Molt-dependent transcriptome analysis of claw muscles in Chinese mitten crab Eriocheir sinensis

Zhihuan Tian,Chuanzhen Jiao 한국유전학회 2019 Genes & Genomics Vol.41 No.5

Background Molting is a critical developmental process for crustaceans, during which the claw muscles undergo periodic atrophy and restoration. But the mechanism underlying this special muscle reshuffle around ecdysis is not yet thoroughly understood. Objective To investigate the molecular mechanism underlying the muscle’s reshuffle over the molting cycle of Chinese mitten crab Eriocheir sinensis. Methods The Illumina high-throughput sequencing technique were used to sequence the transcriptome of the whole claw muscles from Chinese mitten crab Eriocheir sinensis in three molting stages (inter-molt C stage, pre-molt D3− 4 and post-molt A–B stage); the de novo assembly, annotation and functional evaluation of the contigs were performed with bioinformatics tools. Results Totally 129,149 unigenes, 128,190 CDS, 33,770 SSRs and a large number of SNP sites were obtained, and 3700 and 12,771 differentially expressed genes (DEGs) were identified respectively in A–B and D3− 4 stage compared with that in C stage. The identified DEGs were enriched to 746 and 1 408 GO terms respectively in A–B and D3– 4 stage compared with C stage (p ≤ 0.05). KEGG pathway analysis showed that the DEGs were significantly enriched in 14 and 11 pathways in A–B vs C comparison and D3− 4 vs C comparison (p ≤ 0.05), respectively. These DEGs are involved in several biological processes critical for the animal’s growth and development, such as gene expression, protein synthesis, muscle development, new cuticle reconstruction, oxidation–reduction process and glycolytic process. Conclusion The data generated in this study is the first transcriptomic resource from the muscles of Chinese mitten crab, which would facilitate to characterize key molecular processes underlying crab muscle’s growth and development during the molting cycles.

Molecular Characterization of Four Genes Highly Expressed during Megalopa Stage in Chinese Mitten Crab, Eriocheir sinensis

윤태호,배재용,강혜은,최정화,이충일,박원규,김현우 한국해양과학기술원 2015 Ocean science journal Vol.50 No.1

The molecular markers to distinguish different larval stages have various applications in ecological studies. Using the differential display RT-PCR technique, we isolated and characterized four genes, which are expressed predominantly in the megalopa stage of Eriocheir sinensis. The four genes include two cuticular proteins with different domain organization (Ers-CP15 and Ers-CP34) and two skeletal-muscle-specific genes (Ers-SCP and Ers-ActinSK1). The two cuticular protein genes were expressed predominantly in the epidermis and their expression level was significantly higher in the megalopa stage (about 7.0-folds) than it was in the zoea stage. However, their high transcriptional level in zoea IV suggested that these two cuticular protein genes may not be a useful target to discriminate megalopa from zoea. Ers-SCP encoded the invertebratespecific sarcoplasmic calcium binding protein and Ers-ActinSK1 gene encoded the crustacean skeletal muscle actin. Expressions of these two genes were detected only in muscular tissues; leg muscle, claw muscle and thoracic muscle. This suggests that the increased transcription levels of two muscle-specific genes during the megalopa stage are mainly due to increased muscular tissues. Among its three isoforms, Ers-SCPa displayed the highest difference (22.4-folds) between megalopa and zoea suggesting Ers-SCPa is the most reliable marker to distinguish megalopa from zoea. Although Ers-SCPc and Ers-ActinSK1 also showed similar expression profiles to Ers-SCPa and Ers-SCPb, differences in their expression levels were not as high as Ers-SCPa and Ers-SCPb.

