Cloning and expression analyses of sucrose non-fermenting-1-related kinase 1 (SnRK1b) gene during development of sorghum and maize endosperm and its implicated role in sugar-to-starch metabolic transition.

Abstract

A full-length cDNA clone, SbSnRK1b (1530 bp, GenBank accession no. EF544393), encoding a putative serine/threonine protein kinase homologue of yeast (Saccharomyces cerevisiae) SNF1, was isolated from developing endosperm of sorghum [Sorghum bicolor (L.) Moench]. Multiple sequence alignment data showed a phylogenetic affiliation of the sorghum clone with the SnRK1b group of protein kinases that are highly expressed in cereal seed endosperm. The DNA gel blot analyses indicated that SbSnRK1b gene is present as a single- or low copy number gene in sorghum. The RNA and protein gel blot analyses confirmed the expression of SbSnRK1b in developing sorghum caryopses, overlapping with the starch biosynthesis phase, 12-24 days after fertilization. In situ hybridization and immunolocalization data resolved the spatial specificity of SbSnRK1b expression in the basal endosperm transfer cell layer, the unique port of assimilate unloading in the growing sorghum seed. Expression of SbSnRK1b was also evident in the developing sorghum microspores, coincident with the onset of starch deposition phase. As in sorghum, similar spatiotemporal specificity of SnRK1b expression was observed during maize (Zea mays L.) seed development. However, discordant in situ hybridization and immunolocalization data indicated that the expression of SbSnRK1b homologue in maize is under posttranscriptional control during endosperm development.