spinach short root project ista conference 13th june 2018 ... · on germination conditions...
TRANSCRIPT
ENZA ZADEN
For internal use only --
C L I C K T O S TA R T
Spinach short root project – ISTA
conference 13th June 2018 – Sapporo, Japan
Lucile Daron (STR-PR)
20-6-2018
ENZA ZADEN
ENZA ZADEN
Background information
Timeline overview
Validation Study M&M + results
Conclusions
Method Validation report
Outline
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Background information
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Lot 1 - 2011
Field emergence = 50%
Naktuinbouw TG > 85% vs Enza TG = 50%
(TG = Total Germination)
Spinach (SN) conference - October 2011
Results varied between laboratories
Discrepancies ISTA / companies criteria
Project led by Naktuinbouw 2011-2014
Project led by Enza Zaden 2015-2018
SN SR project – Where it all began…
Lot 1 Lot X
Lot 1
Lot X
Lot 1
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Timeline overview
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SN SR project in time
Questionnaires to seed companies
Comparative tests NAKT
Data analysis and results discussion
EZ / NAKT
Batches selection + Experimental
design
EZ preliminary experiments
Data analysis + discussion with ISTA GERCOM + recruiting
laboratories for Validation Study
Test Plan + batches selections
Impact disinfection on SR occurrence + Feedback from
GERCOM reviewers
VRT1 + VRT2 conducted
Comparative tests = RT1
Data analysis + discussion +
experimental design
Side experiments on germination
conditions
Experimental design + samples
sent to labs
RT2 + RT3 conducted
Data analysis + statistical testsReport for GERCOM
+ discussionValidation Report
2011 – 2012 – 2013Preliminary work
2014 – 2015Ring tests between seed
companies
2015 – 2016 – 2017Validation study
2017 – 2018 – 2019Rule validation GERCOM ISTA
Oct.. 2011Spinach Conference NL
Jan. 2012Report of QuestionnairesNAKT
July 2013Report Comparative testsNAKT
Nov. 2013STR involvedEZ
May 2014
Aug. 2014NAKT
Sept. 2014
Oct. 2014EZ (STR) takes the project lead
Feb. 2015EZ
March 2015EZ
July 2015EZ
Sept. 2015
April 2016 July 2016 Nov. 2016Sept. 2015
Nov. 2016 June 2017 Nov. 2017GERCOM
voting = YES
Jan. 2018Report submitted
ISTA Conference Estonia poster
ISTA Conference Denver
First contact with ISTA GERCOM –A. Grim (NAKT)
ENZA ZADEN
SN SR project steps to come
Data analysis + statistical testsReport for GERCOM
+ discussionValidation Report
Nov. 2016 June 2017 Nov. 2017GERCOM voting
Jan. 2018Report submitted
June 2018
ISTA voting = ???Jan. 2019Published in Handbook ?
ISTA Conference Japan
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Validation study 2016 - 2017
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CURRENT VERSION of ISTA RULES
5.2.7.2 Slight defects
“The following defects are considered slight and therefore seedlings are classified as normal:
– primary root with limited damage (e.g. not affecting the conductive tissue) or slight growth retardation;”
Grey area: what is acceptable growth retardation for Spinacia oleracea?
No clear root criterion: causing results variability / discrepancies between laboratories?
GOAL: Testing if standardized root criterion = decreased varibility between
laboratories
Validation Study 2016 + 2017 – Material and Methods
ENZA ZADEN
6 lots from 3 seed companies
7 participating labs
1. GEVES (France)
2. ARO Volcani Center (Israel)
3. SGS Brookings (USA)
4. Landwirtschaftliches Technologie Zentrum Augustenberg - LTZ (Germany)
5. Naktuinbouw (Netherlands)
6. CREA (Italy)
7. Enza Zaden (Netherlands)
Validation Study 2016 + 2017 – Material and Methods
LotSeed size
(mm)Supplier Production year TG (%) SR (%)
Other abnormal
seedlings (%)NG (%)
1 2,75-3,50 Enza Zaden 2009 37 59 3 12 2,50-3,50 Enza Zaden 2013 59 13 18 10
3 2,75-3,50 Pop Vriend 2012 68 21 8 3
4 2,50-3,50 Enza Zaden 2012 80 7 6 7
5 2,60-3,50 Pop Vriend 2015 80 12 4 4
6 x Bayer 2015 88 7 0 5
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High diversity of
germination methods
*lot 2 only 200 seeds tested
2 Validation Ring Tests
(VRT1, VRT2)
Validation Study 2016 + 2017 – Material and Methods
Laboratory MethodTemperature
(°C)
No.
