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In planta Transformation of Ectabcgene Cluster For Salt Tolerance In Rice

By: Contributor(s): Material type: TextTextLanguage: English Publication details: Dharwad University of Agricultural Sciences, Dharwad 2024Edition: M Sc (Agri)Description: 80 32 CmsDDC classification:
  • 630.2742 AJE
Summary: Salt stress significantly impacts global agricultural productivity, particularly in indica rice (Oryza sativa), which is known for its sensitivity to salinity. The popular rice variety BPT-5204, known for its excellent grain quality, matures in 140-150 days and has beenlocally preferred for decades. However, its performance declines significantly under saline conditions, with reduced yield. This study aimed to enhance salt tolerance in rice by introducing the ectABC gene cluster from Halomonas spp, responsible for ectoine biosynthesis, a key osmoprotectant.Osmoprotectant helps to mitigate salt stress bystabilizing cellular structures, maintaining osmotic balance, and protecting against oxidative damage.The ectABC cluster was subcloned into the plant expression vector pRI 101-AN and confirmed through restriction digestion and colony PCR. Using Agrobacterium tumefaciens strain LBA4404, the recombinant vector was successfully introduced into rice via in planta transformation by pricking the embryonic apical meristem with bacterial suspension using a sterileneedle.Transformationefficiency,basedonseedandseedlingregeneration,rangedfrom 1.25 per cent to 1.79 per cent respectively. PCR analysis confirmed transgeneintegration in five T₀ plants. Sequence homology analysis revealed 99.79 per cent identity withHalomonaselongata,affirmingthesuccessfulintegrationofthegenecluster.Thestudy outlines a comprehensive workflow for bypassing tissue culture, reducing associated complications such as somaclonal variation. The in planta approach demonstrated the feasibilityof direct T-DNA deliveryto the rice genome, simplifyingtransformation protocols. This research provides a foundation for engineering salt-tolerant rice, contributing to food security in saline-prone areas. By leveraging the ectABC gene cluster, this work highlights a sustainable solution for improving rice productivity under abiotic stress conditions, aligning with the goals of climate-resilient agriculture.
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THESIS University of Agricultural Sciences, Dharwad 630.2742/AJE 1 Available T14142

Salt stress significantly impacts global agricultural productivity, particularly in indica rice (Oryza sativa), which is known for its sensitivity to salinity. The popular rice variety BPT-5204, known for its excellent grain quality, matures in 140-150 days and has beenlocally preferred for decades. However, its performance declines significantly under saline conditions, with reduced yield. This study aimed to enhance salt tolerance in rice by introducing the ectABC gene cluster from Halomonas spp, responsible for ectoine biosynthesis, a key osmoprotectant.Osmoprotectant helps to mitigate salt stress bystabilizing cellular structures, maintaining osmotic balance, and protecting against oxidative damage.The ectABC cluster was subcloned into the plant expression vector pRI 101-AN and confirmed through restriction digestion and colony PCR. Using Agrobacterium tumefaciens strain LBA4404, the recombinant vector was successfully introduced into rice via in planta transformation by pricking the embryonic apical meristem with bacterial suspension using a sterileneedle.Transformationefficiency,basedonseedandseedlingregeneration,rangedfrom 1.25 per cent to 1.79 per cent respectively. PCR analysis confirmed transgeneintegration in five T₀ plants. Sequence homology analysis revealed 99.79 per cent identity withHalomonaselongata,affirmingthesuccessfulintegrationofthegenecluster.Thestudy outlines a comprehensive workflow for bypassing tissue culture, reducing associated complications such as somaclonal variation. The in planta approach demonstrated the feasibilityof direct T-DNA deliveryto the rice genome, simplifyingtransformation protocols. This research provides a foundation for engineering salt-tolerant rice, contributing to food security in saline-prone areas. By leveraging the ectABC gene cluster, this work highlights a sustainable solution for improving rice productivity under abiotic stress conditions, aligning with the goals of climate-resilient agriculture.

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