The Chinese mitten crab Eriocheir japonica sinensis is one of the most common aquaculture species cultivated in China. The crab is an omnivore, and its hepatopancreas absorbs and stores nutrients. The aim of this study was to elucidate the expressions of the digestive enzyme genes and determine their respective roles in regulating digestive capacity in E. j. sinensis. We sequenced the hepatopancreatic transcriptomes of crabs fed a meat diet (MD), a vegetarian diet (VD), or a mixed diet (MV) and compared the gene expression patterns of these three groups of crabs. A total of 305,887 unigenes were obtained, of which 8747, 10,963, and 8877 were significantly differentially expressed in the comparisons between the MD and MV, VD and MV, and MD and VD diets, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) database-based enrichment analysis revealed that the differentially expressed gene (DEG) responses in the hepatopancreases to the MD mainly involved the “pancreatic secretion,” “glutathione metabolism,” “sphingolipid metabolism,” “fatty acid metabolism,” and “glycerolipid metabolism pathways.” DEG responses to the VD based on KEGG analysis mainly involved the “galactose metabolism,” “starch and sucrose metabolism,” and “fructose and mannose metabolism” pathways. The key digestive enzymes, including trypsin, β-glucosidase, chitinase, and triacylglycerol lipase, were identified. Our results further our understanding of crustacean hepatopancreatic functions during food digestion and provide resources for further studies regarding the molecular basis of omnivorous diets in crustaceans.
Transcriptome Chinese mitten crab Hepatopancreas Omnivore Digestive enzyme
This is a preview of subscription content, log in to check access.
This study was funded by the National Natural Science Foundation of China (Grant Number 31702014), and Doctoral Scientific Research Foundation of Yancheng Teachers University to ZFW, and Open Foundation of Jiangsu Key Laboratory for Bioresources of Saline Soils (Grant Number JKLBS2016007).
HYG, DT, XLS, WQ, RBL, BPT, and ZFW designed and conceived the experiment. ZFW and HYG performed the data analysis and drafted the manuscript. All authors read and approved the final manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare there are no competing interests.
The sampling location was not privately owned or protected, and field sampling did not involve protected species.
Online Resource 4. GO function classifcation of the assembled unigenes in the hepatopancreas of Eriocheir japonica sinensis, including three classes of biological process, cellular process and molecular function. (PDF 291 kb)
Online Resource 8. Gene expression patterns of different digestive enzymes in Eriocheir japonica sinensis fed with three diets (MD, VD and MV). (PDF 205 kb)
Birk RZ, Brannon PM (2004) Regulation of pancreatic lipase by dietary medium chain triglycerides in the weanling rat. Pediatr Res 55:921–926CrossRefGoogle Scholar
Bui THH, Lee SY (2015) Endogenous cellulase production in the leaf litter foraging mangrove crab Parasesarma erythodactyla. Comp Biochem Physiol B Biochem Mol Biol 179:27–36CrossRefGoogle Scholar
Cannicci S, Schubart CD, Innocenti G et al (2017) A new species of the genus Parasesarma De Man 1895 from East African mangroves and evidence for mitochondrial introgression in sesarmid crabs. Zool Anz 269:89–99CrossRefGoogle Scholar
Chu Y, Corey DR (2012) RNA sequencing: platform selection, experimental design, and data interpretation. Nucleic Acid Ther 22:271–274CrossRefGoogle Scholar
Dammannagoda LK, Pavasovic A, Prentis PJ et al (2015) Expression and characterization of digestive enzyme genes from hepatopancreatic transcripts from redclaw crayfish (Cherax quadricarinatus). Aquac Nutr 21:868–880CrossRefGoogle Scholar
Dittel AI, Epifanio CE (2009) Invasion biology of the Chinese mitten crab Eriochier sinensis: a brief review. J Exp Mar Bio Ecol 374:79–92CrossRefGoogle Scholar
Ghosh D, Porter E, Shen B et al (2002) Paneth cell trypsin is the processing enzyme for humandefensin-5. Nat Immunol 3:583–590CrossRefGoogle Scholar
Gross PS, Bartlett TC, Browdy CL et al (2001) Immune gene discovery by expressed sequence tag analysis of hemocytes and hepatopancreas in the Pacific White Shrimp, Litopenaeus vannamei, and the Atlantic White Shrimp L. setiferus. Dev Comp Immunol 25:565–577CrossRefGoogle Scholar
Huang S, Wang J, Yue W et al (2015) Transcriptomic variation of hepatopancreas reveals the energy metabolism and biological processes associated with molting in Chinese mitten crab Eriocheir sinensis. Sci Rep 5:14015CrossRefGoogle Scholar
Jiang H, Yin Y, Zhang X et al (2009) Chasing relationships between nutrition and reproduction: a comparative transcriptome analysis of hepatopancreas and testis from Eriocheir sinensis. Comp Biochem Physiol Part D Genomics Proteomics 4:227–234CrossRefGoogle Scholar
