基于线粒体COI基因序列的壮真蝎与普洱真蝎的分子鉴定

发布时间：2018-12-14 17:17

【摘要】：对蝎类物种的传统分类主要依靠形态和行为特征,但由于该类群种间形态特征极为相似,物种的划分和鉴定困难。为弥补传统分类方法的不足,本研究以线粒体细胞色素氧化酶亚基Ⅰ(COI)基因作为分子标记,对形态相似的壮真蝎Euscorpiops validus和普洱真蝎E.puerensis进行分子水平的物种鉴定。采用PCR扩增测序获得壮真蝎与普洱真蝎共24个样本的COI基因部分片段序列(660 bp),进行了遗传距离、系统发育及单倍型网络图分析。结果显示:壮真蝎15个样本中共检测到4个单倍型,单倍型之间的相似度为99.3%~99.8%;普洱真蝎9个样本中共检测到4个单倍型,单倍型之间的相似度为99.6%~99.8%;2种蝎的种间序列相似度为90.1%~90.6%,单倍型间的稳定差异核苷酸位点数为61个。壮真蝎与普洱真蝎种内平均遗传距离分别为0.004 0、0.002 3,种间平均遗传距离为0.103 9,且种间遗传距离为种内的34.6倍。此外,分子系统发育树显示壮真蝎与普洱真蝎的单倍型序列各自聚为2个单系枝,且具有很高的分枝自举值(100%)。单倍型网络图结果也显示壮真蝎与普洱真蝎8个单倍型明显分为2大类群,且壮真蝎的单倍型HAP2与普洱真蝎的单倍型HAP7之间的突变步数高达62步。上述结果不仅进一步确认壮真蝎与普洱真蝎为2个不同的物种,且表明线粒体COI基因可用于开展真蝎属Euscorpiops物种的分子鉴定。
[Abstract]:The traditional classification of scorpion species mainly depends on morphological and behavioral characteristics, but it is difficult to classify and identify the species due to the similarity of morphological characteristics among species. In order to make up for the deficiency of traditional classification methods, the mitochondrial cytochrome oxidase subunit (COI) gene was used as molecular marker to identify the species of Euscorpiops validus and E.puerensis of Scorpion sinensis. The partial COI gene fragment sequences (660 bp),) of 24 samples from Scorpio and Pu'er Scorpio were obtained by PCR amplification and sequencing. The genetic distance, phylogeny and haplotype network were analyzed. The results showed that 4 haplotypes were detected in 15 Scorpio samples, and the similarity between haplotypes was 99.3 and 99.8, respectively. A total of 4 haplotypes were detected in 9 samples of Pu'er Scorpio, and the similarity between haplotypes was 99.6 and 99.8, respectively. The similarity of interspecific sequences of the two scorpions was 90.1 and 90.6, and the stable difference of nucleotide sites between haplotypes was 61. The average genetic distance between Scorpio and Pu'er Scorpio was 0.004 0.0023, and the average genetic distance between species was 0.103 9, and the genetic distance between species was 34.6 times of that within species. In addition, the molecular phylogenetic tree showed that the haplotypes of Scorpio and Pu'er Scorpio were clustered into two monotypic branches, and had a high branch bootstrap value (100%). The results of haplotype network also showed that 8 haplotypes of Scorpio and Pu'er Scorpio could be divided into two groups, and the number of mutation steps between haplotype HAP2 and haplotype HAP7 of Pu'er scorpion was as high as 62 steps. These results not only confirmed that Scorpio and Pu'er are two different species, but also suggested that mitochondrial COI gene could be used for molecular identification of Euscorpiops species of Scorpio.
【作者单位】：云南大学生命科学学院;
【基金】：云南省大学生创新创业训练计划项目(201310673008) 云南大学引进人才科研启动资金项目(XT412002)
【分类号】：Q953

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