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. 1985 Jun;4(6):1367–1372. doi: 10.1002/j.1460-2075.1985.tb03788.x

Chloroplast transformation by Agrobacterium tumefaciens

Marc De Block 1,3, Jeff Schell 1,2, Marc Van Montagu 1
PMCID: PMC554353  PMID: 16453614

Abstract

A chimeric gene consisting of the promoter region of the nopaline synthase gene (Pnos) fused to the coding sequence of the chloramphenicol acetyltransferase gene (cat gene) of Tn9 was introduced by co-cultivation in tobacco protoplasts followed by selection with 10 μg/ml chloramphenicol. The chloramphenicol-resistant plants derived from these selected calli were unable to transmit the CmR phenotype through pollen. A typically maternal inheritance pattern was observed. Southern blot analysis showed that the chimeric Pnos-cat gene was present in the chloroplasts of these resistant plants. Furthermore, the chloramphenicol acetyltransferase activity was shown to be associated with the chloroplast fraction. These observations are the first proof that the Agrobacterium Ti-plasmid vectors can be used to introduce genes in chloroplasts.

Keywords: Agrobacterium, chloroplast transformation, Ti-plasmid vectors, chimeric genes

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