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Genetic diversity of Pisum sativum wrinkled seed collection for seed sugar composition and concentration . Clarice J. Coyne, Rebecca J. McGee, D. Scott Mattinson, Sam Fuchs, John Fellman. Objective. Genetic diversity of oligosaccharides in USDA pea germplasm - PowerPoint PPT Presentation
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Genetic diversity of Pisum sativum wrinkled seed
collection for seed sugar composition and
concentration
Clarice J. Coyne, Rebecca J. McGee, D. Scott Mattinson, Sam Fuchs, John
Fellman
Objective Genetic diversity of oligosaccharides in
USDA pea germplasm› Set I: wrinkled green pea germplasm› Harvested at green processing stage
Identify diversity for breeders› Soybean (Dierking & Bilyeu 2008)
Pea carbohydrates significance
Pea : inexpensive source of protein, carbohydrate, vitamins and minerals.
Probiotic: summary by Ekvall et al. 2006 Food Chemistry 94: 513–519.
Antinutritional factors Frost tolerance Desiccation protection Carbohydrate transport in the phloem
Introduction to pea LMWC Low molecular weight carbohydrates in
pea seed include:› Sucrose› Fructose› Glucose› Sorbitol› Raffinose› Stachyose› Verbascose
MATERIALS AND M
ETHODS I
Wrinkled pea from the USDA pea core collection and from the VIR collection
Six plants per accession grown in potting mix under greenhouse conditions
Experiment IHarvested at the green pea stage (estimated maturity as no tenderometer available)
Experiment IIHarvest green pea stage using same days after flowering for all accessions (in progress)
MATERIALS AND M
ETHODS II
Green pods were harvested and stored at -20 C until prepared for gas chromotography analysis
TechnicianSam Fuchs
SAMPLING THE ACCESSIONS
Pea seed removed from pod
Uniform seed selectedNote: Ekvall et al 2003
Three subsamples of 1 gram each of uniform color and size pea
Samples ground with a Terex Grinder
ETHANOL EXTRACTION
Samples dried down
CARBOHYDRATES DERIVATIZED
Seed extracts were taken to dryness under reducedpressure.
Derivatized with a mixture of trimethylsilyl imidazole:pyridine(1:1, v/v).
GAS CHROMOTOGRAPHY
Soluble carbohydrates were converted into trimethylsilylderivatives and separated by capillary gas chromatography as described previously (Peterbauer et al .1998).
For quantification, ratios of peak areas to that of the internal standard (phenyl b-D-glucopyranoside) were compared with those of external standards (Sigma).
Results from Experiment I
#574510-USA
#411143-New Zealand
#356974-India
#413684-Hungary
#164971-Turkey
#244150-Netherlands
0.1
1
10
100
1000
10000
Fructose Glucose Sorbitol B-pheyl-Gluco. Sucrose Raffinose Stachyose Verbascose
USDA World Pea Gerplasm Analysis of CarbohydratesC
arbo
hydr
ate,
ug/
g (F
W),
Log 10
Summary Germplasm rich source of diversity in
low molecular weight carbohydrates Methods yield high quality data with on
low standard errors Experiment II: use DAP for uniform
maturity (100 accessions) Experiment III: dry pea accessions
Many thanks USDA Pisum CGC Evaluation funding Greenhouse helpers Landon Charlo,
Alison Hitchcock, Nancy Nydegger-Paulitz
Collaborators Rebecca J. McGee, D. Scott Mattinson, Sam Fuchs, and….
Post-harvest chemist:Professor John Fellman