Mata-Somarribas Carlos, Cardoso das Graças Grazielle, de Oliveira R Pereira Luiza, Côrtes Boité Mariana, Motta Cantanhêde Lilian, Braga Filgueira Camila Patrício, Fallas Adrián, Quirós-Rojas Leana, Morelli Karina A, Ferreira Gabriel Eduardo Melim, Cupolillo Elisa
Centro Nacional de Referencia de Parasitología, Instituto Costarricense de Investigación y Enseñanza en Nutrición y Salud, Cartago, Costa Rica.
Hospital Universitário Gaffrée e Guinle-Universidade Federal do Estado do Rio de Janeiro, Rio de Janeiro, RJ, Brasil.
PLoS One. 2024 Dec 2;19(12):e0309277. doi: 10.1371/journal.pone.0309277. eCollection 2024.
Species delimitation has always been a challenge for taxonomists and for Leishmania studies there is no exception. Herein we attempt to display the usefulness of the mitochondrial gene Cytochrome Oxidase I-coI in classical and barcode-based approaches for Leishmania characterization. A total of 228 samples were analyzed, comprising 28 Leishmania related taxa, mainly from cultures of the Oswaldo Cruz Foundation`s Leishmania Collection. Primers were designed for amplification of coI; sequences were analyzed by distance-based indicators and both the Neighbor Joining and NeighborNet as species grouping techniques. Automatic Barcode Gap Discovery was applied to define species delimitation while for the character-based analysis a software for Barcoding with Logic formulas was employed. Final sequences of 486 bp with 238 parsimonious sites were aligned and edited. Robust groups were formed for most of the genus species, distinctive nucleotide positions in the barcode sequence were observed for 11 of them. A good agreement between the techniques applied and the original characterization was observed. Few species were not distinguished by coI: (i) L. (V.) peruviana, L. (V.) lindenbergi, and L. (V.) utingensis; (ii) L. (L.) venezuelensis and (iii) L. colombiensis and L. equatorensis with identical sequences. Some of these taxa have been, at one time or another, classified as controversial and, for most of them, a higher number of isolates should be studied to properly infer their taxonomic status. CoI represents a mitochondrial target that stands out as a taxonomically important asset with multiple advantages over other genes. This paper corresponds to the first report of coI analysis in Leishmania, a potentially advantageous target for the characterization of this parasite.
物种界定一直是分类学家面临的挑战，利什曼原虫研究也不例外。在此，我们试图展示线粒体基因细胞色素氧化酶I（coI）在利什曼原虫经典鉴定方法和基于条形码技术的鉴定方法中的作用。共分析了228个样本，包括28个与利什曼原虫相关的分类单元，主要来自奥斯瓦尔多·克鲁兹基金会利什曼原虫保藏中心的培养物。设计引物用于扩增coI；通过基于距离的指标以及邻接法和邻域网法这两种物种分组技术对序列进行分析。应用自动条形码间隙发现法来定义物种界定，而基于特征的分析则使用了带有逻辑公式的条形码软件。对486 bp、具有238个简约位点的最终序列进行了比对和编辑。大多数属种形成了稳健的类群，其中11个在条形码序列中观察到了独特的核苷酸位置。观察到所应用的技术与原始鉴定结果之间有很好的一致性。少数物种无法通过coI区分：（i）秘鲁利什曼原虫、林登伯格利什曼原虫和乌廷根利什曼原虫；（ii）委内瑞拉利什曼原虫；（iii）哥伦比亚利什曼原虫和赤道利什曼原虫，它们具有相同的序列。这些分类单元中的一些曾在不同时期被归类为有争议的，并且对于它们中的大多数，应该研究更多的分离株以正确推断其分类地位。CoI代表一个线粒体靶点，作为一种在分类学上重要的资产脱颖而出，相对于其他基因具有多种优势。本文是利什曼原虫coI分析的首次报告，coI是该寄生虫鉴定的一个潜在优势靶点。
