昆虫学报 ›› 2016, Vol. 59 ›› Issue (4): 421-426.doi: 10.16380/j.kcxb.2016.04.007

• 研究论文 • 上一篇    下一篇

假眼小绿叶蝉微卫星位点的生物信息学分析

李倩1, 陈学新2, 韩宝瑜1,*   

  1. (1. 中国计量大学生命科学学院, 浙江省生物计量及检验检疫技术重点实验室, 杭州 310018; 2. 浙江大学昆虫科学研究所, 水稻生物学国家重点实验室, 农业部农业昆虫学重点实验室, 杭州 310058)
  • 出版日期:2016-04-20 发布日期:2016-04-20

Bioinformatic analysis of microsatellites in the tea green leafhopper, Empoasca vitis (Hemiptera: Cicadellidae)

LI Qian1, CHEN Xue-Xin2, HAN Bao-Yu1,*   

  1. (1. Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, College of Life Sciences, China Jiliang University, Hangzhou 310018, China; 2. Institute of Insect Sciences, Zhejiang University; State Key Laboratory of Rice Biology, Ministry of Agriculture Key Laboratory of Agricultural Entomology, Hangzhou 310058, China)
  • Online:2016-04-20 Published:2016-04-20

摘要: 【目的】为开发假眼小绿叶蝉Empoasca vitis分子标记,采用高通量测序技术对假眼小绿叶蝉DNA进行了测序与分析。【方法】本研究基于Illumina HiSeq测序技术,构建了PE文库(~400 bp),对获得的测序数据利用生物信息学分析手段完成全基因组扫描,并进一步使用MISA分析鉴定基因组序列中出现的微卫星序列(SSR)。针对微卫星序列共设计10对引物,并使用3步法进行引物多态性筛选。【结果】共计检测Scaffold数量为183 194条,其中包含SSR的Scaffold共计1 545条,共计筛选出1 569个SSR位点。在假眼小绿叶蝉的微卫星中,共包括87种重复基元类型,二核苷酸与三核苷酸重复序列为主要重复类型,分别占SSRs总数的70.26%和27.84%;二核苷酸重复基元CA/TG和三核苷酸重复基元AAT/ATT是优势重复基元,分别占SSRs总数的33.96%和5.86%。在设计的10对引物中,5对具有多态性,在8个假眼小绿叶蝉个体中共发现16个等位基因。【结论】结果说明假眼小绿叶蝉SSR位点在多态性方面具有极大的可开发性,具有多态性的SSR位点可对假眼小绿叶蝉种群间的分化,种群间的扩散机理和途径及影响因素等问题提供分子视角。

关键词: 假眼小绿叶蝉, 基因组, 微卫星, 生物信息学, 高通量测序

Abstract: 【Aim】 In order to develop the molecular markers of the tea green leafhopper, Empoasca vitis, the high-throughput sequencing technologies were used in sequencing and analysis of DNA of the leafhopper. 【Methods】 Based on Illumina HiSeq sequencing technology, Illumina PE library (~400 bp) was constructed, and then the scanning of the whole genome sequences was finished using bioinformatics methods. Furthermore, MISA was used to identify the microsatellites in the genome. Ten pairs of primers were designed according to the microsatellite loci, and three-step procedure was used to screen the polymorphism of primers. 【Results】 Totally 183 194 scaffolds were examined, among which 1 545 contain microsatellites, and 1 569 microsatellites were identified in all these scaffolds. Among all 87 kinds of repeat motifs, the dominant microsatellite types were dinucleotide and trinucleotide repeats, accounting for 70.26% and 27.84% of the total SSRs, respectively, in E. vitis. CA/TG and AAT/ATTare the most frequent motifs, accounting for 33.96% and 5.86% of the total SSRs, respectively. Five primers among 10 primers designed revealed polymorphism, and 16 alleles were detected from eight individuals of E. vitis. 【Conclusion】 The results suggest that SSRs of E. vitis with polymorphism have potential for further use research. The polymorphism of SSRs may provide a molecular perspective of population divergence, dispersal mechanisms, diffusion path and influencing factors in E. vitis.

Key words: Empoasca vitis, genome, microsatellite, bioinformatics, high-throughput sequencing