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用昆虫血淋巴体外诱导昆虫病原线虫小卷蛾斯氏线虫的转录本分析

Transcripts analysis of the entomopathogenic nematode Steinernema carpocapsae induced in vitro with insect haemolymph.

作者信息

Hao You-Jin, Montiel Rafael, Abubucker Sahar, Mitreva Makedonka, Simões Nelson

机构信息

CIRN and Department of Biology, University of Azores, 9501-801 Ponta Delgada, Azores, Portugal.

出版信息

Mol Biochem Parasitol. 2010 Feb;169(2):79-86. doi: 10.1016/j.molbiopara.2009.10.002. Epub 2009 Oct 27.

Abstract

Steinernema carpocapsae is an insect parasitic nematode widely used in pest control programs. The efficacy of this nematode in controlling insects has been found to be related to the pathogenicity of the infective stage. In order to study the parasitic mechanisms exhibited by this parasite, a cDNA library of the induced S. carpocapsae parasitic phase was generated. A total of 2500 clones were sequenced and 2180 high-quality ESTs were obtained from this library. Cluster analysis generated a total of 1592 unique sequences including 1393 singletons. About 63% of the unique sequences had significant hits (e</=1e-05) to the non-redundant protein database. The remaining sequences most likely represent putative novel protein coding genes. Comparative analysis identified 377 homologs in C. elegans, 431 in C. briggsae and 75 in other nematodes. Classification of the predicted proteins revealed involvement in diverse cellular, metabolic and extracellular functions. One hundred and nineteen clusters were predicted to encode putative secreted proteins such as proteases, proteases inhibitors, lectins, saposin-like proteins, acetylcholinesterase, anti-oxidants, and heat-shock proteins, which can possibly have host interactions. This dataset provides a basis for genomic studies towards a better understanding of the events that occur in the parasitic process of this entomopathogenic nematode, including invasion of the insect haemocoelium, adaptations to insect innate immunity and stress responses, and production of virulence factors. The identification of key genes in the parasitic process provides useful tools for the improvement of S. carpocapsae as a biological agent.

摘要

斯氏线虫是一种昆虫寄生线虫,广泛应用于害虫防治项目。已发现这种线虫在控制昆虫方面的功效与感染阶段的致病性有关。为了研究这种寄生虫表现出的寄生机制,构建了诱导的斯氏线虫寄生阶段的cDNA文库。对总共2500个克隆进行了测序,从该文库中获得了2180个高质量的EST。聚类分析共产生了1592个独特序列,包括1393个单拷贝序列。约63%的独特序列与非冗余蛋白质数据库有显著匹配(e≤1e-05)。其余序列很可能代表推定的新蛋白质编码基因。比较分析在秀丽隐杆线虫中鉴定出377个同源物,在briggsae线虫中鉴定出431个,在其他线虫中鉴定出75个。对预测蛋白质的分类显示其参与多种细胞、代谢和细胞外功能。预测有119个聚类编码推定的分泌蛋白,如蛋白酶、蛋白酶抑制剂、凝集素、类鞘脂激活蛋白、乙酰胆碱酯酶、抗氧化剂和热休克蛋白,这些蛋白可能与宿主相互作用。该数据集为基因组研究提供了基础,有助于更好地理解这种昆虫病原线虫寄生过程中发生的事件,包括侵入昆虫血腔、适应昆虫先天免疫和应激反应以及毒力因子的产生。鉴定寄生过程中的关键基因为改进斯氏线虫作为生物制剂提供了有用的工具。

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