梨小食心虫,中肠,转录组,SSR,分子标记,Illumina HiSeq," /> 梨小食心虫,中肠,转录组,SSR,分子标记,Illumina HiSeq,"/> <span style="font-family:宋体;">梨小食心虫幼虫中肠转录组及</span><span>SSR</span><span style="font-family:宋体;">分子标记分析</span>

昆虫学报 ›› 2018, Vol. 61 ›› Issue (11): 1272-1283.doi: 10.16380/j.kcxb.2018.11.004

• 研究论文 • 上一篇    下一篇

梨小食心虫幼虫中肠转录组及SSR分子标记分析

冷春蒙1,2, 李引1,2, 胡迪1,2, 仵均祥1,2, 李怡萍1,2,*   

  1.  (1. 西北农林科技大学植物保护学院, 植保资源与病虫害治理教育部重点实验室, 陕西杨凌 712100; 2. 西北农林科技大学植物保护学院, 农业部西北黄土高原作物有害生物综合治理重点实验室, 陕西杨凌 712100)
  • 出版日期:2018-11-20 发布日期:2018-11-20

Analysis of the larval midgut transcriptome and SSR markers in Grapholitha molesta (Lepidoptera: Tortricidae)

LENG Chun-Meng1,2, LI Yin1,2, HU Di1,2, WU Jun-Xiang1,2, LI Yi-Ping1,2,*   

  1. (1. Key Laboratory of Plant Protection Resources and Pest Management, Ministry of Education, College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China; 2. Key Laboratory of Integrated Pest Management on Crops in Northwestern Loess Plateau, Ministry of Agriculture, CollegeofPlantProtection,NorthwestA&FUniversity,Yangling,Shaanxi712100,China)
  • Online:2018-11-20 Published:2018-11-20

摘要:  

【目的】梨小食心虫Grapholitha molesta是一种重要的世界性果树害虫。昆虫中肠在食物消化、免疫反应、生长发育等方面发挥着重要作用。本研究旨在建立梨小食心虫幼虫中肠转录组数据库,挖掘其基因信息。【方法】利用高通量测序平台(Illumina HiSeq X Ten)对梨小食心虫幼虫中肠进行转录组测序、组装及生物信息学分析;进而利用转录组数据进行梨小食心虫幼虫中肠SSR分子标记鉴定。【结果】总计获得梨小食心虫幼虫中肠转录组96 419unigenes,与公共数据库比对,共注释到57 300unigenes。通过GO数据库注释,将unigenes的功能分为生物学进程、细胞组分和分子功能三大类55个功能区,其中参与细胞进程、细胞和细胞组分及结合功能的unigenes比例较大。KOG结果显示,10 090unigenes归到25个基因家族,注释到一般功能预测的最多。KEGG数据库中,10 250条序列注释到232个代谢通路,注释到核糖体的基因数最多。此外,用MISH软件,在10 600unigenes中搜索到12 690SSR位点,通过PCR开发出6SSR位点。【结论】本研究成功组装梨小食心虫幼虫中肠参考转录组,为以中肠为靶标的害虫防治提供理论依据。

关键词:  , 梨小食心虫')">梨小食心虫, 中肠')">中肠, 转录组')">转录组, SSR, 分子标记')">分子标记, Illumina HiSeq

Abstract: Aim The oriental fruit moth, Grapholitha molesta is an important worldwide fruit pest. The midgut of insects plays an important role in digestion, immune response, growth and development. The aim of this study is to establish the larval midgut transpcriptome database of G. molesta, and to tap its genetic information. Methods The larval midgut transcriptome of G. molesta was sequenced, assembled and subjected to bioinformatics analysis by using a high throughput sequencing platform (Illumina HiSeq X Ten), and then the SSR molecular markers from the larval midgut were identified by using the transcriptome data. Results A total of 96 419 unigenes in the larval midgut transcriptome of G. molesta were obtained, and 57 300 unigenes were annotated in the public databases. All unigenes were assigned to 55 functional sub-categories of three categories (biological processes, cellular components and molecular function) using Gene Ontology, and a lot of unigenes were annotated to be related to cellular process, cell and cell part, and binding. The results of KOG showed that 10 090 sequences were classified into 25 gene families, with the most annotated to the general function. In the KEGG database, 10 250 unigenes were annotated to 232 metabolic pathways, and the number of genes annotated to ribosomes was the most. In addition, using MISH software, 12 690 SSR loci were searched in 10 600 unigenes, and six SSR loci were developed by PCR. Conclusion In this study we successfully assembled the larval midgut reference transcriptome of G. molesta, which provides a theoretical basis for pest control targeting the midgut.

Key words: Grapholitha molesta, midgut, transpcriptome, SSR, molecular marker, Illumina HiSeq