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青蟹(Scylla olivacea)性成熟过程中睾丸的转录组分析及差异基因表达

Transcriptome Analysis and Differential Gene Expression on the Testis of Orange Mud Crab, Scylla olivacea, during Sexual Maturation.

作者信息

Waiho Khor, Fazhan Hanafiah, Shahreza Md Sheriff, Moh Julia Hwei Zhong, Noorbaiduri Shaibani, Wong Li Lian, Sinnasamy Saranya, Ikhwanuddin Mhd

机构信息

Institute of Tropical Aquaculture, Universiti Malaysia Terengganu, Kuala Terengganu, Terengganu, Malaysia.

School of Fisheries and Aquaculture Sciences, Universiti Malaysia Terengganu, Kuala Terengganu, Terengganu, Malaysia.

出版信息

PLoS One. 2017 Jan 30;12(1):e0171095. doi: 10.1371/journal.pone.0171095. eCollection 2017.

Abstract

Adequate genetic information is essential for sustainable crustacean fisheries and aquaculture management. The commercially important orange mud crab, Scylla olivacea, is prevalent in Southeast Asia region and is highly sought after. Although it is a suitable aquaculture candidate, full domestication of this species is hampered by the lack of knowledge about the sexual maturation process and the molecular mechanisms behind it, especially in males. To date, data on its whole genome is yet to be reported for S. olivacea. The available transcriptome data published previously on this species focus primarily on females and the role of central nervous system in reproductive development. De novo transcriptome sequencing for the testes of S. olivacea from immature, maturing and mature stages were performed. A total of approximately 144 million high-quality reads were generated and de novo assembled into 160,569 transcripts with a total length of 142.2 Mb. Approximately 15-23% of the total assembled transcripts were annotated when compared to public protein sequence databases (i.e. UniProt database, Interpro database, Pfam database and Drosophila melanogaster protein database), and GO-categorised with GO Ontology terms. A total of 156,181 high-quality Single-Nucleotide Polymorphisms (SNPs) were mined from the transcriptome data of present study. Transcriptome comparison among the testes of different maturation stages revealed one gene (beta crystallin like gene) with the most significant differential expression-up-regulated in immature stage and down-regulated in maturing and mature stages. This was further validated by qRT-PCR. In conclusion, a comprehensive transcriptome of the testis of orange mud crabs from different maturation stages were obtained. This report provides an invaluable resource for enhancing our understanding of this species' genome structure and biology, as expressed and controlled by their gonads.

摘要

充足的遗传信息对于可持续的甲壳类渔业和水产养殖管理至关重要。具有重要商业价值的橙膏蟹(Scylla olivacea)在东南亚地区很常见,并且备受追捧。尽管它是适合水产养殖的候选品种,但由于对其性成熟过程及其背后的分子机制缺乏了解,尤其是在雄性个体中,该物种的完全驯化受到了阻碍。迄今为止,尚未有关于橙膏蟹全基因组的数据报道。此前发表的关于该物种的现有转录组数据主要集中在雌性以及中枢神经系统在生殖发育中的作用。对未成熟、成熟和成熟阶段的橙膏蟹睾丸进行了从头转录组测序。总共产生了约1.44亿条高质量 reads,并从头组装成160,569条转录本,总长度为142.2 Mb。与公共蛋白质序列数据库(即UniProt数据库、Interpro数据库、Pfam数据库和黑腹果蝇蛋白质数据库)相比,约15 - 23%的总组装转录本得到了注释,并用GO本体术语进行了GO分类。从本研究的转录组数据中挖掘出总共156,181个高质量单核苷酸多态性(SNP)。不同成熟阶段睾丸之间的转录组比较揭示了一个基因(类β晶状体蛋白基因)具有最显著的差异表达——在未成熟阶段上调,在成熟和成熟阶段下调。这通过qRT-PCR进一步得到验证。总之,获得了不同成熟阶段橙膏蟹睾丸的全面转录组。本报告为增进我们对该物种基因组结构和生物学(由其性腺表达和控制)的理解提供了宝贵资源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4cae/5279790/222a5f921e7c/pone.0171095.g001.jpg

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