Freshwater Perkinsea and marine-freshwater colonizations revealed by pyrosequencing and phylogeny of environmental rDNA
Freshwater Perkinsea and marine-freshwater colonizations revealed by pyrosequencing and phylogeny of environmental rDNA
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DOI：
10.1038/ismej.2010.39
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发表时间：
2010-09-01
期刊：
影响因子：
11
通讯作者：
Shalchian-Tabrizi, Kamran
中科院分区：
文献类型：
--
作者：
Brate, Jon;Logares, Ramiro;Shalchian-Tabrizi, Kamran
原生寄生虫具有重要的生态学意义，因为它们对宿主种群动态和整个生态系统的功能具有重大影响。然而，人们对它们在水生栖息地的流行情况知之甚少。本文研究了原生寄生虫Perkinsus和Parvilucifera （Perkinsea）的多样性和分布格局。我们的方法包括对从高海拔湖泊（lake Finsevatn, Norway）获得的18S rDNA基因进行454焦磷酸测序，并对所有公开的与Perkinsea相关的序列进行系统发育分析。应用的PCR引物定位于包含18S rDNA可变V4区的450bp区域，并针对454技术的Titanium升级进行了优化。从几乎所有真核生物超群中恢复了近5000个长度大于150 bp的序列，其中13个独特的序列与Perkinsea有关。因此，我们对454扩增子测序的新策略能够恢复大量远亲真核生物和以前未知的Perkinsea物种。此外，我们在GenBank中鉴定了40个由其他近期多样性调查产生的Perkinsea序列。重要的是，这些序列的系统发育分析确定了17个栖息地特定的海洋和淡水分支（PERK 1-17）。因此，在整个珀金西的历史上，这些栖息地之间只有少数成功的过渡，这表明海水和淡水之间的边界可能构成了细胞内寄生虫交叉定植的障碍。ISME学报（2010）4,1144-1153；doi: 10.1038 / ismej.2010.39;2010年4月15日在线发布
Protist parasites are ecologically important, as they can have great impact on host population dynamics and functioning of entire ecosystems. Nevertheless, little is known about their prevalence in aquatic habitats. Here, we investigate the diversity and distributional patterns of the protist parasites Perkinsus and Parvilucifera (Perkinsea). Our approach included 454 pyrosequencing of the 18S rDNA gene obtained from a high-altitude lake (Lake Finsevatn, Norway) and phylogenetic analyses of all publicly available sequences related to Perkinsea. The applied PCR primers target a 450 bp region that encompass the variable V4 region of the 18S rDNA gene and have been optimized for the Titanium upgrade of the 454 technology. Nearly 5000 sequences longer than 150 bp were recovered from nearly all eukaryotic supergroups, and of those, 13 unique sequences were affiliated to Perkinsea. Thus, our new strategy for 454 amplicon sequencing was able to recover a large diversity of distantly related eukaryotes and previously unknown species of Perkinsea. In addition, we identified 40 Perkinsea sequences in GenBank generated by other recent diversity surveys. Importantly, phylogenetic analyses of these sequences identified 17 habitat-specific marine and freshwater clades (PERK 1-17). Hence, only a few successful transitions between these habitats have taken place over the entire history of Perkinsea, suggesting that the boundary between marine and fresh waters may constitute a barrier to cross-colonizations for intracellular parasites. The ISME Journal (2010) 4, 1144-1153; doi: 10.1038/ismej.2010.39; published online 15 April 2010