Evaluation of ITS2-28S as a molecular marker for identification of calanoid copepods in the subtropical western North Pacific
Evaluation of ITS2-28S as a molecular marker for identification of calanoid copepods in the subtropical western North Pacific
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DOI：
10.1093/plankt/fbt016
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发表时间：
2013-05-01
影响因子：
2.1
通讯作者：
Tsuda, Atsushi
中科院分区：
文献类型：
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作者：
Hirai, Junya;Shimode, Shinji;Tsuda, Atsushi
我们评价了内转录间隔区2和大亚基核糖体DNA(ITS2-28S)区域作为识别北太平洋西部亚热带甲鱼类桡足类的分子标记。244人中有232人成功扩增出ITS2-28S序列，其成功率远高于细胞色素氧化酶I的扩增成功率(77/244人)。用一对单引物共扩增出194个ITS2-28S序列。通过对ITS2-28S序列的系统发育分析，发现该基因具有高度的遗传变异性，分别存在于黄角夜蛾(N 13)、小褐斑潜蝇(N 9)、腹侧乳杆菌(N 9)和脊椎鱼(N 2)中，并表现出多种序列类型。观察到与两种序列类型相对应的身体大小的差异。利用每个科内810968个碱基的ITS2-28S序列，其他物种的种内遗传变异(00.001)始终低于种间遗传变异(0.005)。虽然有必要对更大的样本量进行进一步的形态和遗传分析，但我们的结果表明，ITS2-28S是一个不需要克隆的核标记，它很容易扩增，并且有足够的可变性，可以作为DNA条形码的标记或识别尺足类甲鱼的遗传群体。
We evaluated the internal transcribed spacer 2 and large subunit ribosomal DNA (ITS2-28S) region as a molecular marker for identifying calanoid copepods in the subtropical western North Pacific. The ITS2-28S sequence was successfully amplified in 232/244 individuals, a much higher rate of success than for cytochrome oxidase I amplification (77/244 individuals). A total of 194 sequences of ITS2-28S were obtained using a single primer pair. A high degree of genetic variability in ITS2-28S was observed in Lucicutia flavicornis (n 13), Nannocalanus minor (n 9), Pleuromamma abdominalis (n 9) and Spinocalanus spinosus (n 2), each showing several sequence types based on phylogenetic analysis of the ITS2-28S. A difference in the body size that corresponded with two sequence types of P. abdominalis was observed. The other species showed consistently lower genetic variability within species (00.001) than between species (0.005) using ITS2-28S of 810968 bp aligned in each family. Although further morphological and genetic analysis of a larger sample size are necessary, our results suggest that ITS2-28S is a nuclear marker that does not need to be cloned and is easily amplified and sufficiently variable as a possible marker for DNA barcoding or to identify genetic groups of calanoid copepods.