Korean Journal of Plant Tissue Culture (식물조직배양학회지)

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Biosynthesis of Sesquiterpene in Hairy Root and Cell Suspension Cultures of Hyoscyamus muticus by Elicitation Using Rhizoctonia solani Extracts

Rhizoctonia solani 추출액 첨가에 의한 Hyoscyamus muticus의 현탁세포배양 및 모상근배양에서 Sesquiterpene 생합성

  • BACK, Kyoungwhan (Department of Agronomy, Kyungpook National University) ;
  • SHIN, Dong Hyun (Department of Agronomy, Kyungpook National University) ;
  • KIM, Kil Ung (Department of Agronomy, Kyungpook National University) ;
  • De HAAS Cynthia R. (Department of Chemical Engineering, The Pennsylvania State University) ;
  • CHAPPELL Joseph (Department of Agronomy, University of Kentucky) ;
  • CURTIS Wayne R. (Department of Chemical Engineering, The Pennsylvania State University)
  • Published : 1997.09.01
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Abstract

The extracellular sesquiterpenoids were accumulated in cell and hairy root cultures of Hyoscyamus muticus by elicitation using extracts of Rhizoctonia solani. The vetispiradiene synthase (VS) which is the first committed step in biosynthetic pathway leading to formation of solavetivone, lubimin, and rishitin from isoprenoid intermediate farnesyl pyrophosphate was induced upon elicitation, whereas no sesquiterpenoids and VS activity were detected in both control cell and hairy root cultures. VS activity increased rapidly and reached its maximum 12 h in both cell and hairy root cultures upon elicitor treatment. VS activities were paralleled with the absolute levels of VS polypeptide(s). Interestingly, the profiles of sesquiterpenoid accumulation in hairy root cultures were different from those in cell cultures. The hairy root culture seemed to fail to metabolize solavetivone further to lubimin.

Hyoscyamus muticus의 phytoalexin으로 알려진 solavetivone, lubimin, rishitin 생합성에 결정적 역할을 담당하는 isoprenoid pathway의 첫 분지효소인 vetispiradiene synthase의 유도특성과 solavetivone, lubimin phytoalexin 생합성과의 관련성을 구명하기 위하여 H. muticus 현탁세포배양 및 모상근배양에 Rhizoctonia solani 추출액을 elicitor로 처리하였다. Elicitor을 처리하지 않은 현탁세포 및 모상근배양에서는 효소활성, 효소단백질 및 solavetivone, lubimin등이 전혀 검출되지 않았으나, elicitor을 처리한 세포에서는 효소활성이 급속히 유도되어 처리 12시간 후에 최고의 활성을 보였으며 그 후 점점 감소하였다. 효소활성의 정도는 면역반응을 이용하여 측정한 효소 단백질의 함량과 밀접한 상관관계를 보였으며, phytoalexin 총 함량은 모상근배양에서 2배이상 높았다. 특히 solavetivone 및 lubimin 생합성은 현탁 및 모상근배양에서 서로 다른 특이성을 보였는데, 현탁배양에서는 lubimin을, 모상근배양에서는 solavetivone을 선택적으로 다량 생산하였다.

Keywords

References

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