Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Molecular Cloning and Characterization of Sesquiterpene Cyclase cDNAs from Pepper Plant Infected with Phytophthora capsici

  • Kim, Jong-Bum (Division of Biochemistry, Department of Biological Resources, National Institute of Agricultural Science and Technology) ;
  • Lee, Sung-Gon (Division of Biochemistry, Department of Biological Resources, National Institute of Agricultural Science and Technology) ;
  • Ha, Sun-Hwa (Division of Biochemistry, Department of Biological Resources, National Institute of Agricultural Science and Technology) ;
  • Lee, Myung-Chul (Division of Cellular Genetics, Department of Biological Resources, National Institute of Agricultural Science and Technology) ;
  • Ye, Wan-Hye (Division of Plant Pathology, Department of Crop Protection, National Institute of Agricultural Science and Technology) ;
  • Lee, Jang-Yong (Division of Rice Breeding, Crop Experimental Station) ;
  • Lee, Shin-Woo (Division of Biochemistry, Department of Biological Resources, National Institute of Agricultural Science and Technology) ;
  • Kim, Jung-Bong (Division of Biochemistry, Department of Biological Resources, National Institute of Agricultural Science and Technology) ;
  • Cho, Kang-Jin (Division of Biochemistry, Department of Biological Resources, National Institute of Agricultural Science and Technology) ;
  • Hwang, Young-Soo (Division of Biochemistry, Department of Biological Resources, National Institute of Agricultural Science and Technology)
  • Received : 2001.05.07
  • Published : 2001.06.30
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Abstract

Pepper plants (Nogkwang, 60-day old) were inoculated with Phytophthora capsici to induce sesquiterpene cyclase associated with the biosynthesis of phytoalexin (capsidiol), a substance related to the defense against pathogens in plants. One day after inoculation, mRNA was isolated from the root, cDNA synthesized, and a library constructed in a ZAP express XR vector. The efficiency was $2{\times}10^6pfu/{\mu}g$. Sesquiterpene cyclase cDNA from Hyoscyamus muticus was labeled with $^{32}P$ and used as a probe for screening the cDNA library. After the third screening, 25 positive clones were selected. Through restrictive digestion and DNA gel-blot analysis, six different cyclase gene expressions were identified. PSC1B sequences of the six clones were determined, which were 1966 base pairs encoded 556 amino acids with an expected molecular weight of 63.8 kDa. Response against the pathogen was different between the resistant and susceptible peppers. After the infection of the pathogen, the expression of PSC genes continued in the resistant peppers while the plants were alive. The expression in the susceptible peppers lasted for only 4 days.

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