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<p>Water deficit remains a major constraint to improving mungbean growth and yield. Advances in genetic engineering have facilitated the development of drought-tolerant mungbean cultivars, often by introducing stress-responsive genes under the control of strong constitutive promoters such as <i>CaMV35S</i>. However, constitutive promoters derived from viruses or bacteria are associated with negative effects on plant growth and development, as well as biosafety concerns. In contrast, plant-specific promoters offer controlled gene expression, avoid systemic side-effects and generally perform better in crops, owing to their higher compatibility with the host transcriptional machinery. Root-specific promoters provide a powerful tool for targeted gene expression in roots, facilitating improved nutrient acquisition and water absorption in plants for improving drought and salt resilience. In the present study, we isolated a root-preferential promoter from mungbean root tissues that regulates the transcription factor <i>VrWRKY32</i>. The promoter was cloned and functionally characterized in mungbean root tissues using <i>Agrobacterium rhizogenes</i>-mediated transformation. To assess its activity, the <i>VrWRKY32</i> promoter was fused with GFP (<i>eGFP</i>) and GUS (<i>uidA</i>) reporter genes and introduced into the mungbean cultivar cv. <i>K-851</i>. Successful integration of the transgenes in hairy roots was confirmed by PCR and Southern hybridization. Semi-quantitative RT-PCR further verified expression of GUS and GFP driven through <i>VrWRKY32</i> promoter in transgenic hairy roots. In addition, histochemical staining demonstrated strong promoter activity in transgenic hairy roots. Collectively, the results indicate that the <i>VrWRKY32</i> promoter exhibits strong activity in mungbean root tissues and holds potential for driving root-preferential expression of target gene(s), particularly those associated with drought response and tolerance.</p>

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Isolation and Functional Characterization of a Root-preferential Stress Responsive VrWRKY32 Gene Promoter in Mungbean using Agrobacterium Rhizogenes-mediated Stable Transformation with GFP and GUS Reporters

  • 期刊论文

  • 发表日期：

    2026年3月3日

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  • Sanjeev Kumar,
  • Priti Kumari,
  • Babatunde Olanrewaju Okunlola,
  • Neeti Sanan-Mishra

摘要

Water deficit remains a major constraint to improving mungbean growth and yield. Advances in genetic engineering have facilitated the development of drought-tolerant mungbean cultivars, often by introducing stress-responsive genes under the control of strong constitutive promoters such as CaMV35S. However, constitutive promoters derived from viruses or bacteria are associated with negative effects on plant growth and development, as well as biosafety concerns. In contrast, plant-specific promoters offer controlled gene expression, avoid systemic side-effects and generally perform better in crops, owing to their higher compatibility with the host transcriptional machinery. Root-specific promoters provide a powerful tool for targeted gene expression in roots, facilitating improved nutrient acquisition and water absorption in plants for improving drought and salt resilience. In the present study, we isolated a root-preferential promoter from mungbean root tissues that regulates the transcription factor VrWRKY32. The promoter was cloned and functionally characterized in mungbean root tissues using Agrobacterium rhizogenes-mediated transformation. To assess its activity, the VrWRKY32 promoter was fused with GFP (eGFP) and GUS (uidA) reporter genes and introduced into the mungbean cultivar cv. K-851. Successful integration of the transgenes in hairy roots was confirmed by PCR and Southern hybridization. Semi-quantitative RT-PCR further verified expression of GUS and GFP driven through VrWRKY32 promoter in transgenic hairy roots. In addition, histochemical staining demonstrated strong promoter activity in transgenic hairy roots. Collectively, the results indicate that the VrWRKY32 promoter exhibits strong activity in mungbean root tissues and holds potential for driving root-preferential expression of target gene(s), particularly those associated with drought response and tolerance.