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Identification and Biological Activity of Two New Phytotoxins Isolated from Botrytis cinerea

Botrytis cinerea로부터 분리한 두 개의 새로운 phytotoxin의 구조 결정 및 생물활성

  • Kim, Geum-Jung (Chemical Biotechnology Research Center, Korea Research Institute of Chemical Technology) ;
  • Yoon, Mi-Young (Chemical Biotechnology Research Center, Korea Research Institute of Chemical Technology) ;
  • Kim, Heung-Tae (Department of Plant Medicine, Chungbuk National University) ;
  • Choi, Gyung-Ja (Chemical Biotechnology Research Center, Korea Research Institute of Chemical Technology) ;
  • Jang, Kyoung-Soo (Chemical Biotechnology Research Center, Korea Research Institute of Chemical Technology) ;
  • Choi, Yong-Ho (Chemical Biotechnology Research Center, Korea Research Institute of Chemical Technology) ;
  • Park, Myung-Soo (Chemical Biotechnology Research Center, Korea Research Institute of Chemical Technology) ;
  • Cha, Byeong-Jin (Department of Plant Medicine, Chungbuk National University) ;
  • Kim, Jin-Cheol (Chemical Biotechnology Research Center, Korea Research Institute of Chemical Technology)
  • 김금정 (한국화학연구원 산업바이오화학연구센터) ;
  • 윤미영 (한국화학연구원 산업바이오화학연구센터) ;
  • 김홍태 (충북대학교 식물의학과) ;
  • 최경자 (한국화학연구원 산업바이오화학연구센터) ;
  • 장경수 (한국화학연구원 산업바이오화학연구센터) ;
  • 최용호 (한국화학연구원 산업바이오화학연구센터) ;
  • 박명수 (한국화학연구원 산업바이오화학연구센터) ;
  • 차병진 (충북대학교 식물의학과) ;
  • 김진철 (한국화학연구원 산업바이오화학연구센터)
  • Published : 2009.08.01

Abstract

We discovered two novel phytotoxins produced by the pathogenic fungus, Botrytis cinerea. Among the twenty-five B. cinerea isolates, which were obtained from various host plants in 1994 and 1996, twenty-two showed strong or moderate pathogenicity on five plants such as cucumber, tomato, red pepper, tobacco and Chinese cabbage. The culture filtrate of the B. cinerea 2-16 strain showed the most potent phytotoxic activity in a tobacco leaf-wounding assay. Two novel phytotoxins were isolated from the liquid cultures of B. cinerea 2-16 by ethyl acetate extraction, flash silica gel column chromatography, silica gel column chromatography, Sephadex LH-20 column chromatography, preparative TLC and subsequently preparative HPLC. Their chemical structures were determined to be 3-O-acetyl botcinol and 3-O-acetyl botcinolide, respectively, by mass and NMR spectral analyses. These two phytotoxins caused leaf necrosis in a leaf-wounding bioassay, and significant electrolyte leakage from leaf tissues of tobacco. In the two bioassays tested, 3-O-acetyl botcinol exhibited stronger phytotoxic activity than 3-O-acetyl botcinolide. This is the first report on the production of both 3-O-acetyl botcinol and 3-O-acetyl botcinolide from B. cinerea.

Botrytis cinerea가 생산하는 phytotoxin이 병원성에 미치는 영향을 조사하던 중에 두 개의 새로운 phytotoxin을 발견하였다. 1994년과 1996년에 여러 식물체로부터 분리한 Botrytis cinerea 균주들을 기주별, 지역별, 형태적 특성별로 25개의 균주를 선별한 후 단포자 분리를 실시하였다. 22균주는 오이, 토마토, 고추 담배, 배추 등 5가지 식물에 대해 강하거나 중간 정도의 병원성을 보였다. 그러나 KJ 균주를 포함한 3균주는 병원성이 매우 약하였다. 식물에 대한 phytotoxicity를 조사하기 위하여 25균주의 배양 여액을 담배잎에 leaf-wounding assay를 실시한 결과 2-16균주가 가장 강한 활성을 보였다. 따라서 2-16 균주의 배양액을 ethyl acetate로 추출한 후 flash silica gel column chromatography, silica gel column chromatography, Sephadex LH-20 column chromatography, preparative TLC 및 preparative HPLC 등을 통하여 두 개의 phytotoxin을 분리하였다. 분리한 phytotoxin을 질량분석과 핵자기 공명분석을 통해 구조동정을 실시한 결과 3-O-acetyl botcinol과 3-O-acetyl botcinolide로 동정하였다. 두 개의 물질은 leaf-wounding bioassay에서 잎에 괴사를 일으켰으며, 또한 담배 잎에서 심각한 전해질 누출을 일으켰다. 두 생물검정에서 3-O-acetyl botcinol이 3-O-acetyl botcinolide보다 강한활성을보였다. B. cinerea가 생산한 두 개의 phytotoxin에 대해서는 본 논문에서 처음으로 보고하는 바이다.

