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Salinity of cultivable land is a growing global concern that has been affecting the yields of major crops worldwide such as pigeon pea. In the current study, transgenic pigeon pea plants were developed using an in-planta Agrobacterium-mediated genetic transformation method wherein OsRuvB, a rice DNA helicase gene, was incorporated to induce salt tolerance in pigeon pea plants. Transformation efficiency of 35.7% was achieved with stable insertion of OsRuvB in transgenic lines. When subjected to salinity stress induced by 75 mM NaCl increase in chlorophyll content, relative water content, peroxidase and catalase activity in transgenic lines was observed over the wild type plants. Membrane injury index and lipid peroxidation were significantly reduced in transgenic lines. Proline and Total Soluble Sugar content were enhanced in both transgenic plants and wild type strains. It was inferred that transgenic lines were tolerant to salinity stress and tolerance may be imparted through an alternative unknown pathway. ARTICLE HISTORY

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Sensing stress and activating the downstream signaling pathways is the imperative step for stress response. Lectin receptor‐like kinase (LecRLK) is an important family that plays a key role in sensing stress conditions through lectin receptor and activates downstream signaling by kinase domain. We identified the role of OsLecRLK gene for salinity stress tolerance and hypothesized its role in Na+ extrusion from cell. OsLecRLK overexpression and downregulation (through artificial miRNA) transgenic lines were developed and its comparison with wild‐type (WT) plants were performed overexpression transgenic lines showed better performance, whereas downregulation showed poor performance than WT. Lower accumulation of reactive oxygen species (ROS), malondialdehyde and toxic ion, and a higher level of proline, RWC, ROS scavengers in overexpression lines lead us to the above conclusion. Based on the relative expression of stress‐responsive genes, ionic content and interactome protein, working model highlights the role of OsLecRLK in the extrusion of Na+ ion from the cell. This extrusion is facilitated by a higher expression of salt overly sensitive 1 (Na+/K+ channel) in overexpression transgenic line. Altered expression of stress‐responsive genes and changed biochemical and physiological properties of cell suggests an extensive reprogramming of the stress‐responsive metabolic pathways by OsLecRLK under stress condition, which could be responsible for the salt tolerance capability.

Copy number of Cry1Ac in transgenic pigeonpea plants was determined by quantitative real time PCR using Syber Green as fluorescence indicator. Gene specific primers designed to amplify relatively long amplicons (400 -600 bp), for Cry1Ac was used to increase specificity and sensitivity of Real time PCR. Estimated copy number in transgenic lines using realtime quantitative PCR and southern hybridization was correlated and found to be same i.e. single copy number. This study shows effectivness of real-time PCR method for estimating the transgene copy number in transgenic pigeonpea plants by a simple, accurate and cost effective manner.

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