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[摘要]
目的 通过分析不同居群草珊瑚Sarcandra glabra和金粟兰属5种近缘植物的核糖体基因内部转录间隔区(rDNA-ITS)序列,为草珊瑚鉴定和品种鉴别提供模式识别与思路。方法 采集不同产地的草珊瑚18份、金粟兰6份,所有样品提取总DNA,并对其ITS片段进行扩增、测序,并搜索GenBank数据库中收录的其他4种金粟兰属植物的ITS序列,运用ClustalX 2.1软件比较不同产地草珊瑚与其他金粟兰属植物的ITS序列差异,并对其进行聚类分析。结果 草珊瑚样品不同居群间的ITS序列相似度达99%,其中位点总突变率ITS1(2.7%)>ITS2(1.4%),而与金粟兰属其他植物相比,位点总突变率ITS2(20.3%~22.7%)>ITS1(15.9%~18.3%)。聚类分析表明18个不同种群的草珊瑚居群间变异极小,且与金粟兰属5种近缘植物具有显著差异的聚类识别。结论 草珊瑚ITS1与ITS2分别为不同居群间及与不同属近缘植物间比较有效的鉴别序列,具有较多个特异性信息位点的草珊瑚ITS序列用于鉴别金粟兰属5种近缘植物,具显著差异的聚类识别,是草珊瑚和金粟兰属近缘植物鉴定和品种鉴别的有效分子标记。
[Key word]
[Abstract]
Objective To study the ribosomal DNA internal transcribed spacer (rDNA-ITS) sequences of Sarcandra glabra from different areas and five plants in Chloranthus Swartz, and to provide pattern recognition and thread for the species identification of S. glabra. Methods The ITS sequences of 18 populations of S. glabra and six populations of Chloranthus spicatus were amplified by PCR with universal primer of ITS and sequenced, and the ITS sequences of the other plants in Chloranthus Swartz were searched in GeneBank. Data were analyzed by ClustalX 2.1, and a cluster analysis was presented by UPGMA. Results The semblance of ITS sequences of S. glabra from different areas was 99%. The total mutation rate of ITS1 sequence (2.7%) was higher than that of ITS2 sequence (1.4%). However, compared with other plants in Chloranthus Swartz, the total mutation rate of ITS2 sequence (20.3%—22.7%) was higher than that of ITS1 sequence (15.9%—18.3%). The cluster analysis showed that there was little variation among the 18 populations of S. glabra, but there was significant difference with the five plants of Chloranthus Swartz. Conclusion ITS sequence can be used to identify the plants from different areas and in different genus, ITS sequence of S. glabra has several specified information sites to identify the five plants in Chloranthus Swartz, with significantly different cluster analyseis, and is the active molecular marker for the species identification of S. glabra and plants in Chloranthus Swartz.
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[基金项目]
福建省教育厅A类项目(JA11138);福建省科技重大项目(2011Y4005)