Plectranthus scutellarioides (L.) R. Br. (previously Coleus blumei Benth.) Cymbalaria muralis G. Gaertn. et al. Geranium hybrids Gerbera jamesonii Adlam (Helichrysum bracteatum (Vent.) Andrews see Xerochrysum bracteatum (Vent.) Tzvelev) Xerochrysum bracteatum (Vent.) Tzvelev (previously Helichrysum bracteatum (Vent.) Andrews) (Helipterum humboldtianum (Gaudich.) DC. see Rhodanthe humboldtiana (Gaudich.) Paul G. Wilson) Rhodanthe humboldtiana (Gaudich.) Paul G. Wilson (previously Helipterum humboldtianum (Gaudich.) DC.) (Helipterum manglesii (Lindl.) F. Muell. ex Benth. see Rhodanthe manglesii Lindl.) Rhodanthe manglesii Lindl. (previously Helipterum manglesii (Lindl.) F. Muell. ex Benth.) (Helipterum roseum (Hook.) Benth. see Rhodanthe chlorocephala (Turcz.) Paul G. Wilson) Rhodanthe chlorocephala (Turcz.) Paul G. Wilson (includes Helipterum roseum (Hook.) Benth.) (Kochia scoparia (L.) Schrad. see Bassia scoparia (L.) A. J. Scott) Bassia scoparia (L.) A. J. Scott (previously Kochia scoparia (L.) Schrad.)

2013-04-12 12:40 Page 8/106

CURRENT VERSION

PROPOSED VERSION

Leontopodium alpinum Cass.

(Leontopodium alpinum Cass. see Leontopodium nivale (Ten.) Hand.Mazz.) Leontopodium nivale (Ten.) Hand.Mazz. (previously Leontopodium alpinum Cass.) Lobelia fulgens Humb. & Bonpl. ex Willd. Lupinus hybrids (Matricaria recutita L. see Matricaria chamomilla L.) Matricaria chamomilla L. (previously Matricaria recutita L.) Myosotis hybrids (Petunia ×hybrida hort. ex E. Vilm. see Petunia ×atkinsiana (Sweet) D. Don ex W. H. Baxter) Petunia ×atkinsiana (Sweet) D. Don ex W. H. Baxter (previously Petunia ×hybrida hort. ex E. Vilm.) (Scabiosa caucasica M. Bieb. see Lomelosia caucasica (M. Bieb.) Greuter & Burdet) Lomelosia caucasica (M. Bieb.) Greuter & Burdet (previously Scabiosa caucasica M. Bieb.) (Senecio cineraria DC. see Jacobaea maritima (L.) Pelser & Meijden) Jacobaea maritima (L.) Pelser & Meijden (previously Senecio cineraria DC.) (Senecio cruentus (Masson ex L’Hér.) DC. see Pericallis cruenta (Masson ex L’Hér.) Bolle) Pericallis cruenta (Masson ex L’Hér.) Bolle (previously Senecio cruentus (Masson ex L’Hér.) DC.) Sinningia speciosa (Lodd. et al.) Hiern (Solanum diflorum Vell. see Solanum pseudocapsicum L.) Solanum pseudocapsicum L. (previously Solanum diflorum Vell.) (Tripleurospermum perforatum (Mérat) M. Laínz see Tripleurospermum inodorum (L.) Sch. Bip.) Tripleurospermum inodorum (L.) Sch. Bip. (previously Tripleurospermum perforatum (Mérat) M. Laínz)

Lobelia fulgens Willd. Lupinus hybrids* Matricaria recutita L.

Myosotis hybrids* Petunia ×hybrida hort. ex E. Vilm.

Scabiosa caucasica M. Bieb.

Senecio cineraria DC.

Senecio cruentus (Masson ex L’Hér.) DC.)

Sinningia speciosa (G. Lodd.) Hiern Solanum diflorum Vell. . Tripleurospermum perforatum (Mérat) M. Laínz

2. Changes to assignments of genera to families CURRENT VERSION

PROPOSED VERSION

Portulacaceae

Montiaceae

Taxodiaceae

Cupressaceae

a. Agricultural and vegetable Claytonia b. Tree and shrub Cryptomeria OGM13-05 Proposed Changes to the ISTA Rules Edition 2014 Approved by ECOM and RUL on 3 April 2013