independent
samples
Repetitions
no. seeds
Total no. seeds
tested *
Water supply
(mL)
Light regime
(light/dark)
(h)
First-Final
germination
assessment (days
after sowing)
Root:shoot
threshold
GEVES PP 15 2 2 * 100 400 32 8h/16h 7-21 1:2LTZ PP 15 2 4 * 50 400 35 8h/16h 7-14 2:3
NAKTUINBOUW PP 15 2 2 * 100 400 40 12h/12h 7-21 1:2
ARO VOLCANI BP 15 2 4 * 50 400
Not exact
amount:
spraying
several times
0h/24h 7-21
No defined
threshold: if
no change in
root length
during testCREA BP 15 2 2 * 100 400 7 0h/24h 6-21 1:2
SGS BROOKINGS BP 15 2 2 * 100 400
Towels soaked
and excess
water removed
8h/16h 7-21 1
ENZA ZADEN PP 15 2 4 * 50 400 40 8h/16h 7-14 1
Ring test no.Germination
method
Repetitions
and no. of
seeds
Pre- treatment
Germination
temperature
(°C)
Water supply
(mL)
Light regime
(light/dark)
Germination
assessment
(days after
sowing)
Root:shoot
threshold
1 Variable Variable Variable 15 Variable Variable Variable Variable2 Variable Variable Variable 15 Variable Variable Variable 0,5
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Defining R:S threshold for lab tests
Available germination results:
Pleated Paper (PP) at 15 °C to assess normal seedlings %
Grow Out (GO) at 15 °C to assess normal plants %
Validation Study 2016 + 2017 – Results
Prediction normal plants % by normal seedlings %
R:S = 1 under estimates
R:S = 0.5 estimates accurately
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Per Seed Lot
Per Method
Per Lab * Method
(Charts from Statistical tool ‘ISTAgermMV’, Jean-Louis Laffont, 2017)
Validation Study 2016 + 2017 – results
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Per Method * Lot
Repeatability & Reproducibility
Repeatability improved !
Reproducibility very much improved !
Conclusion
Standardized root criterion = more uniform results within and between labs
Validation Study 2016 + 2017 – results
Method ...p rS
rf RS
2
Run̂
2
Lot Run̂
VRT1 81 4.96 1.07 11.07 7.94 5.91
VRT2 82 4.74 1.04 8.37 5.14 4.61
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Conclusions
ENZA ZADEN
SN SR project - Conclusions
CURRENT VERSION PROPOSED VERSION
5.2.7.2 Slight defects
The following defects are considered slight and
therefore seedlings are classified as normal:
– primary root with limited damage (e.g. not affecting
the conductive tissue) or slight growth retardation;
5.2.7.2 Slight defects
The following defects are considered slight and
therefore seedlings are classified as normal:
– primary root with limited damage (e.g. not affecting
the conductive tissue) or slight growth retardation (for
Spinacia oleracea, the length of the primary root has to
be equal to or greater than the half of the length of the
hypocotyl);
“Results from validation studies showed that a root criterion (with root:shoot ratio = 0.5) for the assessment of spinach seedlings led to a more uniform evaluation between laboratories.It is therefore proposed by the Germination Committee to include this criterion into the Rules.”From ‘Rules Proposals GERCOM 2018’, Sylvie Ducournau, November 2017
Final voting 14th of June 2018…
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Method Validation report
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Validation Study report available on ISTA’s website:
‘OGM18-07 ISTA Method Validation Reports’, pages 32-50
(https://www.seedtest.org/en/preparatory-documents-_content---1--3402.html)
Available online
ENZA ZADEN
- ISTA- GEVES- ARO VOLCANI CENTER- CREA- SGS BROOKINGS- LTZ Center for Agricultural Technology- NAKTUINBOUW- POP VRIEND- BAYER
Thank you to…
ENZA ZADEN
Thanks to the staff of the participating ISTA laboratories: CREA, GEVES-SNES, NAKTUINBOUW, ARO VOLCANI CENTER, LTZ and SGS
BROOKINGS. Thanks in particular to the reference persons of those laboratories: Rita Zecchinelli, Sylvie Ducournau, Anton Grim/Marcel
Toonen, Miriam Dekalo Keren, Andrea Jonitz and Sarah Dammen (respectively).
Thank you to the Germination Committee members for their constructive input during our debates.
Thanks to Nadine Ettel, TCOM coordinator at the ISTA Secretariat and to Andreea Militaru, TCOM administrator for their support.
Thanks to the technical reviewers Harry Nijenstein and Christine Herzog.
Thanks to Jean-Louis Laffont and the ISTA Statistics Committee for running the statistical analyses.
A special thanks to Sylvie Ducournau for her patience, support and guidance during this project.
Thank you to…
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Any question?