目的基于核糖体18S r DNA基因对基氏蠊螨进行分子鉴定。方法采集和分离储藏物样本,进行螨类的形态学鉴定。提取单个螨的基因组DNA,经PCR扩增、克隆和测序获得COⅠ基因和18S r DNA基因,将所获序列进行Blast对比。检索Gen Bank数据库中蠊...目的基于核糖体18S r DNA基因对基氏蠊螨进行分子鉴定。方法采集和分离储藏物样本,进行螨类的形态学鉴定。提取单个螨的基因组DNA,经PCR扩增、克隆和测序获得COⅠ基因和18S r DNA基因,将所获序列进行Blast对比。检索Gen Bank数据库中蠊螨属18S r DNA基因序列,利用Clustal X 1.83软件进行多序列比对,基于MEGA X软件进行序列分析并以邻接法(Neighbor-Joining,NJ)构建系统发育树。结果采集的样本经形态和COⅠ基因双重鉴定为基氏蠊螨。同时,所选取的10个基氏蠊螨的18S r DNA基因序列完全一致,均表现出A/T碱基偏向性,与同属的Blattisocius tarsalis和Blattisocius everti分别有98.73%和98.94%的同源性。基于18S r DNA基因序列的系统进化树显示,基氏蠊螨与Blattisocius tarsalis和Blattisocius everti聚为一支。结论本研究建立了基氏蠊螨基于18S r DNA基因序列的分子鉴定方法,为基氏蠊螨的准确鉴定奠定基础。展开更多
Previous studies found intraspecific diversity in Scrippsiella trochoidea A. R. Loeblich Ill, a widely distributed calcareous cyst-producing dinoflagellate. In this study, three strains （ST-I, ST-D6 and ST-K） of S. ...Previous studies found intraspecific diversity in Scrippsiella trochoidea A. R. Loeblich Ill, a widely distributed calcareous cyst-producing dinoflagellate. In this study, three strains （ST-I, ST-D6 and ST-K） of S. trochoidea isolated from the East Asian waters were studied, together with other geographical strains, to resolve their phylogenetic relationships. For the three East Asian isolates, two highly diverse regions of nuclear-encoded ribosomal DNA （rDNA）, the 5.8S rDNA and its flanking internal transcribed spacers 1 and 2, and the 5＇ portion of the large-subunit rDNA （encompassing the ＂DI＂ and ＂D2＂ domains） were sequenced. Homologous sequences from other geographical isolates were selected from the GenBank database and the phylogenetic relationships were inferred from the molecular data of these strains. Strains of S. trochoidea were found to cluster into three major clades （STR1, STR2 and STR3）, as reported in previous studies. Two of the three strains ST-1 and ST-K, were grouped in clade STR2, the other strain, ST-D6, belonged to clade STR3. The intraspecific diversity ofS. trochoidea in East Asian waters makes it necessary to carry out phylogenetic investigations in combination with data of biogeography, population dynamics, and life cycle on the ecophysiology of a region.展开更多
文摘目的基于核糖体18S r DNA基因对基氏蠊螨进行分子鉴定。方法采集和分离储藏物样本,进行螨类的形态学鉴定。提取单个螨的基因组DNA,经PCR扩增、克隆和测序获得COⅠ基因和18S r DNA基因,将所获序列进行Blast对比。检索Gen Bank数据库中蠊螨属18S r DNA基因序列,利用Clustal X 1.83软件进行多序列比对,基于MEGA X软件进行序列分析并以邻接法(Neighbor-Joining,NJ)构建系统发育树。结果采集的样本经形态和COⅠ基因双重鉴定为基氏蠊螨。同时,所选取的10个基氏蠊螨的18S r DNA基因序列完全一致,均表现出A/T碱基偏向性,与同属的Blattisocius tarsalis和Blattisocius everti分别有98.73%和98.94%的同源性。基于18S r DNA基因序列的系统进化树显示,基氏蠊螨与Blattisocius tarsalis和Blattisocius everti聚为一支。结论本研究建立了基氏蠊螨基于18S r DNA基因序列的分子鉴定方法,为基氏蠊螨的准确鉴定奠定基础。
基金Supported by the Knowledge Innovation Program of Chinese Academy of Sciences （No. KZCX2-YW-208） and National Natural Science Foundation of China （No. 40676072）
文摘Previous studies found intraspecific diversity in Scrippsiella trochoidea A. R. Loeblich Ill, a widely distributed calcareous cyst-producing dinoflagellate. In this study, three strains （ST-I, ST-D6 and ST-K） of S. trochoidea isolated from the East Asian waters were studied, together with other geographical strains, to resolve their phylogenetic relationships. For the three East Asian isolates, two highly diverse regions of nuclear-encoded ribosomal DNA （rDNA）, the 5.8S rDNA and its flanking internal transcribed spacers 1 and 2, and the 5＇ portion of the large-subunit rDNA （encompassing the ＂DI＂ and ＂D2＂ domains） were sequenced. Homologous sequences from other geographical isolates were selected from the GenBank database and the phylogenetic relationships were inferred from the molecular data of these strains. Strains of S. trochoidea were found to cluster into three major clades （STR1, STR2 and STR3）, as reported in previous studies. Two of the three strains ST-1 and ST-K, were grouped in clade STR2, the other strain, ST-D6, belonged to clade STR3. The intraspecific diversity ofS. trochoidea in East Asian waters makes it necessary to carry out phylogenetic investigations in combination with data of biogeography, population dynamics, and life cycle on the ecophysiology of a region.
