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Additional virulence genes in conjunction with efficient selection scheme, and compatible culture regime enhance recovery of stable transgenic plants in cowpea via Agrobacterium tumefaciens-mediated transformation

Paper ID Volume ID Publish Year Pages File Format Full-Text
24878 43542 2008 8 PDF Available
Title
Additional virulence genes in conjunction with efficient selection scheme, and compatible culture regime enhance recovery of stable transgenic plants in cowpea via Agrobacterium tumefaciens-mediated transformation
Abstract

A critical step in the development of robust Agrobacterium tumefaciens-mediated transformation system in recalcitrant grain legume, cowpea is the establishment of optimal conditions for efficient T-DNA delivery into target tissue and recovery of transgenic plants. A dramatic increase in efficiency of T-DNA delivery was achieved by constitutive expression of additional vir genes in resident pSB1 vector in Agrobacterium strain LBA4404. A geneticin based selection system permitted rapid and efficient identification of transgenic shoots without interfering with their regeneration, and eliminated the bulk of escapes. Supplementation of 0.5 μM kinetin to medium containing 5.0 μM benzyl aminopurine after 1 week of culture followed by 3 weeks of culture were found critical for optimal multiplication and elongation of transformed shoots from cotyledonary node explants. Combining these three developments, we recovered fertile transgenic plants at a frequency of 1.64%, significantly higher than previous reports. The presence, integration, expression and inheritance of transgenes were confirmed by molecular analysis. The protocol developed for cultivar Pusa Komal will facilitate the transfer of desirable traits into cowpea.

Keywords
Transgenic cowpea; Vigna unguiculata; Agrobacterium tumefaciens; Virulence genes; Geneticin selection
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Additional virulence genes in conjunction with efficient selection scheme, and compatible culture regime enhance recovery of stable transgenic plants in cowpea via Agrobacterium tumefaciens-mediated transformation
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 135, Issue 1, 20 May 2008, Pages 97–104
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us