In silico analysis and gene expression of heat stress responses genes in Hordeum vulgare L.

Abstract

Barley has been selected or bred for specific adaptation to abiotic stresses in geographically distinct areas of the world. The purpose of this study is to identify differentially expressed genes and analyze biological processes related to heat stress in barley. The GEO dataset of heat-shock (HS) treated barley was statically analyzed using the integrated GEO2R tool in the package of Gene Expression Omnibus dataset. Cytoscape software and STRING website were used for Gene Ontology (GO) enrichment, protein-protein interaction analysis and network visualization. Our bioinformatics analysis displayed genes that were consistently differentially expressed in HS in barley plant. The results show that biological pathways and protein-protein interaction networks are associated with these genes. Based on the bioinformatics analysis results, NF-YA2 gene was selected as the hub gene responsible for HS. A qRT-PCR was designed to approve the in silico results and analyze the expression of NF-YA2. Barley plants were grown in the greenhouse, heat treated at 42 °C and then harvested. Identified genes were validated by means of qRT-PCR. According to the qRT-PCR results, NF-YA2 was up-regulated in sensitive plants and down-regulated in the tolerant plants. The results showed that NF-YA2 is a negative regulator in abiotic stress tolerance in barley.