Transgenic tobacco plants constitutively expressing peanut BTF3 exhibit increased growth and tolerance to abiotic stresses.

Abiotic stresses limit crop growth and productivity worldwide. Cellular tolerance, an important abiotic stress adaptive trait, involves coordinated activities of multiple proteins linked to signalling cascades, transcriptional regulation and other diverse processes. Basal transcriptional machinery is considered to be critical for maintaining transcription under stressful conditions. From this context, discovery of novel basal transcription regulators from stress adapted crops like peanut would be useful for improving tolerance of sensitive plant types. In this study, we prospected a basal transcription factor, BTF3 from peanut (Arachis hypogaea L) and studied its relevance in stress acclimation by over expression in tobacco. AhBTF3 was induced under PEG-, NaCl-, and methyl viologen-induced stresses in peanut. The constitutive expression of AhBTF3 in tobacco increased plant growth under non stress condition. The transgenic plants exhibited superior phenotype compared to wild type under mannitol- and NaCl-induced stresses at seedling level. The enhanced cellular tolerance of transgenic plants was evidenced by higher cell membrane stability, reactive oxygen species (ROS) scavenging activity, seedling survival and vigour than wild type. The transgenic lines showed better in vitro regeneration capacity on growth media supplemented with NaCl than wild type. Superior phenotype of transgenic plants under osmotic and salinity stresses seems to be due to constitutive activation of genes of multiple pathways linked to growth and stress adaptation. The study demonstrated that AhBTF3 is a positive regulator of growth and stress acclimation and hence can be considered as a potential candidate gene for crop improvement towards stress adaptation.

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.

Expression analysis of drought stress specific genes in Peanut (Arachis hypogaea , L.).

Improving drought tolerance through gene manipulation has been of importance for modern agriculture, which requires identification and validation of candidate genes. Prospecting candidate genes from drought adapted crop species is of immense significance. To identify candidate stress responsive genes from adapted crop, we carried out expression analysis of a few drought responsive ESTs from Arachis hypogaea L. (peanut). The expression patterns of nine AhDR (Arachis hypogea drought responsive) clones were analysed under drought. Quantitative reverse transcription PCR analysis revealed stress responsive nature of the selected genes. The clones AhDR 118 (putative cyclin T-like), AhDR185 (aldehyde reductase-like), AhDR193 (cholin kinase-like) and AhDR 76 (proline amino peptidase-like) showed more than five fold increase in expression. Highly upregulated genes analysed for expression pattern against salinity at seedling level indicated that these genes provide cross protection. This paper is the first report indicating the association of peanut genes cyclin T, proline amino peptidase and choline kinase to drought tolerance, and the possible roles of these genes are discussed.