Effects of Two Starch Synthase IIa Isoforms on Grain Components and Other Grain Traits in Barley

Starch biosynthesis in cereal crops is a complex pathway regulated by multiple starch synthetic enzymes. Starch synthase IIa (SSIIa) is well-known to be one of the major starch synthases and is very important in amylopectin biosynthesis. It has significant effects on grain composition and kernel traits. However, there are few reports on the association of natural variation of SSIIa in barley and grain composition and characteristics. In this work, two SSIIa isoforms were first identified as SSIIa^H and SSIIa^L by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis, mass spectrometry, and Western blotting. Sequence analysis of the SSIIa gene demonstrated that a 33 bp insertion coding a peptide of APPSSVVPAKK caused different SSIIa, e.g., SSIIa^H and SSIIa^L. Based on this molecular difference, a polymerase chain reaction marker was developed, which could be used to screen different SSIIa genotypes easily. Kernel hardness of SSIIa genotypes was significantly higher than that of SSIIa Chinese barley cultivars. The proportion of SSIIa genotypes was extremely low in Australian barley cultivars (5/24) and much higher in Tibetan hull-less barley cultivars (46/74), consistent with the end-use requirements of barley grain. This study provided new information in barley endosperm starch synthesis and indicated that it is valuable for choosing the preferred SSIIa genotype according to the end-use requirements.