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MITE-AFLP를 이용한 자포니카 벼의 다양성 검정 KCI 등재
Diversity Analysis of Japonica Rice using MITE-transposon Display
pp.259-268
1. 자포니카 벼 114 계통에 대해 다양성과 근연관계를 확인하고자 MITE 중에서 mPing family를 이용하여 MITE-TD 기법으로 분석하여 품종간의 다양성 정도를 산출한 결과 마커들의 PIC 값이 0.293~0.499 범위로 나타났다. 2. 두 개의 mPing primer와 selective primer인 BfaI+G 와 BfaI+C의 조합을 이용하였을 때, 공시계통인 114개의 자포니카 벼 전체를 구분할 수 있었다. 3. NTSYS-pc를 이용한 근연관계 분석 결과, 유사계수의 범위는 0.802에서 부터 0.081까지였고, 자포니카 벼 114 품종은 크게 5 개의 그룹으로 분류되었다. 4. 8 개의 MITE-AFLP marker 연관분석을 밀양 23호/합천앵미 3호 조합 RIL을 이용하여 실시한 결과, 이들은 염색체 l번, 2번, 4번, 5번, 7번 그리고 9번에 각각 위치함을 확인하였다.
Miniature inverted transposable elements (MITEs) are abundant genomic components in plant including rice. MITE-transposon display (MITE-TD) is an Amplified Fragment Length Polymorphism (AFLP)-related technique based on MITE sequence. In this study, we used the MITE-AFLP for the analysis of diversity and relation-ship of the 114 japonica accessions. Of the several MITEs, the mPing family was applied to detect polymorphisms based on PCR amplification. The BfaI adaptor primer and the specific primer derived from mPing terminal inverted repeat (TIR) region were used to PCR amplification of 114 accessions. Nine primer pairs produced a total of 160 polymorphic bands. PIC values of the polymorphic bands generated by nine primer pairs ranged from 0.269 (BfaI + ACT) to 0.426 (BfaI + T). Each accession revealed a distinct fingerprint with two primer combinations, BfaI + G and BfaI + C. Cluster analysis using marker-based genetic similarity classified 114 accessions into five groups. MITE-AFLP markers were genetically mapped using a population of 80 BILs (BC1F7) derived from a cross between the rice accessions, Milyang 23 and Hapcheonaengmi 3. Eight of the markers produced with the primer pair BfaI + 0 were mapped on chromosomes 1, 2, 4, 5, 7, and 9. Considering that one MITE-AFLP marker on chromosome 7 was tightly linked to the Rc gene, the MITE-AFLP markers will be useful for gene tagging and molecular cloning.
