Co-expression of AtbHLH17 and AtWRKY28 confers resistance to abiotic stress in Arabidopsis

Stress adaptation in plants involves altered expression of many genes through complex signaling pathways. To achieve the optimum expression of downstream functional genes, we expressed AtbHLH17 (AtAIB) and AtWRKY28 TFs which are known to be upregulated under drought and oxidative stress, respectively in Arabidopsis. Multigene expression cassette with these two TFs and reporter gene GUS was developed using modified gateway cloning strategy. The GUS assay and expression analysis of transgenes in transgenic plants confirmed the integration of multigene cassette. The transgenic lines exhibited enhanced tolerance to NaCl, Mannitol and oxidative stress. Under mannitol stress condition significantly higher root growth was observed in transgenics. Growth under stress and recovery growth was substantially superior in transgenics exposed to gradual long term desiccation stress conditions. We demonstrate the expression of several downstream target genes under various stress conditions. A few genes having either WRKY or bHLH cis elements in their promoter regions showed higher transcript levels than wild type. However, the genes which did not have either of the motifs did not differ in their expression levels in stress conditions compared to wild type plants. Hence co-expressing two or more TFs may result in upregulation of many downstream target genes and substantially improve the stress tolerance of the plants.