Zhang Hanyong, Okii Erika, Gotoh Eiji, Shiraishi Susumu
Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
J Nematol. 2018;50(3):281-302. doi: 10.21307/jofnem-2018-034.
Mitogenomic diversity and genetic population structure of the pinewood nematode (PWN) Bursaphelenchus xylophilus inhabiting Kyushu, Japan were analyzed. A method for performing long PCR using single nematodes and sequencing nematode mitochondrial genomes individually is presented here. About 8 kb (∼55%) of the complete mitochondrial genome was successfully obtained from 285 individuals collected from 12 populations. The 158 single nucleotide polymorphisms detected corresponded to 30 haplotypes, clearly classified into two clades. Haplotype diversity was 0.83, evidencing a remarkable high diversity within Kyushu. The high genetic differentiation among the 12 populations (0.331) might be due to past invasion and expansion routes of PWN in northeastern and southeastern Kyushu. The distinct genetic composition of populations within the northwestern, central western, and southwestern Kyushu seems to be mostly related to the extinction of pine forests and long-range migration of PWN due to human activity. Overall, direct long PCR and sequencing of single nematode individuals are effective methods for investigating mitochondrial polymorphisms, and these are effective tools for PWN population genetics and other intraspecific studies.
分析了栖息于日本九州的松材线虫（PWN）松材线虫（Bursaphelenchus xylophilus）的线粒体基因组多样性和遗传种群结构。本文介绍了一种使用单个线虫进行长片段PCR并单独对线虫线粒体基因组进行测序的方法。从12个种群收集的285个个体中成功获得了约8 kb（约55%）的完整线粒体基因组。检测到的158个单核苷酸多态性对应于30个单倍型，明显分为两个分支。单倍型多样性为0.83，表明九州地区具有显著的高多样性。12个种群之间的高遗传分化（0.331）可能是由于松材线虫过去在九州东北部和东南部的入侵和扩张路线所致。九州西北部､中西部和西南部种群独特的遗传组成似乎主要与松林的灭绝以及由于人类活动导致的松材线虫远距离迁移有关。总体而言，直接对单个线虫个体进行长片段PCR和测序是研究线粒体多态性的有效方法，也是松材线虫种群遗传学和其他种内研究的有效工具。
