PDF(Pubmed)
Abstract：
UNASSIGNED: Barbronia, a genus of freshwater macrophagous leeches, belongs to Erpobdelliformes (Salifidae: Clitellata: Annelida), and B. weberi, a well-known leech within this genus, has a worldwide distribution. However, the systematics of Barbronia have not yet been adequately investigated, primarily due to a few molecular markers, and only 20 Barbronia sequences available in the GenBank database. This gap significantly limits our understanding of the Barbronia species identification, as well as the phylogenetic placement of the genus Barbronia within Salifidae.
UNASSIGNED: Next-generation sequencing (NGS) was used to simultaneously capture the entire mitochondrial genome and the full-length 18S/28S rDNA sequences. The species boundary of Barbronia species was estimated using bGMYC and bPTP methods, based on all available Barbronia COI sequences. Uncorrected COI p-distance was calculated in MEGA. A molecular data matrix consisting of four loci (COI, 12S, 18S, and 28S rDNA) for outgroups (three Haemopis leeches) and 49 erpobdellid leeches, representing eight genera within the Suborder Erpobdelliformes was aligned using MAFFT and LocARNA. This matrix was used to reconstruct the phylogenetic relationship of Barbronia via Bayesian inference (BI) and the maximum likelihood (ML) method.
UNASSIGNED: The full lengths of the mitochondrial genome, 18S and 28S rDNAs of B. cf. gwalagwalensis, are 14847 bp, 1876 bp 1876 bp, and 2863 bp, respectively. Both bGMYC and bPTP results based on COI data are generally congruent, suggesting that the previously proposed taxa (B. arcana, B. weberi formosana, and B. wuttkei or Erpobdella wuttkei) are synonyms of B. weberi. The specimens listed in the B. gwalagwalensis group, however, are split into at least two Primary Species Hypotheses (PSHs). The p-distance of the first PSH is less than 1.3% but increased to 4.5% when including the secondary PSH (i.e., B. cf. gwalagwalensis). In comparison, the interspecific p-distance between the B. weberi group and the B. gwalagwalensis group ranged from 6.4% to 8.7%, and the intraspecific p-distance within the B. weberi group is less than 0.8%. Considering the species delimitation results and the sufficient large p-distance, the specimen sampled in China is treated as B. cf. gwalagwalensis. The monophyly of the four Erpobdelliformes families Salifidae, Orobdellidae, Gastrostomobdellidae sensu stricto and Erpobdellidae is well supported in ML and BI analysis based on a data of four markers. Within the Salifidae, a well-supported Barbronia is closely related to a clade containing Odontobdella and Mimobdella, and these three genera are sister to a clade consisted of Salifa and Linta. According to the results of this study, the strategy of simultaneous obtaining both whole mitochondria and nuclear markers from extensively sampled Salifids species using NGS is expected to fathom both the species diversity of B. gwalagwalensis and the evolutionary relationship of Salifidae.
UNASSIGNED: Next-generation sequencing (NGS) was used to simultaneously capture the entire mitochondrial genome and the full-length 18S/28S rDNA sequences. The species boundary of Barbronia species was estimated using bGMYC and bPTP methods, based on all available Barbronia COI sequences. Uncorrected COI p-distance was calculated in MEGA. A molecular data matrix consisting of four loci (COI, 12S, 18S, and 28S rDNA) for outgroups (three Haemopis leeches) and 49 erpobdellid leeches, representing eight genera within the Suborder Erpobdelliformes was aligned using MAFFT and LocARNA. This matrix was used to reconstruct the phylogenetic relationship of Barbronia via Bayesian inference (BI) and the maximum likelihood (ML) method.
UNASSIGNED: The full lengths of the mitochondrial genome, 18S and 28S rDNAs of B. cf. gwalagwalensis, are 14847 bp, 1876 bp 1876 bp, and 2863 bp, respectively. Both bGMYC and bPTP results based on COI data are generally congruent, suggesting that the previously proposed taxa (B. arcana, B. weberi formosana, and B. wuttkei or Erpobdella wuttkei) are synonyms of B. weberi. The specimens listed in the B. gwalagwalensis group, however, are split into at least two Primary Species Hypotheses (PSHs). The p-distance of the first PSH is less than 1.3% but increased to 4.5% when including the secondary PSH (i.e., B. cf. gwalagwalensis). In comparison, the interspecific p-distance between the B. weberi group and the B. gwalagwalensis group ranged from 6.4% to 8.7%, and the intraspecific p-distance within the B. weberi group is less than 0.8%. Considering the species delimitation results and the sufficient large p-distance, the specimen sampled in China is treated as B. cf. gwalagwalensis. The monophyly of the four Erpobdelliformes families Salifidae, Orobdellidae, Gastrostomobdellidae sensu stricto and Erpobdellidae is well supported in ML and BI analysis based on a data of four markers. Within the Salifidae, a well-supported Barbronia is closely related to a clade containing Odontobdella and Mimobdella, and these three genera are sister to a clade consisted of Salifa and Linta. According to the results of this study, the strategy of simultaneous obtaining both whole mitochondria and nuclear markers from extensively sampled Salifids species using NGS is expected to fathom both the species diversity of B. gwalagwalensis and the evolutionary relationship of Salifidae.
