rDNA

rDNA
  • 文章类型: Journal Article
    全球范围内,家禽业受到各种艾美耳球虫引起的球虫病的严重威胁。这种原生动物寄生虫栖息在全球家禽胃肠道的上皮内衬中,并可引起严重的临床疾病。本研究是在克什米尔各个地区的家禽养殖场进行的,印度,调查影响肉鸡的艾美球虫种类的患病率和系统发育关系。在一年的时间里,从克什米尔的60个家禽养殖场收集粪便样本,并采用形态学和分子技术进行艾美球虫物种鉴定。结果显示球虫病的患病率很高,58.3%(35/60)的农场对艾美球虫呈阳性。最流行的物种是E.tenella(31/35,88.6%),其次是E.acervulina(25/35,71.4%),E.最大值(19/35,54.3%),E.米蒂斯(18/35,51.4%),和E.necatrix(9/35,25.7%)。还观察到患病率的季节性变化,秋季(86.7%)和夏季(66.7%)比率最高。此外,较年轻的鸟类(3-4周)的感染率(85.7%)高于较年长的鸟类(57.9%)(5-6周)。在94.2%(33/35)的阳性农场发现混合感染。使用ITS1序列进行的系统发育分析证实了物种聚类,并揭示了艾美球虫物种之间的进化关系。E.tenella和E.necatrix形成了一个不同的进化枝,而E.acervulina形成了另一个。该研究强调了分子技术在准确物种鉴定中的重要性,并为克什米尔家禽球虫病的流行病学提供了宝贵的见解。有效的控制策略,包括疫苗接种和改进的管理实践,有必要减轻与这种广泛的家禽疾病相关的经济损失。
    Globally, the poultry industry is seriously threatened by coccidiosis caused by various species of Eimeria. This protozoan parasite inhabits the epithelial lining of the gastrointestinal tract of poultry globally and can cause serious clinical disease. The present study was carried out on poultry farms located in various regions of Kashmir, India, to investigate the prevalence and phylogenetic relationships of Eimeria species affecting broiler chickens. Over a period of one year, fecal samples were collected from 60 poultry farms in Kashmir and morphological and molecular techniques were employed for Eimeria species identification. Results revealed a high prevalence of coccidiosis, with 58.3% (35/60) of farms positive for Eimeria. The most prevalent species were E. tenella (31/35, 88.6%) followed by E. acervulina (25/35, 71.4%), E. maxima (19/35, 54.3%), E. mitis (18/35, 51.4%), and E. necatrix (9/35, 25.7%). Seasonal variation in prevalence was also observed, with the highest rates in autumn (86.7%) and summer (66.7%). Additionally, younger birds (3-4 weeks) exhibited higher infection rates (85.7%) compared to older birds (57.9%) (5-6 weeks). Mixed infection was found in 94.2% (33/35) of positive farms. Phylogenetic analysis using ITS1 sequences confirmed species clustering and revealed evolutionary relationships among Eimeria species. E. tenella and E. necatrix formed a distinct clade, while E. acervulina formed another. The study underscores the importance of molecular techniques in accurate species identification and provides valuable insights into the epidemiology of coccidiosis in poultry in Kashmir. Effective control strategies, including vaccination and improved management practices, are necessary to mitigate the economic losses associated with this widespread poultry disease.
