clarice j. coyne, rebecca j. mcgee, d. scott mattinson, sam fuchs, john fellman

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