泽兰实蝇雄性附腺高通量转录组测序数据组装和分析

De novo assembly and transcriptome characterization of male accessory gland in Procecidochares utilis Stone

泽兰实蝇Procecidochares utilis Stone是恶性杂草紫茎泽兰Eupatorium adenophorum Spreng重要的专食性天敌。为进一步开发泽兰实蝇的基因资源,深入了解其遗传信息,本研究采用Illumina HiSeq 2000高通量测序技术对泽兰实蝇的雄性附腺组织进行了转录组测序,构建了转录组数据库,获得62 684 346条Clean Reads数据;拼接组装后获得89 195条Unigene数据,平均长度为898 bp;与NR、NT、KO、SwissProt、PFAM、GO、KOG七大数据库进行Blast信息比对(E-value为10-5),共获得52 743个注释基因;与NR数据库比对发现,泽兰实蝇雄性附腺转录组基因序列与瓜实蝇Bactrocera cucurbitae具有较高的同源性,为29.1%;将泽兰实蝇转录组的Unigene的功能通过与KOG数据库进行注释比对划分为25类;GO数据库注释可分为3类,即细胞组分、生物过程和分子功能,共包括65个分支;KEGG分析发现,泽兰实蝇转录组数据中按照代谢通路可分为92类,利用Blast蛋白库比对和Estscan软件进行CDS预测,获得长度大于300 nt的CDS共48 509个;通过SSR分析,共获得69 352个SSR标记,数量最高的SSR类型为单碱基重复,为47 139条,出现频率为67.97%,最少的是五碱基重复SSR,只有27条,出现频率仅为0.039%。本研究中获得的转录组信息可为今后进行泽兰实蝇分子标记的开发和关键基因的克隆及功能分析等研究提供基础数据。

Procecidochares utilis Stone is an important predatory natural enemies of the virulent weed Eupatorium adenophorum Spreng. In order to further develop genetic resources of P. utilis and investigate the profile of gene expression and elucidate the functional genes, In this study,by performing Illumina Hiseq 2000 and de novo assembly, the transcriptome of male accessory gland was sequenced, the data were filtered and assembled, and the unigenes were compared and annotated. Totally, 62 684 346 valid short sequences were obtained, and 89 195 unigenes were spliced by de novo, the average length was 898 bp. Also, blast information was compared with NR, NT, KO, SwissProt, PFAM, GO, KOG, etc.(E-value was 10-5), a total of 52 743 annotation genes were obtained. Compared with the NR database, the genomic sequence of the pheromone transcriptome was highly homologous to Bactrocera cucurbitae, which was 29.1%. Unigene function of P. utilis transcriptome was divided into 25 categories by annotated comparison with the KOG database. According to GO annotation database, unigene function can be divided into three categories, i.e., cellular components, biological processes and molecular functions, including a total of 65 branches. KEGG analysis found that the transcriptome data were divided into 92 types according to the metabolic pathways. Using Blast protein library comparison and Estscan software to predict CDS, a total of 48 509 CDS with a length of more than 300 nt were obtained. Using MISA software, the results showed that there were 69 352 SSR from the 89 195 unigenes, and the most type of SSR was mononucleotide 47 139 with the frequency of 67.97%. Moreover, the five hexanucleotide only had 27 repeat SSR and the frequency was only 0.039%. The transcriptome information obtained in this study can provide basic data for the future development of the molecular markers of P. utilis molecule and the clone and functional analysis of key genes.