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  • 文章类型: Journal Article
    Stenocranius亚属包含两个隐秘的物种:Lasiopodomysgregalis(细分为三个异样分布和遗传分离良好的谱系A,B,和C)和Lasiopodomysraddei。为了确定这种研究较少的隐匿物种复合体的核型特征,我们对来自南西伯利亚和蒙古41个地区的138个个体进行了比较细胞遗传学分析.首次提供了RaddeiL.核型和GregalisL.谱系的核型的详细描述。所有检查过的窄头田鼠的A染色体互补物由2n=36组成,常染色体臂的基本数量(FNa)为50。物种之间,差异染色的模式相似,尽管在L.gregalis谱系中发现了其他C-异色块;Ag阳性核仁组织者和核糖体DNA(rDNA)簇位于8个和9个肢端中心对上,分别。在早更新世遗物L.raddei中未发现B染色体(Bs),虽然在所有L.gregalis谱系中检测到一到五个小的异色顶心Bs;Bs的数量和频率在谱系内变化很大,但没有观察到个体内变异。在这两个物种中,端粒重复在所有染色体的末端可见,包括B.Bs上rDNA簇的数量和定位因B携带者而异。对几种减数分裂蛋白的免疫检测表明,meio-Bs在转录上无活性,并且具有与性染色体相似的减数分裂行为模式(假定Bs与性染色体具有某些同源性)。大自然,L.gregalis中Bs的遗传和稳定性机制需要进一步研究。
    The subgenus Stenocranius contains two cryptic species: Lasiopodomys gregalis (subdivided into three allopatrically distributed and genetically well-isolated lineages A, B, and C) and Lasiopodomys raddei. To identify karyotype characteristics of this poorly studied cryptic species complex, we used comparative cytogenetic analysis of 138 individuals from 41 localities in South Siberia and Mongolia. A detailed description of the L. raddei karyotype and of the L. gregalis lineage С karyotype is presented for the first time. The A chromosome complement of all examined narrow-headed voles consisted of 2n = 36 and a fundamental number of autosomal arms (FNa) of 50. Between species, patterns of differential staining were similar, though additional C-heterochromatic blocks were found in L. gregalis lineages; Ag-positive nucleolar organizers and ribosomal DNA (rDNA) clusters are located on eight and nine acrocentric pairs, respectively. No B chromosomes (Bs) were found in the Early Pleistocene relic L. raddei, while one to five small heterochromatic acrocentric Bs were detected in all L. gregalis lineages; the number and frequency of Bs varied considerably within lineages, but no intraindividual variation was observed. In both species, telomeric repeats were visualized at termini of all chromosomes, including Bs. The number and localization of rDNA clusters on Bs varied among B-carriers. Immunodetection of several meiotic proteins indicated that meio-Bs are transcriptionally inactive and have a pattern of meiotic behavior similar to that of sex chromosomes (some homology of Bs to sex chromosomes is supposed). The nature, mechanisms of inheritance and stability of Bs in L. gregalis require further investigation.
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  • 文章类型: Journal Article
    微自噬通过直接吞噬液泡膜降解液泡中的货物。微核自噬选择性地降解出芽酵母中的一部分细胞核。液泡通过核-液泡连接(NVJ)与细胞核接触,在微核吞噬中,含有核仁蛋白的一部分细胞核突入NVJ的液泡中,其次是脱落和退化。营养饥饿后雷帕霉素复合物1(TORC1)蛋白激酶靶蛋白失活可诱导微自噬和微核吞噬。这里,我们表明,VAMP相关蛋白(VAP)Scs2及其旁系Scs22是NVJ完整性和核仁蛋白微核吞噬降解所必需的。另一方面,在VAP突变细胞中,微核自噬的核仁动力学前提没有受到损害。最后,酵母VAP对长期营养饥饿期间的生存能力至关重要。这项研究揭示了VAP在适应营养饥饿反应中的新兴作用。
    Microautophagy degrades cargos in the vacuole by direct engulfment of the vacuolar membrane. Micronucleophagy selectively degrades a portion of the nucleus in budding yeast. The vacuole contacts the nucleus via the nucleus-vacuole junction (NVJ), and in micronucleophagy a portion of the nucleus containing nucleolar proteins is made to protrude into the vacuole at the NVJ, followed by abscission and degradation. Microautophagy and micronucleophagy are induced by inactivation of target of rapamycin complex 1 (TORC1) protein kinase after nutrient starvation. Here, we show that the VAMP-associated proteins (VAPs) Scs2 and its paralog Scs22 are required for NVJ integrity and micronucleophagic degradation of nucleolar proteins. On the other hand, nucleolar dynamics prerequisite for micronucleophagy were not impaired in VAP mutant cells. Finally, yeast VAPs were critical for viability during prolonged nutrient starvation. This study sheds light on the emerging role of VAP in adaptation in responses to nutrient starvation.
