In-silico characterization and gene expression analysis of HSP101 under heat stress conditions

Abstract

Abstract To understand the genetic and molecular mechanisms that regulate heat stress responses in wheat genotypes, gene expression analysis and in-silico characterization of heat shock proteins 101 (HSP101) was performed. Three isoforms of HSP101 genes (HSP101a, HSP101b, and HSP101c), known for their roles in plant growth responses and adaptation to heat stress were examined. qRT-PCR analysis revealed constitutive expression of HSP101 transcripts in wheat plants, with maximum expression observed at 40°C heat shock treatment. Heat-tolerant genotypes (DBW 90, RAJ 4083, and DBW 71) exhibited a progressive upregulation of HSP101 gene transcripts with increasing heat shock treatments. For the in-silicocharacterization of HSP101a, HSP101b, and HSP101c transcripts gene sizes were found 240bp, 214bp, and 183bp through automated sequencing. The BLAST analysis confirmed their high homology with chaperone proteins and other HSP101-related genes in wheat. Phylogenetic comparisons, and amino acid translations provided insights into their evolutionary relationships and similarities with proteins from different plant species. Predictions of physio-chemical properties indicated that the proteins were basic in nature, and subcellular localization of HSP101a and HSP101b was extracellular, and HSP101c found mitochondrial in nature. 3D model of protein was also generated using SWISS MODEL, for understanding their molecular functions and active site identification. The Ramachandran analysis confirmed the structural quality of the protein models, with most residues falling within favored regions. This study provides valuable insights into the expression and molecular characteristics of HSP101 heat shock proteins in heat-tolerant wheat genotypes, contributing to a better understanding of the mechanisms involved in heat tolerance of wheat.