Sequence polymorphisms in Zmisa2 gene are significantly associated with starch pasting and gelatinization properties in maize (Zea mays L.)

Researches on the genetic basis of starch pasting and gelatinization properties will provide foundation of maize improvement for quality, feed and industrial applications. Maize gene Zmisa2, encoding an isoamylase-type starch debranching enzyme, plays important roles in starch biosynthesis. In this study, the genomic sequences of the gene Zmisa2 in 72 elite maize inbred lines were obtained, and the nucleotide polymorphisms and haplotype diversity were detected. In addition, seven pasting and four gelatinization properties of maize were measured for the tested inbred lines using rapid visco analyzer and differential scanning calorimeter, respectively. A total of 99 sequence variants, including 91 SNPs and 8 indels, were identified at the promoter and coding regions of this gene. Although the frequency of polymorphism in promoter region is much higher than that of coding region, the SNPs in the coding region of maize gene Zmisa2 classified this gene into 21 haplotypes, which encode 11 different ISA2 proteins. Furthermore, the association of the variants of Zmisa2 gene with maize starch pasting and gelatinization properties was estimated, and the results revealed that seven SNPs in coding region, including four nonsynonymous sites, were significantly associated with phenotypic variations of pasting time and enthalpy of transition (ΔH). These results suggested that the polymorphism in maize Zmisa2 locus could be used in molecular marker-assisted selection for improvement of quality in maize breeding programs.