De novo Transcriptome Analysis of Chinese Citrus Fly, Bactrocera minax (Diptera: Tephritidae), by High-Throughput Illumina Sequencing.
De novo Transcriptome Analysis of Chinese Citrus Fly, Bactrocera minax (Diptera: Tephritidae), by High-Throughput Illumina Sequencing.
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DOI：
10.1371/journal.pone.0157656
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Liu YH
中科院分区：
文献类型：
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作者：
Wang J;Xiong KC;Liu YH
中国柑橘蝇（Bactrocera minax, Enderlein）是亚洲温带地区柑橘最具破坏性的害虫之一。迄今为止，由于缺乏基因组信息和无法在实验室中培育这种昆虫，涉及到最小裂裂螨分子生物学和生理学的研究仍然很少。在本研究中，使用Illumina测序技术进行转录组的从头组装。共获得20,928,907条clean reads，组装成33,324条unigenes，平均长度为908.44 bp。通过比对NCBI非冗余蛋白（Nr）、Swiss-Prot、Clusters of Orthologous Groups （COG）、Gene Ontology （GO）和Kyoto Encyclopedia of Genes and Genomes Pathway （KEGG）数据库对Unigenes进行注释。鉴定出可能参与胁迫耐受的基因，包括20个热休克蛋白（Hsps）基因、26个谷胱甘肽s -转移酶（GSTs）基因和2个铁蛋白亚基基因。这些基因可能与最小二代白僵虫滞育期的抗逆性有关。此前已有研究发现，20E可避免小蠹蛹的滞育，但其作用机制尚不清楚。因此，我们确定了编码20E生物合成途径酶的基因，包括Neverland、Spook、Phantom、Disembodied、Shadow、Shade和Cyp18a1，以及编码20E受体蛋白、ecdysone受体（EcR）和ultraspiracle （USP）的基因。20e相关基因在不同发育阶段以及20e处理和未处理蛹之间的表达模式表明它们在滞育程序中起作用。此外，还检测到1,909个简单重复序列（SSRs），这将有助于分子标记的开发。本研究的发现极大地提高了我们对该物种的遗传学认识，为今后对该物种的进一步研究奠定了基础。
The Chinese citrus fly, Bactrocera minax (Enderlein), is one of the most devastating pests of citrus in the temperate areas of Asia. So far, studies involving molecular biology and physiology of B. minax are still scarce, partly because of the lack of genomic information and inability to rear this insect in laboratory. In this study, de novo assembly of a transcriptome was performed using Illumina sequencing technology. A total of 20,928,907 clean reads were obtained and assembled into 33,324 unigenes, with an average length of 908.44 bp. Unigenes were annotated by alignment against NCBI non-redundant protein (Nr), Swiss-Prot, Clusters of Orthologous Groups (COG), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes Pathway (KEGG) database. Genes potentially involved in stress tolerance, including 20 heat shock protein (Hsps) genes, 26 glutathione S-transferases (GSTs) genes, and 2 ferritin subunit genes, were identified. These genes may play roles in stress tolerance in B. minax diapause stage. It has previously been found that 20E application on B. minax pupae could avert diapause, but the underlying mechanisms remain unknown. Thus, genes encoding enzymes in 20E biosynthesis pathway, including Neverland, Spook, Phantom, Disembodied, Shadow, Shade, and Cyp18a1, and genes encoding 20E receptor proteins, ecdysone receptor (EcR) and ultraspiracle (USP), were identified. The expression patterns of 20E-related genes among developmental stages and between 20E-treated and untreated pupae demonstrated their roles in diapause program. In addition, 1,909 simple sequence repeats (SSRs) were detected, which will contribute to molecular marker development. The findings in this study greatly improve our genetic understanding of B. minax, and lay the foundation for future studies on this species.