벼멸구(Nilaparvata lugens)에서 마이크로새털라이트 마커의 분리 및 특성검정

Isolation and Characterization of Microsatellites in the Brown Planthopper, Nilaparvata lugens $St{\aa}l$

  • 문점희 (캘리포니아대학 버클리캠퍼스 곤충/환경 정책관리학과) ;
  • 송유한 (경상대학교 농업생명과학연구원) ;
  • Mun Jeomhee (Division of Insect Biology, Environmental Science, Policy and Management, University of California) ;
  • Song Yoo Han (Institute of Agriculture and Life Science, Gyeong-Sang National University) ;
  • Roderick George K. (Division of Insect Biology, Environmental Science, Policy and Management, University of California)
  • 발행 : 2004.12.01

초록

벼멸구(Nilaparvata lugens)는 벼에 가장 큰 피해를 주는 해충 중의 하나로서, 마이토콘드리아 DNA를 분석한 선행 연구결과에 의하면 북 베트남의 홍하유역을 중심으로 남쪽과 북쪽의 개체군이 유전적으로 뚜렷한 차이를 보이고 있다. 그러나 이러한 마이토콘드리아 DNA의 변이로는 좀더 상세한 지역간 개체군의 유전적 변이를 검정할 수 없으므로, 마이크로새털라이트 마커를 이용할 수 있는 방법을 모색하였다. 총 37개 마이크로새털라이트 위치를 분석한 결과 5개 위치에서 성공적으로 라벨을 할 수 있었으며, 그 중 2개 위치에서 유용한 개체군 변이정보를 얻을 수 있었다. 이러한 두 위치에서 벼멸구의 생태형(1, 2, 3형)에 따른 변이를 검정할 수 있는지의 여부를 검정한 결과, 두 위치 중에서 한 곳(27035)에서는 생태형간의 차이를 나타내지 않았으나, 다른 한 곳(7314)에서는 생태형 간에 차이를 보였다. 따라서 마이크로새털라이트 마커를 이용하면 좀 더 상세한 벼멸구 지역 개체군의 차이를 검정하여 이동과 분산의 근원과 경로를 알아내는데 유용한 방법이 될 것으로 생각된다.

The brown planthopper, Nilaparvata lugens, is among the most serious insect pests of rice. It is widely distributed in Asia, Australia and Pacific islands. An earlier mitochondrial DNA study revealed that there exist significant genetic differences between populations north and south of the Red River Delta region in Vietnam. However the mitochondrial DNA was not sufficiently variable to examine the sources of immigration. For a more detailed analysis of geographic population structure of N. lugens, we developed microsatellite markers. Thirty-seven putative microsatellite loci were isolated using a magnetic biotin method, and five primer pairs designed from the flanking regions of sequenced microsatellite clones were labeled with fluorescent. Of these five primer sets, two have proven to be useful across all the samples we used in this study. We used variation at these two microsatellite loci to test the hypothesis that N. lugens biotypes (1, 2, and 3) sampled from laboratory selection constituted distinct genetic units. Allele frequency differences among the three major biotype categories were not significantly different at one locus (27035). However, the other (7314) did show differences among the major three biotypes. The methods we describe here will be useful for studying population structure of crop pest and for tracking the patterns of migratory pest like the rice planthoppers.

키워드

참고문헌

  1. Kisimoto, R. 1976. Synoptic weather conditions inducing long distance migration of planthoppers, Sogatella furcifera Horvath and Nilaparvata lugens St$\aa$l. Ecol. Entomol. 1: 95-109 https://doi.org/10.1111/j.1365-2311.1976.tb01210.x
  2. Kisimoto, R and K Sogawa. 1995. In Insect Migration: Tracking resources through space and time. V.A. Drake and A.G. Gatehouse eds, pp. 67-91. Cambridge, Cambridge University Press
  3. Mun, J.H., Y.H. Song, K.L. Heong and G.. K. Roderick. 1999. Genetic variation among Asian populations of rice planthoppers, Nilaparvata lugens (St$\aa$l) and Sogatella furcifera Horvath (Homoptera: Delphacidae): Mitochondrial DNA sequences. Bull. of Entomol. Res. 89: 245-253
  4. Pathak, M.D., C.H. Cheng and M.E. Fortuno. 1969. Resistance to Nilaparvata impicticeps and N. lugens in varieties of rice. Nature 223: 502-504 https://doi.org/10.1038/223502a0
  5. Roderick, G.K. 1996. Geographic structure of insect populations: gene flow, phylogeography, and their uses. Ann. Rev. of Entomol. 41: 263-90
  6. Saxena, R.C. and A.A. Barrion. 1985. Biotypes of the brown planthopper Nilaparvata lugens (St$\aa$l) and strategies in deployment of host plant resistance. Insect Sci. Applic. 6: 271- 289
  7. Schneider, S., J-M. Kueffer, D. Roessli and L. Excoffier. 2001. Arlequin ver 2.1. Geneva: http://anthropologie.unige.ch/arlequin, University of Geneva
  8. Sogawa, K. 1997. Overseas immigration of rice planthoppers into Japan and associated meteorological systems. Pages 13-35 in Proceedings of China-Japan Joint Workshop on Migration and Management of Insect Pests of Rice in Monsoon Asia, China National Rice Research Institute, Hangzhou, China
  9. Song, Y.H. 1995. Risk Assessment for the monsoonic migratory pests of rice in Korea. Pages 101-112 in Proceedings of International Workshop on the Pest Management Strategies in Asian Monsoon Agroecosystems, eds. N. Hokyo and G. Norton, Kumamoto, Japan: Kyushu National Agricultural Experiment Station
  10. Tanaka, K. 1999. Quantitative genetic analysis of biotypes of the brown planthopper N. lugens. Entomol. Exp. et Appl. 90: 279-287 https://doi.org/10.1023/A:1026456230292
  11. Uhm, K.B., J.S. Park, Y.I. Lee, K.M. Choi, M.H. Lee and J.O. Lee. 1988. Relationship between some weather conditions and immigration of the brown planthopper, Nilaparvata lugens St$\aa$l. Kor. J. Appl. Eetomol. 27: 200-210
  12. Vos, P., R. Hogers, M. Bleeker, M. Reijans, T. van de Lee, M. Hornes, A. Frijters, J. Pot, J. Peleman, M. Kuiper and M. Zabeau. 1995. AFLP: a new technique for DNA fingerprinting. Nucleic Acid Res. 23: 4407-4414 https://doi.org/10.1093/nar/23.21.4407
  13. Wright, S. 1931. Evolution in Mendelian populations. Genetics 16: 97-159
  14. Zane, L., L. Bargelloni and T. Patarnello. 2002. Strategies for microsatellite isolation: a review. Mol. Ecol. 11: 1-16 https://doi.org/10.1046/j.0962-1083.2001.01418.x