Department of Ecology and Parasitology, Zoological Institute, Karlsruhe Institute of Technology, Kornblumenstrasse 13, Karlsruhe, Germany.
BMC Genomics. 2013 Feb 8;14:87. doi: 10.1186/1471-2164-14-87.
Anguillicola crassus is an economically and ecologically important parasitic nematode of eels. The native range of A. crassus is in East Asia, where it infects Anguilla japonica, the Japanese eel. A. crassus was introduced into European eels, Anguilla anguilla, 30 years ago. The parasite is more pathogenic in its new host than in its native one, and is thought to threaten the endangered An. anguilla across its range. The molecular bases for the increased pathogenicity of the nematodes in their new hosts is not known.
A reference transcriptome was assembled for A. crassus from Roche 454 pyrosequencing data. Raw reads (756,363 total) from nematodes from An. japonica and An. anguilla hosts were filtered for likely host contaminants and ribosomal RNAs. The remaining 353,055 reads were assembled into 11,372 contigs of a high confidence assembly (spanning 6.6 Mb) and an additional 21,153 singletons and contigs of a lower confidence assembly (spanning an additional 6.2 Mb). Roughly 55% of the high confidence assembly contigs were annotated with domain- or protein sequence similarity derived functional information. Sequences conserved only in nematodes, or unique to A. crassus were more likely to have secretory signal peptides. Thousands of high quality single nucleotide polymorphisms were identified, and coding polymorphism was correlated with differential expression between individual nematodes. Transcripts identified as being under positive selection were enriched in peptidases. Enzymes involved in energy metabolism were enriched in the set of genes differentially expressed between European and Asian A. crassus.
The reference transcriptome of A. crassus is of high quality, and will serve as a basis for future work on the invasion biology of this important parasite. The polymorphisms identified will provide a key tool set for analysis of population structure and identification of genes likely to be involved in increased pathogenicity in European eel hosts. The identification of peptidases under positive selection is a first step in this programme.
Anguillicola crassus 是一种经济和生态上重要的鳗鱼寄生线虫。A. crassus 的原生范围在东亚,在那里它感染日本鳗 Anguilla japonica。30 年前,A. crassus 被引入欧洲鳗 Anguilla anguilla 中。这种寄生虫在新宿主中的致病性比在原生宿主中更强,被认为威胁到其在整个范围内濒危的 An. anguilla。线虫在新宿主中致病性增加的分子基础尚不清楚。
我们从 Roche 454 焦磷酸测序数据为 A. crassus 组装了一个参考转录组。从 An. japonica 和 An. anguilla 宿主的线虫中过滤出可能的宿主污染物和核糖体 RNA 的原始读数(总共 756,363 个)。剩余的 353,055 个读数组装成 11,372 个高置信度组装的 contigs(覆盖 6.6 Mb)和另外 21,153 个低置信度组装的 singletons 和 contigs(覆盖另外 6.2 Mb)。高置信度组装 contigs 的大约 55% 被注释了具有域或蛋白质序列相似性的功能信息。仅在线虫中保守的序列或 A. crassus 特有的序列更有可能具有分泌信号肽。鉴定了数千个高质量的单核苷酸多态性,并且个体线虫之间的差异表达与编码多态性相关。鉴定为正选择的转录本在肽酶中富集。参与能量代谢的酶在欧洲和亚洲 A. crassus 之间差异表达的基因集中富集。
A. crassus 的参考转录组质量很高,将成为该重要寄生虫入侵生物学未来工作的基础。鉴定的多态性将为分析种群结构和鉴定可能与欧洲鳗宿主致病性增加相关的基因提供关键工具集。正选择下的肽酶鉴定是该计划的第一步。