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  • 文章类型: Journal Article
    淡水蟹(Potamiscusmanipuriensis),通常在曼尼普尔邦被当地人作为当地美食消费,被发现藏有Microphallussp。(微叶科),它们在形态上与早期报道的不同地区的小phi属的后cer虫不同。所以,利用rDNA标记区:较大的亚基(LSU)或28S(D1-D3区)和转录间间隔区2(ITS2)进行了基于PCR的这种后a的分子表征。序列和系统发育分析证实,所研究的分类单元属于小虫属的小虫科。
    Freshwater crabs (Potamiscus manipuriensis), commonly consumed as local delicacies by the native people in the state of Manipur, were found to harbour metacercariae of Microphallus sp. (Family Microphyllidae), which were morphologically different from metacercariae of Microphallus spp reported earlier from different regions. So, PCR-based molecular characterization of this metacercaria was done utilizing rDNA marker regions: larger subunit (LSU) or 28S (D1-D3 region) and inter-transcribed spacer 2 (ITS2). Sequence and phylogenetic analyses confirmed that the taxon under study belonged to family Microphyllidae of genus Microphallus.
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  • 文章类型: Journal Article
    有数百个rDNA拷贝,尚不清楚它们是否具有形成不同类型核糖体的序列变异。这里,我们开发了一种长读变体调用算法,称为RGA,这表明人类rDNA基因座的变异主要是插入缺失(indel)变体。我们开发了全长rRNA测序(RIBO-RT)和原位测序(SWITCH-seq),这表明翻译核糖体具有rRNA的变异。超过1,000个变体被低表达。然而,数十种变体丰富,并形成不同的rRNA亚型,在indel附近具有不同的结构,如通过长阅读rRNA结构探测与硫酸二甲酯测序相结合所揭示的。rRNA亚型在内胚层/外胚层来源的组织中显示差异表达,在癌症中,低丰度rRNA变体可以变得高度表达。一起,这项研究在rRNA变体水平上鉴定了核糖体的多样性,它们的染色体位置,和独特的结构以及核糖体变异与组织特异性生物学和癌症的联系。
    With hundreds of copies of rDNA, it is unknown whether they possess sequence variations that form different types of ribosomes. Here, we developed an algorithm for long-read variant calling, termed RGA, which revealed that variations in human rDNA loci are predominantly insertion-deletion (indel) variants. We developed full-length rRNA sequencing (RIBO-RT) and in situ sequencing (SWITCH-seq), which showed that translating ribosomes possess variation in rRNA. Over 1,000 variants are lowly expressed. However, tens of variants are abundant and form distinct rRNA subtypes with different structures near indels as revealed by long-read rRNA structure probing coupled to dimethyl sulfate sequencing. rRNA subtypes show differential expression in endoderm/ectoderm-derived tissues, and in cancer, low-abundance rRNA variants can become highly expressed. Together, this study identifies the diversity of ribosomes at the level of rRNA variants, their chromosomal location, and unique structure as well as the association of ribosome variation with tissue-specific biology and cancer.
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  • 文章类型: Journal Article
    核糖体DNA(rDNA)构成了物种内非常保守的DNA序列,位于核仁区域,负责编码三种主要类型的rRNA(18S,5.8S和28S)。虽然对rDNA的历史研究集中在其结构和编码能力上,最近的研究已经转向探索其在各种生物过程中的功能作用。在这次审查中,我们总结了rDNA甲基化与胚胎发育的主要发现,多种物种的衰老和疾病,包括表观遗传改变,rDNA甲基化的相关生物学过程和潜在应用。我们概述了当前的相关研究,并指出了该领域的差距。
    The ribosomal DNA (rDNA) constitutes a remarkably conserved DNA sequence within species, located in the area of the nucleolus, and responsible for coding three major types of rRNAs (18S, 5.8S and 28S). While historical investigations into rDNA focused on its structure and coding capabilities, recent research has turned to explore its functional roles in various biological processes. In this review, we summarize the main findings of rDNA methylation with embryonic development, aging and diseases in multiple species, including epigenetic alterations, related biological processes and potential applications of rDNA methylation. We present an overview of current related research and identify gaps in this field.
