Expression of Escherichia coli branching enzyme in caryopses of transgenic rice results in amylopectin with an increased degree of branching

Abstract

Physiochemical properties of starch are dependent on several factors including the relative abundance of amylose and amylopectin, and the degree of branching of amylopectin. Utilizing Agrobacterium-mediated transformation, a construct containing the coding region of branching enzyme of Escherichia coli, under transcriptional control of the rice (Oryza sativa L.) starch-branching enzyme promoter was introduced into rice cv. Nakdong. To enhance glgB expression, the first intron of rice starch-branching enzyme and the matrix attachment region (MAR) sequence from chicken lysozyme were included in the expression vector. Eleven independent transgenic rice plants were generated. Southern blot analysis indicated that the copy number of glgB integrated into transgenic rice varied from one to five. High-performance liquid chromatographic analysis of starch from transgenic lines revealed that amylopectin from transgenic lines exhibited greater branching than that of non-transgenic rice. The A/B1 ratio in amylopectin increased from 1.3 to 2.3 and the total branching ratio, A+B1/B-rest, increased from 6 to 12 in transgenic rice. The observed increase in the short-chain fractions with a degree of polymerization between 6 and 10 is expected to have a significant effect on retrogradation. Our study demonstrates that amylopectin branching can be altered in vivo, thus changing the physicochemical properties of starch.