昆虫学报 ›› 2019, Vol. 62 ›› Issue (6): 694-702.doi: 10.16380/j.kcxb.2019.06.005

• 研究论文 • 上一篇    下一篇

基于转录组数据的齿缘刺猎蝽微卫星分子标记开发

黎东海1,2, 赵萍1,2,*   

  1. (1. 广东省生物资源应用研究所, 广东省动物保护与资源利用重点实验室, 广东省野生动物保护和利用公共实验室, 广州 510260; 2. 凯里学院植物保护系, 贵州凯里 556011)
  • 出版日期:2019-06-20 发布日期:2019-06-04

Development of microsatellite markers based on the transcriptome data of Sclomina erinacea (Heteroptera: Reduviidae)

LI Dong-Hai1,2, ZHAO Ping1,2,*   

  1. (1. Guangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Institute of Applied Biological Resources, Guangzhou 510260, China; 2. Department of Plant Protection, Kaili University, Kaili, Guizhou 556000, China)
  • Online:2019-06-20 Published:2019-06-04

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摘要：

【目的】齿缘刺猎蝽Sclomina erinacea是我国猎蝽科天敌昆虫常见种类，其不同地理种群存在明显形态差异。本研究旨在利用已经获得的齿缘刺猎蝽转录组数据筛选微卫星位点，为齿缘刺猎蝽种群遗传多样性和遗传分化研究开发可靠的微卫星分子标记。【方法】基于高通量测序技术平台Illumina HiSeqTM 2000获得齿缘刺猎蝽转录组数据(42 215条unigenes)，利用MISA软件进行搜索发掘SSR标记；利用Primer Premier 3软件设计SSR引物，从中随机选取54对SSR引物，利用PCR技术在中国齿缘刺猎蝽9个地理种群上进行验证。【结果】利用MISA软件搜索到微卫星位点2 395个，它们分布在2 107条unigenes上，其主要重复类型是三核苷酸重复(占总SSR的44.43%)，其次是二核苷酸重复(占总SSR的40.08%)，再次是四核苷酸重复(占总SSR的12.94%)。利用Primer Premier 3 软件成功设计出2 000余对SSR引物。随机选取的54对引物对9个齿缘刺猎蝽不同地理种群进行的SSR位点PCR扩增结果表明，共有16对引物较好地扩增出目的片段。【结论】研究表明利用齿缘刺猎蝽转录组数据可以大量发掘微卫星分子标记。本研究为进一步开展齿缘刺猎蝽的种群遗传学研究奠定了基础。

关键词: 齿缘刺猎蝽, 转录组, 微卫星, 引物设计, 高通量测序

Abstract: 【Aim】 Sclomina erinacea is a common natural enemy insect in forest ecological system of China, and there are obvious morphological differences among its different geographical populations. This research aims to screen microsatellite loci based on the transcriptome data of S. erinacea, so as to develop reliable microsatellite markers for the further study of the genetic diversity and genetic differentiation of S. erinacea populations. 【Methods】 The SSR markers were searched by MISA software among 42 215 unigenes of the transcriptome data previously obtained by high-throughput sequencing platform Illumina HiSeqTM 2000. SSR primers were designed by using Primer Premier 3 software. Fifty-four SSR primers randomly selected were tested in nine geographical populations of S. erinacea from China by PCR. 【Results】 A total of 2 395 SSR loci were found in 2 107 unigenes by MISA software. The main repeat types were trinucleotide repeats (44.43%), followed by dinucleotide repeats (40.08%) and tetranucleotide repeats (12.94%). More than 2 000 pairs of SSR primers were successfully designed by using Primer Premier 3 software. The PCR results of SSR loci of nine different geographical populations of S. erinacea with 54 pairs of primers randomly selected showed that 16 pairs of SSR primers could successfully amplify target fragments. 【Conclusion】 The results show that it is feasible to exploit microsatellite markers from the transcriptome data of S. erinacea. This study lays a foundation for the further study of population genetics of S. erinacea.

Key words: Sclomina erinacea, transcriptome, microsatellite, primer design, high-throughput sequencing

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