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用3个与水稻温敏雄性核不育(TGMS)基因紧密连锁的PCR标记C365-1、S2-40和In Del37,对云南低海拔(260 m)早季种植的2个籼稻TGMS株系配制的杂交组合的F2和F3代分离群体中筛选出的146个单株(126个TGMS单株和20个可育株)及经多代自交获得的15个TGMS株系的基因型进行鉴别,结果表明:只有标记C365-1在来自两个群体的不育株和可育株DNA池间显示出多态性。不育株DNA池产生两个PCR片段,分别为418 bp(定名为C365-418 bp)和344 bp(定名为C365-344 bp),而可育株DNA池只产生C365-418 bp片段。经克隆测序比对,显示多态性的C365-344 bp片段与水稻第2染色体上的OJ1001_D05克隆的DNA序列相似性为84%,与TGMS基因ptgms2-1和tms5的距离约为19.1 kb;与水稻第6染色体上的OJ1001_B06.3和OJ1001_B06.4两个克隆间隔区的DNA序列的相似性为98%,与TGMS基因TMS相距约1 c M。在146个单株中,标记C365-1的基因型值与单株育性表型值间的相关系数为0.944,达极显著水平。来自15个TGMS株系的每一个株系都显示出与不育株DNA池相同的带型。以上结果说明,PCR标记C365-1可较为有效地鉴别水稻TGMS株携带的TGMS基因,该标记可作为水稻TGMS株基因型分子标记辅助选择的有效标记。
Two indica rice TGMS lines planted in low-altitude (260 m) Yunnan early season were prepared by using three PCR markers C365-1, S2-40 and In Del37 which were closely linked to the TGMS gene in rice (126 TGMS single plants and 20 fertile plants) and 15 TGMS lines obtained from multiple generations of selfing were selected from the segregation population of F2 and F3, The results showed that only the marker C365-1 showed polymorphism among the sterile DNA and fertile DNA pools from two populations. The sterile DNA pool produced two PCR fragments, 418 bp (designated C365-418 bp) and 344 bp (designated C365-344 bp), respectively, while the fertile DNA pool produced only C365-418 bp fragments. The DNA sequence of the C365-344 bp fragment of the polymorphism and the OJ1001_D05 clone on chromosome 2 of rice was 84%, and the distance from the TGMS genes ptgms2-1 and tms5 was about 19.1 kb by cloning and sequencing. The DNA sequence similarity of OJ1001_B06.3 and OJ1001_B06.4 on the chromosome 6 of rice was 98% and about 1 cM away from the TGMS gene TMS. In 146 plants, the correlation coefficient between the genotype value of marker C365-1 and the phenotypic value of single fertility was 0.944, reaching a very significant level. Each of the lines from the 15 TGMS lines showed the same band pattern as the sterile DNA pool. The above results indicate that the PCR marker C365-1 can effectively identify the TGMS gene carried by the rice TGMS strain and can be used as an effective marker for molecular marker assisted selection of the rice TGMS strain.