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东方果实蝇睾丸转录组分析揭示了参与精子发生的基因以及标记物发现。

Transcriptome profiling of the testis reveals genes involved in spermatogenesis and marker discovery in the oriental fruit fly, Bactrocera dorsalis.

作者信息

Wei D, Li H-M, Yang W-J, Wei D-D, Dou W, Huang Y, Wang J-J

机构信息

Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing, China.

出版信息

Insect Mol Biol. 2015 Feb;24(1):41-57. doi: 10.1111/imb.12134. Epub 2014 Sep 25.

Abstract

The testis is a highly specialized tissue that plays a vital role in ensuring fertility by producing spermatozoa, which are transferred to the female during mating. Spermatogenesis is a complex process, resulting in the production of mature sperm, and involves significant structural and biochemical changes in the seminiferous epithelium of the adult testis. The identification of genes involved in spermatogenesis of Bactrocera dorsalis (Hendel) is critical for a better understanding of its reproductive development. In this study, we constructed a cDNA library of testes from male B. dorsalis adults at different ages, and performed de novo transcriptome sequencing to produce a comprehensive transcript data set, using Illumina sequencing technology. The analysis yielded 52 016 732 clean reads, including a total of 4.65 Gb of nucleotides. These reads were assembled into 47 677 contigs (average 443 bp) and then clustered into 30 516 unigenes (average 756 bp). Based on BLAST hits with known proteins in different databases, 20 921 unigenes were annotated with a cut-off E-value of 10(-5). The transcriptome sequences were further annotated using the Clusters of Orthologous Groups, Gene Orthology and the Kyoto Encyclopedia of Genes and Genomes databases. Functional genes involved in spermatogenesis were analysed, including cell cycle proteins, metalloproteins, actin, and ubiquitin and antihyperthermia proteins. Several testis-specific genes were also identified. The transcripts database will help us to understand the molecular mechanisms underlying spermatogenesis in B. dorsalis. Furthermore, 2913 simple sequence repeats and 151 431 single nucleotide polymorphisms were identified, which will be useful for investigating the genetic diversity of B. dorsalis in the future.

摘要

睾丸是一种高度特化的组织,通过产生精子在确保生育能力方面发挥着至关重要的作用,精子在交配时会传递给雌性。精子发生是一个复杂的过程,会产生成熟精子,涉及成年睾丸生精上皮显著的结构和生化变化。鉴定参与橘小实蝇精子发生的基因对于更好地理解其生殖发育至关重要。在本研究中,我们构建了不同年龄雄性橘小实蝇成虫睾丸的cDNA文库,并使用Illumina测序技术进行从头转录组测序以生成一个全面的转录数据集。分析产生了52016732条clean reads,包括总共4.65Gb的核苷酸。这些reads被组装成47677个重叠群(平均443bp),然后聚类成30516个单基因(平均756bp)。基于在不同数据库中与已知蛋白质的BLAST比对,20921个单基因被注释,截止E值为10^(-5)。转录组序列进一步使用直系同源群簇、基因直系同源和京都基因与基因组百科全书数据库进行注释。分析了参与精子发生的功能基因,包括细胞周期蛋白、金属蛋白、肌动蛋白、泛素和抗热蛋白。还鉴定了几个睾丸特异性基因。转录本数据库将有助于我们了解橘小实蝇精子发生的分子机制。此外,鉴定出2913个简单序列重复和151431个单核苷酸多态性,这将有助于未来研究橘小实蝇的遗传多样性。

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