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栽培稻 (OryzasativaL .)杂种F1花粉不育性至少由 6个基因座位所控制。为了定位其中的一个基因座位S b ,选用了 15 8个微卫星标记对粳型品种“台中 6 5”及其近等基因系TISL2之间的多态性进行了分析。结果发现第 5染色体短臂上的RM13在亲本间存在多态性。连锁分析表明 ,RM13与S b座位紧密连锁。根据微卫星标记分析的结果 ,在RGP(RiceGenomeResearchProgramofJapan)发表的遗传图谱的相应位置上选取了 11个RFLP标记 ,利用这些标记的末端序列设计特异PCR引物 ,进行ALP (ampliconlengthpolymorphism)分析 ,结果 11对引物均没有ALP。用6种四碱基识别位点的限制性内切酶进行PBR(PCR_basedRFLP)分析 ,发现由R2 2 13S和R830两克隆设计的STS(sequence_taggedsite)标记在T6 5与TISL2之间存在酶切多态性。R830STS、RM13和R2 2 13SSTS与S b座位的遗传距离分别为 3.3cM (centiMorgan)、5 .2cM和 5 .5cM。这些以PCR为基础的分子标记可以直接应用于分子标记辅助选择中 ,从而建立了基因定位与分子标记辅助选择一体化的技术体系 ,使S b座位在育种中可以得到有效的利用
Cultivated rice (Oryzasativa L.) hybrid F1 pollen sterility is controlled by at least 6 loci. In order to locate one of these loci, S b, 15 8 microsatellite markers were selected to analyze the polymorphism of japonica rice “Taichung 65” and its near isogenic TISL2. The results showed that RM13 on the short arm of chromosome 5 had polymorphism in the parents. Linkage analysis showed that RM13 was closely linked to SB seat. According to the results of microsatellite marker analysis, eleven RFLP markers were selected at the corresponding positions in the genetic map published by Rice Genome Research Program of Japan (RGP). Based on the end sequences of these markers, specific PCR primers were designed and ampliconlength polymorphism analysis was performed. As a result, 11 pairs of primers No ALP. Using PCR-based restriction fragment length polymorphism (PCR-RFLP) analysis of six restriction sites at the four-base recognition sites, it was found that the sequence-tagged site (STS) tag designed by both R2 2 13S and R830 clones had an enzymatic digestion polymorphism between T6 5 and TISL2 Sex. The genetic distances for R830STS, RM13 and R2 2 13SSTS and SB loci were 3.3 cM (centiMorgan), 5.2 cM and 5.5 cM, respectively. These PCR-based molecular markers can be directly applied to molecular marker-assisted selection, thus establishing a technology system integrating gene mapping with molecular marker-assisted selection, which makes S b seat effectively utilized in breeding