A Myostain-like Gene Expressed Highly in the Muscle Tissue of Chinese mitten crab, Eriocheir sinensis

김경선,전정민,김현우 한국수산과학회 2009 Fisheries and Aquatic Sciences Vol.12 No.3

A complete cDNA, which encodes for a myostatin-like protein (Es-MSTN), was isolated from the Chinese mitten crab, Eriocheir sinensis. Es-MSTN was composed of 2,397 nucleotides and the open reading frame (ORF) specified a protein containing 468 amino acids. Es-MSTN exhibited 32% amino acid sequence identity and 52% similarity to human myostatin. Multiple sequence alignment analysis indicated that Es-MSTN possessed the conserved proteolytic cleavage site (RXXR) for maturation of the protein and nine cysteine residues for disulfide bridges. Besides the conserved structural features, Es-MSTN also exhibits its unique characters; a longer N-terminal domain which is involved in protein folding and latent form of myostatin and absence of the cleavage site for BMP-1/tolloid family of metalloproteinase to activate mature myostatin. Phylogenetic analysis suggests that Es-MSTN showed the closely related to both vertebrate myostatin and GDF11. Es-MSTN is expressed highly in the claw muscle, leg muscle, thoracic muscle and heart, and moderately in the hindgut suggesting that Es-MSTN may play important roles in the muscle tissues. As homolog of mammalian myostatin and GDF11, Es-MSTN may be involved in development of muscular tissue and further study will help to produce high-quality seafood.

A Myostain-like Gene Expressed Highly in the Muscle Tissue of Chinese mitten crab, Eriocheir sinensis

Kim, Kyoung-Sun,Jeon, Jeong-Min,Kim, Hyun-Woo The Korean Society of Fisheries and Aquatic Scienc 2009 Fisheries and Aquatic Sciences Vol.12 No.3

A complete cDNA, which encodes for a myostatin-like protein (Es-MSTN), was isolated from the Chinese mitten crab, Eriocheir sinensis. Es-MSTN was composed of 2,397 nucleotides and the open reading frame (ORF) specified a protein containing 468 amino acids. Es-MSTN exhibited 32% amino acid sequence identity and 52% similarity to human myostatin. Multiple sequence alignment analysis indicated that Es-MSTN possessed the conserved proteolytic cleavage site (RXXR) for maturation of the protein and nine cysteine residues for disulfide bridges. Besides the conserved structural features, Es-MSTN also exhibits its unique characters; a longer N-terminal domain which is involved in protein folding and latent form of myostatin and absence of the cleavage site for BMP-1/tolloid family of metalloproteinase to activate mature myostatin. Phylogenetic analysis suggests that Es-MSTN showed the closely related to both vertebrate myostatin and GDF11. Es-MSTN is expressed highly in the claw muscle, leg muscle, thoracic muscle and heart, and moderately in the hindgut suggesting that Es-MSTN may play important roles in the muscle tissues. As homolog of mammalian myostatin and GDF11, Es-MSTN may be involved in development of muscular tissue and further study will help to produce high-quality seafood.

Population genetic analysis reveals secondary contact between Eriocheir sinensis and E. japonica in South Korea

Jung Jibom,Wu Xugan,Park Taeseo,Lee Seok-hyun,Hwang Hee-seung,Jung Juwon,Jung Jongwoo 한국유전학회 2022 Genes & Genomics Vol.44 No.5

Background: The Chinese mitten crab (Eriocheir sinensis) and the Japanese mitten crab (E. japonica) of the family Varunidae, which are also critical fishery species in their native habitats, including China, Japan, and South Korea, exhibit a rare migration life history for the decapod life cycle. Eriocheir sinensis and E. japonica in South Korea may have originated from speciation or secondary contact in South Korea after speciation of these two species; however, the genetic relationship between these South Korean populations remains unclear, and need to be clarified. Objective: This study examined the population genetic properties of E. sinensis and E. japonica in South Korea to reveal the origin of their co-existence. Methods: Mitochondrial DNA cytochrome c oxidase I sequences of 120 E. sinensis and E. japonica from 6 sampling sites in South Korea were sequenced. Their genetic diversity and haplotype data were compared to previously reported data from the Chinese mainland, Taiwan, Japan, and Russia. Results: The haplotype network, analysis of molecular variance, FST values, Mantel test, and Bayesian skyline plot results of South Korean E. sinensis and E. japonica demonstrated that E. sinensis and E. japonica were isolated from their central populations, Chinese and Japanese populations, by the distance due to the temperature, salinity, ocean current flow, or a geohistorical event. In addition, genetic analysis indicated that the South Korean populations of the two species were marginal populations, exhibiting low genetic diversity, significant negative neutrality values, and radial haplotype networks. The current results also suggest that the Geum River is an overlapping habitat for the natural populations of E. sinensis and E. japonica in South Korea. Further research is required to examine the geohistorical and evolutionary event between E. sinensis and E. japonica in South Korea. Conclusions: The co-existence of E. sinensis and E. japonica in South Korea may have originated from secondary contact after their speciation.

