Corn Germination 2016 Melissa Phillips Monsanto Seed Technology Center Waterman Illinois USA Purpose The purpose of the referee was to introduce Agar plant culturing media as a potential candidate for use in germination testing for corn ID: 659720
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National RefereeAgar MediaCorn Germination2016
Melissa Phillips
Monsanto
Seed Technology Center
Waterman, Illinois USASlide2
PurposeThe purpose of the referee was to introduce Agar (plant culturing media) as a potential candidate for use in germination testing for corn.
Participants
16 Labs in total signed up to participate
Agar was provided upon request to 7 labs
Seed was sent out June 2016
Data was returned August- Sept
10 labs returned dataSlide3
Previous StudiesProtocolKept with more traditional studies of “open protocols”
Agar requires more specific requirements
6 corn samples
Samples tested using in-house AOSA approved method and agar media
Process was to be repeated 5-7 days later to establish lab repeatabilityAgar concentration to be used 0.8%-1%Slide4
Statistics supplied by Paul Cannon and Kirk Remund- Monsanto
Variance
Method
Mean Germ
Median Germ
Repeatability*
Reproducibility*
Part to Part
Interaction
TotalAgar84.38828.9 (17%)29.0 (17%)99.2 (58%)15.1 (9%)172.2Current87.19024.2 (19%)4.3 (3%)89.0 (70%)10.5 (8%)128.0
*Adequate measurement systems have total Repeatability + Reproducibility less than 10% of total variation
Repeatability is the closeness of the agreement between the results of the same measure and carried out in the same conditions.Reproducibility is the precision under reproducibility conditions, i.e. conditions where test results are obtained with the same method on identical test items in different laboratories with different operators using different equipment.
Agar results are on par with current methods with in each lab.
Agar results are less reproducible than current methods when compared across multiple labs.Slide5
Corn Germination ReproducibilitySlide6
Significant Outliers; Why?
What is Occurring?
Some labs were in alignment,
others were not
Post referee survey
Current Method
CCP or CCP/Sand vs. Rolled Towel
Experience with Agar
First Time to Experienced
Equipment used for agar preparationStoveMicrowaveHotplateAutoclaveSlide7
Corn Germination based on agar experience
* sample numbers are not large enough to be significant
No conclusions can be made based on experienceSlide8
Corn germination based on media
* sample numbers are not large enough to be significantSlide9
ConclusionStudy results were not supportive of a 2016 rule proposal for corn germinationResults show that there is still reproducibility issues across some labsIn-lab repeatability is on par with current approved methods
The use of specific protocols increases the repeatability of the agar methodSlide10
Further analysis of factors impacting repeatability and reproducibilityFuture Targeted/ Strategic StudyProtocol of corn agar that will allow us to control critical to quality control variables
Moving Forward
Thank you!
Special thanks to all of the referee participants!