Wang J X, Yang G S, Fu T D, Meng J L
National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China.
Yi Chuan Xue Bao. 2000;27(11):1012-6.
Bulked segregant analysis was used to identify RAPD markers linked to the Rfp of pol cms in rapeseed (Brassica napus L.) from the fertile and sterile DNA bulks. DNA bands amplified from 1040 random 10-mer primers were screened. Two polymorphic bands S1019(720) and S1036(810) were found linked to the Rfp locus at the same side with a map distance of 5.8 cM and 12.3 cM respectively. These polymorphic fragments were cloned and sequenced. Nucleotide sequence information was used to design 20-24-mer oligo nucleotide primers for PCR amplification. The SCAR markers that generated from the long primers showed the same pattern of segregation as the original RAPD markers in the backcross population. The SCAR markers would facilitate selection on the Pol CMS restorer lines in rapeseed.
利用混合分离群体分析法,从油菜(甘蓝型油菜)可育和不育DNA混合池中鉴定与油菜pol细胞质雄性不育恢复基因(Rfp)连锁的随机扩增多态性DNA(RAPD)标记。对从1040个随机10聚体引物扩增出的DNA条带进行筛选。发现两个多态性条带S1019(720)和S1036(810)与Rfp基因座位于同一侧,图距分别为5.8厘摩和12.3厘摩。对这些多态性片段进行克隆和测序。利用核苷酸序列信息设计20 - 24聚体寡核苷酸引物用于聚合酶链反应(PCR)扩增。由长引物产生的序列特异性扩增区域(SCAR)标记在回交群体中显示出与原始RAPD标记相同的分离模式。这些SCAR标记将有助于油菜Pol细胞质雄性不育恢复系的选择。