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| 绿鳍马面鲀雌雄性腺转录组比较分析 |
| Comparative analysis of male and female gonadal transcriptome of Thamnaconus septentrionali |
| 投稿时间：2023-04-26 修订日期：2023-05-22 |
| DOI： |
| 中文关键词: 绿鳍马面鲀 转录组 性腺 差异表达基因 |
| 英文关键词: Thamnaconus septentrionalis Transcriptome Gonads Differentially expressed gene |
| 基金项目:中国水产科学研究院黄海水产研究所基本科研业务费资助(20603022022014)；烟台市科技创新发展计划项目(2022XCZX078)；福建省区域发展项目(2022N3015) |
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| 中文摘要: |
| 为探明绿鳍马面鲀性腺发育相关基因表达特征，优化亲本生殖调控技术，本研究对绿鳍马面鲀雌雄亲体的精巢和卵巢进行了转录组测序分析，经de novo拼装，最终获得119,219个单基因（unigene），N50长度为1,255 bp。在NR、NT、KO、SwissProt、PFAM、GO及KOG数据库分别注释到24,009、35,057、18,453、26,971、30,294、11,420和21,613个unigene。绿鳍马面鲀精巢和卵巢转录组中存在18,954个差异表达基因（DEG），相对于精巢，在卵巢中上调表达的基因有11,265个，下调表达的有7,689个。选取bmp2、sox3、figla、hsd17b1、cyp19a、cyp17、foxl2、star和amh 9个DEGs进行实时荧光定量PCR（qPCR）验证，结果显示，qPCR结果与RNA-Seq分析一致。GO和KEGG富集结果分析，发现amh、cyp17和star可能在绿鳍马面鲀雄性精子发生过程起关键作用；bmp2、foxl2、cyp19a、figla和hsd17b1在雌性卵子发生和卵巢类固醇生成过程发挥重要作用。本研究通过比较绿鳍马面鲀精巢和卵巢的转录组表达差异，初步阐明了精巢和卵巢的基因表达特征，为进一步研究绿鳍马面鲀的性腺发育机制奠定基础。 |
| 英文摘要: |
| Thamnaconus septentrionali has excellent breeding characteristics, is omnivorous and easy to domesticate, especially suitable for netting, can clean the netting and effectively reduce labour costs. The T. septentrionali is available in various sizes and can be raised to market at 100g. Under the water temperature of 18~25℃, it can reach the commercial size after 5~6 months of breeding. As the Yellow and Bohai Seas are cold in winter, the suitable breeding cycle for green fins is from May to November every year. Under natural conditions, the spawning period of T. septentrionali in the Yellow Bohai Sea is from early May to early June, which cannot make good use of the suitable breeding cycle of net box culture. Therefore, reproductive regulation is needed to advance the reproductive period of T. septentrionali. Most of the existing studies are on the expression and functional analysis of individual genes, while the overall expression analysis of gonad-related genes based on histology has not been reported. In this experiment, the first transcriptome sequencing analysis of the spermatophore and ovary of T. septentrionali was performed using the Illumina high-throughput sequencing platform. 165,981,523 raw reads were sequenced from cDNA, and 161,234,846 clean reads were obtained after quality control. The Q20 of each sample was above 98.43% and Q30 was above 95.25%, and the GC content of the sample bases was not less than 52.31%. The above data indicate that the sequencing data are accurate and of good quality, and can be used for subsequent analysis. The unigene obtained above was annotated in the NR, NT, KO, SwissProt, PFAM, GO and KOG databases and 24,009, 35,057, 18,453, 26,971, 30,294, 11,420 and 21,613 unigene were annotated respectively. The KEGG annotation results can be divided into five major categories: the organic systems branch, with 2,115 genes in nine pathways, followed by the metabolism branch, with 1,444 genes in 12 pathways, environmental information processing, with 1,200 genes in three pathways, and genetic information processing, with 1,128 genes in four pathways. environmental information processing (1,200 genes in 3 pathways); genetic information processing (1,128 genes in 4 pathways); and cellular processes (1,128 genes in 4 pathways). The number of genes enriched in cellular processes-related pathways was 927, distributed in four related pathways. The signal transduction pathway was the most annotated KEGG pathway, with 804 genes. The experiment has 24,546 unigenes annotated in the KOG database, which are classified into 26 categories according to their functions. The largest number of unigene was annotated in the signal transduction mechanisms category (5,411), followed by the general function prediction only category (4,252 unigene) and the posttranslational modification, protein folding and chaperone category (1,713 unigene). unigene posttranslational modification, protein turnover, chaperones; transcription, 1,613; and other related functionsThere were 18,954 differentially expressed genes (DEGs) in the spermatophore and ovary transcriptomes of the T. septentrionali, with 11,265 genes up-regulated and 7,689 down-regulated in the ovary relative to the spermatophore. The GO functional enrichment analysis revealed that DEGs were most enriched in the cellular component of biological processes, the intracellular component of the cellular component subclass, and the nucleic acid binding of the molecular functional subclass.GO analysis of the differentially expressed genes in males and females gave us a partial list of genes related to reproductive process (GO:0022414), sexual reproduction (GO:0019953), gamete formation (GO:0007276,), sex differentiation(GO:0007548) and gonad development(GO:00084060). To further characterise the specific functions of DEGs in the spermatophore and ovary of the T. septentrionali, the enriched signalling pathways were further analysed through the KEGG database. A total of 154 KEGG pathways were included in this study, from which the top 30 KEGG pathways were selected. The functional pathways with the highest number of expressed genes are closely related to gonadal sex differentiation and gonadal development, including the Insulin signaling pathway, Steroid hormone biosynthesis, FoxO signaling pathway, M-TOR signaling pathway, and Progesterone-mediated oocyte maturation. Among these are FoxO signalling pathway regulatory genes that can be expressed in multiple processes such as cell cycle control, apoptosis and gluconeogenesis. The insulin signalling pathway plays an important role in regulating developmental, metabolic and lifespan physiological processes,and the insulin signalling pathway is involved in gonadal development and maturation. Nine DEGs, bmp2, sox3, figla, hsd17b1, cyp19a, cyp17, foxl2, star and amh, were selected for real-time fluorescence quantitative PCR (qPCR) validation. qPCR results were consistent with RNA-Seq analysis. GO and KEGG enrichment results were analysed and revealed that amh, cyp17 and star may play a key role in male spermatogenesis of Thamnaconus septentrionali; bmp2, foxl2, cyp19a, figla and hsd17b1 play important roles in female oogenesis and ovarian steroidogenesis. By comparing the transcriptome expression differences between the spermatophore and ovary of Thamnaconus septentrionali, this study initially elucidated the gene expression characteristics of the spermatophore and ovary, laying the foundation for further research on the reproductive development mechanism of Thamnaconus septentrionali and providing theoretical support for optimizing reproductive regulation techniques. |
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