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转录组测序揭示了招潮蟹(Charybdis feriatus)睾丸和卵巢之间性别表达偏好模式的基因表达特征。

Transcriptome-seq provides insights into sex-preference pattern of gene expression between testis and ovary of the crucifix crab ( Charybdis feriatus).

机构信息

Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University , Shantou , China.

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

出版信息

Physiol Genomics. 2018 May 1;50(5):393-405. doi: 10.1152/physiolgenomics.00016.2018. Epub 2018 Mar 23.

Abstract

The crucifix crab, Charybdis feriatus, which mainly inhabits Indo-Pacific region, is regarded as one of the most high-potential species for domestication and incorporation into the aquaculture sector. However, the regulatory mechanisms of sex determination and differentiation of this species remain unclear. To identify candidate genes involved in sex determination and differentiation, high throughput sequencing of transcriptome from the testis and ovary of C. feriatus was performed by the Illumina platform. After removing adaptor primers, low-quality sequences and very short (<50 nt) reads, we obtained 80.9 million and 66.2 million clean reads from testis and ovary, respectively. A total of 86,433 unigenes were assembled, and ~43% (37,500 unigenes) were successfully annotated to the NR, NT, Swiss-Prot, KEGG, COG, GO databases. By comparing the testis and ovary libraries, we obtained 27,636 differentially expressed genes. Some candidate genes involved in the sex determination and differentiation of C. feriatus were identified, such as vasa, pgds, vgr, hsp90, dsx-f, fem-1, and gpr. In addition, 88,608 simple sequence repeats were obtained, and 61,929 and 77,473 single nucleotide polymorphisms from testis and ovary were detected, respectively. The transcriptome profiling was validated by quantitative real-time PCR in 30 selected genes, which showed a good consistency. The present study is the first high-throughput transcriptome sequencing of C. feriatus. These findings will be useful for future functional analysis of sex-associated genes and molecular marker-assisted selections in C. feriatus.

摘要

铁脚寄居蟹,主要栖息于印度洋-太平洋地区,被认为是最具潜在驯化和纳入水产养殖的物种之一。然而,该物种的性别决定和分化的调控机制尚不清楚。为了鉴定参与性别决定和分化的候选基因,我们通过 Illumina 平台对铁脚寄居蟹的睾丸和卵巢进行了转录组高通量测序。去除接头引物、低质量序列和非常短(<50nt)的reads 后,我们分别从睾丸和卵巢获得了 8090 万和 6620 万条清洁reads。共组装了 86433 条unigenes,其中约 43%(37500 条)成功注释到 NR、NT、Swiss-Prot、KEGG、COG、GO 数据库。通过比较睾丸和卵巢文库,我们获得了 27636 个差异表达基因。鉴定到了一些参与铁脚寄居蟹性别决定和分化的候选基因,如 vasa、pgds、vgr、hsp90、dsx-f、fem-1 和 gpr。此外,还获得了 88608 个简单序列重复,分别从睾丸和卵巢检测到 61929 和 77473 个单核苷酸多态性。30 个选定基因的定量实时 PCR 验证显示结果具有良好的一致性。本研究是首次对铁脚寄居蟹进行高通量转录组测序。这些发现将有助于未来对铁脚寄居蟹性别相关基因的功能分析和分子标记辅助选择。

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