Analysis and manipulation of the starch biosynthesis pathway in hexaploid spring wheat (Triticum aestivum L.)

Abstract

Starch is an important component of a wheat grain, comprising 50-70% of its dry weight. Its biosynthesis involves a complex pathway mediated by several enzymes, each of which is encoded by genes that have more than one family member. To better understand starch synthesis in wheat grains, this study characterized the sucrose-starch metabolic pathway using physiological, molecular, biochemical and metabolic approaches. These analyses led to the identification of genes that appear to have predominant expression during grain development in wheat including, TaSUT1, TaSuSy2, AGPL1, SSI, SSIIIa and SBEIIa, suggesting that these genes play a regulatory role in starch accumulation. This was further confirmed by comparative analyses of starch synthesis between cultivars with contrasting thousand kernel weights, which revealed a closer association of the expression of the same set of genes with starch accumulation in developing wheat grains. The effect on starch yield of one of the candidate genes identified, AGPase, was examined through a transgenic approach, which involved expression of a gene encoding modified version of maize AGPase large subunit, designated as Sh2r6hs, in wheat under the control of maize’s constitutive Ubiquitin1 promoter. This manipulation of the wheat AGPase activity produced wheat lines with increased AGPase activity, grain weight and grain starch level, suggesting that the wheat grain size can be enhanced through increasing the capacity of starch synthesis both in the source and sink tissues. The study also identified and characterized a partial fragment of wheat rbcS promoter, and indicated that the promoter fragment can potentially be used as a tool for targeting the expression of genes of interest in photosynthetic source tissues.