벼멸구(Nilaparvata lugens)에서 마이크로새털라이트 마커의 분리 및 특성검정 Isolation and Characterization of Microsatellites in the Brown Planthopper, Nilaparvata lugens $St{\\aa}l$원문보기
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.
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 Rive...
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.
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.
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제안 방법
Results of this experiment using two microsatellite loci were inconclusive for documenting different categories of biotypes. However, one microsatellite locus proved discriminating among three biotype str ains, showing that this approach is promising for the development of genetic makers to track the migration patterns of the N.
The sequences that contained repeat sequences were used for designing microsatellite primer with Oligo (MBI) and the primers were tested for ampli fications against all planthopper individuals of field population. Ones successfully amplifying planthop per sequence with repeat region were labeled with fluorescent dyes and used for AFLP an시ysis on an ABI 377.
대상 데이터
5%) contained repet itive motifs with flanking region suitable for design ing PCR primers. Thirty-seven experimental primer pairs were designed. Of these, only five pairs (2.
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