梨小食心虫雄成虫、蛹和幼虫的转录组比较分析

Comparative analysis of transcriptomes of male adult,pupa and the 4th-instar larva of the oriental peach moth Grapholita molesta

为挖掘杀虫剂靶标及解毒代谢和抗药性相关基因,利用高通量测序技术对梨小食心虫Grapholita molesta雄成虫、蛹和幼虫进行转录组测序和数据组装,并对转录组数据进行功能注释和比较分析。结果表明,对梨小食心虫雄成虫、蛹和幼虫测序所得高质量reads组装得到195473个转录本,含有158206条unigene,其中有71762条unigene在至少1个数据库中能获得功能注释,有8186条unigene在所有数据库中均能获得注释,分别占unigene总数的45.36%和5.17%。在GO数据库中共有45810条unigene的注释划分到56个不同功能区中,鉴定到240个与解毒代谢相关的基因,包括61个羧酸酯酶(carboxylesterases,CarE)基因、38个谷胱甘肽-S-转移酶(glutathione S-transferase,GST)基因、92个细胞色素P450酶基因和49个ABC转运蛋白基因。雄成虫和蛹之间共注释到262个代谢通路,包括42143个基因,其中差异表达基因有1561个;蛹和4龄幼虫之间共注释到232个代谢通路,包括39127个基因,其中差异表达基因有1095个;雄成虫和4龄幼虫之间共注释到235个代谢通路,包括41436个基因,其中差异表达基因有1582个。选择ABC转运蛋白基因进行深入分析,可划分为8个亚家族;对6个ABC转运蛋白基因进行实时荧光定量PCR验证,显示其表达情况与转录组分析结果基本一致,表明转录组分析结果可靠。

In order to explore the genes associated with insecticide targets,detoxification and insecticide resistance,the high-throughput sequencing technique was used to sequence and assemble the transcriptomes of male,pupa and the 4 th-instar larva of the oriental peach moth Grapholita molesta.The results showed that the high-quality reads assembled from sequencing of male,pupa and larva of the G.molesta to 195473 transcripts containing 158206 unigene,of which 71762 unigene(45.36%)could be functionally annotated in at least one database and 8186 unigene(5.17%)could be annotated in all databases.There were 45810 unigenes annotated in the GO database,divided into 56 different functional groups.Two hundred and forty genes related to detoxification metabolism were identified,including 61 carboxylesterases(CarE),38 glutathione S-transferases(GST),92 cytochrome P450 enzymes(cytochrome P450)and 49 ABC transporter protein genes.A total of 262 metabolic pathways were annotated between adult males and pupae,including 42143 genes,of which 1561 were differentially expressed;232 metabolic pathways were annotated between pupae and 4 th-instar larvae,including 39127 genes,of which 1095 were differentially expressed;235 metabolic pathways were annotated between adult males and 4 th-instar larvae,including 41436 genes,of which 1582 were differentially expressed genes.The ABC transporter protein genes were selected for in-depth analysis and could be classified into eight subfamilies;real-time fluorescence quantitative PCR validation of six ABC transporter protein genes showed that their expression profiles were generally consistent with the results of transcriptome sequencing,indicating that the results of transcriptome sequencing was reliable.