Keywords

References

  1. Collado, I. G, Hemandex-Galan, R., Duran-Patron, R. and Cantoral, J M. 1995. Metabolites from a shake culture of Botrytis cinerea. Phytochemistry 38: 647-650 https://doi.org/10.1016/0031-9422(94)00690-U
  2. Collado, I. G, Aleu, J, Hernandez-Galan, R. and Hanson, J R. 1996. Some metabolites of Botrytis cinerea related to botcinolide. Phytochemistry 42: 1621-1624 https://doi.org/10.1016/0031-9422(96)00164-1
  3. Cutler, H. G, Jacyno, J M., Harwood, J. S., Dulik, D., Goodrich, P. D. and Roberts, R. G 1993. Botcinolide: A biologically active natural product from Botrytis cinerea. Biosci. Biotech. Biochem. 57: 1980-1982 https://doi.org/10.1271/bbb.57.1980
  4. Cutler, H. G, Parker, S. R., Ross, S. A., Crumley, F. G and Schpeiner, P. R. 1996. Homobotcinolide: A biologically active natural homolog of botcinolide from Botrytis cinerea. Biosci. Biotech. Biochem. 60: 656-658 https://doi.org/10.1271/bbb.60.656
  5. Deighton, N., Muchenschnabel, I. and Colmenares, A. J. 2001. Botrydial is produced in plant tissues infected by Botrytis cinerea. Phytochemistry 57: 689-692 https://doi.org/10.1016/S0031-9422(01)00088-7
  6. Dickinson, S. 1969. Studies in the physiology of obligate parasitism. VI. Directed growth. Phytopathol. Z. 66: 38-49 https://doi.org/10.1111/j.1439-0434.1969.tb03084.x
  7. Edlich, w., Lorenz, G, Lyr, H. and Pommer, E.-H. 1988. Studies on the biochemical basis of resistance against dicarboximide fungicides. Brighton Crop Protection Coriference-Pests and Diseases 1: 391-396
  8. Elad, Y. and Evensen, K. 1995. Physiological aspects of resistance to Botrytis cinerea. Phytopathology 85: 637-643
  9. Jacyno, J. M., Harwood, J. S., Cutler, H. G and Dulik, D. M. 1994. Structure and solution-state confonnation of botcinolide, a new biologically active metabolite from the fungus Botrytis cinerea. Tetrahedron 50: 11585-11592 https://doi.org/10.1016/S0040-4020(01)85653-2
  10. Kim, J.-C., Choi, G J., Kim, H. T., Kim, H.-J. and Cho, K. Y. 2000. Pathogenicity and pyrenocine production of Curvularia inaequalis isolated from zoysia grass. Plant Dis. 84: 684-688 https://doi.org/10.1094/PDIS.2000.84.6.684
  11. 김종진, 김재원, 이창원, 정영륜, 1997. Botrytis cinerea 균주들이 생산하는 polygalacturonase, laccase, $\beta$-glucosidase의 균주간 활성 및 병원성과의 상관관계. 한국식물병리학회지 13:255-231
  12. Rebordinos, L., Cantoral, J. M., Prieto, M. V, Hanson, J. R. and Collado, I. G 1996. The phytotoxic activity of some metabolites of Botrytis cinerea. Phytochemistry 42: 383-387 https://doi.org/10.1016/0031-9422(95)00909-4
  13. Salinas, J., Warnaar, F. and Verhoeff, K. 1986. Production of cutin hydrolyzing enzymes by Botrytis cinerea in vitro. Phytopathol. Z. 116: 299-307 https://doi.org/10.1111/j.1439-0434.1986.tb00924.x
  14. Sasaki, I. and Nagayama, H. 1994. $\beta$-glucosidase from Botrytis cinerea: Its relation to the pathogenicity ofthis fungus. Biosci. Biotech. Biochem. 58: 616-620 https://doi.org/10.1271/bbb.58.616
  15. Tonukari, N. J., Scott-Craig, J. S. and Walton, J. D. 2000. The Cochliobolus carbonum SNFI gene is required for cell walldegrading enzyme expression and virulence on maize. Plant Cell 12: 237-247 https://doi.org/10.1105/tpc.12.2.237
  16. Van Kan, J. A. L., Van't Klooster, J. W., Wagemakers, C. A. M., Dees, D. C. T. and Van der Vlugtbergmans, C. J. B. 1997. Cutinase A of Botrytis cinerea is expressed, but not essential, during penetration of gerbera and tomato. MPMI 10: 30-38 https://doi.org/10.1094/MPMI.1997.10.1.30
  17. Wasty, E. H., Farag, S. A., Tarabieh, M. A. and Abd-Elmoety, S. H. 1978. Studies on enzymes of different strains of Botrytis cinerea. Phytopathology 92: 168-179 https://doi.org/10.1111/j.1439-0434.1978.tb03598.x