Jiankai W, Xiaojun Z, Yang Y et al (2014) Comparative transcriptomic characterization of the early development in Pacific white shrimp Litopenaeus vannamei. PLoS ONE 9:e106201CrossRefGoogle Scholar
Li X, Cui Z, Liu Y et al (2013) Transcriptome analysis and discovery of genes involved in immune pathways from hepatopancreas of microbial challenged mitten crab Eriocheir sinensis. PLoS ONE 8:e68233CrossRefGoogle Scholar
Li X, Xu Z, Zhou G et al (2015a) Molecular characterization and expression analysis of five chitinases associated with molting in the Chinese mitten crab, Eriocheir sinensis. Comp Biochem Physiol B Biochem Mol Biol 187:110–120CrossRefGoogle Scholar
Li Y, Hui M, Cui Z (2015b) Comparative transcriptomic analysis provides insights into the molecular basis of the metamorphosis and nutrition metabolism change from zoea to megalopae in Eriocheir sinensis. Comp Biochem Physiol Part D Genomics Proteomics 13:1–9CrossRefGoogle Scholar
Merzendorfer H (2013) Insect-derived chitinases. Adv Biochem Eng Biotechnol 136:19–50PubMedGoogle Scholar
Michiels MS, Valle JCD, Mañanes AAL (2017) Trypsin and N-aminopeptidase (APN) activities in the hepatopancreas of an intertidal euryhaline crab: biochemical characteristics and differential modulation by histamine and salinity. Comp Biochem Physiol A Mol Integr Physiol 204:228–235CrossRefGoogle Scholar
R Development Core Team (2015) R: A language and environment for statistical computing. R Foundation for Statistical Computing, ViennaGoogle Scholar
Rao R, Zhu YB, Alinejad T et al (2015) RNA-Seq analysis of Macrobrachium rosenbergii hepatopancreas in response to Vibrio parahaemolyticus infection. Gut Pathog 7:6CrossRefGoogle Scholar
Rivera-Pérez C, García-Carreño F (2011) Effect of fasting on digestive gland lipase transcripts expression in Penaeus vannamei. Mar Genom 4:273–278CrossRefGoogle Scholar
Rojo L, Garcíacarreño F (2012) Cold-adapted digestive aspartic protease of the clawed lobsters Homarus americanus and Homarus gammarus: biochemical characterization. Mar Biotechnol 15:87–96CrossRefGoogle Scholar
Roux MM, Pain A, Klimpel KR et al (2002) The lipopolysaccharide and beta-1,3-glucan binding protein gene is upregulated in white spot virus-infected shrimp (Penaeus stylirostris). J Virol 76:7140–7149CrossRefGoogle Scholar
Rudnick DA, Hieb K, Grimmer KF et al (2003) Patterns and processes of biological invasion: the Chinese mitten crab in San Francisco Bay. Basic Appl Ecol 4:249–262CrossRefGoogle Scholar
Salma U, Uddowla MH, Kim M et al (2012) Five hepatopancreatic and one epidermal chitinases from a pandalid shrimp (Pandalopsis japonica): cloning and effects of eyestalk ablation on gene expression. Comp Biochem Physiol B Biochem Mol Biol 161:197–207CrossRefGoogle Scholar
Shi Y, Burn P (2004) Lipid metabolic enzymes: emerging drug targets for the treatment of obesity. Nat Rev Drug Discov 3:695–710CrossRefGoogle Scholar
Wang L, Yan B, Liu N et al (2009) Effects of cadmium on glutathione synthesis in hepatopancreas of fresh water crab, Sinopotamon yangtsekiense. Chemosphere 74:51–56CrossRefGoogle Scholar
Wang W, Wu X, Liu Z et al (2014) Insights into hepatopancreatic functions for nutrition metabolism and ovarian development in the crab Portunus trituberculatus: gene discovery in the comparative transcriptome of different hepatopancreas stages. PLoS ONE 9:e84921CrossRefGoogle Scholar
Wang Z, Xu S, Du K et al (2016) Evolution of digestive enzymes and RNASE1 provides insights into dietary switch of cetaceans. Mol Biol Evol 33:3144–3157CrossRefGoogle Scholar
Wang Z, Bai Y, Zhang D et al (2018) Adaptive evolution of osmoregulatory-related genes provides insight into salinity adaptation in Chinese mitten crab, Eriocheir sinensis. Genetica 146:303–311CrossRefGoogle Scholar
Watanabe H, Tokuda G (2010) Cellulolytic systems in insects. Annu Rev Entomol 55:609–632CrossRefGoogle Scholar
Wei W, Wu X, Liu Z et al (2014) Insights into hepatopancreatic functions for nutrition metabolism and ovarian development in the crab Portunus trituberculatus: gene discovery in the comparative transcriptome of different hepatopancreas stages. PLoS ONE 9:e84921CrossRefGoogle Scholar
Wei B, Yang Z, Wang J et al (2017) Effects of dietary lipids on the hepatopancreas transcriptome of Chinese mitten crab (Eriocheir sinensis). PLoS ONE 12:e182087Google Scholar
Zhu B, Tang L, Yu Y et al (2017) Identification of ecdysteroid receptor-mediated signaling pathways in the hepatopancreas of the red swamp crayfish, Procambarus clarkii. Gen Comp Endocrinol 246:372–381CrossRefGoogle Scholar
1.Jiangsu Key Laboratory for Bioresources of Saline Soils, Jiangsu Synthetic Innovation Center for Coastal Bio-agriculture, Jiangsu Provincial Key Laboratory of Coastal Wetland Bioresources and Environmental Protection, School of Ocean and Biological EngineeringYancheng Teachers UniversityYanchengChina
2.College of Biotechnology and Pharmaceutical EngineeringNanjing Tech UniversityNanjingChina