Cloning, expression analysis and In silico characterization of HSP101: a potential player conferring heat stress in Aegilops speltoides (Tausch) Gren

Pratibha Jakhu; Priti Sharma; Inderjit Singh Yadav; Parampreet Kaur; Satinder Kaur; Parveen Chhuneja; Kuldeep Singh

doi:10.1007/s12298-021-01005-2

Cloning, expression analysis and In silico characterization of HSP101: a potential player conferring heat stress in Aegilops speltoides (Tausch) Gren

Physiol Mol Biol Plants. 2021 Jun;27(6):1205-1218. doi: 10.1007/s12298-021-01005-2. Epub 2021 Jun 9.

Authors

Pratibha Jakhu¹, Priti Sharma¹, Inderjit Singh Yadav¹, Parampreet Kaur¹, Satinder Kaur¹, Parveen Chhuneja¹, Kuldeep Singh^{1

2}

Affiliations

¹ School of Agricultural Biotechnology, Punjab Agricultural University, Ludhiana, 141 004 India.
² National Bureau of Plant Genetic Resources, Delhi, 110012 India.

Abstract

Heat shock protein (HSP101) function as molecular chaperones and confer thermotolerance to plants. In the present investigation, identification, comprehensive expression analysis, phylogeny and protein modelling of HSP101 gene has been done in Aegilops speltoides accession Pau3583. In the present study, we cloned and in silico characterized a HSP101C gene designated as AsHSP101C-Pau3583. AsHSP101C-Pau3583 is 4180 bp long with seven exons and six introns and encoded a polypeptide of 910 amino acids predicted by FGENESH. We have identified 58 SNPs between the AsHSP101C-Pau3583 and reference gene sequence extracted from Ae. speltoides TGAC assembly. Real-time RT-PCR analysis of expression levels of HSP101 gene in two wheat genotypes under heat stress revealed that gene namely HSP101C was up-regulated in Aegilops speltoides acc. Pau3583 by > fourfold in comparison to Triticum aestivum cv. PBW343 under heat stress signifies that it plays a role in conferring heat tolerance. Sequence comparison and phylogenetic analysis of AsHSP101C-Pau3583 with seven wheat homologs Triticum aestivum, Aegilops speltoides (TGAC), Triticum durum cv Cappelli, Triticum durum cv Strongfield, Triticum monococcum, Aegilops tauschii and Triticum urartu showed significant similarities with highly conserved coding regions and functional domains (AAA, AAA + 2, ClpB domains), suggesting the conserved function of HSP101C in different species. The illustration of the protein models of HSP101C in homologs provided information for the ATP-binding motifs within the nucleotide binding domains (NBD), specific for the chaperone activity. These findings are important and identified SNPs could be used for designing markers for ensuring the transfer of AsHSP101C-Pau3583 gene into hexaploid wheat and its role in heat tolerance.

Supplementary information: The online version contains supplementary material available at 10.1007/s12298-021-01005-2.

Keywords: Aegilops speltoides; Expression analysis; HSP101; Heat shock protein; Heat tolerance; Protein models.