Korean journal of applied entomology (한국응용곤충학회지)

Korean Society of Applied Entomology (한국응용곤충학회)
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Characterization of New Bacillus thuringiensis Isolated with Bioactivities to Tobacco Cutworm, Spodoptera litura (Lepidoptera: Noctuidae)

담배거세미나방에 살충효과를 나타내는 새로운 Bacillus thuringiensis 균주의 특성

  • Kim, Da-A (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Kim, Jin-Su (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Kil, Mi-Ra (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Paek, Seung-Kyoung (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Choi, Su-Yeon (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Jin, Da-Yong (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Youn, Young-Nam (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Hwang, In-Cheon (Central Research Institute, Kyung Nong Co.) ;
  • Yu, Yong-Man (Dept. Applied Biology, College of Agriculture and Life Sciences, Chungnam National University)
  • 김다아 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 김진수 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 길미라 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 백승경 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 최수연 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 김대용 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 윤용남 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 황인천 (㈜경농 중앙연구소) ;
  • 유용만 (충남대학교 농업생명과학대학 응용생물학과)
  • Published : 2008.03.30
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Abstract

Bacillus thuringiensis with selected high toxicities against tobacco cutworm, Spodoptera litura were isolated from domestic soils. When being observed under a phase-contrast microscope, the insecticidal crystal proteins were showed a bipyramidal crystal types. New CAB 109 isolate was identified to B. thuringiensis subsp. aizawai in the H serotype. As a results of insecticidal activities between CAB 109 isolate and 3 existing ready-made products against 3rd larva of S. litura, CAB 109 isolate showed 100% mortality with spore concentration $(1.3{\times}10^7cfu/ml)$. It was a very high insecticidal activity compared with a existing ready-made B. t. products. $LD_{50}$ values of CAB 109 isolate was $9.78{\times}10^5,\;6.87{\times}10^6\;and\;1.83{\times}10^7cfu/ml$ spore concentration against 2nd, 3rd and 4th larva of S. litura, respectively. Unlike Plutella xylostella, S. litura was slowly died after application up to 7 days. The weight of S. litura larva applied with CAB 109 isolate were 6-7 times less than controlled group. Even though it didn't die, it did not grow into next larva. The result observed with scanning electron microscope was that CAB 109 isolate of B. t. aizawai formed a typical bipyramidal crystal protein type. Otherwise, when CAB 109 isolate was examined with SDS-PAGE and with trypsin, there was no difference between CAB 109 strain and ready-made products of B. thuringiensis.

국내 토양으로부터 분리한 Bacillus thuringiensis 균주에서 담배거세미나방(Spodoptera litura)에 선택적으로 살충효과를 나타내는 새로운 균주를 선발하였다. 이 균주의 결정성 독소단백질은 위상차 현미경과 주사전자현미경으로 관찰한 결과 전형적인 이중피라미드 형태이며 H serotype으로 동정한 결과 Bacillus thuringiensis subsp. CAB 109로 동정되었다. 난방제 해충인 담배거세미나방 3령충에 대해 B. t. subsp. aizawai CAB 109균주외 분리된 다른 3균주의 생물활성을 비교 검토하였다. 살충활성 결과에서 B. t. subsp. aizawai CAB 109 균주가 $1.3{\times}10^7$ (cfu/ml)에서 100% 사망률로 다른 균주에 비해 높은 활성을 나타내었다. 담배거세미나방은 배추좀나방과 다르게 7일 이상 경과한 뒤에야 100% 사충률을 나타내었다. 한편, 담배거세미나방 2령, 3령, 4령충에 대한 살충활성의 검정에서 B. t. aizawai subsp. CAB 109 균주의 $LD_{50}$값이 각각 $9.78{\times}10^5,\;6.87{\times}10^6,\;1.83{\times}10^7$ (cfu/ml)으로 살충활성을 나타내었다. B. t.를 섭식한 담배거세미나방 유충의 사망 현상은 지효성이며 대조군보다 무게가 $6{\sim}7$배정도 적으며 다음 령기로 성장하지 못하고 죽었다. 또한 B. t. subsp. aizawai CAB 109 균주의 SDS-PAGE에서의 단백질 패턴과 trypsin을 처리한 단백질의 결과에서 기존의 B. t. subsp. aizawai와 CAB 109균주와의 차이점은 발견할 수 없었다.

