Screening of seeds from wild pennycress (Thlaspi arvense L.) populations for sinigrin levels using a blood glucose meter and test strips

Authors: PHIPPEN, MARY - Western Illinois University; BERHOW, MARK - Retired ARS Employee; Brownstein, Korey; WESLEY, TAD - Western Illinois University; PHIPPEN, WINTHROP - Western Illinois University

Submitted to: Industrial Crops and Products
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/25/2024
Publication Date: 7/9/2024
Citation: Phippen, M.E., Berhow, M.A., Brownstein, K.J., Wesley, T.L., Phippen, W.B. 2024. Screening of seeds from wild pennycress (Thlaspi arvense L.) populations for sinigrin levels using a blood glucose meter and test strips. Industrial Crops and Products. https://doi.org/10.1016/j.indcrop.2024.119082.
DOI: https://doi.org/10.1016/j.indcrop.2024.119082

Interpretive Summary: Field pennycress is a new oil seed cover crop being commercialized in the United States. Pennycress seed is abundant in sinigrin, a glucosinolate, which has anti-nutritional factors making the seed and meal less palatable to animals. A rapid and inexpensive method using a blood glucose meter and test strips was developed to screen seeds of 429 wild pennycress lines for sinigrin. While HPLC/MS is the most accurate analysis for sinigrin in pennycress seed, determining sinigrin levels with a glucose meter can provide an inexpensive screening method to identify populations of interest with low sinigrin levels. In fact, the glucose meter results correlated to the HPLC/MS analysis results. The screening of the wild populations for sinigrin levels in the seed will help researchers who would like to identify low glucosinolate lines for potential food markets or perhaps increase sinigrin levels to improve biotic stress tolerance and potential industrial markets for pennycress seed meal.

Technical Abstract: Field pennycress, Thlaspi arvense L., is a new oil seed cover crop being commercialized as a source of biofuel in the United States. Along the path to commercialization, pennycress seed and meal will be marketed as animal feed. As a member of the Brassicacaeae family, pennycress seed is abundant in glucosinolates which are anti-nutritional factors that make the seed and meal less palatable to animals. More specifically, sinigrin has been identified as the only glucosinolate present in pennycress seed. As work continues to develop varieties low in glucosinolates, a rapid assay is needed to determine the sinigrin level in pennycress seed. The objective of this study was to develop and utilize a high throughput, rapid assay to screen pennycress seeds from wild populations for total sinigrin content. Pennycress seeds were ground in potassium phosphate buffer with endogenous myrosinase to catalyze the enzymatic hydrolysis of sinigrin to equimolar amounts of glucose. Glucose content was determined using a blood glucose meter and test strips. From the molar concentration of glucose, the molar concentration of sinigrin was known and the sinigrin level in seeds was calculated in µmol/g. The results were confirmed by HPLC/MS analysis. A rapid and inexpensive method was developed and used to screen seeds of 429 wild pennycress lines for sinigrin. Sinigrin levels in seed ranged from 25 to 85 µmol/g in the pennycress lines screened. Environmental factors also seemed to contribute to the levels of sinigrin in seeds found within the line ‘tt8-t/ARV1’ grown at ten different locations in the Midwest, USA. This rapid and inexpensive assay will allow breeders to efficiently screen lines and select for varieties that produce seeds with low sinigrin levels for improved feed value.