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“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
M Sc
 
Creator RAJENDRA PRASAD MEENA
 
Contributor Anita Grover
 
Subject planting, genes, biological phenomena, transgenics, pathogens, diseases, transgenic plants, fungi, brassica juncea, pcr
 
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 [e.g. NoPR1 (NPR1) gene] is one of the
strategies being followed internationally. In order to develop resistance against Alternaria
brassicae, nineteen putative transgenic lines of B. juncea having NPR1 gene under control of
35S promoter were developed earlier. In the present study, these transgenic lines were confirmed
for gene integration, expression and its effect on resistance against Alternaria. Gene integration
was confirmed by 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 with 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 by
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, lesions number and lesion size. The symptoms of infection
were observed on day 3-4 after inoculation in control plants whereas those in transgenic plants
symptoms were observed on day 6-7 after inoculation. The levels of resistance in-vitro varied
from 2.0 to 2.5 fold as compared to the control. In-vivo assays revealed 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.
 
Date 2016-10-19T11:20:06Z
2016-10-19T11:20:06Z
2010
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/80989
 
Format application/pdf
 
Publisher IARI, NATIONAL RESEARCH CENTRE ON PLANT BIOTECHNOLOGY