Keywords

References

  1. Ahn, S.B., I.S. Kim, W.S. Cho, M.H. Lee and K.M. Choi. 1989. The Occurance of the crop insect pests from Korea in 1988. Korean J. Appl. Entomol. 28(4): 246-253
  2. Apaydin, O., A.F. Yenidunya, S. Harsa and H. Gunes. 2005. Isolation and characterization of Bacillus thuringiensis strains from different grain habitats in Turkey. World J. Microbiol. Biotechnol. 21: 285-292 https://doi.org/10.1007/s11274-004-3633-y
  3. Aronson, A.I., W. Beckman and P. Dunn. 1986. Bacillus thuringiensis and related insect pathogens. Microbil. Rev. 50: 1-24
  4. Bae, S.D., B.R. Choi, Y.H. Song and H.J. Kim. 2003. Insecticide susceptibility in the different larva of tobacco cutworm, Spodoptera litura Fabricius (Lepidoptera: Noctuidae) collected in the soybean fields of Milyang, Korea. Korean J. Appl. Entomol. 42(3): 225-231
  5. Bae, S.D., H.J., Kim, G.H., Lee and S.T., Park. 2007. Seasonal occurrence of tobacco cutworm, Spodoptera litura Fabricius and beet armyworm, Spodoptera exigua hubner using sex pheromone traps at different locations and regions in Yeongnam district. Korean J. Appl. Entomol. 46(1): 27-35 https://doi.org/10.5656/KSAE.2007.46.1.027
  6. Bae, S.D., K.B. Park and Y.J. Oh. 1997. Effect of temperature and food source on the egg and larval development of tobacco cutworm, Spodoptera litura Fabricius. Korean J. Appl. Entomol. 36(1): 48-54
  7. Cho, J.R., W.R. Song., S.Y., Hwang, H.S. Kim and J.O. Lee. 1996. Age-related susceptibilty of Spodoptera litura larvae to some insecticides. Korean J. Appl. Entomol. 35(3): 249-253
  8. Choi, J.Y., H.S. Kim, B.R. Jin, K.Y. Seol, H.Y. Park and S.K. Kang. 1996. Pathogenicity and production of Spodoptera exigua nuclear polyhedrosis virus. Korean J. Appl. Entomol. 35(3): 228-231
  9. Crickmore, N., D.R. Zeigler, J. Feitelson, E. Schnepf, J. Van rie, D. Lereclus, J. Baum and D.H. Dean. 1998. Revision of the nomenclature for the Bacillus thuringiensis pesticidal crystal proteins. Microbiol. Mol. Biol. Rev. 62(3): 807-813
  10. Da Silva, S.M.B., J.O. Silva-Werneck, R. Falcao, A.C. Gomes, R.R. Fragoso, M.T. Quezado, O.B.O. Neto, J.B. Aguiar, M.F. G. de Sa, A. Bravo and R.G. Monnerat. 2004. Characterization of novel Brazilian Bacillus thuringiensis strains active against Spodoptera frugiperda and other insect pests. J. Appl. Ent. 128: 102-107 https://doi.org/10.1046/j.1439-0418.2003.00812.x
  11. Finney, D.J. 1971. Probit analysis, estimation of the median effective dose. Cambridge University Press. London. 19-47
  12. Gill, S.S., E.A. Cowles and V. Francis. 1995. Identification, isolation, and cloning of a Bacillus thuringiensis CryIAc toxin binding protein from the midgut of the lepidopteran insect Heliothis virescens. J. Biol. Chem. 270: 27277-27282 https://doi.org/10.1074/jbc.270.45.27277
  13. Goh, H.G., S.G. Lee, B.P. Lee, K.M. Choi and J.H. Kim. 1990. Simple mass-rearing of beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), on an artificial diet. Korean J. Appl. Entomol. 29: 180-183