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CURRENT VERSION

PROPOSED VERSION

Liquidambar

Hamamelidaceae

Altingiaceae

Nothofagus

Fagaceae

Nothofagaceae

Sequoia

Taxodiaceae

Cupressaceae

Sequoiadendron

Taxodiaceae

Cupressaceae

Taxodium

Taxodiaceae

Cupressaceae

Asclepias

Asclepiadaceae

Apocynaceae

Cleome

Capparaceae

Cleomaceae

Nemophila

Hydrophyllaceae

Boraginaceae

Phacelia

Hydrophyllaceae

Boraginaceae

Pholistoma

Hydrophyllaceae

Boraginaceae

c. Flower, spice, herb and medicinal

VOTE TO ACCEPT ITEM

CURRENT VERSION

PROPOSED VERSION

2.8 Tables for lot size and sample sizes Table 2A

2.8 Tables for lot size and sample sizes Table 2A

… the 2007 ISTA Congress …

… the 2013 ISTA Congress …

… to 2007 Congress changes; …

… to 2013 Congress changes; …

YES VOTES

NO VOTES

RESULT

B.2

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PART C. RULES CHANGES AND NEW METHODS REQUIRING A VOTE Chapter 1: Certificates C.1.1. Changes to reporting for the tetrazolium test results for coated seed, seed tapes and mats. New text for reporting Tetrazolium test resuts for coated seed, seed mats and seed tapes, see proposal under Chapter 11.

Note not for voting now but for reference only. See after Chapter 11 for voting. C.1.2. Inclusion of reporting requirements for tetrazolium testing of seed mixtures Chapter 18. Changes to Chapter 1 are required due the acceptance of the text for Tetrazolium testing of seed mixtures in Chapter 18. See voting record following proposal C.18.1.

Note not for voting now but for reference only. See after Chapter 18 for voting. C.1.3. Inclusion of reporting requirements for new Chapter 19. Changes to Chapter 1 are required due the acceptance of the new Chapter 19 in the ISTA Rules. See voting record following proposal C.19.1.

Note not for voting now but for reference only. See after Chapter 19 for voting.

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Chapter 2: Sampling C.2.1. Storage of samples after testing The storage time of submitted samples is proposed to be moved from 2.5.4.6 to 2.5.3. As a consequence the storage time should be applied also to the submitted samples on which ISTA Blue Certificates have been issued. Also due to practical reasons it is proposed that the storage time of one year should be counted from the receipt of samples and not from the issuance of ISTA Certificates. It is also proposed that in the cases where storage time of one year is expected to affect test results though samples are preserved in appropriate conditions the requirement of storage time of one year is not a requirement for moisture proof containers and samples of recalcitrant or intermediate species. The following proposal was developed by the Bulking and Sampling Committee and has been discussed with the Moisture and Storage Committees. This proposal has been approved by a vote of the Bulking and Sampling Committee. CURRENT VERSION

PROPOSED VERSION

2.5.3. Storage of samples after testing … Protection against insects and rodents may be necessary.

2.5.3. Storage of samples after testing … Protection against insects and rodents may be necessary. To provide for re-testing by the original or by another seed testing laboratory, samples on which ISTA Certificates have been issued must be stored at least for one year from the receipt of the sample. Submitted samples in moisture proof containers, and samples of recalcitrant or intermediate species, must be stored under appropriate conditions for as long as it can be expected that the results of a re-test are not affected by the storage.

VOTE TO ACCEPT ITEM

When a re-test in a different testing laboratory is required, a portion shall be drawn from the stored sample in accordance with 2.5.2.2, and submitted to the designated laboratory. The remainder shall be retained in store.

When a re-test in a different testing laboratory is required, a portion must be drawn from the stored sample in accordance with 2.5.2.2, and submitted to the designated laboratory. The remainder must be retained in store.

…

…

2.5.4.6 Storage of submitted samples after testing

2.5.4.6 Storage of submitted samples after testing

To provide for re-testing by the original or by another seed testing laboratory, submitted samples on which ISTA Certificates have been issued shall be stored for one year from the date of issue of the certificate. Only in the case of very expensive seeds …

Submitted samples on which ISTA Certificates have been issued must be stored. Only in the case of very expensive seeds …

YES VOTES

NO VOTES

RESULT

C.2.1

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C.2.2. Changes to the minimum submitted sample sizes of coated seeds Bulking and Sampling Committee (BSC) proposes that the minimum submitted sample sizes of coated seeds are decreased. In the current Rules the requirements for submitted sample sizes are inconsistent between non-coated seeds and coated seeds. For non-coated seeds the submitted sample size can be equal to the working sample size for purity (i.e. to be 2500 seeds) if determination of other seeds by number is not requested. This is not possible for coated seeds. In the current Rules the submitted sample size for coated seeds is always 7500 seeds for purity and germination tests though e.g. the minimum working sample size for purity test is 2500 seeds. For coated seeds, the submitted samples shall contain at least the number of pellets or seeds indicated in column 2 of Table 2B Part 1 and 2. A survey was sent to six laboratories. Five laboratories were of the opinion that sample sizes can be decreased and one laboratory hesitated due to possible heterogeneity in seed lots. However, according to the opinion of the BSC possible heterogeneity problem should be smaller in coated seeds than in non-coated seeds and on the otherhand the submitted sample size does not solve heterogeneity problems. The following proposal was therefore developed by the Bulking and Sampling Committee and approved by a vote. CURRENT VERSION