Genetic Differences and Variations in Freshwater Crab(Eriocheir sinensis) and Swimming Crab(Portunus trituberculatus)

윤종만 한국발생생물학회 2006 발생과 생식 Vol.10 No.1

참게(Eriocheir sinensis)와 꽃게(Portunus trituberculatus)의 2종으로부터 genomic DNA를 분리 추출하였다. 선택된 7개의 OPA-05, OPA-13, OPA-16, OPB-06, OPB-15, OPB-17 and OPD-10의 RAPD primer를 이용하여 identical, polymorphic 그리고 specific fragment를 얻어냈다. 본 연구에서 부안산 참게 집단에서는 505개의 fragment Genomic DNA isolated from two species of Korean freshwater crab(Eriocheir sinensis) and swimming crab(Portunus trituberculatus) was amplified several times by PCR reactions. The seven arbitrarily selected primers OPA-05, OPA-13, OPA-16, OPB-06, OPB-15, OPB-17 and OPD-10 were used to generate the identical, polymorphic, and specific fragments. 505 fragments were identified in the freshwater crab species, and 513 in the swimming crab from Buan: 81 specific fragments(16.0%) in the freshwater crab species and 100(19.5%) in the swimming crab. 165 identical fragments, with an average of 23.6 per primer, were observed in the freshwater crab species. 66 fragments, with an average of 9.4 per primer, were identified in the swimming crab species. The numbers of polymorphic fragments in the freshwater crab and swimming crab were 50 and 14, respectively. The oligonucleotides decamer primer OPB-17 generated identical DNA fragments, approximately 300 bp, in both the freshwater crab and swimming crab species. Compared separately, the average genetic difference was higher in the swimming crab than in the freshwater crab species. The average genetic difference was between the freshwater crab and swimming crab species. The dendrogram obtained by the seven primers indicates four genetic clusters: cluster 1(FRESHWATER 01), cluster 2(FRESHWATER 02, 03, 04, 05 and 06), cluster 3(FRESHWATER 07, 08, 09, 10 and 11), and cluster 4(SWIMMING 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 and 22). The shortest genetic distance displaying significant molecular difference was between individuals SWIMMING no. 18 and SWIMMING no. 17 from swimming crab(0.096). Ultimately, individual no. 02 of the freshwater crab was most distantly related to freshwater crab no. 03(genetic distance = 0.770). As stated above, the potential of RAPD-PCR to identify diagnostic markers for the identification of two crab species has been demonstrated.

카드뮴과 수은 폭로시 참게(mitten crab, Eriocheir sinensis) 유생의 사망률과 생물농축

이복규 ( Bok Kyu Lee ),허만규 ( Man Kyu Huh ) 한국환경생물학회 2004 환경생물 : 환경생물학회지 Vol.22 No.3

참게(E. sinensis)의 유생 생장에 미치는 기초적인 환경 및 생물학적 정보를 얻기 위해 본 실험을 실시하였다. 참게 유생을 카드뮴과 수은의 0.1, 0.2, 0.3 ppm농도에 처리하였다. 참게의 생존율과 카드뮴과 수은의 중금속 농도와 양의 상관을 나타내었다. 그런데 96시간 사육시 카드뮴과 수은의 반수치사농도는 유생 단계간 유의한 차이를 나타내었다. 반수치사농도는 카드뮴이 수은보다 높았으므로 수은이 더 치명적이었다. 중금속의 축적은 카드뮴과 수은 모두 메가로파 시기부터 급격히 증가하였다. 따라서 참게의 유생의 생육에서 카드뮴과 수은은 참게 유생의 생존에 치명적이며 생물농축이 일어났다. This experiment was studied to obtain basic environmental effects and biological information on the early growth of larval mitten crab (Eriocheir sinensis). Mitten crabs were maintained for each larval stage with solutions containing 0.1, 0.2, and 0.3 ppm of cadmium (Cd) and mercury (Hg). The relationship between survival rate of mitten crabs and metal content in the growth chamber showed a positive correlations. However, it showed a significant difference in the 96 hr-LC_(50) values of Cd and Hg for the E. sinensis larvae from the first zoea larva to juvenile. Hg was more toxic to E. sinensis larval than Cd. When Cd and Hg are released into the water, they enter E. sinensis larval and are biological magnified. These results of survival rate and bioaccumulation are very important when considering the survival of the mitten crab.

