摘要
为从分子水平上鉴定稻属,对稻属9种32份植物材料DNA条形码基因atpF-atpH进行PCR扩增并对PCR扩增产物测序,以近缘种植物为外类群建立系统进化树及序列差异分析,设计了鉴定稻属的特异性引物。结果表明,atpF-atpH扩增产物为1条500bp左右的目标条带,对于供试的稻属材料,扩增效率和测序成功率均达到100%,应用该特异性引物对稻属材料进行扩增,均获得与预期大小一致的113bp特异性目的片段,而对非稻属材料的扩增结果均呈阴性。经序列差异分析,稻属植物在UPGMA聚类树上聚成一个单系类群,节点支持率为97%,显示atpF-atpH序列可用于稻属的分类鉴定。这说明在条码基因差异位点分析的基础上,设计特异分子标记可用于稻属的鉴定,为珍贵物种的口岸鉴定提供了检测方法。
In order to identify Oryza species at the molecular level, a total nine species of thirty-two plant materials were amplified the barcode gene atpF-atpH and sequenced the PCR products. Comparative sequencing of the atpF-atpH gene was carried out in order to design specific primers for Oryza species. A phylogenetic tree which related species was outgroup was created. The results were as follows： The atpF-atpH PCR product was about 500 bp target fragment. Efficiency of amplification and sequencing success rates were up to 100 percent for tested Oryza species materials. The 113 bp target fragment was amplified by the PCR from the Oryza species materials, but other plant materials not Oryza species were not amplified. According to phylogenetic analysis, Oryza species materials were clustered a single group on UPGMA phylogenetic tree and node support rate was up to 97 percent. It showed that based on the analysis of sequence difference of barcode genes, specific molecular markers could be used for the identification of Oryza species. It provided detection methods for precious species identification at port.
出处
《中国农学通报》
CSCD
2012年第24期173-178,共6页
Chinese Agricultural Science Bulletin
基金
质检公益性行业科研专项"防止水稻重要资源流失的DNA条形码技术研究"(10-45)
中国检科院基础科研业务费"几种云南珍稀植物种质资源的分子鉴定"(2009JK029)
