Studies on the construction of Bt gene expression vector and its transformation in tea plant

Luo YingYing; Liang YueRong

Journal of Tea Science 20(2): 141-147


Accession: 003570112

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The pGA471 Bt [Bacillus thuringiensis] gene DNA was digested with Hind III and Bgl II and inserted into the vector pCAMBIA2301. The constructed plasmid, with Bt gene, intron-GUS gene and NPT II gene, was transformed into Escherichia coli and introduced into the Agrobacterium strains LBA4404, EHA105 and pRi15834 through a triparental cross method. The Bt gene was transformed into the tea leaves and callus by an Agrobacterium-mediated method. The GUS gene was successfully expressed in both the callus and leaves of tea. Hygromycin was identified as a more effective screening agent for tea callus than kanamycin, and exhibited an optimum concentration of 20 micro l/ml; kanamycin was more effective for tea leaves, exhibiting an optimum concentration of 50 micro l/ml.