High Plasticity of the Gut Microbiome and Muscle Metabolome of Chinese Mitten Crab (Eriocheir sinensis) in Diverse Environments

( Xiaowen Chen ),( Haihong Chen ),( Qinghua Liu ),( Kangda Ni ),( Rui Ding ),( Jun Wang ),( Chenghui Wang ) 한국미생물생명공학회(구 한국산업미생물학회) 2021 Journal of microbiology and biotechnology Vol.31 No.2

Phenotypic plasticity is a rapid response mechanism that enables organisms to acclimate and survive in changing environments. The Chinese mitten crab (Eriocheir sinensis) survives and thrives in different and even introduced habitats, thereby indicating its high phenotypic plasticity. However, the underpinnings of the high plasticity of E. sinensis have not been comprehensively investigated. In this study, we conducted an integrated gut microbiome and muscle metabolome analysis on E. sinensis collected from three different environments, namely, an artificial pond, Yangcheng Lake, and Yangtze River, to uncover the mechanism of its high phenotypic plasticity. Our study presents three divergent gut microbiotas and muscle metabolic profiles that corresponded to the three environments. The composition and diversity of the core gut microbiota (Proteobacteria, Bacteroidetes, Tenericutes, and Firmicutes) varied among the different environments while the metabolites associated with amino acids, fatty acids, and terpene compounds displayed significantly different concentration levels. The results revealed that the gut microbiome community and muscle metabolome were significantly affected by the habitat environments. Our findings indicate the high phenotypic plasticity in terms of gut microbiome and muscle metabolome of E. sinensis when it faces environmental changes, which would also facilitate its acclimation and adaptation to diverse and even introduced environments.

Geographic Variations in Four Freshwater Crab (Eriocheir sinensis) Populations throughout Its Distribution Range

윤종만 한국발생생물학회 2009 발생과 생식 Vol.13 No.2

Genomic DNA samples isolated from four geographical freshwater crab (Eriocheir sinensis) populations collected in the inland of the Korean Peninsula (Gunsan, Paju, and Nampo) and a Chinese site, were used for PCR amplification. Seven decamer primers generated 19 specific loci (19/243 loci, 7.81%) in the Gunsan population, 32 (32/215 loci, 14.88%) in the Paju population, 19 (19/231 loci, 8.23%) in the Nampo population and 62 (62/340 loci, 18.24%) in a Chinese population. The average 8.9 specific loci exhibited inter-individual-specific characteristics, thus revealing DNA polymorphisms in the Chinese population. The number of unique shared loci to each population and number of shared loci by the four populations were generated by molecular analysis using seven primers in four populations. 35 unique shared loci to each population, with an average of 5.0 per primer, were observed in the Gunsan population, and 50 loci, with an average of 7.1 per primer, were observed in the Chinese population. The hierarchical dendrogram indicates three main branches: cluster 1 (GUNSAN 01 ~ GUNSAN 05, PAJU 06 ~ PAJU 10 and NAMPO 11 ~ NAMPO 15) and cluster 2 (CHINESE 16, 17, 18, 19 and 20). Conclusively individual no. 20 of the PAJU 10 freshwater crab was most distantly related to CHINESE no. 20 (genetic distance = 0.667). Taken together, these results demonstrate the potential of RAPD analysis to identify diagnostic markers for the identification of four freshwater crab populations.