PROPOSED VERSION

2.5.4.4 Submitted sample

2.5.4.4 Submitted sample

Minimum size of submitted samples are as follows:

The minimum size of submitted samples are as follows:

a) For moisture determination, 100 g for species that have to be ground (see Table 9A) and 50 g for all other species. When moisture meters are to be used for testing, a larger sample size may be necessary. Contact the ISTA seed testing laboratory for specific instructions.

a) For moisture determination, 100 g for species that must be ground (see Table 9A) and 50 g for all other species. When moisture meters are to be used for testing, a larger sample size may be necessary. Contact the ISTA seed testing laboratory for specific instructions.

b)…

b)…

c) For all other tests, at least the weight prescribed in column 3 of Table 2A. As long as a determination of other seeds by number is not requested, the submitted sample shall weigh at least the amount indicated for the working sample for purity analysis in column 4 of Table 2A. In the case of coated seeds, the submitted samples shall contain not less than the number of pellets or seeds indicated in column 2 of Table 2B, Part 1 and Part 2.

c) For all other tests, at least the weight prescribed in column 3 of Table 2A. As long as a determination of other seeds by number is not requested, the submitted sample must weigh at least the amount indicated for the working sample for purity analysis in column 4 of Table 2A. In the case of coated seeds, the submitted samples must contain not less than the number of pellets or seeds indicated in column 2 of Table 2B, Part 1 and Part 2. As long as a determination of other seed by number or size grading is not requested, the submitted sample need only contain as a minimum, the number of seeds indicated for the working sample for purity analysis in column 3 of Table 2B Parts 1 and 2.

If the submitted sample is smaller than prescribed, …

If the submitted sample is smaller than prescribed, …

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Table 2B Part 1. Sample sizes (number of seeds) for pelleted seeds, encrusted seed and seed granules Determinations

Purity analysis (including verification of species) Weight determination Germination Determination of other seeds Determination of other seeds (encrusted seeds and seeds granules) Size grading

Minimum submitted sample 2500 2500 2500 10000 25000

Minimum working sample 2500 Pure pellet fraction 400 7500 25000

5000

1000

Table 2B Part 2. Sample sizes (number of seeds) for seed tapes and mats Determinations

Minimum submitted sample Verification of species 300 Germination 2000 Purity analysis (if required) 2500 Determination of other 10000 seeds

Minimum working sample 100 400 2500 7500

C.2.3. Consequential changes affecting Chapter 11: Testing of Coated Seeds In the Chapter 11: Testing of Coated Seeds there are Tables 11A and 11B that are copies of the Tables 2B Part 1 and Part 2. In the case that the proposal concerning Tables 2B Part 1 and Part 2 is accepted then also Tables 11A and 11B should be changed. Table 11A. Sample sizes of pelleted seeds in number of pellets. Note: this table is a copy of Table 2B Part 1 Determinations Purity analysis (including verification of species) Weight determination Germination Determination of other seeds Determination of other seeds (encrusted seeds and seeds granules) Size grading

OGM13-05 Proposed Changes to the ISTA Rules Edition 2014 Approved by ECOM and RUL on 3 April 2013

Minimum submitted sample 2500

Minimum working sample 2500

2500 2500 10000 25000

Pure pellet fraction 400 7500 25000

5000

1000

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Table 11A. Sample sizes of seed tapes Note: this table is a copy of Table 2B Part 2 Determinations

Minimum submitted sample Verification of species 300 Germination 2000 Purity analysis (if required) 2500 Determination of other 10000 seeds VOTE TO ACCEPT ITEM

YES VOTES

NO VOTES

Minimum working sample 100 400 2500 7500

RESULT

C.2.3

C.2.4. Consequential changes affecting Chapter 16: Rules for size and grading of seeds Current Rules for size grading of seeds for Beta seeds and pelleted seeds indicate the same working sample weights (two samples of 50 grams) regardless of seed species or coating material. In Chapter 16 there is no reference to Chapter 2, Table 2B Part 1 where the submitted sample and working sample sizes for coated seeds are described. The following proposal was developed by the Bulking and Sampling Committee and approved by a vote. CURRENT VERSION

PROPOSED VERSION

16.1 For Beta seeds and pelleted seeds

16.1 For Beta seeds and pelleted seeds

The control of size grading is carried out on a sample, weighing at least 250g, which must be sent must be sent to the testing laboratory in an airtight container. Two working samples of about 50 g (not less than 45g and not more than 55g) each are used. Each sample is subjected to a screening analysis. ….

