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Plant and Cell Physiology, 1999, Vol. 40, No. 12 1232-1242
© 1999

Molecular Cloning of a Defense-Response-Related Cytochrome P450 Gene from Tobacco

Daigo Takemoto1, Makoto Hayashi2,3, Noriyuki Doke1, Mikio Nishimura2 and Kazuhito Kawakita1

1 Plant Pathology Laboratory, Graduate School of Bioagricultural Sciences, Nagoya University Chikusa, Nagoya, 464-8601 Japan
2 Department of Cell Biology, National Institute for Basic Biology Okazaki, 444-0867 Japan

Plant defenses against pathogen attack involve a series of inducible responses that contribute to resistance. Tobacco leaves injected with HWC (hyphal wall components prepared from Phytophthora infestans) elicitor showed typical defense responses, including the induction of localized necrosis and the accumulation of pathogenesis-related proteins. In order to elucidate the molecular mechanisms by which plant defense systems are activated, we screened tobacco plants for genes differentially expressed in response to HWC. We performed differential screening by RT-PCR with random primers and obtained PCR products specific to HWC-treated leaf RNA. Northern hybridization using the PCR products as probes confirmed that one transcript was actually induced by HWC treatment. As the deduced amino acid sequence of this clone showed the highest degree of similarity to elicitor-induced soybean cytochrome P450 CYP82A4, it was designated CYP82E1. The expression of CYP82E1 was strongly induced in tobacco by the soybean pathogen Pseudomonas syringae pv. glycinea (nonpathogenic on tobacco), but it was activated only slightly and in a delayed fashion by the tobacco pathogen P. syringae pv. tabaci (pathogenic on tobacco), implying that the product of CYP82E1 may be involved in disease resistance in tobacco.

3 Present address: Department of Applied Plant Science, Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai, 981-8555 Japan.

(Received May 6, 1999; Accepted October 8, 1999)
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