Germination Committee - ISTA · 2012-06-21 · • 1 fast germinating variety • 2 levels of germination quality • 1 slow germinating variety • 2 levels of germination quality

Germination Committee Progress Report

Sylvie Ducournau

Agenda

• Germination committee members• Validation studies

– New methods– New species

• Rules proposals for voting 2012 and future• Publications• Workshops

Committee MembershipChair : 1 Sylvie Ducournau France

Vice-chair : 2 Anny Van Pijlen Netherlands

Members : 3 Ignacio Aranciaga Argentina

4 Ronald Don United Kingdom

5 Kari Fiedler United States

6 Fabio Gorian Italy

7 Andrea Jonitz Germany

8 Krystyna Kolasinska Poland Retired 2012

9 Augusto Martinelli Argentina

10 Lea Mazor Israel

11 Gillian McLaren United Kingdom

12 Harry Nijenstein Netherlands

13 Takayuki Okuda Japan

14 Hakon Tangeras Norway

15 Rita Zecchinelli Italy

Joël Léchappé France ECOM representative

Validation studiesNew methods

• Shortening the duration of the germination test for Lolium, Festuca and Poa species – Harry Nijenstein

• Use of BP substrate for germination method of Festuca and Lolium - Zita Ripka

• Other studies in progress

Shortening the duration of the germination test for Lolium,

Festuca and Poa speciesTest plan of the comparative test 8 participating laboratories

Species tested• Lolium perenne• Festuca rubra• Poa pratensis

4 seed lots per species• 1 fast germinating variety

• 2 levels of germination quality

• 1 slow germinating variety• 2 levels of germination quality

Germination tests

• On pure seeds• On non dormant seeds• With all the temperatures available for each species • Counts after:

• 7, 10 and 14 days for Lolium perenne• 7, 14 and 21 days for Festuca rubra• 7, 14, 21 and 28 days for Poa pratensis

Shortening the duration of the germination test for Lolium,

Festuca and Poa species

• Germination tests are completed

• Several statistical studies

– Repeatability and reproducibility– Statistical comparison between temperature and test

duration for each species

• A draft validation report has been circulated between the participants and a presentation and a discussion of the results is scheduled before final conclusion

Use of BP substrate for germination method of

Festuca and Lolium

Test plan of the peer validation study

3 participating laboratories

Species tested• Lolium perenne• Festuca rubra

3 seed lots with different levels of germination, per species

Germination tests

• Non dormant seeds

• With TP and BP substrates

• At one alternating temperature 2030°C

• Addition of KNO3

Repeatability resultsLo

lium

- %

Nor

mal

See

dlin

gs

BP TP

70

75

80

85

90

95

100

70

75

80

85

90

95

100

Lolium - % Normal Seedlings

Fest

uca

- % N

orm

al S

eedl

ings

BP TP

80

85

90

95

80

85

90

95

Festuca - % Normal Seedlings

Lolium Festuca

BP BP

Sr fr Sr fr89.17 3.68 1.18 91.39 3.04 1.09

TP TP

Sr fr Sr fr90.06 2.76 0.92 92.92 1.8 0.7

...p...p...p

fr2 > 1 overdispersion

Use of BP substrate for germination method of Festuca

and Lolium

• Due to the results of repeatability the committee decided not to validate the use of BP for the germination method of these species

• A proposal is to repeat the validation study with more laboratories involved

• A detailed presentation will be given by Zita Ripka during the side meeting of the germination committee

Other studies for new methods in progress

• Use of GA3 for Panicum maximum and Panicum coloratum germination test

(Test organizer: Augusto Martinelli)

• Use of KNO3 for lettuce germination test (Test organizer: Gillian Mc Laren)

• Test plan in preparation and search for seed sample

Validation studiesNew species

• Chenopodium quinoa

• Bambusa bambos and Bambusa vulgaris

• Cleome gynandra

Chenopodium quinoa

• Test organizers: Ignacio Aranciaga and Lesly González

• Test plan has been validated

6 participating laboratories 3 seed lots with different levels of germination Substrates : TP and BP Temperatures : 20°C and 2030°C Dormancy breaking treatments : None, Prechilling, Prechilling + KNO3 2 counts after 7 and 10 days

