CNRS-UPS-ENFA, UMR5174, EDB (Laboratoire Évolution & Diversité biologique), 118, route de Narbonne, 31062 Toulouse, France.
University of Leipzig, Interdisciplinary Center for Bioinformatics, Härtelstraße 16-18, 04107 Leipzig, Germany.
C R Biol. 2014 May;337(5):295-301. doi: 10.1016/j.crvi.2014.03.003. Epub 2014 Apr 4.
Little is known about the variations of nematode mitogenomes (mtDNA). Sequencing a complete mtDNA using a PCR approach remains a challenge due to frequent genome reorganizations and low sequence similarities between divergent nematode lineages. Here, a genome skimming approach based on HiSeq sequencing (shotgun) was used to assemble de novo the first complete mtDNA sequence of a root-knot nematode (Meloidogyne graminicola). An AT-rich genome (84.3%) of 20,030 bp was obtained with a mean sequencing depth superior to 300. Thirty-six genes were identified with a semi-automated approach. A comparison with a gene map of the M. javanica mitochondrial genome indicates that the gene order is conserved within this nematode lineage. However, deep genome rearrangements were observed when comparing with other species of the superfamily Hoplolaimoidea. Repeat elements of 111 bp and 94 bp were found in a long non-coding region of 7.5 kb, as similarly reported in M. javanica and M. hapla. This study points out the power of next generation sequencing to produce complete mitochondrial genomes, even without a reference sequence, and possibly opening new avenues for species/race identification, phylogenetics and population genetics of nematodes.
关于线虫线粒体基因组(mtDNA)的变异,人们知之甚少。由于基因组频繁重组以及不同线虫谱系之间的序列相似度低,使用 PCR 方法对完整 mtDNA 进行测序仍然是一项挑战。在这里,我们采用基于 HiSeq 测序(鸟枪法)的基因组掠取方法,从头组装了根结线虫(Meloidogyne graminicola)的首个完整 mtDNA 序列。获得了一个富含 AT(84.3%)的基因组,大小为 20,030 bp,平均测序深度超过 300。使用半自动方法鉴定了 36 个基因。与 M. javanica 线粒体基因组的基因图谱进行比较表明,该基因在这个线虫谱系内的排列是保守的。然而,与超级家族 Hoplolaimoidea 的其他物种相比,观察到了深度的基因组重排。在 7.5 kb 的长非编码区中发现了 111 bp 和 94 bp 的重复元件,这与 M. javanica 和 M. hapla 中的报道类似。这项研究表明,即使没有参考序列,下一代测序也有能力产生完整的线粒体基因组,并可能为线虫的物种/种群鉴定、系统发育和群体遗传学开辟新的途径。