Soybean Cultivar Identification Using High Performance Liquid Chromatography of Chromatography of Seed Proteins

Abstract

A substantial increase in the number of soybean [Glycine max (L.) Merr.J cultivars has occurred in recent years. Many of these are derived from a narrow genetic base, which makes traditional cultivar identification increasingly difficult. As a result, new techniques that detect minor biochemical differences need to be evaluated for their ability to differentiate among cultivars. This study examines the applicability of reversed phase high performance liquid chromatography (RP‐HPLC) for soybean cultivar identification. Salt soluble proteins of 14 cultivars were extracted from unimbibed seeds, eluted from a reversed phase C4 column with increasing concentrations of acetonitrile, and monitored at 280 nm. Seeds were obtained for study from three differing growing locations in Ohio. While no qualitative differences of elution profiles were detected within cultivars and locations, comparisons of quantitative RP‐HPLC seed protein profiles permitted separation of genetically diverse genotypes. Retention time ratios for selected peaks were not significantly different among cultivars, but peak area percentage showed that quantitative differences were useful criteria for distinguishing closely related lines. This study documents that RP‐HPLC analysis of soybean seed proteins can distinguish divergent soybean genotypes and suggests that peak area percentage of RP‐HPLC profiles is a superior method of cultivar analysis.