摘要：
■巴布罗尼亚，淡水巨噬细胞水蛭属，属于Erpobdelliformes（Salifidae：Clitellata：Annelida），B.Weberi,这个属中著名的水蛭，具有全球分布。然而,Barbronia的系统尚未得到充分的研究，主要是由于一些分子标记，GenBank数据库中只有20个Barbronia序列。这种差距极大地限制了我们对Barbronia物种鉴定的理解，以及Barbronia属在Salifidae中的系统发育位置。
■下一代测序（NGS）用于同时捕获整个线粒体基因组和全长18S/28SrDNA序列。使用bGMYC和bPTP方法估算了Barbronia物种的物种边界，基于所有可用的BarbroniaCOI序列。在MEGA中计算未校正的COIp-距离。由四个基因座（COI，12S,18S,和28SrDNA）用于外群（三个Haemopis水ches）和49个erpobdellid水ches，使用MAFFT和LocARNA对代表亚阶Erpobdelliformes中的八个属进行了比对。该矩阵用于通过贝叶斯推理（BI）和最大似然（ML）方法重建Barbronia的系统发育关系。
■线粒体基因组的全长，B.cf的18S和28SrDNA。gwalagwalensis,是14847个基点，1876bp1876bp，和2863个基点，分别。基于COI数据的bGMYC和bPTP结果通常是一致的，表明先前提出的分类单元(B.arcana,B.weberiformosana,和B.wuttkei或Erpobdellawuttkei）是B.weberi的同义词。B.gwalagwalensis组中列出的标本，然而,分为至少两个主要物种假设(PSHs)。第一个PSH的p距离小于1.3%，但在包括次要PSH时增加到4.5%(即，B.cf.gwalagwalensis）。相比之下,B.weberi组和B.gwalagwalensis组之间的种间p距离为6.4％至8.7％，并且B.weberi组内的种内p距离小于0.8%。考虑到物种定界结果和足够大的p距离，在中国采样的标本被视为B.cf。gwalagwalensis.四个Erpobdelliformes家族的单系，Orobdellidae,基于四个标记物的数据，在ML和BI分析中很好地支持了严格的胃杆菌和Erpobdellidae。在Salifidae中，一个支持良好的巴布罗尼亚与一个含有奥登托贝拉和米诺贝拉的进化枝密切相关，这三个属是由Salifa和Linta组成的进化枝的姐妹。根据这项研究的结果,使用NGS从广泛采样的Salifids物种中同时获得整个线粒体和核标记的策略有望同时了解B.gwalagwalensis的物种多样性和Salifidae的进化关系。
■下一代测序（NGS）用于同时捕获整个线粒体基因组和全长18S/28SrDNA序列。使用bGMYC和bPTP方法估算了Barbronia物种的物种边界，基于所有可用的BarbroniaCOI序列。在MEGA中计算未校正的COIp-距离。由四个基因座（COI，12S,18S,和28SrDNA）用于外群（三个Haemopis水ches）和49个erpobdellid水ches，使用MAFFT和LocARNA对代表亚阶Erpobdelliformes中的八个属进行了比对。该矩阵用于通过贝叶斯推理（BI）和最大似然（ML）方法重建Barbronia的系统发育关系。
■线粒体基因组的全长，B.cf的18S和28SrDNA。gwalagwalensis,是14847个基点，1876bp1876bp，和2863个基点，分别。基于COI数据的bGMYC和bPTP结果通常是一致的，表明先前提出的分类单元(B.arcana,B.weberiformosana,和B.wuttkei或Erpobdellawuttkei）是B.weberi的同义词。B.gwalagwalensis组中列出的标本，然而,分为至少两个主要物种假设(PSHs)。第一个PSH的p距离小于1.3%，但在包括次要PSH时增加到4.5%(即，B.cf.gwalagwalensis）。相比之下,B.weberi组和B.gwalagwalensis组之间的种间p距离为6.4％至8.7％，并且B.weberi组内的种内p距离小于0.8%。考虑到物种定界结果和足够大的p距离，在中国采样的标本被视为B.cf。gwalagwalensis.四个Erpobdelliformes家族的单系，Orobdellidae,基于四个标记物的数据，在ML和BI分析中很好地支持了严格的胃杆菌和Erpobdellidae。在Salifidae中，一个支持良好的巴布罗尼亚与一个含有奥登托贝拉和米诺贝拉的进化枝密切相关，这三个属是由Salifa和Linta组成的进化枝的姐妹。根据这项研究的结果,使用NGS从广泛采样的Salifids物种中同时获得整个线粒体和核标记的策略有望同时了解B.gwalagwalensis的物种多样性和Salifidae的进化关系。
