Comprehensive analysis of Dendrobium catenatum HSP20 family genes and functional characterization of DcHSP20–12 in response to temperature stress

Heat shock proteins (HSPs) are a class of protective proteins in response to abiotic stress in plants, and HSP20 plays an essential role in response to temperature stress. However, there are few studies on HSP20 in Dendrobium catenatum. In this study, 18 DcHSP20 genes were identified from the D. catenatum genome. Phylogenetic analysis showed that DcHSP20s could be classified into six subgroups, each member of which has similar conserved motifs and gene structures. Gene expression analysis of 18 DcHSP20 genes revealed that they exhibited variable expression patterns in different plant tissues. Meanwhile, all 18 DcHSP20 genes were induced to be up-regulated under high temperature, while six genes (DcHSP20–2/9/10/12/16/17) were significantly up-regulated under low temperature. Moreover, combining gene expression under high and low temperature stress, the DcHSP20–12 gene was cloned for functional analysis. The germination ratios, fresh weights, root lengths of two DcHSP20–12-overexpressing transgenic Arabidopsis thaliana lines were significantly higher, but MDA contents were lower than that of wild-type (WT) plants under heat and cold stresses, displayed enhanced thermotolerance and cold-resistance. These results lay a foundation for the functional characterization of DcHSP20s and provide a candidate gene, DcHSP20–12, for improving the tolerance of D. catenatum to temperature stress in the future.