Identification of novel mutations in the rice starch branching enzyme I gene via TILLING by sequencing

Starch branching enzymes (SBEs) incorporate α-1,6-branched glucosidic linkages into α-1,4-glucan chains. Rice has three SBE isoforms (SBEI, SBEIIa, and SBEIIb), which differ in the amylose chain lengths that they transfer to amylopectin. SBEI mutants characterized to date have had little or no effect on starch structure or content. In this study, TILLING by sequencing was employed to identify SBEI mutations in a population of chemically induced mutants of the cultivar Nipponbare (n?=?4096), resulting in the detection of 37 putative mutations. After removing mutations in introns or predicted to be synonymous, Sanger sequencing of M2 and M3 generation mutants confirmed six mutations and homozygous lines for four of these mutations were isolated. Only one line, NM-4936, exhibited a visible grain phenotype and produced sufficient M4 seeds for morphological and physicochemical analyses. In contrast to Nipponbare, NM-4936 grains are opaque due to rounded, loosely packed starch granules, and consequently, grain hardness was significantly reduced. NM-4936 grains exhibited significantly reduced width and thickness, which is reflected in reduced grain weight. Physicochemical analysis revealed significant differences in apparent amylose content (15 vs. 19%) and protein content (9.2 vs. 4.8%). Amylopectin chain-length distribution analysis indicated that the mutant has a slight but significant decrease in B₃ chains [degree of polymerization (DP)?≥?37, P?<?0.01] and a very slight but significant increase in B₁ chains (DP 13–24; P?<?0.05). Identification of novel SBEI mutations will be useful in elucidating its function and may provide useful germplasm for breeding novel quality traits.     

Analysis of QTL responsible for grain iron and zinc content in doubled haploid lines of rice (Oryza sativa) derived from an intra-japonica cross

Quantitative trait loci (QTL) related to the grain iron and zinc contents of brown rice were mapped by using a doubled haploid population derived from an intra-japonica cross between 'Hwaseonchal' and 'Goami 2'. QTL–QTL, background–QTL, and background–background interactions and candidate genes that affect grain iron and zinc contents were preliminarily identified. Twenty-one iron- and zinc-related QTL were found. The major-effect QTL qFe7 and qZn7 provided the highest contribution to phenotypic variance for grain iron and zinc contents. The colocation of zinc- and iron-related QTL on chromosomes 1, 4, 7 and 11 may account for the strong correlation between iron and zinc contents. A region on chromosome 7 and epistatic interaction between loci on chromosomes 2 and 10 affected iron content. qZn7 and qZn11.3 exerted additive effects on zinc content. Eleven iron- and zinc-related candidate genes colocated with qFe7, qZn7 and the region on chromosome 7 with an additive effect on iron content. The major-effect QTL identified here may be useful for breeding biofortified rice.     

Quality of head and chalky rice and deterioration of eating quality by chalky rice

Chalk, an opaque area in the rice grain, is an important quality characteristic in rice and occurs most commonly when grains are exposed to high temperatures during development. Chalky rice decreases the value of rice because of its undesirable appearance and eating quality for consumers. We investigated the chemical composition, morphological structure, cooking, texture properties of cooked rice, and pasting and gelatinization properties to evaluate the reason for the deterioration in eating quality of chalky rice.