Project description:microRNAs (miRNAs) are an evolutionarily conserved class of non-coding RNA molecules, which regulate kinds of biological processes at post-transcriptional level. Investigation of miRNAs expression profiles using high-throughput strategies is efficiently conductive to identify and characterize miRNAs. In this study, through Solexa deep sequencing approach, we obtained 115 orange spotted grouper (Epinephelus coioides) encoded miRNAs. Among them, 107 miRNAs shared high similarity with miRNAs encoded by zebrafish (Danio rerio) and other four vertebrates, indicating that cellular miRNAs are highly conserved between species. 18-26 nt small RNAs from GS cells were sequenced in one Solexa lane
Project description:microRNAs (miRNAs) are an evolutionarily conserved class of non-coding RNA molecules, which regulate kinds of biological processes at post-transcriptional level. Investigation of miRNAs expression profiles using high-throughput strategies is efficiently conductive to identify and characterize miRNAs. In this study, through Solexa deep sequencing approach, we obtained 115 orange spotted grouper (Epinephelus coioides) encoded miRNAs. Among them, 107 miRNAs shared high similarity with miRNAs encoded by zebrafish (Danio rerio) and other four vertebrates, indicating that cellular miRNAs are highly conserved between species.
Project description:Sex differentiation is a complex process that requires many factors to regulate proliferation, differentiation, development, and organization of the gonads. In teleosts, the molecular mechanisms of sex differentiation are diverse and unclear, especially in hermaphrodite fish. In the present study, 15 cell types, including germ cells, follicle cells, and theca cells, and so on, were identified using single-cell RNA sequencing (scRNA-seq) in the gonads during sex differentiation in a hermaphrodite fish, orange-spotted grouper (Epinephelus coioides). Two pre-follicle cell types were defined, and the differentiating trajectories of the follicle cells were outlined. Notably, both pre-follicle and follicle cells highly expressed male-related genes synchronously, such as amh, sox9, and dmrt3 in pre-follicle cells, as well as dmrt1 in follicle cells. Oocyte possessed two distinct states, with high expression of oocyte development related genes in one state and spermatogenesis related genes in the other state, respectively. Our results provide novel insights into cell types and lineage tracing in the gonads during sex differentiation in hermaphrodite species.
Project description:Grouper is an important commercial maricultural fish, which suffer viral nervous necrosis (VNN) disease at the larval and juvenile stages, but the changes of transcriptomics and proteomics during viral infection remain unknown. In this study, we applied RNA-seq and label-free mass spectrum for the first time to depict the map of transcriptomics and proteomics in non-infected, susceptible-infected and tolerate-infected grouper in larval stage. Further analyses showed that the transcriptome and proteome change dramatically among 3 distinct groups, indicating that different immune response for infected and perststent grouper in larval stage. These valuable transcriptomics and proteomics datasets enable the investigation of molecular mechanism in nervous necrosis (VNN) virus infection, thus helps the further development of molecular breeding and marine fishery
Project description:Cortisol was injected into the protogynous epinephelus coioides to investigate the role of this hormone on sex change. Following injection, we evaluated sex-related gene expression during the processes of cortisol-induced sex change in epinephelus coioides.
Project description:The purpose of subject was to explore the optimum protein requirement of juvenile grouper (Epinephelus coioides). In the test, 450 juveniles with an average weight (10.02 ± 0.22) g were randomly divided into six groups with triplicate, and were fed with 350, 400, 450, 500, 550 and 600 g/kg iso-lipid test diet twice 1 day for 8 weeks, respectively. The results showed that: (1) With the increase of protein level, the body weight gain rate and specific growth rate first increased and then reduced, while the feed coefficient rate first decreased and then increased, while the protein efficiency significantly decreased (P < 0.05). (2) With the increase of protein level, the condition factor, hepaticsomatic index and visceralsomatic index significantly reduced (P < 0.05). (3) With the increase of protein level, the crude protein content of whole fish and muscle gradually increased, while the crude lipid content gradually decreased. (4) High-protein diet (550-600 g/kg) significantly increased the plasma total protein content and decreased the triglyceride content of orange-spotted grouper (P < 0.05). (5) Compared with the 350 g/kg group, 500, 550, 600 g/kg groups significantly increased the activities of glutamic-pyruvic transaminase and glutamic oxaloacetic transaminase in liver (P < 0.05). (6) With the increase of protein level, the protease activity of intestine first increased and then decreased, and reached the maximum at the protein level of 500 g/kg, while lipase and amylase decreased significantly (P < 0.05). (7) The activities of acid phosphatase, superoxide dismutase and lysozyme in liver increased first and then decreased with the increase of protein level, and reached the maximum in the 400 g/kg protein group. According to the analysis specific growth rate, the optimum protein level of juvenile orange-spotted grouper is 521.84 g/kg.
Project description:BACKGROUND: Orange-spotted grouper (Epinephelus coioides) is an economically important marine fish cultured in China and Southeast Asian countries. The emergence of infectious viral diseases, including iridovirus and betanodavirus, have severely affected food products based on this species, causing heavy economic losses. Limited available information on the genomics of E. coioides has hampered the understanding of the molecular mechanisms that underlie host-virus interactions. In this study, we used a 454 pyrosequencing method to investigate differentially-expressed genes in the spleen of the E. coioides infected with Singapore grouper iridovirus (SGIV). RESULTS: Using 454 pyrosequencing, we obtained abundant high-quality ESTs from two spleen-complementary DNA libraries which were constructed from SGIV-infected (V) and PBS-injected fish (used as a control: C). A total of 407,027 and 421,141 ESTs were produced in control and SGIV infected libraries, respectively. Among the assembled ESTs, 9,616 (C) and 10,426 (V) ESTs were successfully matched against known genes in the NCBI non-redundant (nr) database with a cut-off E-value above 10-5. Gene ontology (GO) analysis indicated that "cell part", "cellular process" and "binding" represented the largest category. Among the 25 clusters of orthologous group (COG) categories, the cluster for "translation, ribosomal structure and biogenesis" represented the largest group in the control (185 ESTs) and infected (172 ESTs) libraries. Further KEGG analysis revealed that pathways, including cellular metabolism and intracellular immune signaling, existed in the control and infected libraries. Comparative expression analysis indicated that certain genes associated with mitogen-activated protein kinase (MAPK), chemokine, toll-like receptor and RIG-I signaling pathway were alternated in response to SGIV infection. Moreover, changes in the pattern of gene expression were validated by qRT-PCR, including cytokines, cytokine receptors, and transcription factors, apoptosis-associated genes, and interferon related genes. CONCLUSION: This study provided abundant ESTs that could contribute greatly to disclosing novel genes in marine fish. Furthermore, the alterations of predicted gene expression patterns reflected possible responses of these fish to the virus infection. Taken together, our data not only provided new information for identification of novel genes from marine vertebrates, but also shed new light on the understanding of defense mechanisms of marine fish to viral pathogens.