烟台海域暴发浒苔ITS及5.8S rDNA的克隆及序列分析

Cloning and sequence analysis of the ITS and 5.8S rDNA of Enteromorpha prolifera forming green tide around the coast of Yantai

以烟台海域引发"绿潮"的浒苔为研究对象,采用PCR技术扩增出浒苔的ITS-1、5.8SrDNA及ITS-2片段,将扩增出的片段纯化后克隆至pGEM-TEasy载体,筛选阳性克隆进行序列测定。结果表明,浒苔的ITS-1序列长度为195bp,5.8S序列为155bp,ITS-2序列为181bp,该序列与浒苔属的多种物种ITS序列具有很高的同源性,在ITS-1区、5.8SrDNA区和ITS-2区仅存在4个转换/颠换位点。结合GenBank注册序列和本研究的结果发现,单纯依靠ITS序列并不能对浒苔属种类进行有效的分类鉴定。

The internal transcribed spacer and 5.8S rDNA of Enteromorpha prolifera forming green tide around the coast of Yantai were amplified by polymerase chain reaction （PCR） . The resultant PCR product was cloned into pGEM-T Easy vector and subjected to nucleotide sequencing. Sequence analysis revealed that the ITS-1, 5.8 S and ITS-2 rDNA ofE. prolifera were of 195 bp, 155 bp and 181 bp in length, respectively. The Enteromorphaprolifera ITS sequence shared significant homology with other reported ITS sequences of Enteromorpha with only one transition site and three transversion sites. These results suggested that analysis of ITS sequence could not effectively distinguish the species of Enteromorpha.