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  • 文章类型: Journal Article
    编码核糖体RNA的基因在整个生命中高度保守,并且在几乎所有的真核生物中都存在于称为rDNA的大型串联重复序列阵列中。rDNA重复单元大小在大多数真核生物中是保守的,但在哺乳动物中却急剧扩张,主要通过分离相邻rRNA编码区的基因间间隔区的扩展。在这里,我们使用来自主要羊水谱系代表的长读序列数据来确定羊水进化中rDNA单位大小增加的位置。我们发现,羊膜动物rDNA单位大小分为两个狭窄的大小类别:除单调外,所有羊膜动物中的“正常”(〜11-20kb),有袋动物和异形哺乳动物,具有“大”(35-45kb)尺寸。我们证实,基因间间隔区长度的增加解释了这种哺乳动物大小的增加,但是,与哺乳动物rDNA单位大小的一致性形成鲜明对比,哺乳动物基因间间隔区序列差异很大。这些结果表明,在哺乳动物祖先中发生了基因间间隔区大小的大量增加,尽管在哺乳动物进化过程中序列发生了实质性变化,但仍得以维持。这指向先前未认识到的对基因间间隔区长度的约束,一个被认为基本上是中立的地区。我们通过推测这种约束的可能原因来完成。
    The genes encoding ribosomal RNA are highly conserved across life and in almost all eukaryotes are present in large tandem repeat arrays called the rDNA. rDNA repeat unit size is conserved across most eukaryotes but has expanded dramatically in mammals, principally through the expansion of the intergenic spacer region that separates adjacent rRNA coding regions. Here, we used long-read sequence data from representatives of the major amniote lineages to determine where in amniote evolution rDNA unit size increased. We find that amniote rDNA unit sizes fall into two narrow size classes: \"normal\" (∼11-20 kb) in all amniotes except monotreme, marsupial, and eutherian mammals, which have \"large\" (∼35-45 kb) sizes. We confirm that increases in intergenic spacer length explain much of this mammalian size increase. However, in stark contrast to the uniformity of mammalian rDNA unit size, mammalian intergenic spacers differ greatly in sequence. These results suggest a large increase in intergenic spacer size occurred in a mammalian ancestor and has been maintained despite substantial sequence changes over the course of mammalian evolution. This points to a previously unrecognized constraint on the length of the intergenic spacer, a region that was thought to be largely neutral. We finish by speculating on possible causes of this constraint.
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  • 文章类型: Journal Article
    核糖体DNA(rDNA)拷贝存在于多个染色体中,并且推测拷贝数的个体间差异会影响对抵抗训练的肥大反应。因此,我们检测了rDNA拷贝数是否与抗阻训练诱导的骨骼肌肥大相关.参与者(n=53名男性,21±1岁;n=29名女性,21±2岁)进行了10-12周的全身阻力训练。确定肥大结果,干预前的股外侧肌(VL)活检的相对rDNA拷贝数也是如此。在所有参与者中检测干预前后VL活检总RNA,和核糖体含量和生物发生的mRNA/rRNA标记也在训练前的29名女性中进行了测定,训练1周后24小时,训练10周后处于基础状态。在所有参与者中,相对rDNA拷贝数与训练诱导的全身瘦体重变化之间无明显关联(r=-0.034,p=0.764),股外侧肌厚度(r=0.093,p=0.408),平均肌纤维横截面积(r=-0.128,p=0.259),或肌肉RNA浓度的变化(r=0.026,p=0.818),在检查每个性别时,这些趋势是相似的。然而,所有Pol-I调节子mRNA以及45S前rRNA,28SrRNA和18SrRNA在雌性第一次训练后24小时增加。使用LHCN-M2肌管的后续研究表明,双酚A(BPA)诱导的相对rDNA拷贝数减少并未显着影响胰岛素样生长因子诱导的肌管肥大。这些发现表明相对rDNA拷贝数与肌纤维肥大无关。
    Ribosomal DNA (rDNA) copies exist across multiple chromosomes, and interindividual variation in copy number is speculated to influence the hypertrophic response to resistance training. Thus, we examined if rDNA copy number was associated with resistance training-induced skeletal muscle hypertrophy. Participants (n = 53 male, 21 ± 1 yr old; n = 29 female, 21 ± 2 yr old) performed 10-12 wk of full-body resistance training. Hypertrophy outcomes were determined, as was relative rDNA copy number from preintervention vastus lateralis (VL) biopsies. Pre- and postintervention VL biopsy total RNA was assayed in