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林业科学 ›› 2019, Vol. 55 ›› Issue (5): 104-113.doi: 10.11707/j.1001-7488.20190512

• 论文与研究报告 • 上一篇    下一篇

星天牛转录组及三大解毒酶家族相关基因系统发育分析

韩小红1, 卢赐鼎1, 华银1, 林浩宇1, 是雨霏1, 吴松青1,2, 张飞萍1,2, 梁光红1,2   

  1. 1. 福建农林大学林学院 福州 350002;
    2. 生态公益林重大有害生物防控福建省高校重点实验室 福州 350002
  • 收稿日期:2018-08-27 修回日期:2019-03-13 出版日期:2019-05-25 发布日期:2019-05-20
  • 基金资助:
    国家重点研发项目(2017YFD0600105);国家自然科学基金项目(31870641);福建省科技厅引导性项目(2016N0002);福建省高校产学合作项目(2018N5101)。

Phylogenetic Analysis of Transcriptome and Three Detoxification Enzyme Families Related Genes in Anoplophora chinensis (Coleoptera: Cerambycidae)

Han Xiaohong1, Lu Ciding1, Hua Yin1, Lin Haoyu1, Shi Yufei1, Wu Songqing1,2, Zhang Feiping1,2, Liang Guanghong1,2   

  1. 1. College of Forestry, Fujian Agriculture and Forestry University Fuzhou 350002;
    2. Key Laboratory of Integrated Pest Management in Ecological Forests, Fujian Province University, Fujian Agriculture and Forestry University Fuzhou 350002
  • Received:2018-08-27 Revised:2019-03-13 Online:2019-05-25 Published:2019-05-20

摘要: [目的]研究星天牛转录组信息及体内细胞色素P450单加氧酶(cytochrome P450 monooxygenase,CYP)、羧酸酯酶(carboxylesterase,CarE)和谷胱甘肽S-转移酶(glutathione S-transferase,GST)三大解毒酶系的表达谱情况,通过构建系统进化树,探析其这3种酶系基因家族的类别及其系统演化关系,为星天牛对多寄主种类的适应性与杀虫剂的抗药性机制研究提供依据。[方法]采用新一代高通量测序技术Illumina HiSeqTM2500对星天牛进行转录组测序与数据分析,通过数据筛选并进一步与鞘翅目近缘物种的解毒酶同源蛋白进行系统发育分析。[结果]经测序、序列拼接得到55 260条unigenes,将其与公共数据库进行同源比对,注释了32 247条unigenes,在Swiss-Prot数据库注释的unigenes的数量最多(99.11%);注释到Nr数据库时与赤拟谷盗的unigenes同源性最高(60.25%);在GO数据库中,共有8 083个unigenes被成功注释,根据其功能大致可分为3类47亚类;在KEGG数据库中,4 600条unigenes与162个预测路径可以匹配。通过基因注释发现248条解毒酶系基因在星天牛体内表达,包括139条CYP基因、72条CarE基因、37条GST基因。系统发育分析发现,星天牛的大部分CYP基因都至少与一种鞘翅目近缘物种的CYP基因聚类在一起,P450基因中的CYP6家族数量与其他近缘物种(鞘翅目)类似,包含的基因数量最多;CarE解毒酶蛋白基因主要为β-酯酶、神经趋化蛋白、外源性代谢酶、未知等4种类型,分别可能参与星天牛的一些激素以及信息素的加工、神经和发育过程、消化与解毒等生理过程;星天牛的GST基因聚类主要包含Microsomal GST、Sigma GST、Omega GST、Delta GST、Theta GST等亚家族,具有保护生物大分子免受氧化损伤、催化活力等功能。[结论]本研究获得星天牛的转录组信息,初步探明星天牛体内三大解毒酶系的表达谱及分化情况,可为星天牛对多寄主种类的适应性与杀虫剂抗药性的产生及遗传机制研究提供基础数据和参考。

关键词: 星天牛, 转录组, 高通量测序, 系统发育, 解毒酶蛋白基因

Abstract: [Objective]This study aimed to investigate the transcriptome information and the expression profiles of cytochrome P450 monooxygenase (P450) (CYP), carboxylesterase (CarE) and glutathione S-transferase (GST) detoxification enzymes in Anoplophora chinensis. The phylogenetic tree was constructed to explore the types and phylogenetic relationships of the three enzymatic gene families, and to provide a basis for the study of the adaptability of A. chinensis to multiple host species and the mechanism of insecticide resistance.[Method]The high-throughput sequencing technique, Illumina HiSeqTM2500, was employed for transcriptome sequencing of A. chinensis, and the data analysis. The phylogenetic analysis of detoxifying enzymes proteins with their homologous protein from close relative species of Coleoptera was conducted.[Result]The clean reads were then de novo assembled into 55 260 unigenes, and 32 247 unigenes were annotated by homologous comparison with public databases. Most unigenes were annotated in the Swiss-Prot database (99.11%), the unigenes of A. chinensis had the highest homology (60.25%) to those of Tribolium castaneum when annotated to Nr database. In the GO database, 8 083 unigenes were successfully annotated and was able to be broadly divided into 3 categories and 47 sub-types, and 4 600 unigenes matched with 162 prediction paths in KEGG database. Using gene annotation, 248 detoxification enzyme genes were found to be expressed in A. chinensis, including 139 CYP genes, 72 CarE genes, and 37 GST genes. Phylogenetic analysis showed that most of the CYP genes of A. chinensis were clustered with at least one of the related species of Coleoptera. In the P450 gene family, the number of CYP6 family was similar to other related species (Coleoptera), and contained the largest number of genes. The CarE detoxification enzyme genes were mainly composed of four types:β-esterase, neurochemotactic protein, exogenous metabolic enzyme and unknown category, and they may be involved in several processes (hormones and pheromones, nerve and developmental processes, digestion and detoxification, respectively). The GST expression mainly consisted of Microsomal GST, Sigma GST, Omega GST, Delta GST, Theta GST subfamilies, which have the function of protecting biomolecules from oxidative damage and catalytic activity.[Conclusion]This study presented the data of transcriptome, and preliminarily analyzed the expression and differentiation profiles of three detoxifying enzymes in A. chinensis, which could provide fundamental information and reference for adaptability to multi-host species, resistance of pesticides, and genetic mechanism of A. chinensis.

Key words: Anoplophora chinensis, transcriptome, high-throughput sequencing technology, phylogeny, detoxifying enzyme gene

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