Revision of the barley WRKY gene family phylogeny and expression analysis of the candidate genes in response to drought

The WRKY belongs to an important plant specific transcription factor families which are involved in response to various environmental stresses, as well as in growth and developmental processes. In the present report, a genome-wide identification and characterization of WRKY gene family in barley led to revision of HvWRKYs to 93 members. The phylogenetic tree was also reconstructed based on the full-length WRKY protein sequences in barley and Arabidopsis. HvWRKYs were classified into three major groups (I, II, and III) and group II was further divided to 5 subgroups (a to e). HvWRKYs were named after this classification. Interestingly, some specific motifs were discovered in subgroups IIa, IIb, and III. Analyzing the available microarray data revealed eight candidate WRKY genes which were up-regulated under drought and salinity stresses compared to the optimum conditions at seedling stage in barley. Expression profiles of these WRKY genes were validated by quantitative real-time PCR. All the investigated candidate genes (HvWRKY_IIc2, HvWRKY_III11, HvWRKY_ IIb2, HvWRKY_IId4, HvWRKY_III23, HvWRKY_IIa5 and HvWRKY_IIc19) except for HvWRKY_I8 were significantly up-regulated by drought stress at the seedling stage in drought-tolerant genotype, indicating their role in drought tolerance. We hope the presented information would be helpful toward achieving drought tolerant cultivars through genetic engineering or molecular breeding.