Analysis of the expression, subcellular and tissue localisation of phosphoglucan, water dikinase (PWD/GWD3) in Solanum tuberosum L.: a bioinformatics approach for the comparative analysis of two α-glucan, water dikinases (GWDs) from Solanum tuberosum L.

Abstract

There are several important factors affecting the rate of starch decomposition in plants, including the circadian clock, the regulation of gene expression, the regulation of enzyme activities and starch phosphorylation by glucan, water dikinase activities (GWDs). One isoform of glucan, water dikinase named GWD3 or PWD (EC 2.7.9.5) was isolated for the first time from Arabidopsis thaliana, and now we report its isolation and identification in Solanum tuberosum L. leaves and tubers. We compare StGWD3 sequence to the other GWDs sequences using bioinformatics tools and propose also structural models for the starch-binding domains in StGWD3 and StGWD1. The StGWD3 gene expression and protein were localised in different heterotrophic and autotrophic potato tissues and organs using in situ RT-PCR and immunolocalisation methods, respectively. Diurnal changes in the transcript abundance of StGWD3 in leaves were analysed using quantitative real-time PCR and they appeared to be typical for most genes involved in starch degradation in chloroplasts.