Department of Helminthology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
CNRS ISEM-CIRAD ASTRE, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand.
Parasit Vectors. 2020 Jul 20;13(1):364. doi: 10.1186/s13071-020-04242-8.
Molecular advances have accelerated our understanding of nematode systematics and taxonomy. However, comparative analyzes between various genetic markers have led to discrepancies in nematode phylogenies. This study aimed to evaluate the suitability of using mitochondrial 12S and 16S ribosomal RNA genes for nematode molecular systematics.
To study the suitability of mitochondrial 12S and 16S ribosomal RNA genes as genetic markers for nematode molecular systematics, we compared them with the other commonly used genetic markers, nuclear internal transcribed spacer 1 and 2 regions, nuclear 18S and 28S ribosomal RNA genes, and mitochondrial cytochrome c oxidase subunit 1 gene. After that, phylum-wide primers for mitochondrial 12S and 16S ribosomal RNA genes were designed, and parasitic nematodes of humans and animals from 75 taxa with 21 representative species were inferred through phylogenetic analyzes. Phylogenetic analyzes were carried out using maximum likelihood and Bayesian inference algorithms.
The phylogenetic relationships of nematodes based on the mitochondrial 12S rRNA gene supported the monophyly of nematodes in clades I, IV, and V, reinforcing the potential of this gene as a genetic marker for nematode systematics. In contrast, the mitochondrial 16S rRNA gene only supported the monophyly of clades I and V, providing evidence that the 12S rRNA gene is more suitable for nematode molecular systematics. In this study, subclades of clade III containing various nematode families were not monophyletic when the 16S or 12S rRNA gene was used as the genetic marker. This is similar to the phylogenetic relationship revealed by previous studies using whole mitochondrial genomes as genetic markers.
This study supports the use of the 12S rRNA gene as a genetic marker for studying the molecular systematics of nematodes to understand intra-phyla relationships. Phylum-wide primers for nematodes using mitochondrial ribosomal genes were prepared, which may enhance future studies. Furthermore, sufficient genetic variation in the mitochondrial 12S and 16S rRNA genes between species also allowed for accurate taxonomy to species level, revealing the potential of these two genes as genetic markers for DNA barcoding.
分子进展加速了我们对线虫系统发生和分类学的理解。然而,各种遗传标记的比较分析导致了线虫系统发育的差异。本研究旨在评估线粒体 12S 和 16S 核糖体 RNA 基因用于线虫分子系统发生的适宜性。
为了研究线粒体 12S 和 16S 核糖体 RNA 基因作为线虫分子系统发生遗传标记的适宜性,我们将其与其他常用遗传标记(核内转录间隔区 1 和 2 区、核 18S 和 28S 核糖体 RNA 基因以及线粒体细胞色素 c 氧化酶亚基 1 基因)进行了比较。之后,设计了用于线粒体 12S 和 16S 核糖体 RNA 基因的门-wide 引物,并通过系统发育分析推断了来自 75 个分类群的 21 个代表性物种的人类和动物寄生线虫。系统发育分析采用最大似然法和贝叶斯推断算法进行。
基于线粒体 12S rRNA 基因的线虫系统发育关系支持线虫在类群 I、IV 和 V 中的单系性,这增强了该基因作为线虫系统发生遗传标记的潜力。相比之下,线粒体 16S rRNA 基因仅支持类群 I 和 V 的单系性,这表明 12S rRNA 基因更适合线虫分子系统发生。在本研究中,当使用 16S 或 12S rRNA 基因作为遗传标记时,包含各种线虫科的类群 III 的亚群不是单系的。这与之前使用全线粒体基因组作为遗传标记进行的系统发育研究的结果相似。
本研究支持使用 12S rRNA 基因作为遗传标记来研究线虫的分子系统发生,以了解属内关系。本研究还制备了使用线粒体核糖体基因的线虫门-wide 引物,这可能会促进未来的研究。此外,种间线粒体 12S 和 16S rRNA 基因的充分遗传变异也允许对物种水平进行准确的分类,揭示了这两个基因作为 DNA 条形码遗传标记的潜力。