Shoot regeneration in stem expiants and its amenability to Agrobacterium tumefaciens mediated gene transfer in Brassica carinata

Narasimhulu, S. B. ; Kirti, P. B. ; Mohapatra, T. ; Prakash, Shyam ; Chopra, V. L. (1992) Shoot regeneration in stem expiants and its amenability to Agrobacterium tumefaciens mediated gene transfer in Brassica carinata Plant Cell Reports, 11 (7). pp. 359-362. ISSN 0721-7714

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Official URL: http://www.springerlink.com/content/a80c7047c1045f...

Related URL: http://dx.doi.org/10.1007/BF00233366

Abstract

Immature stem segments of seven different genotypes of Brassica carinata produced shoots with variable frequencies when cultured in MS medium with BAP and picloram at 0.2 mg/l each. Line 171, which produced shoots with 100% efficiency from both cut ends of the expiant, was selected for testing the amenability of this regeneration protocol for genetic transformation. A non-oncogenic Agrobacterium tumefaciens containing plasmid PCV 730, a binary vector carrying resistance genes for kanamycin and hygromycin, was used. A cocultivation period of 4 d with a bacterial concentration of approximately 2.5×10 cells/ml, followed by a recovery period of 2 d, produced transformed shoots that could be selected and rooted in the presence of kanamycin at 15 mg/l. Transformation was confirmed by neomycin phospho-transferase assay and Southern blot analysis. Seed analysis of transformed plants indicated that kanamycin resistance was inherited in the progeny.

Item Type:Article
Source:Copyright of this article belongs to Springer-Verlag.
Keywords:Brassica carinata; Agrobacterium tumefaciens; Shoot Regeneration; Gene Transfer
ID Code:6521
Deposited On:20 Oct 2010 10:57
Last Modified:28 May 2011 06:46

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