论文部分内容阅读
采用PCR技术对中国烟台、威海和莱州三个地点仿刺参的16S rRNA、COI、IrRNA-COI三个mtDNA基因片段进行了扩增和测序,分别得到了长度约为570bp、640bp和900bp的三段序列,分析了三个采样点样品的遗传多样性。结果表明,每段基因序列扩增均获得两种单倍型,已在Genbank中注册(注册号码:AY852278—AY852283)。通过比较,三个地点样品的序列差异不显著,无缺失、插入、颠换的现象,只有个别位点出现转换,其中16S rRNA序列最为保守,16S rRNA、COI、IrRNA-COI三段序列A+T的平均含量为56·2%、59·2%、61·8%,均大于G+C含量。三个采样点样品同源性为99·84%—99·96%。将获得的仿刺参DNA序列和Genbank中的来自东太平洋8个外群进行基因序列比较,结果发现,不同目、科的海参的序列差异显著。采用DNAsp统计了各类位点数;MEGA2·1分析了碱基组成和遗传距离,构建了系统树。系统树体现的分类关系与这些物种的形态学分类关系完全一致。
Three mtDNA fragments of 16S rRNA, COI and IrRNA-COI were amplified and sequenced by PCR from three sites of Yantai, Weihai and Laizhou in China. Three mtDNA fragments with length of 570bp, 640bp and 900bp were obtained respectively Segment sequences were used to analyze the genetic diversity of the three sampling spots. The results showed that two haplotypes were obtained for each gene sequence amplification and have been registered in Genbank (Accession No. AY852278-AY852283). By comparison, there was no significant difference in the sequence of the three loci samples, no deletion, insertion, transversion, only the conversion of individual loci, of which the 16S rRNA sequence was the most conservative. The 16S rRNA, COI, IrRNA- The average contents of T were 56.2%, 59.2% and 61.8%, both higher than the G + C content. The homology of the three sampling points was 99.84% -99.96%. Comparing the DNA sequence of the obtained sea cucumber with that of GenBank in eight outlying populations of the East Pacific, it was found that the sequences of sea cucumber from different orders and families were significantly different. DNAsp was used to calculate the number of each loci. MEGA2.1 was used to analyze the base composition and genetic distance, and a phylogenetic tree was constructed. The phylogenetic class relationships are exactly the same as the morphological classification of these species.