Bambusa bambos and Bambusa vulgaris

Species Prescriptions for: Additional directions incl. recommendations for breaking dormancy

Substrate Temperature (°C) First count (d) Final count (d)

Bambusa bambosPaper (BP)

25 ±1°C (to be confirmed)

7 – 9 (to be confirmed)

21 – 24 (to be confirmed)

To be studiedBambusa vulgaris

Information has been given from Dr. K. Ramamoorthy (India)

Test plan is not yet set up and seeds are waited

Need also for participants for comparative testing

Cleome gynandra

• Test organizer: Lea Mazor

• Preliminary germination tests

• Search for seeds

Rules proposals

• Amendment to the description of seedling abnormalities1 Abnormalities of the root system

11 The primary root

…

11/05 is deeply cracked or broken

11/06 is split from the tip or split right through

Rules proposals

Ending a germination test at a pre-determined germination level

• Objective: to allow finalizing germination tests as soon as a required level of germination is reached.

• Current situation– When a customer requires information on the germination level before

the date of the final count, a PROVISIONAL certificate can be issued. – However, if a PROVISIONAL certificate is issued, then it has to be

indicated on the PROVISIONAL certificate that a FINAL certificate will be issued later, once the germination test has been completed.

Rules proposalsEnding a germination test at a pre-determined germination level

• Proposal: indicating in chapter 1 that it is not necessary to issue a FINAL certificate when the customer does not need to have it

• Consequences for the Certificate– No changes to the germination chapter; the germination results are included

in the germination box of the certificate – The test has been concluded, but we are not sure that no new seeds will

germinate. Some categories may not be reported

• Advantages– Seed trade is speeded up, as seed lots can be shipped earlier.– Less work in the lab: less counts.– Less administrative work and costs involved as only one certificate will be issued.

Rules proposals


• Changes in Chapter 1– Add to paragraph 1.2.4 “Provisional certificate”

However, if an applicant instructs the laboratory to terminate a germination test once a predetermined germination percentage has been reached, the certificate must include the statement under “Other determinations” that a final certificate will not be issued

Rules proposals


• Changes in Chapter 1 and Chapter 5– Add to paragraphs 1.5.2.6 and 5.9 “Reporting results”

The following additional information must be reported under “Other determinations”

If an applicant requests that the test be terminated when the sample reaches a predetermined germination percentage without issuing a final certificate, the following statement: “Instructions have been received from the applicant to end the testing of the sample at this stage. A final certificate will not be issued.”

In-house germination methodsFor discussion

• Objectives– To prevent the proliferation of different germination

methods published in the Rules and the increasing number of validation studies that are required

– To provide methods easier to carry out, faster or cheaper, while keeping the required level of reliability

Rules proposals

• Proposals of the committeeGeneral principals– In-house germination methods or variations of ISTA germination

methods can be used as alternative germination methods, provided that these methods are validated by comparison with current ISTA germination methods.

– Alternative methods must be statistically validated using a peer validation study

– Alternative methods can be used only for species that are already included in the ISTA Rules.


• Proposals of the committee

Conditions of germination test that could be modified– Growing media

– Temperature

– Duration

– Dormancy breaking treatments


• Proposals of the committee

Reporting results– When an in-house method has been validated, then the results

must be reported in the germination boxes of the ISTA certificate with the following statement being added under “Other determinations”:

• “The germination method used is an in-house method validated by comparison with an ISTA standard method”.

– The germination conditions of the alternative method must also be described under “Other determinations”.


PublicationsAmendments to the ISTA

Handbook for Seedling Evaluation

• Amendments to Section 4 Laboratory

• Conditions for Seedling Evaluation – Containers

– Counting equipment

Proposal for a method to be used in order to evaluate counting equipment

– Germination apparatus



• Amendments to Section 12: Seedling Type D –Seedling Group A-1-2-3-1

Seedling as a wholethe seedling is abnormal if it

• is deformed…• has the endosperm detached from the scutellum

Lolium sp.



