Calli from mature embryo of “Jimai-24” wheat ( Triticum aestivum L.) were induced on medium containing Zhengdingmycin then continuously cultured on medium containing 0.5% NaCl till to regenerate plants named 8901-17 salt-tolerant mutant. “Jimai-24” was compared with 8901-17 by using the technique of RAPD. Thirty-five out of 280 random primers could detect DNA polymorphism. The similarity index was 0.978, indicating that they were NILs (near-isogenic lines). Two F2 populations (“Jimai-24”×8901-17 and 8901-17דZhongmai-9”) had been constructed using the method of half-division. The two relative DNA pools (salt tolerant DNA pool and susceptible DNA pool) which come from the two F2 populations, respectively, had been made according to the method of BSA (bulked segregant analysis). RAPD analysis between the two DNA relative pools was carried on with above 35 random primers which could detect DNA polymorphism definitely. The identical polymorphism between the two sets of DNA pools come from the two F2 populations could be determined only by OperonQ4 primer. This result implied that the polymorphic fragment amplified by OperonQ4 primer was the molecular marker of RAPD closely linked to the salt tolerant mutation locus.
利用RAPD-BSA技术筛选小麦耐盐突变位点的分子标记
索广力1 黄占景1 何聪芬1 沈银柱1* Jian WANG2（1.河北师范大学生命科学学院，石家庄050016;2. The John Curtin School of Medical Research,PO Box 334,Canberra ACT2601,Australia)
摘要：以耐盐性差的"冀麦24” (Triticum aestivum L.)和其经正定霉素诱变后获得的耐盐突变体8901-17为材料,用280个引物在两者之间进行RAPD分析,其中35个引物扩增出DNA多态性 ,其相似性系数为0.978,证明二者为近等基因系(near_isogenic line,NIL).用分株法建立两个F2群体("冀麦24”×8901_17和8901_17×"中麦9”),在两个群体中按照BSA (bulke d segregant analysis)方法分别构建两个对应DNA池(耐盐池和不耐盐池),用上述能扩增出明显多态性的35个引物在对应的耐盐和不耐盐DNA池之间进行RAPD分析,发现只有OperonQ 4引物在对应的两个DNA池扩增出的多态性在两个F2群体之间是一致的,说明其扩增产物是与耐盐突变位点紧密连锁的RAPD分子标记.
关键词： 小麦；分子标记；RAPD-BSA；耐盐突变体
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