Genetic Transformation of Tobacco with Rat Cytochrome P4501A1 Monooxygenase Gene

E. YORDANOVA¹, N. GORINOVA¹, R. BACHVAROVA¹, M. NEDKOVSKA¹, A. ATANASSOV¹ and H. OHKAWA²

¹Institute of Genetic Engineering, BG-2232 Kostinbrod – 2, Bulgaria
²Department of Biological and Environmental Science, Faculty of Agriculture, Kobe University, Kobe 657-8501, Japan

Abstract

YORDANOVA, E., N. GORINOVA, R. BACHVAROVA, M. NEDKOVSKA, A. ATANASSOV and H. OHKAWA, 2000. Genetic transformation of tobacco with rat cytochrome P4501A1 monooxygenase gene. Bulg. J. Agric. Sci., 6: 1-7

Five economically important Bulgarian tobacco cultivars were successfully transformed with fused rat cytochrome P4501A1 and yeast reductase genes, conferring resistance to herbicide chlortoluron. The genes were introduced into tobacco plants by Agrobacterium tumefaciens transformation. The transformation efficiency varied from 32.4 to 57.1% in regard of the genotype. Under green house conditions transgenic tobacco plants showed high level of resistance to herbicide chlortoluron. Resistant plants were self-pollinated and the segregation of the trait was followed in T1 generation. The transgenic nature of transformed plants was confirmed by GUS, ELISA and PCR analyses. Engineering of transgenic tobacco plants, expressing rat P4501A1/yeast reductase genes and exhibiting herbicide resistance appear to be important for parasites management programs as well as phytoremediation. The resulting transgenic plants with higher herbicide tolerance to chlortoluron will help to study the effects of the expression of rat cytochrome P4501A1 on plant – parasite (like Orobanche ramosa as a main root parasite at tobacco) interactions.

Key words: cytochrome P450, chlortoluron, herbicide resistance