Abstract
BackgroundThe Chinese mitten crab Eriocheir sinensis is an important economic crustacean and has been seriously attacked by various diseases, which requires more and more information for immune relevant genes on genome background. Recently, high-throughput RNA sequencing (RNA-seq) technology provides a powerful and efficient method for transcript analysis and immune gene discovery.Methods/Principal FindingsA cDNA library from hepatopancreas of E. sinensis challenged by a mixture of three pathogen strains (Gram-positive bacteria Micrococcus luteus, Gram-negative bacteria Vibrio alginolyticus and fungi Pichia pastoris; 108 cfu·mL−1) was constructed and randomly sequenced using Illumina technique. Totally 39.76 million clean reads were assembled to 70,300 unigenes. After ruling out short-length and low-quality sequences, 52,074 non-redundant unigenes were compared to public databases for homology searching and 17,617 of them showed high similarity to sequences in NCBI non-redundant protein (Nr) database. For function classification and pathway assignment, 18,734 (36.00%) unigenes were categorized to three Gene Ontology (GO) categories, 12,243 (23.51%) were classified to 25 Clusters of Orthologous Groups (COG), and 8,983 (17.25%) were assigned to six Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Potentially, 24, 14, 47 and 132 unigenes were characterized to be involved in Toll, IMD, JAK-STAT and MAPK pathways, respectively.Conclusions/SignificanceThis is the first systematical transcriptome analysis of components relating to innate immune pathways in E. sinensis. Functional genes and putative pathways identified here will contribute to better understand immune system and prevent various diseases in crab.
Highlights
Chinese mitten crab Eriocheir sinensis (Crustacea: Decapoda: Grapsidae, Eriocheir) (Henri Milne Edwards, 1854) is one of the important economic aquaculture species in China
With rapid development of large-scale culture, frequent outbreaks of diseases caused by viruses, bacteria and rickettsia-like organisms have led to catastrophic economic losses in cultured E. sinensis stocks [1,2,3]
We prepared a mixture of three pathogen strains (Gram-positive bacteria Micrococcus luteus, Gram-negative bacteria Vibrio alginolyticus and fungi Pichia pastoris), which were suspended in 0.1 mol/L PBS with the final pathogens concentration of 108 cfu?mL21
Summary
Chinese mitten crab Eriocheir sinensis (Crustacea: Decapoda: Grapsidae, Eriocheir) (Henri Milne Edwards, 1854) is one of the important economic aquaculture species in China. Innate immune system provides a first line for host to defense against invading pathogens. It is composed of cellular responses like phagocytosis and encapsulation, and humoral responses that produce immunerelated factors. Immune relevant genes, such as crustin [4], cathepsin L [5], prophenoloxidase (proPO) [6], C-type lectin [7]. The Chinese mitten crab Eriocheir sinensis is an important economic crustacean and has been seriously attacked by various diseases, which requires more and more information for immune relevant genes on genome background. High-throughput RNA sequencing (RNA-seq) technology provides a powerful and efficient method for transcript analysis and immune gene discovery
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