The use of micro-beam X-ray diffraction for the characterization of starch crystal structure in rice mutant kernels of waxy, amylose extender, and sugary1

Micro-beam X-ray diffraction analyses were carried out on slices from wild-type rice and mutants. The method permitted the crystalline structure to be analyzed with less destruction of cells and fewer artificial effects that occur during the purification of starch from tissue and provided information concerning the localization and crystalline structure of starches distributed in the endosperm. The starch on kernel slices from the wild-type and waxy mutant, carrying a defect for the Granule-bound starch synthase I (GBSSI) gene, displayed an A-type of diffraction pattern; no difference in the crystalline patterns between starches located in the inner and outer region in a kernel were observed. A double mutant of the waxy and amylose extender (ae) mutant carrying a defect for the Starch branching enzyme IIb gene accumulated amylose-free B-type starches. The kernel slice from the double mutant of waxy and sugary1, mutated on the Isoamylase I locus, displayed an A-type diffraction pattern in the outer region and was amorphous in the inner region. A chain-length distribution analysis of polyglucans in kernels from wild-type and mutants showed that ae amylopectin had more long chains and less short chains than the wild-type and waxy amylopectin. On the other hand, the water-soluble polysaccharide in the inner region of the sugary1 endosperm had more short chains than the amylopectin in the outer region counterparts. These results indicate that branch chain length in amylopectin is crucial in determining the formation of A- and B-type starches.