Gelatinisation temperature (GT) is one of the key traits measured in programs for breeding rice (Oryza sativa L.). It is commonly estimated by the alkali spreading value (ASV), and less commonly by differential scanning calorimetry (DSC). Using a diverse set of germplasm, it was determined that DSC values associate poorly with ASV, are not correlated with amylose content but correlate with cooking time. Rice varieties are traditionally grouped into three classes of GT based on ASV: high, intermediate and low. However, the distribution of DSC values of 4000 samples shows only two classes: high and low. Large differences in the distributions of chain lengths synthesised by starch synthase IIa (SSIIa) support the two classes as the major grouping, two haplotypes associating with each peak. Each peak of DSC values spanned 10 C. The chain length distribution of the amylopectin molecules from varieties at the upper boundary of each peak showed significantly more chains that span both the crystalline and amorphous lamellae of a cluster than varieties at the other end of that distribution. Improved varieties, classified as intermediate GT by ASV, belong to both of the classes defined by DSC, implying that some enzyme, other than SSIIa is involved in intermediate GT.
Gel consistency (GC) is a standard assay used in rice improvement programmes to determine whether rice cultivars/breeding lines of high amylose content are soft or firm textured when cooked. In this study, we show that sequence variation in exon 10 of the Waxy (Wx) gene associates with GC using RILs derived from parents with high amylose content that differ in GC. The association was validated using a diverse set of traditional varieties, selected on the basis of amylose content, from the generation challenge programme. Structural investigations to explain how the mutation leads to differences in GC showed a strong association between GC and the proportion of amylose that leaches. It was shown that cooked rices of hard GC do not change in hardness over 24 h, whereas rices of soft GC retrograde significantly over 24 h. This leads to the conclusion that the mutation on exon 10 of the Wx gene affects the proportion of amylose bound to amylopectin and the proportion able to leach, and these structural differences alter the composition of the gel, which affects the amount of time the gel takes to reach a final hardness. The SNP described here completes the set of markers required to genotype for the current traits of cooking quality, but selecting the allele for soft texture has the negative result of also selecting for retrogradation potential.
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