The control of size grading is carried out on a sample, weighing at least 250g or for pelleted seeds, a sample consisting of the number of seeds indicated in the Table 2B Part 1. The sample must be sent to the testing laboratory in an airtight container. Two working samples of about 50 g (not less than 45g and not more than 55g) each are used. For pelleted seeds, two working samples of about 1000 seed each are used. Each sample is subjected to a screening analysis. … If this tolerance is exceeded, a further sample of 50g or 1000 pelleted seeds (and if necessary a fourth sample) must be analysed. …

If this tolerance is exceeded, a further sample of 50g (and if necessary a fourth sample) must be analysed. ….

VOTE TO ACCEPT ITEM

YES VOTES

NO VOTES

RESULT

C.2.4

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Chapter 3: The Purity Analysis C.3.1. Adding the use of an anemometer for the uniform blowing method To use an anemometer to monitor the equivalent air velocity (EAV) value of the optimum blowing point obtained using the ISTA calibration samples. There is no existing procedure for using the EAV value to monitor the calibration points for General blowers in the ISTA Rules. Expected benefits Currently, ISTA requires a regular calibration of the blowers using the ISTA uniform calibration sample (UCS). However, the frequent use of a UCS can cause it to rapidly deteriorate. Using simply the air gate opening between calibrations is reliable in some blowers but not in others. Using a desired air velocity point regardless of gate opening, motor conditions, etc. is a more reliable way to reproduce the correct point as it is determined with a calibration sample. The EAV of the optimum blowing point is the air velocity value of the air gate opening at the optimum blowing point for a specific calibration sample. The casual factor in seed separations in any blowing procedure is the velocity of air that flows through the working sample. Therefore, using the EAV directly to monitor the procedure allows the analyst to detect any undesirable variation. In addition, using the EAV makes the application of the blowing procedure simple and verifiable at any time. Using an anemometer will minimize the use of the UCS, and thus protect the integrity of the calibration samples. This approach is critical to maintain the integrity of the UCS in all future blowing procedures. This technology in combination with the use of uniform calibration samples should increase the uniformity within and across laboratories. Measuring the EAV takes only a few minutes, thus it will be easy for laboratories to keep records of the equivalent air velocity value of the optimum blowing point for quality control purposes. The following proposal was developed by the Blowing Procedure Working Group of the Purity Committee and approved by vote. CURRENT VERSION

PROPOSED VERSION

3.4 Apparatus

3.4 Apparatus

Aids …

3.4.1 Magnifiers, reflected light and sieves Aids …

Hand lenses … Sieves …

Hand lenses … Sieves …

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CURRENT VERSION

PROPOSED VERSION 3.4.2 Seed blowers

Seed blowers can be used to separate light-weight material such as chaff and empty florets in grasses from the heavier seeds.

Seed blowers can be used to separate light-weight material such as chaff and empty florets from the heavier seeds for all species as a tool for purity analysis.

Blowers that will give the most accurate separations normally handle only small samples (up to 5g). A good blower should provide a uniform flow of air, be capable of standardization and retain all the particles which it separates.

Blowers that will give the most accurate separations normally handle only small samples (up to 5g). A good blower should provide a uniform flow of air, be capable of standardization and retain all the particles which it separates. For certain species and varieties of Poaceae, seed blowers must be used by the uniform blowing method (3.5.2.5) to separate light-weight material such as chaff and empty florets from the heavier seeds.

In order to maintain a uniform flow of air the blower should have one or more air compression chambers and a fan driven by a uniform speed motor. The diameter of the blowing tube should be in proportion to the size of the working sample and the tube should be long enough to allow satisfactory separation of the sample. The valve or gate that regulates the air flow should be capable of precise adjustment, should be calibrated and marked to permit easy reading, and its construction and location should prevent areas of strong and weak currents in the blowing tube.

In order to maintain a uniform flow of air, the blower should have one or more air compression chambers and a fan driven by a uniform-speed motor. The diameter of the blowing tube should be in proportion to the size of the working sample, and the tube should be long enough to allow satisfactory separation of the sample. The valve or air gate that regulates the air flow should be capable of precise adjustment, should be calibrated and marked to permit easy reading, and its construction and location should prevent areas of strong and weak currents in the blowing tube.

