昆虫学报 ›› 2019, Vol. 62 ›› Issue (5): 547-560.doi: 10.16380/j.kcxb.2019.05.003

• 研究论文 • 上一篇    下一篇

花椒窄吉丁转录组及化学感受相关基因的分析

杨平, 谢寿安*, 巩雪芳, 车显荣, 王延来, 吕淑杰   

  1. (西北农林科技大学林学院, 陕西杨陵 712100)
  • 出版日期:2019-05-20 发布日期:2019-05-16

Analysis of the transcriptome and chemoreception-related genes of Agrilus zanthoxylumi (Coleoptera: Buprestidae)

YANG Ping, XIE Shou-An*, GONG Xue-Fang, CHE Xian-Rong, WANG Yan-Lai, LÜ Shu-Jie   

  1. (College of Forestry, Northwest A&F University, Yangling, Shaanxi 712100, China)
  • Online:2019-05-20 Published:2019-05-16

摘要:

【目的】建立花椒窄吉丁Agrilus zanthoxylumi转录组数据库,挖掘其基因组数据。【方法】采用高通量测序平台Illumina NovaSeq6000对花椒窄吉丁成虫不同组织(触角、头、胸、腹、足、翅)进行转录组测序、序列组装,使用BLAST软件将花椒窄吉丁unigenes序列与公共数据库进行比对,BLAST同源性搜索的方法从中筛选出与其化学感受相关的基因,并与其他已研究发表的鞘翅目昆虫化学感受相关基因的核酸序列比对,进行系统发育分析。利用RPKM值对这些基因在花椒窄吉丁成虫不同组织中的表达量进行分析。【结果】经测序及序列拼接后共得到80 320条unigenes和169 398条contigs,其G+C比例分别是37.63%和39.18%,平均长度分别为828.75和1 084.33 bp; unigenes长度主要分布在200~400 bp的共有37 374条,占全部unigenes的46.53%。在NR数据库中注释的与花椒窄吉丁转录组相似基因所属物种分布为赤拟谷盗Tribolium castaneum所占比例26.80%,其后依次是小家鼠Mus musculus(13.87%),蛀犀金龟 Oryctes borbonicus(5.26%),山松甲虫Dendroctonus ponderosae(4.10%),黑蚁Lasius niger(2.36%),其他物种所占比例是47.61%。在GO数据库中比对到27 488个unigenes,可分为分子功能、细胞组分和生物学进程三大类共59个过程;利用KOG数据库功能注释分为25类,注释到unigenes 最多功能类别的是普通功能,共4 666个,最少的是与细胞移动相关,共40个;将80 320个unigenes映射到KEGG数据库中,21 104条unigens注释到代谢通路35个,占26.27%,注释最多的代谢途径是转录途径,共涉及到2 037条unigenes。从NR数据库中共注释到8个气味受体(odorant receptor, OR)基因、7个离子型受体(ionotropic receptor, IR)基因和6个味觉受体(gustatory receptor, GR)基因,各组织中的基因表达量分析发现,这3类化学感受相关基因在成虫触角中的表达量显著高于其他组织中的表达量,雌、雄触角中表达量差异最显著的为AzanOR1,而在雌、雄成虫足中这3类化学感受相关基因表达量没有差异。【结论】本研究首次获得了花椒窄吉丁转录组数据,为进一步研究花椒窄吉丁的基因功能奠定了分子基础。

关键词: 花椒窄吉丁, 转录组, 高通量测序, 基因注释, 生物信息学

Abstract: 【Aim】 To establish the transcriptome database of the Agrilus zanthoxylumi, and to obtain its genomic data. 【Methods】 The transcriptome of the different tissues (antennae, head, thorax, abdomen, leg, wing) of A. zanthoxylumi adults was sequenced using an Illumina NovaSeq 6000 platform, and the unigene sequences were compared with the homologous sequences in public databases by BLAST. Chemoreception-related genes were screened out by BLAST homology search, and their nucleotide sequences were compared with those of other reported chemoreception-related genes of Coleoptera insects for phylogenetic analysis. The expression levels of these genes in different tissues of A.zanthoxylumi adults were analyzed based on the RPKM value. 【Results】 After sequencing and sequence splicing, a total of 80 320 genes and 169 398 contigs were obtained. The G+C proportions of contigs and unigenes were 37.63% and 39.18%, respectively, and the average length of contigs and the average length of unigenes were 1 084.33 and 828.75 bp, respectively. There were 37 374 unigenes with a length distribution mainly in 200-400 bp, accounting for 46.53% of the total unigenes. Origin species distribution of similar genes with unigenes of A. zanthoxylumi annotated in the NR database was Tribolium castaneum (26.80%), Mus musculus (13.87%), Oryctes borbonicus (5.26%), Dendroctonus ponderosae (4.10%), Lasius niger (2.36%), and other species (47.61%). There were 27 488 unigenes aligned to the GO database, which can be divided into three functional categories including molecular function, cellular component and biological process in a total of 59 processes. According to the gene annotation against the  KOG database, these unigenes can be divided into 25 categories. The most annotated functional category was the common function (4 666 unigenes), while the least annotated was cell movement-related (only 40 unigenes). When 80 320 unigenes were mapped to the KEGG database, 21 104 unigenes were grouped into 35 metabolic pathways, accounting for 26.27% of the total. The most annotated metabolic pathway was the transcription pathway (2 037 unigenes). By gene annotation against the NR database, we found eight odorant receptor (OR) genes, seven ionotropic receptor (IR) genes and six gustatory receptor (GR) genes. Tissue expression profiles showed that the three kinds of chemoreception-related genes (OR, IR and GR genes) had significantly higher expression levels in the antennae than in other tissues of adults, and the expression level of AzanOR1 in the antennae showed the most significant difference between female and male, while the expression levels of the three kinds of chemoreception-related genes in the legs of adults showed no difference between female and male. 【Conclusion】 This study obtained the transcriptome data of A. zanthoxylumi. The results provide a molecular foundation for further studying the gene function in A. zanthoxylumi.

Key words: Agrilus zanthoxylumi, transcriptome, HiSeq, gene annotation, bioinformatics