【作者】 刘雪； 李明顺； 李新海； 田清震； 白丽； 张世煌；

【Author】 LIU Xue;LI Ming-Shun;LI Xin-Hai;TIAN Qing-Zhen;BAI Li;ZHANG Shi-Huang (The Institute of Crop Breeding and Cultivation CAAS/Key Lab. of Crop Genetic & Breeding, Ministry of Agriculture, Beijing 100081, China)

【机构】 中国农业科学院作物育种栽培研究所/农业部作物遗传育种重点开放实验室；

【摘要】 针对四种多株叶片混合DNA取样方法,利用SSR标记分析了Pob69和Pob70群体的遗传变异,探讨建立玉米群体遗传多样性分析的技术体系。从两个群体中分别提取50个单株叶片DNA和5、10、15、20个单株叶片混合样本DNA,用均匀分布在玉米染色体上的17对SSR引物对不同处理的混合DNA样本进行扩增,通过聚丙烯酰胺凝胶电泳,比较50个单株DNA与四种混合叶片的DNA扩增的等位基因数目、多态性信息量和遗传多样性指数,表明用10株叶片混合提取的DNA样本可以代替相同数目(10个)的单株DNA。同时比较不同数量单株DNA的混合样本和相同数量多株叶片混合提取的DNA扩增的等位基因数目,证实可以用混合叶片提取的DNA来代替同等数量单株DNA的混合样本。这样可以进一步减少工作量,提高效率。该技术策略已用于研究大量玉米群体的遗传关系。更多还原

【Abstract】 Diallel analysis was commonly used to detect the genetic variation among or in maize (Zea mays L.) populations, which makes great efforts to facilitate hybrids development. In recent years, molecular markers are convinced as a powerful tool in detecting the genetic diversity of maize inbreds, while it remains under investigation and optimization in analyzing maize populations. In this paper, two populations, Pob69 and Pob70, and simple sequence repeat (SSR) markers were adopted to establish the technique procedure to detect genetic variation in maize populations. The modified CTAB method was used to extract DNA of 50 individuals of each population and DNA from leaf mixture of 5, 10, 15 and 20 individuals, respectively. 17 SSR primers normally selected from 10 chromosomes of maize were employed for PCR amplification of 50 individual DNAs and 4 kinds of bulk DNA samples from leaf mixture. The bulk DNA from leaf mixture of 10 individuals was identified to be the best choice in replacing the same number of individuals through comparing the number of the alleles, polymorphism information content and diversity index. Furthermore, the bulk DNA extracted from leaf mixture of 10 individuals was confirmed as a labor saving and efficient approach. The bulking fingerprinting method has been being adopted to analyze the relationships among maize populations with polyacrylamide gel electrophoresis.更多还原