As post-transcriptional regulators, microRNAs (miRNAs) play an important role in growth and reproductive processes. So far, there is limited information regarding crustacean miRNAs. To explore the potential role of miRNAs in the gonadal development of the prawn Macrobrachium rosenbergii, we constructed seven small RNA libraries from ovarian and testicular tissues at various stages using somatic tissue as the control. A total of 1,954 known and 129 novel miRNAs were retrieved. By comparing differentially expressed miRNAs (DEMs) between testes and ovaries, forty-one miRNAs were identified with sex-biased expression patterns, including 17 ovary-biased and 24 testis-biased patterns. Furthermore, the putative target genes of the sex-biased miRNAs, such as cyclin L1, mitogen-activated protein kinase 7 (MAPK 7), heat shock protein (HSP), and zinc finger protein, were significantly enriched in many reproduction-related pathways including the Gonadotropin-releasing hormone (GnRH) pathway, glycolysis, gluconeogenesis pathway, ovarian steroidogenesis, estrogen signaling pathway, MAPK pathway, Wnt pathway, and insulin signaling pathway, implicating potential regulatory roles of miRNAs in reproduction. These data aid in the further investigation of the mechanism of gonadal development and reproductive regulation mediated by miRNA in M. rosenbergii.

MicroRNA (miRNA) is a posttranscriptional downregulator that plays a vital role in a wide variety of biological processes. In this study, we constructed five ovarian and testicular small RNA libraries using two somatic libraries as reference controls for the identification of sex‐biased miRNAs and gonadal differentially expressed miRNAs (DEMs) of the Chinese mitten crab, Eriocheir sinensis. A total of 535 known and 243 novel miRNAs were identified, including 312 sex‐biased miRNAs and 402 gonadal DEMs. KEGG pathway analysis showed that DEM target genes were statistically enriched in MAPK, Wnt, and GnRH signaling pathway, and so on. A number of the sex‐biased miRNAs target genes associated with sex determination/differentiation, such as IAG, Dsx, Dmrt1, and Fem1, while others target the genes related to gonadal development, such as P450s, Wnt, Ef1, and Tra‐2c. Dual‐luciferase reporter assay in vitro further confirmed that miR‐34 and let‐7b can downregulate IAG expression, miR‐9‐5p, let‐7d, let‐7b, and miR‐8915 can downregulate Dsx. Taken together, these data strongly suggest a potential role for the sex‐biased miRNAs in sex determination/differentiation and gonadal development in the crab.

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Red pigment concentrating hormone (RPCH) and pigment dispersing hormone (PDH) are crustacean neuropeptides involved in broad physiological processes including body color changes, circadian rhythm, and ovarian growth. In this study, the full-length cDNA of RPCH and PDH were identified from the brain of the Chinese mitten crab Eriocheir sinensis. The deduced RPCH and PDH mature peptides shared identical sequence to the adipokinetic hormone/RPCH peptides family and the β-PDH isoforms and were designated as Es-RPCH and Es-β-PDH, respectively. Es-RPCH and Es-β-PDH transcripts were distributed in the brain and eyestalks. The positive signals of Es-RPCH and Es-β-PDH were localized in the neuronal clusters 6, 8, 9, 10, and 17 of the brain as revealed by in situ hybridization. The expression level of Es-RPCH and Es-β-PDH mRNA in nervous tissues were all significantly increased at vitellogenic stage, and then decreased at the final meiotic maturation stage. The administrated with synthesized Es-RPCH peptide results in germinal vesicles shift toward the plasma membrane in vitellogenic oocyte, and significant decrease of the gonad-somatic index (GSI) and mean oocyte diameter as well as the expression of vitellogenin mRNA at 30 days post injection in vivo. Similar results were also found when injection of the Es-β-PDH peptide. In vitro culture demonstrated that Es-RPCH and Es-β-PDH induced germinal vesicle breakdown of the late vitellogenic oocytes. Comparative ovarian transcriptome analysis indicated that some reproduction/meiosis-related genes such as cdc2 kinase, cyclin B, 5-HT-R and retinoid-X receptor were significantly upregulated in response to Es-RPCH and Es-β-PDH treatments. Taken together, these results provided the evidence for the inductive effect of Es-RPCH and Es-β-PDH on the oocyte meiotic maturation in E. sinensis.

Recently, green tide outbreaks have resulted in severe coastal ecology and economic effects in China. Jiangsu coastal areas are usually the site of early green tide outbreaks. To clarify the effects of green tide outbreaks in Jiangsu coastal areas, this study analyzed microbial communities during green tide‐free and green tide outbreak periods (May and July, respectively) through 16S rDNA sequencing. Sequences were clustered into 4117 operational taxonomic units (OTUs), 1044 and 3834 of which were obtained from the May and July groups, respectively. Redundancy analysis indicated that green tide occurrence was closely associated with the temperature, pH, and concentrations of various nutrients. Diversity analysis revealed that the July group had a richer microbial community than the May group, in agreement with the results of propagule culture. Moreover, comparative analysis revealed that samples in the May and July groups clustered together. According to Megan analysis, the May group had much more Psychrobacter, Sulfitobacter, and Marinomonas than the July group, whereas the other genera were predominantly found in July, such as Ascidiacerhabitans, Synechococcus Hydrotalea, and Burkholderia‐Paraburkholderia. These findings suggest that green tide outbreaks affect marine microbial communities, and detecting the changes in the identified genera during green tide outbreaks may contribute to green tide forecasting. Practitioner Points Jiangsu coastal areas are usually the site of early green tide outbreaks. Green tide occurrence was related to the concentrations of various nutrients. Microbial species and community structure significantly changed after green tide outbreak.

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