Genetic Transformation of Coleus blumei Benth. Using Agrobacterium

Nataša Bauer¹, Dunja Leljak Levanić¹, Snježana Mihaljević² and Sibila Jelaska¹*

¹University of Zagreb, Faculty of Science, Department of Molecular Biology, Rooseveltov trg 6, HR-10000 Zagreb, Croatia
²Institute Ruđer Bošković, Bijenička 54, HR-10000 Zagreb, Croatia

Article history:
Received: March 8, 2002
Accepted: June 18, 2002

Key words:
callus culture, Coleus blumei, transformation, Agrobacterium

Summary:
Efficiency in transformation of three hybrids of Coleus blumei using wild and mutant strains of Agrobacterium sp. was evaluated. Successful transformation depended on specific bacterial strain-plant genotype combination and co-cultivation treatment. The wild type B6S3 of the bacterial strains employed gave the maximum efficiency. Strains C58C1(pArA4abc), GV3101(pGV2215), 8196 and A281 were also effective. Among several co-cultivation conditions tested, the most efficient treatment was as follows: excised leaf explants were incubated immeadiately in bacterial suspension for 5 min and were co-cultured with Nicotiana tabacum crown gall callus for two days. Plant hormone autotrophy, PCR analyses and hybridization analysis confirmed genetic transformation of the cultures. Sixteen different transgenic callus and cell cultures were maintained for more than four years in the absence of exogenous growth regulators and antibiotics without deterioration in the growth rate.  

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