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Transformation and Analysis of Transgentic Pigeonpea Tolerant to Pod Borer

KrishiKosh

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Title Transformation and Analysis of Transgentic Pigeonpea Tolerant to Pod Borer
 
Creator Savita G.Kanthi
 
Contributor Sumangala Bhat
 
Subject Plant Bio-Technology
 
Description In this study, attempts were made to refine the protocol for transformation in
pigeonpea (Cajanus cajan L.). Cotyledonary node explants (taken from 15 days old
seedlings) of Asha and Maruti genotypes were co-cultivated with Agrobacterium strain
LBA4404 harbouring pCAMBIA 1305.1 containing cry1Ac and gus and hpt genes as reporter
and selectable marker respectively. Hygromycin level of 7.5 mg/l was used for the selection
of transformants. Presence of 0.02 per cent Triton x100 in Agrobacterium suspension gave
maximum transformants (24.1%). Among different acetosyringone levels (0, 100, 150, 200,
250 μM) tried, 100 μM was found better and gave 25.05 per cent putative transformants. In
addition, vaccum infiltration for 20 minutes increased the recovery of putative transformants
(32.87%). Putative transformants were further tested for the presence of transgene using hpt
and cry1Ac specific primers. Of the 405 plants tested, only three were positive and which
were also positive for the expression when tested using Bt expression strips. Seeds (T2)
collected from 10 T1 plants were sown and T2 plants were analyzed. None of the T1 plants
gave 100 per cent positive progenies. T2 plants, which were positive for PCR, were also
positive for expression.
 
Date 2016-10-17T13:41:11Z
2016-10-17T13:41:11Z
2006
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/80707
 
Format application/pdf
 
Publisher UAS, Dharwad