MOLECULAR AND PHENOTYPIC ANALYSIS OF Brassica juncea TRANSGENICS HAVING DEFENSE REGULATORY NPR1 GENE
KrishiKosh
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Title |
MOLECULAR AND PHENOTYPIC ANALYSIS OF Brassica juncea TRANSGENICS HAVING DEFENSE REGULATORY NPR1 GENE
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Creator |
RAJENDRA PRASAD MEENA
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Contributor |
Anita Grover
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Subject |
marketing, grain, economics, wheats, distribution systems, supply, sugar, wells, hydrocarbons, grain legumes
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Description |
T-8304
Alternaria blight is one of the serious diseases of Brassica juncea causing 45-58% loss in the yield. Manipulating defense regulatory genes [eg. No PR1 (NPR1) gene] is one of the strategies being followed internationally. In order to develop resistance against Alternaria brassicae, nineteen putative transgenic lines of Brassica juncea having NPR1 gene under control of 35S promoter were developed earlier. In the present study, these transgenic lines were confirmed for gene integration and expression and its effect on resistance against Alternaria. Gene integration was confirmed by doing PCR with NPT II primers. In order to reconfirm the NPR1 gene integration, PCR was done using 35S forward primer and NPR1 reverse primer. All the nineteen putative lines were found to be positive. RT-PCR using NPR1 primers were done to check levels of gene expression. Eleven lines (1,5,6,7,8,9,11,12,14,16,18) were found to be over expressing NPR1 about 5 fold over nontransgenic control while other eight lines (2,3,4,10,13,15,17,19) did not show such high expression (only 2 fold). Seven lines encompassing both high NPR1 expressing and not so high NPR1 expressing, were analyzed for disease resistance. Resistance was scored in terms of time of onset of symptoms and size and number of lesions. The symptoms of infection appeared after day 3 and 4 of post inoculation in control plants while it was after day 6 and 7 of post inoculation in transgenic plants. The levels of resistance in-vitro varied from 2 to 2.5 fold as compared to the control. In-vivo assays showed 2 to 3 fold resistance in transgenic plants as compared to control plants. It was found that expression level of NPR1 is directly related to levels of resistance |
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Date |
2016-10-19T10:57:03Z
2016-10-19T10:57:03Z 2010 |
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Type |
Thesis
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Identifier |
http://krishikosh.egranth.ac.in/handle/1/80975
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Format |
application/pdf
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Publisher |
IARI, NATIONAL RESEARCH CENTRE ON PLANT BIOTECHNOLOGY
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