Development of transgenic pigeon pea (Cajanus cajan L.) plants containing Lectine receptor like kinase gene for improving abiotic stress tolerance
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Date
2019-05
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CCSHAU, Hisar
Abstract
Pigeon pea (Cajnus cajan L.) is a nutrient rich, sixth most important and second most
important legume pulse crop of the world and India respectively after chickpea. Production and
productivity of pigeon pea is affected by various abiotic stresses but salinity is the most deleterious
stress which causes considerable loss in crop’s productivity. In the present study, transgenic pigeon pea
plants (cv.. Manak) carrying OsLecRLK gene, were developed for salinity tolerance using
Agrobacterium tumefaciens strain LBA4404 with pCAMBIA1300- OsLecRLK gene (Kharb et al., 2018
Patent Application No. 201811012099). Putative transformants were screened through PCR
amplification using gene specific primers and out of 86 plants 16 plants were found positive giving a
transformation efficiency of 18.6%. Six higher yielding lines were selected and checked for stable
transgene integration. Southern hybridization and Real-time PCR analysis was done to find out the
copy no. of transgene in selected transgenic lines. Seeds from T0 pigeon pea plants were sown to raise
T1 generation and a total of 52 plants (out of 120) showed the presence of OsLecRLK gene. Physiobiochemical
analysis was done to assess the efficacy of transgene via subjecting wild type and selected
T1 transgenic plants to 75 mM salt stress. The results showed that transgenic line performed better in
terms of maintaining higher chlorophyll content, relative water content, proline content, total soluble
sugar content, peroxidase and catalase activity in comparison to the wild type plants. Moreover, MDA
content and membrane injury index were significantly reduced in transgenic lines when compared to
the wild type plants indicating that the transgenic lines were less affected by salt stress. Among all
transgenic lines, L-19 performed well with respect to all the parameters studied and can be taken
further for the development of transgenic pigeon pea plants for salt stress tolerance.