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Drought stress has adverse effects on growth, water relations, photosynthesis and yield of groundnut. WRKY transcription factors (TFs) are the plant-specific TFs which regulate several down-stream stress-responsive genes and play an essential role in plant biotic and abiotic stress responses. We found that  WRKY3  gene is highly up-regulated under drought stress conditions and therefore isolated a new  WRKY3TF  gene from a drought-adapted horsegram ( Macrotyloma uniflorum  Lam. Verdc.). Conserved domain studies revealed that protein encoded by this gene contains highly conserved regions of two WRKY domains and two C2H2 zinc-finger motifs. The fusion protein localization studies of transient  MuWRKY 3-YFP revealed its nuclear localization. Overexpression of  MuWRKY3  TF gene in groundnut ( Arachis hypogaea  L.) showed increased tolerance to drought stress compared to wild-type (WT) plants.  MuWRKY3  groundnut transgenics displayed lesser and delayed wilting symptoms than WT plants after 10-days of drought stress imposition. The transgenic groundnut plants expressing  MuWRKY3  showed less accumulation of malondialdehyde, hydrogen peroxide (H 2 O 2 ), and superoxide anion (O 2  ∙- ), accompanied by more free proline, total soluble sugar content, and activities of antioxidant enzymes than WT plants under drought stress. Moreover, a series of stress-related  LEA, HSP, MIPS, APX, SOD , and  CAT  genes found up-regulated in the transgenic groundnut plants. The study demonstrates that nuclear-localized  MuWRKY3  TF regulates the expression of stress-responsive genes and the activity of ROS scavenging enzymes which results in improved drought tolerance in groundnut. We conclude that  MuWRKY3  may serve as a new putative candidate gene for the improvement of stress resistance in plants.

frontiers in plant science

A Novel WRKY Transcription Factor, MuWRKY3 (Macrotyloma uniflorum Lam. Verdc.) Enhances Drought Stress Tolerance in Transgenic Groundnut (Arachis hypogaea L.) Plants