Agrobacterium-mediated transformation and plant regeneration of Brassica oleracea var. capitata |
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Authors: | P K Pius P N Achar |
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Institution: | (1) Department of Microbiology, University of Durban Westville, Durban 4000, South Africa Fax: 31-204 4809 e-mail: pachar@pixie.udw.ac.za, ZA |
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Abstract: | An efficient protocol for Agrobacterium tumefaciens-mediated transformation of four commercial cultivars of Brassica oleracea var. capitata is described. A strain of A. tumefaciens LBA4404 with the neomycin phosphotransferase gene (nptII) and a CaMV 35S-peroxidase gene cassette were used for co-cultivation. Preliminary selection of regenerated transgenic plants
was performed on kanamycin-containing medium. The frequency of transgenic plants was calculated on the basis of GUS (β-glucuronidase)
activity detected by the histochemical X-gluc test. Tissue-specific GUS expression driven by the peroxidase gene promoter
in transgenic plants was analysed by GUS staining. The transformation rates of the commercial cultivars of B. oleracea was higher than in previous reports. Southern blot analysis revealed that integration of marker genes occurred in single
and multiple loci in the genome. All transgenic plants grew normally after a brief vernalization period and showed stable
inheritance of the marker gene. The present study demonstrates that morphologically normal, fertile transgenic plants of B. oleracea can be obtained.
Received: 24 August 1999 / Revision received: 23 November 1999 / Accepted: 3 December |
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Keywords: | Agrobacterium tumefaciens NPT II genes β -Glucuronidase Transgenic plants |
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