  14. Goh, H.G., J.D. Park, Y.M. Choi, K.M. Cho and I.S. Park. 1991. The host plants of beet armyworm, Spodoptera exigua (Hübner), (Lepidoptera: Noctuidae) and its occurrence. Korean J. Appl. Entomol. 30(2): 111-116
  15. Goldberg, L.J. and J. Margalit. 1977. A bacterial spore demonstration rapid larvicidal activity against Anopheles serengotii, Uranotaenia unguiculata, Culex univittatus, Aedes aegypti and Culex pipiens. Mosq. News. 37: 355-358
  16. Gough, J.M., D.H. Kemp, R.J. Akhurst, R.D. Pearson and K. Kongsuwan. 2005. Identification and characterization of proteins from Bacillus thuringiensis with high toxic activity against the sheep blowfly, Lucilia cuprina. J. Invertebr. Pathol. 90: 39-46 https://doi.org/10.1016/j.jip.2005.05.012
  17. Hofte, H. and H.R. Whiteley. 1989. Insecticidal crystal proteins of Bacilllus thuringiensis. Microbiol. Rev. 53(2): 242-255
  18. Kim, D.A., J.S. Kim, M.R. Kil, Y.N. Youn, D.S. Park and Y.M. Yu. 2006. Isolation and activity of insect pathogenic Bacillus thuringiensis strain from soil. Korean J. Appl. Entomol. 45(3): 357-362
  19. Kim, S.G., J.D. Park, D.I. Kim., D.J. Im., K.C. Kim and Y.M. Yu. 2003. Effects of field application of Spodoptera litura nucleopolyhedrovirus to control S. litura in chrysanthemum. Korean J. Appl. Entomol. 42(2): 153-157
  20. Laemmli, U. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 277: 680-685
  21. Lee, G.H., S.D. Bae, H.J. Kim, S.T. Park and M.Y. Choi. 2006. Economic injury levels for the common cutworm, Spodoptere litura (Fabricius) (Lepidoptera: Noctuidae) on soybean. Korean J. Appl. Entomol. 45(3): 333-337
  22. Ohba, M. and K. Aizwa. 1978. Serological identification of Bacillus thuringiensis and related bacteria isolated in Japan. J. Invertebr. Pathol. 32: 303-309 https://doi.org/10.1016/0022-2011(78)90193-3
  23. Park, H.W., H.S. Kim, D.W. Lee, Y.M. Yu, B.R. Jin and S.K. Kang. 1995. Expression and synergistic effect of three types of crystal protein genes in Bacillus thuringiensis. Biochem. Biophys. Res. Commun. 214(2): 602-607 https://doi.org/10.1006/bbrc.1995.2328
  24. Raymond, M. 1985. Presentation d'un programme d'analyse logprobit pour micro-ordinnateur. Cah. ORSTOM, Ser. Ent. Med. et Parasitol. 22: 117-121
  25. Tamez-Guerra, P., A.A. Iracheta, B. Pereyra-Alferez, L.J. Galan- Wong, R. Gomez-Flores, R.S. Tamez-guerra, and C. Rodriguez- Padilla. 2004. Characterization of Mexican Bacillus thuringiensis strains toxic for lepidopteran and coleopteran larvae. J. Invertebr. Pathol. 8: 7-18
  26. Yasutake, K., N.D. Binh, K. Kagoshima, A. Uemori, A. Ohgushi, M. Maeda, E. Mizuki, Y.M. Yu and M. Ohba. 2006. Occurrence of parasporin-producing Bacillus thuringiensis in Vietnam. Can. J. Microbiol. 52: 365-372 https://doi.org/10.1139/W05-134
  27. Zheng, S.J., B. Henken, R.A. de Maagd, A. Purwito, F.A. Krens and C. Kik. 2005. Two different Bacillus thuringiensis toxin genes confer resistance to beet armyworm (Spodoptera exigua Hubner) in transgemic Bt-shallots (Allium cepa L.). Transgemic Research, 14: 261-272 https://doi.org/10.1007/s11248-005-0109-2

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