摘要
以1对近等基因系(NIL)及其回交群体(BC1)为材料,采用BSA法,利用AFLP技术,筛选与Rf3基因连锁的分子标记。在筛选的128个AFLP引物组合中,有2个能在NIL及其可育池、不育池间扩增出多态性条带RR6和RR7。100个BC1个体验证结果表明,AFLP标记RR6扩增产物中仅出现2个重组体,重组率2%,由此估测RR6距Rf6基因约2.0cM。并成功地将此标记转化为 SCAR标记,进行了NIL和 BC1个体的特异性扩增。在来自综 3 × P138的F2:3的群体上经RFLP分析后,将RR6定位于第二染色体的长臂上。不仅为辅助育种奠定了基础,而且为克隆Rf3基因提供了有益的信息。
The maize CMS-S near isogenic line (NIL) developed by auther and the backcross progeny (BC1) derived from it were used to identify molecular markers linked to the Rf3 gene and subsequently determine its chromosomal location on the linkage map of maize. Bulk segreant analysis was performed using AFLP technique. From the survey of AFLP primer combination, two AFLP markers, (EcoRI -AGG/ Mse I -CAC and EcoRI -AAC/Mse I -CAG), which were named RR6 and RR7 respectively, linked to the Rf3 gene were identified. However, AFLP marker RR6 showed polymorphism between parents, and bulks were used to survey the available 100 individuals of the BC1 population, 2 out of 100 shed recombination. The recombination-rate was 2%. The genetic distance between Rf3 gene and AFLP marker RR6, was approximately 2.0cM. And then, the RR6 was successfully cloned and sequenced, primer synthesized and converted to SCAR marker so that PCR marker can be developed for the marker-assisted selection. In RFLP analysis, marker RR6 linked to Rf3 was found to be located between RFLP loci asg20 and php20581b, and mapped on chromosome 2L.
基金
"九五"国家重点科技攻关项目!(96-002-02-05-1)