【摘要】 采用BSA法 ,应用RAPD技术筛选 5 0 0个随机引物 ,共扩增出 16 14条DNA谱带 ,其中有OPA 0 1、OPP 0 9、OPP 14、OPH 19、OPN 0 8、OPN 0 7、OPN 2 0、OPY 14、OPS 2 0、OPJ 0 6十个引物在抗感池间扩增出了多态性谱带。分析表明 ,132株后代中 ,OPN 0 772 7有 4株重组 ,OPN 0 83 73 有 8株重组 ,OPY 14 60 0 有 12株重组 ,重组率为 3 0 %、6 1%和 9 1% ,换算为图距单位为 3 2cM、6 5cM和 11 7cM。其中OPN 0 83 73 、OPN 0 772 7与抗蚜基因紧密连锁。将这两个引物扩增出的RAPD片段回收、纯化、克隆、测序。结果表明 :两片段长分别为 72 7bp和 373bp。将这两个标记命名为OPN 0 772 7和OPN 0 83 73 。根据测序结果设计 2对特异性引物 ,进行特异性扩增 ,将RAPD标记成功地转化为SCAR标记。同时构建了抗蚜基因的连锁群。更多还原

【Abstract】 Molecular markers of RAPD linked to the single dominant gene of aphid resistance on sorghum were detected in BTAM 428×ICS 12B F 2 bulked segregation populations. In total, 500 random primer were used to screen the markers linked to the resistance gene, and 1614 DAN bands were amplified by using the primers of OPA 01, OPP 09,OPP 14,OPH 19,OPN 08,OPN 07,OPN 20,OPY 14,OPS 20,OPJ 06.The co segregation analysis of 132 F 2 individuals showed that four recombinants by OPN 07 727 were found, eight recombinants by OPN 08 373 , twelve recombinants by OPY 14 600 . The frequency of recombinants were 3 0%, 6 1%,9 1%, and the linkage map distance were 3 2 cM, 6 5 cM, 11 7 cM respectively. OPN 07 727 and OPN 08 373 markers were the most tightly linked to the resistant gene. The fragments amplified by OPN 07 727 and OPN 08 373 were cleaned and cloned into pMD 18 T Vector, and sequenced the length to be 727 bp and 373 bp. According to the sequences, two specific primers were designed for perform PCR and the RAPD markers were succeeded conversing to SCAR markers.更多还原