NGS 기반 밀양23호/기호벼 재조합자식 유전집단(MGRIL)을 이용한 CAPS 마커 개발 및 줄기 굵기 형질 관련 QTL 분석
Development of NGS-based New CAPS Markers and QTL Analysis of Stem Diameter using Milyang23/Gihobyeo Recombinant Inbred Lines (MGRIL)
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초록
차세대 염기서열 분석방법을 통해 밀양23호와 기호벼의 유전체 서열을 분석하여 변이를 탐색하였다. 대량으로 생산된 SNP을 기반으로 Lee et al. (2014)이 선발한 후보 CAPS 마커 중에서4개의 제한효소(EcoRⅠ, HindⅢ, PstⅠ, XhoⅠ) 자리에 위치한특이적인 SNP을 후보 CAPS 마커로 개발하였다. 후보 CAPS 마커 7,843개 중 101개가 안정적인 CAPS 마커로 개발되었다 이들 CAPS 마커를 PCR 기반 마커들로 구성된 Lee et al. (2014) 연구의 유전지도와 통합하여 보다 정밀해진 분자유전지도를작성하였다. 총 유전거리는 1,696.97cM였으며, 마커 간 평균거리는 3.64cM로 이전 연구에 비해 거리가 줄어든 것을 확인하였다. 또한, 벼의 줄기 굵기 형질에 대해 QTL을 탐색한 결과, 총 9개의 유의성이 있는 QTL을 찾을 수 있었으며 모두 기존에보고되지 않은 새로운 QTL이었다. 간장, 수장 그리고 수수 형질에 관한 새로운 QTL은 발견되지 않았지만, 7가지 형질에 대한주동 및 미동 QTL 중 일부는 본 연구에서 새롭게 개발한 CAPS 마커로 확인되었으며 보다 정밀한 QTL 유전지도 작성이 가능하였다. NGS 기반 CAPS 마커로 작성된 분자유전지도를 통해높은 정확성과 신규로 개발된 QTL의 신뢰성을 확인하였으며, 이를 이용하여 향후 대량의 마커를 이용하여 고밀도 유전지도를작성하고 이에 대한 QTL 분석을 통해 농업적으로 유용하게쓰일 수 있을 것이다.
Abstract
The rice recombinant inbred lines derived from Milyang23 and Gihobyeo cross were used in genetic mapping and QTL analysis studies. In this study, we developed a new 101 CAPS markers based on the SNPs in the whole genome region between these varieties. As a result, the total genetic distance and average distances were 1,696.97 cM and 3.64 cM, respectively. In comparison to the distance of the previous genetic map constructed based on 365 DNA markers, the new genetic map was found to have a decreased distance. The map was applied for the detection of QTLs on all seven traits relevant to diameter of stem internode, length of culms, length of panicles and the number of panicles including the correlation analysis between each trait. The QTLs results were similar to the report in previous studies, whereas the distance between the markers was narrowed and accuracy increased with the addition of 101 CAPS markers. A total of 9 new QTLs were detected for stem internode traits. Among them, qI1D-6 had higher LOD of 5.1 and phenotype variation of 50.92%. In this experiment, a molecular map was constructed with CAPS markers using next generation sequencing showing high accuracy for markers and QTLs. In the future, developing more accurate QTL information on stem internode diameters with various agriculturally important traits will be possible for further rice breeding.
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