Transformation and Analysis of Transgentic Pigeonpea Tolerant to Pod Borer
KrishiKosh
View Archive InfoField | Value | |
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
|
|