Genetic transformation of Indica rice for sheath blight resistance using chitinase gene
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Date
2021
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Punjab Agricultural University, Ludhiana
Abstract
Trichoderma chitinase enzymes hydrolyze β-1, 4-glycosidic bonds of chitin found in the cell wall of Rhizoctonia solani, the causal agent of rice sheath blight. In this study Tvchit42 gene was introduced into rice using Agrobacterium-mediated genetic transformation. The gene construct, carrying Tvchit42 gene under the control of the CaMV35S promoter and the NOS terminator, was mobilised into two Agrobacterium tumefaciens strains, LBA4404 and EHA105, which were used to agro-infect PR121 and Kitaake rice calli, respectively. In PR121 and Kitaake, a total of 99 and 414 plants were regenerated from transformed calli, respectively. The presence of transgene was confirmed by PCR, revealing amplicon of size 1293 bp corresponding to Tvchit42 with transformation efficiency of 0 % and 0.64 % in
PR121 and Kitaake, respectively. Real-Time PCR was used to confirm transgene integration and expression in T0 transformants, revealed 2.94-6.14 fold increase in Tvchit42 expression in six Kitaake plants as compared to non-transgenic controls. Moderate resistance was observed in five transgenic plants after inoculation with RS-1 isolate of R. solani, indicating role of Tvchit42 to provide resistance against rice sheath blight disease.
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Sharma, Manveer (2021) Genetic transformation of Indica rice for sheath blight resistance using chitinase gene (Unpublished Ph.D. Dissertation). Punjab Agricultural University, Ludhiana, Punjab, India.