Development and characterization of transgenic pigeon pea (Cajanus cajan (L.) Millsp.) with OsRuvB gene for salinity tolerance
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Date
2018
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CCSHAU
Abstract
Salinity is a major stress which causes considerable loss in crop productivity. In this study, OsRuvB, a DNA helicase, under the control of CaMV35S promoter, is used to develop salinity tolerant transgenic pigeon pea plants (cv. Manak) using Agrobacterium-mediated transformation method. The transgene integration in 70 putative T0 plants was checked using PCR analysis and 25 plants showed amplification for the OsRuvB gene using specific primers, giving a transformation efficiency of 35.7 %. Out of these, 6 comparatively higher yielding plants were selected in the T1 generation for segregation analysis and to screen the plants for stable transgene integration. Further, Southern hybridization and real time PCR were done to determine the copy number of the transgene in these selected transgenic lines. To assess the efficacy of transgene, the wild type, and transgenic plants of T1 generation were subjected to 75mM NaCl stress and analysis was done based on various physiological parameters. The results showed that in addition to more chlorophyll and relative water content under salinity, the transgenic plants also showed higher activity of peroxidase and catalase. Level of proline and total soluble sugar was elevated in T1 transgenic plants when grown under salt stress, but the increase was lower than in wild type plants under salt stress. Moreover, membrane injury index and lipid peroxidation were found to be significantly reduced in transgenic plants in comparison to the wild type plants. These results indicate that transgenic pigeon pea lines were less affected by salt stress. The transgenic lines didn‘t have significant increase in osmolytes proline and total soluble sugar, which indicates that the tolerance is being imparted either by some other osmolytes or some entirely different mechanism yet to be uncovered might be working in these plants.
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