烟蚜若蚜及有翅和无翅成蚜转录组及翅型相关基因的表达差异分析

摘要

[目的]旨在了解烟蚜Myzus persicae不同翅型(有翅和无翅)的转录组特征,丰富烟蚜转录组数据信息,为探明烟蚜翅型分化的分子机制打下基础.[方法]利用高通量测序技术IlluminaHiSeqTM2000分别检测烟蚜若蚜、无翅成蚜和有翅成蚜转录组表达差异;通过实时荧光定量PCR(qPCR)方法,分析翅型相关基因表达模式.[结果]经过测序获得质量值高于20的碱基比例(Q20)均不低于94％的数据(GenBank登录号:SRR6112438,SRR6112436和SRR6112437);所有reads组装成128 065个unigenes,平均长度607.08 bp;将所得序列注释到NR,NT,KOG,GO和KEGG等数据库进行比对,共有128 065个unigenes被注释.基因表达分析发现,与无翅成蚜和若蚜两者相比,有翅成蚜翅形成、肽类激素和受体蛋白相关基因的表达量存在显著差异,表明这些差异表达基因在烟蚜生长发育过程中和烟蚜翅的形成有关.qPCR分析结果显示,在有翅成蚜与无翅成蚜中,Bursicon(bur-α和bur-β),flightin(Fln),Wntless(Wnt),Distal-less(Dll),Decapentaplegic (Dpp),Juvenile hormone epoxide hydrolase基因(JHEH)和Ecdysone receptor基因(EcR)表达量存在显著差异,表明这些基因可能与翅型发育有关.[结论]烟蚜翅形成的过程中体内bur-α和bur-β,Fln,Wnt,Dll,Dpp,JHEH和EcR基因表达发生了变化.该研究结果揭示了烟蚜翅形成相关基因的总体表达特征,为进一步深入开展烟蚜翅形成的研究奠定基础.%[Aim] This study aims to understand the transcriptome features of Myzus persicae of different wing types (winged and apterous),to enrich its transcriptome data,and to reveal the molecular mechanisms of wing dimorphism of this insect.[Methods] The differences in transcriptome expression between nymphs,apterous adults,and winged adults of M.persicae were detected by high-throughput sequencing technology Illumina HiSeqTM2000.The expression patterns of wing type related genes were analyzed by real-time quantitative PCR (qPCR).[Results] The percentage of nucleotides with the quality score over 20 was not less than 94％ (GenBank accession numbers:SRR6112438,SRR6112436 and SRR6112437).All reads were assembled into 128 065 unigenes with an average length of 607.08 bp.The resulting sequences were annotated to databases NR,NT,KOG,GO,KEGG,etc.,and a total of 128 065 unigenes were annotated.Gene expression analysis showed that the genes of flight proteins,peptide hormones,receptor proteins,etc.,which are related to wing formation during growth,were differentially expressed between winged adults and apterous adults or nymphs,qPCR analysis showed that there were significant differences in the expression levels of wing type differentiation related genes including Bursicon (bur-α and bur-β),flightin (Fln),Wntless (Wnt),Distal-less (Dll),Decapentaplegic (Dpp),juvenile hormone epoxide hydrolase gene (JHEH) and ecdysone receptor gene (EcR) between winged adults and apterous adults.[Conclusion] The expression levels of bur-α,bur-β,Fln,Wnt,Dll,Dpp,JHEH and EcR change during wing formation in M.persicae.Our results show the general expression characteristics of wing formation genes and provide a basis for further research of wing formation in M.persicae.