Optimization of protein extraction and proteomic studies in Cenchrus polystachion (L.) Schult

Abstract

Apomicts have been studied at their genetic levels, but there are no any direct evidence of its mechanism. In order to understand the mechanism involved, a close relative of Pennisetum, Cenchrus polystachion, an apomictic species was explored for more insights into protein expression in reproductive structures. Optimization of protein extraction was studied with the leaf tissue and optimized protocol was extrapolated to other five tissues. The phenol-based protein extraction emerged as the best method for plant leaf tissue providing a better protein yield, separation of bands, removal of non-protein components like polyphenolic compounds and nucleic acids. The proteome analysis of leaf, stigma, immature ovary, seed, anther sac and pollen tissues of Cenchrus polystachion were carried out identifying a total of 135407 proteins against the Poaceae database from UNIPROT/TrEMBL. The target candidate proteins found in all the tissues were identified and mainly comprised of Actin Protein, PIP, Starch Synthase, ATP Synthase, Glutathione S Transferase, Dehydroascorbate reductase, Ascorbate peroxidase and heat shock proteins. Visualization and descriptive statistics conveyed all the necessary information to understand the differential expression of proteins in Cenchrus polystachion. This study forms a base to understand the role of tissue specific expressed proteins in an apomictic plant.