• Amendments to Section 13: Seedling Type D –Seedling Group A-1-2-3-2

Seedling as a wholethe seedling is abnormal if it

• is deformed…• has the endosperm detached from the scutellum

Zea mays

Sorghum sp

• The objective was to give more precision in the Rules and the Handbook on what is a "retarded primary root“

• An experiment was performed in 2010/2011 by Takayuki Okuda to study the effect of test germination conditions on root/shoot ratio– Many factors influence the ratio (duration, light, moisture)– The proposal at the end of the study was to “set a minimum

ratio and let the labs decide according to their germination conditions”

– With regards of the results of the experiment the committee decided not to include more precise guideline on retarded primary root in the Handbook

PublicationsGuidance on

“retarded primary root”

Workshops

ISTA Workshop on Flower Seed Testing

• Organized by ISTA Naktuinbouw

• From 06.06.2012 - 09.06.2012

• In Roelofarendsveen, The Netherlands

• Contribution of the Germination Committee to the lecture and practical sessions on germination of Pelargonium and Zinnia

Pelargonium

Zinnia

ISTA Germination Workshop17-19 September 2012, Saskatoon

Lecturers: Sylvie Ducournau and Kari Fiedler Local organisers: Steve Jones and Brenda Baergen

Hosted by: Seed Science & Technology Section, SaskatoonCanadian Food Inspection Agency – Agence Canadienne d’inspection des aliments

Questions to the committee

Subject of question From (country)Seedling evaluation and germination method of Vicia faba Portugal

Excel sheet for water retention calculation Ireland

Rounding up germination results Australia

Retesting due to fresh seeds Canada

Germination conditions of Senna occidentalis (L.) Link, Senna sophera (L.) Roxb. and Senna surattensis (Burm. f.) Irwib and BarnebyEmbryo classification Peru

Soybean seedling evaluation Argentina

Germination method for Swertia Chiraita Nepal

Helianthus seedling evaluation Portugal

Zea mays seedling evaluation Argentina

Paper for germination testing UgandaAssessment of root system depending on the germination medium USAHelianthus seedling evaluation Switzerland


Subject of question From (country)Difference between ISTA and AOSA regarding the evaluation of root system of Helianthus annuus

USA

Germination method for Cannabis sativa Spain

Sorghum seedling evaluation Argentina

Pictures of normal and abnormal seedlings IrakWater retention calculation New Zealand

Spinach germination: evaluation of root system The Netherlands

Quality characterisctics of organic growing media Japan

Using more than 400 seeds when delivering an ISTA Certificate CanadaLettuce cotyledon necrosis USATerminal bud evaluation Argentina

Secondary roots in Lupinus ItalyBlended seed germination test IranAddress of a company that supplies good quality paper Zambia


Subject of question From (country)Double seedlings and Fused seedlings IndiaTolerance tables for germination tests in different laboratories Belgium

ISTA Hndbook on Seedling Evaluation - Seedling Type D USAEquipment of light for germination testing JapanGermination of spinach The NetherlandsList of the germination temperatures for vegetable seed ZambiaGermination of grasses The NetherlandsPhotos of maize seedlings IrakCalculation and reporting of germination results New ZealandBeet washer USARounding tool FranceManufacturer producing Jacobsen Germinator MoldaviaTreated seeds laboratory germination test BrasilSand for germination testing USAStandardized germination testing method of tomatoe seeds according to ISTA Germany

Main questions Germination Methods New species Seedling evaluation Substrate suppliers and checks Equipment Calculations and tolerance tables Report of germination results ISTA / AOSA evaluation

Future work

• Introduction of new methods– Shortening the duration of germination test for Trifolium

repens, Trifolium pratense and Medicago sativa from 10 days to 7 days

– Add the temperature of 20°C for the germination of Spinacia oleracea

• Introduction of new species– Eustoma sp.– Brassica carinata

Future work

• Publications– Handbook: improvement of tree and shrubs photos of

seedlings– Handbook: guidance on assessment of Glycine max.

seedlings• Questionnaire on:

– Light used for germination testing– Particle size of sand – Seed disinfection prior to germination test

Thanks to all the members of the committee, the Statistics committee

and the ISTA SecretariatThank you for your attention

Documents

Germination Committee - ISTA · 2012-06-21 · • 1 fast germinating variety • 2 levels of germination quality • 1 slow germinating variety • 2 levels of germination quality