A manometer is desirable for standardizing the blower. A blower to be used for the uniform blowing method must be capable of:

A seed blower to be used for the uniform blowing method must be capable of:

a) blowing at different pressures (determined by the use of the calibration samples) to suit different species;

a) blowing at different air velocities (determined by the use of the calibration samples) to suit different species;

b) maintaining a uniform flow of air along the tube at any required pressure;

b) maintaining a uniform flow of air at the velocity required by the crop species under test;

c) rapid adjustment to any pressure likely to be required. The setting to provide each pressure should be checked periodically by blowing a calibration sample issued under the authority of ISTA;

c) rapid adjustment to any velocity likely to be required. The setting to provide each velocity should be checked annually by blowing a calibration sample issued under the authority of ISTA;

d) accurate time setting.

d) accurate time setting.

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CURRENT VERSION

PROPOSED VERSION 3.4.2.1 Calibration of the seed blower The air gate openings and the Equivalent Air Velocity (EAV) value (see 3.4.2.2) of the optimum blowing point for a General type seed blower are determined by using the uniform calibration samples. Calibration samples are issued under the authority of ISTA and are available for Dactylis glomerata and Poa pratensis. Prior to calibration, the calibration samples must be exposed to room conditions overnight. For those not having a General type seed blower, please contact the ISTA Secretariat. The air gate opening for the varieties of Poa pratensis listed in Table 3A, with an average thousand-seed weight less than 0.35 g, and for Poa trivialis is obtained by multiplying the value of the air gate setting for Poa pratensis by 0.82 (applies only for General type seed blowers). 3.4.2.2 Determination of the equivalent air velocity After a General type seed blower has been calibrated according to 3.4.2.1, the EAV of the air gate opening must be measured using an anemometer. The following procedure must be used: 1. Set the blower at the optimum blowing point, i.e. the air gate opening, obtained with the ISTA uniform calibration sample for the relevant species, e.g. Dactylis glomerata or Poa pratensis. Do not change that air gate opening. 2. Remove the sample cup from the cup holder, insert the anemometer with digital display facing up, and align the fan of the anemometer over the blower opening where the air flows from the chamber into the sample cup holder. 3. Turn on the anemometer and select metres per second (m/s), hold the anemometer in a steady position and then turn on the blower.

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CURRENT VERSION

PROPOSED VERSION 4. Read the air velocity value after the digital display of the anemometer reaches a steady reading (typically about 30 seconds after the blower was turned on). Example: If the anemometer indicates 2.3 m/s most frequently and fluctuates between 2.2 and 2.4 m/s, the EAV value of that specific airgate opening would be recorded as 2.3 ± 0.1 m/s. Once the optimum air velocity has been measured, the seed blower can be recalibrated using the anemometer, by adjusting the blower setting until the optimum air velocity for the blower and species or variety is reached. The EAV for one blower is not transferable to another blower. The optimum blowing point must be verified using the ISTA uniform calibration sample after major servicing of the blower, such as changing parts of the motor or the glass column. In general, it is strongly recommended that the blowing point be verified annually using the ISTA uniform calibration sample. Laboratories that can not, or do not, use the EAV to determine the blowing point must calibrate the blower with the ISTA uniform calibration sample. Note: Frequent use of the ISTA uniform calibration sample can cause a shift in blowing point due to deterioration and monitoring the blowing point simply by air gate opening may be reliable in some blowers and not in others. 3.4.2.3 Anemometer type Any suitable anemometers can be used as long as the anemometer fits in the sample cup holder compartment of the blower and has a scale calibrated in metres per second for reading the air velocity value. 3.4.2.4 Calibration of the anemometer The anemometer should be calibrated at the set intervals set by the laboratory. In addition, the batteries should be replaced once a year.

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CURRENT VERSION

PROPOSED VERSION

3.5.2.5 Uniform blowing method This method is obligatory for Poa pratensis, Poa trivialis and Dactylis glomerata.

3.5.2.5 Poa pratensis, Poa trivialis and Dactylis glomerata For Poa pratensis, Poa trivialis and Dactylis glomerata, the uniform blowing method (see 3.4) is obligatory.

The working sample size is 1 g for Poa pratensis and Poa trivialis, and 3 g for Dactylis glomerata.

The working sample size is 1 g for Poa pratensis and Poa trivialis, and 3 g for Dactylis glomerata.

The blowing pressure is determined for Poa pratensis and Dactylis glomerata by means of a calibration sample issued under the authority of ISTA. The blowing pressure for the varieties of Poa pratensis listed in Table 3A with an average weight of 1000 seeds