Tartary buckwheat (Fagopyrum tataricum Gaertn.) is increasingly considered as an important functional food material because of its rich nutraceutical compounds. Reserve starch is the major component of tartary buckwheat seed. However, the gene sequences and the molecular mechanism of tartary buckwheat starch synthesis are unknown so far. In this study, the complete genomic sequence and full-size cDNA coding tartary buckwheat granule-bound starch synthase I (FtGBSSI), which is responsible for amylose synthesis, were isolated and analyzed. The genomic sequence of the FtGBSSI contained 3947 nucleotides and was composed of 14 exons and 13 introns. The cDNA coding sequence of FtGBSSI shared 63.3%-75.1% identities with those of dicots and 56.6%-57.5% identities with monocots (Poaceae). In deduced amino acid sequence of FtGBSSI, eight motifs conserved among plant starch synthases were identified. A cleavage at the site IVC↓G of FtGBSSI protein produces the chloroplast transit sequence of 78 amino acids and the mature protein of 527 amino acids. The FtGBSSI mature protein showed an identity of 73.4%-77.8% with dicot plants, and 67.6%-70.4% with monocot plants (Poaceae). The mature protein was composed of 20 α-helixes and 16 β-strands, and folds into two main domains, N- and C-terminal domains. The critical residues which are involved in ADP and sugar binding were predicted. These results will be useful to modulate starch composition of buckwheat kernels with the aim to produce novel improved varieties in future breeding programs.
Keywords: ADP; ADP glucose pyrophosphorylase; AGPase; Adenosine diphosphate; Agrobacterium tumefaciens glycogen synthase; Amylose; AtGS; BE; Branching enzyme; CDS; Coding sequence; DBE; Debranching enzyme; EcGS; Escherichia coli glycogen synthase; FtGBSSI; GBSSI; GP; GS; GT; Glycogen phosphorylase; Glycogen synthase; Glycosyltransferase; Granule-bound starch synthase I; MalP; Maltodextrin phosphorylase; NJ method; Neighbor joining method; ORF; Open reading frame; Oryza sativa japonica GBSSI; OsGBSSI; PCR; Phyre2; Polymerase chain reaction; Protein homology/analogy recognition engine version 2.0; RACE cloning; Rapid amplification of cDNA ends cloning; Reserve starch; SDS-PAGE; SS; Sodium dodecyl sulfate polyacrylamide gel electrophoresis; Starch synthase; Tartary buckwheat; Tartary buckwheat granule-bound starch synthase I; UDP; UTR; Untranslated region; Uridine diphosphate.
© 2013.