all participants, and mRNA/rRNA markers of ribosome content and biogenesis were also assayed in the 29 female participants before training, 24 h following training bout 1, and in the basal state after 10 wk of training. Across all participants, no significant associations were evident between relative rDNA copy number and training-induced changes in whole body lean mass (r = -0.034, P = 0.764), vastus lateralis thickness (r = 0.093, P = 0.408), mean myofiber cross-sectional area (r = -0.128, P = 0.259), or changes in muscle RNA concentrations (r = 0.026, P = 0.818), and these trends were similar when examining each gender. However, all Pol-I regulon mRNAs as well as 45S pre-rRNA, 28S rRNA, and 18S rRNA increased 24 h following the first training bout in female participants. Follow-up studies using LHCN-M2 myotubes demonstrated that a reduction in relative rDNA copy number induced by bisphenol A did not significantly affect insulin-like-growth factor-induced myotube hypertrophy. These findings suggest that relative rDNA copy number is not associated with myofiber hypertrophy.NEW & NOTEWORTHY We examined ribosomal DNA (rDNA) copy numbers in men and women who resistance trained for 10-12 wk and found no significant associations with skeletal muscle hypertrophy outcomes. These data, along with in vitro data in immortalized human myotubes whereby rDNA copy number was reduced, provide strong evidence that relative rDNA copy number is not associated with anabolism.
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  • 文章类型: Journal Article
    核仁与控制分化和癌变的基因形成染色体间接触。DUX4基因指定具有两个同源结构域的转录因子。以前,使用圆形染色体构象捕获(4‰)方法对细胞群体,证明DUX4基因簇与核仁形成频繁接触。还发现这些接触在热休克处理后几乎完全消失。4C方法作为所有连接介导的方法能够检测核中染色质环之间相当紧密的相互作用。为了独立地确认单细胞中接触的形成和频率,我们使用FISH方法。这里,我们显示在单个细胞中的DUX基因在所有测试的HEK293T细胞中形成稳定的接触。在热休克期间,DUX4基因可逆地远离细胞核1-3μm。我们得出结论,核仁形成的染色体间接触很强,动态,和可逆的,提供差异化状态的启动和维持。
    Nucleoli form interchromosomal contacts with genes controlling differentiation and carcinogenesis. DUX4 genes specify transcription factor possessing two homeodomains. Previously, using Circular Chromosome Conformation Capture (4С) approach on population of cells, it was demonstrated that DUX4 gene clusters form frequent contacts with nucleoli. It was found also that these contacts are almost completely abolished after heat shock treatment. 4C approach as all ligation-mediated methods is capable to detect rather close interactions between chromatin loops in nuclei. In order to independently confirm the formation and the frequency of the contacts in single cells we used FISH approach. Here, we show that DUX genes in single cells form stable contacts in all tested HEK293T cells. During heat shock, DUX4 genes reversibly move 1-3 µm away from the nuclei. We conclude that interchromosomal contacts formed by nucleoli are strong, dynamic, and reversible, providing both the initiation and maintenance of a differentiated state.
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  • 文章类型: Journal Article
    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。
    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.
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