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New Genera of RNA Viruses in Subtropical Seawater, Inferred from Polymerase Gene Sequences▿ †

机译:从聚合酶基因序列推断亚热带海水中RNA病毒的新属Gene

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摘要

Viruses are an integral component of the marine food web, contributing to the disease and mortality of essentially every type of marine life, yet the diversity of viruses in the sea, especially those with RNA genomes, remains very poorly characterized. Isolates of RNA-containing viruses that infect marine plankton are still rare, and the only cultivation-independent surveys of RNA viral diversity reported so far were conducted for temperate coastal waters of British Columbia. Here, we report on our improvements to a previously used protocol to investigate the diversity of marine picorna-like viruses and our results from applying this protocol in subtropical waters. The original protocol was simplified by using direct filtration, rather than tangential flow filtration, to harvest viruses from seawater, and new degenerate primers were designed to amplify a fragment of the RNA-dependent RNA polymerase gene by reverse transcription-PCR from RNA extracted from the filters. Whereas the original protocol was unsuccessful in a preliminary test, the new protocol resulted in amplification of picorna-like virus sequences in every sample of subtropical and temperate coastal seawater assayed. These polymerase sequences formed a diverse, but monophyletic cluster along with other sequences amplified previously from seawater and sequences from isolates infecting marine protists. Phylogenetic analysis suggested that our sequences represent at least five new genera and 24 new species of RNA viruses. These results contribute to our understanding of RNA virus diversity and suggest that picorna-like viruses are a source of mortality for a wide variety of marine protists.
机译:病毒是海洋食物网中不可或缺的组成部分,实质上导致了每种海洋生物的疾病和死亡,但是海洋中病毒的多样性,尤其是具有RNA基因组的病毒,仍然很难被鉴定。分离出感染海洋浮游生物的含RNA的病毒仍​​然很少,并且迄今为止,仅报告了不列颠哥伦比亚省温带沿海水域进行的唯一与培养无关的RNA病毒多样性调查。在这里,我们报告了我们对先前使用的协议进行的改进,以研究海洋小or类病毒的多样性,以及在亚热带水域中应用该协议的结果。通过使用直接过滤而不是切向流过滤从海水中收集病毒简化了原始操作流程,并设计了新的简并引物,以通过逆转录PCR扩增RNA提取的RNA来扩增RNA依赖性RNA聚合酶基因的片段。过滤器。原始协议在初步测试中失败,而新协议导致在所分析的亚热带和温带沿海海水的每个样本中都扩增了小核糖核酸样病毒序列。这些聚合酶序列与先前从海水中扩增出的其他序列以及感染海洋原生生物的分离株中的序列一起,形成了一个多样化的单系统簇。系统发育分析表明,我们的序列至少代表5个新属和24个新种RNA病毒。这些结果有助于我们对RNA病毒多样性的理解,并表明像picorna一样的病毒是导致各种各样的海洋生物死亡的原因。

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