基于微卫星标记的中国枣食芽象甲地理种群遗传多样性分析
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摘要
【目的】枣食芽象甲Scythropus yasumatsui是我国北方枣树Zizyphus jujuba上的一种重要的灾害性害虫,在陕西、山西、河北、河南等枣树主产区普遍发生。本研究旨在揭示我国枣食芽象甲不同种群间的遗传分化和基因交流规律。【方法】利用枣食芽象甲转录组测序的SSR序列,使用荧光标记PCR及毛细管电泳分型方法,筛选出8个微卫星位点,对我国5个省份(山西、陕西、宁夏、河北和河南)10个地理种群共308头枣食芽象甲样本进行种群遗传多样性分析。【结果】8个SSR位点均存在无效等位基因且偏离哈迪-温伯格平衡。各位点的有效等位基因数(Ne)为2.113～8.016,多态信息含量(PIC)为0.561～0.908,期望杂合度(He)为0.476～0.865;种群间遗传分化系数(Fst)平均值为0.151;基因流(Nm)平均值为1.594。枣食芽象甲种群间遗传分化系数与地理距离之间显著正相关(r=0.596,P=0.0035),基于Nei’s遗传距离和Cavalli-Sforza&Edwards余弦遗传距离的系统进化树将10个地理种群均聚为3个相同的分支。【结论】结果说明,枣食芽象甲种群遗传多样性较高,不同地理种群间存在高度的遗传分化,且有一定的基因交流;地理隔离是影响枣食芽象甲地理种群遗传分化和基因交流的重要因素。
【Aim】Scythropus yasumatsui, a major pest of Chinese jujube(Zizyphus jujuba) trees in northern China, is widely distributed in the main producing areas of jujube in provinces including Shaanxi, Shanxi, Hebei and Henan in China. This study aims to elucidate the genetic differentiation and gene exchange among S. yasumatsui populations in China. 【Methods】 Based on simple sequence repeats(SSRs) from the previously established transcriptome database, eight SSR loci were screened by PCR with fluorescence labeling and capillary electrophoresis genotyping, and the population genetic diversity was analyzed based on 308 individuals of 10 geographic populations of S. yasumatsui from five provinces(Shanxi, Shaanxi, Ningxia, Hebei and Henan). 【Results】 All of the eight microsatellite loci had null alleles and deviated from Hardy-Weinberg equilibrium. Among the loci, the effective number of alleles(Ne) ranged from 2.113 to 8.016, the polymorphic information content(PIC) ranged from 0.561 to 0.908, and the expected heterozygosity(He) was between 0.476 and 0.865. The average index of genetic differentiation(Fst) among populations was 0.151, and the average gene flow(Nm) was 1.594. There was a significantly positive correlation between the genetic differentiation index and the geographic distance among different populations of S. yasumatsui(r=0.596, P=0.0035), and the phylogenetic trees based on Nei's genetic distance and Cavalli-Sforza & Edwards genetic distance revealed that the 10 populations were all clustered into the three identical branches. 【Conclusion】 The results suggest that S. yasumatsui populations have a high level of genetic diversity, and there are a high genetic differentiation and a certain gene flow among different geographic populations. Geographic division is a critical factor which affects the genetic differentiation and gene exchange among S. yasumatsui populations.
【Aim】Scythropus yasumatsui, a major pest of Chinese jujube(Zizyphus jujuba) trees in northern China, is widely distributed in the main producing areas of jujube in provinces including Shaanxi, Shanxi, Hebei and Henan in China. This study aims to elucidate the genetic differentiation and gene exchange among S. yasumatsui populations in China. 【Methods】 Based on simple sequence repeats(SSRs) from the previously established transcriptome database, eight SSR loci were screened by PCR with fluorescence labeling and capillary electrophoresis genotyping, and the population genetic diversity was analyzed based on 308 individuals of 10 geographic populations of S. yasumatsui from five provinces(Shanxi, Shaanxi, Ningxia, Hebei and Henan). 【Results】 All of the eight microsatellite loci had null alleles and deviated from Hardy-Weinberg equilibrium. Among the loci, the effective number of alleles(Ne) ranged from 2.113 to 8.016, the polymorphic information content(PIC) ranged from 0.561 to 0.908, and the expected heterozygosity(He) was between 0.476 and 0.865. The average index of genetic differentiation(Fst) among populations was 0.151, and the average gene flow(Nm) was 1.594. There was a significantly positive correlation between the genetic differentiation index and the geographic distance among different populations of S. yasumatsui(r=0.596, P=0.0035), and the phylogenetic trees based on Nei's genetic distance and Cavalli-Sforza & Edwards genetic distance revealed that the 10 populations were all clustered into the three identical branches. 【Conclusion】 The results suggest that S. yasumatsui populations have a high level of genetic diversity, and there are a high genetic differentiation and a certain gene flow among different geographic populations. Geographic division is a critical factor which affects the genetic differentiation and gene exchange among S. yasumatsui populations.
引文
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Chapuis MP, Loiseau A, Michalakis Y, Lecoq M, Estoup A, 2005. Characterization and PCR multiplexing of polymorphic microsatellite loci for the locust Locusta migratoria. Mol. Ecol. Notes, 5(3): 554-557.
Dieringer D, Schl?tterer C, 2003. MICROSATELLITE ANALYSER (MSA): a platform independent analysis tool for large microsatellite data sets. Mol. Ecol. Notes, 3(1): 167-169.
Du XD, 2007. Biological characteristics and control of Scythropus yasumatsui. Shanxi Fruits, (2): 43. [杜仙当, 2007. 枣飞象生物学特性及防治. 山西果树, (2): 43]
Duan XL, 2015. Genetic Diversity and Genetic Structure of Bird Cherry-oat Aphid, Rhopalosiphum padi (L.) in China and Inductive Effect of Pesticide. PhD Dissertation, Northwest A&F University, Yangling, Shaanxi. [段辛乐, 2015. 我国禾谷缢管蚜种群遗传多样性和遗传结构及农药对其胁迫效应研究. 陕西杨凌: 西北农林科技大学博士学位论文]
Excoffier L, Lischer HEL, 2010. Arlequin suite ver 3.5: a new series of programs to perform population genetics analyses under Linux and Windows. Mol. Ecol. Resour., 10(3): 564-567.
Fang B, Duan XL, Zhang YL, Yang ZQ, Chen MH, 2017. Microsatellite development and genetic diversity analysis of apple buprestid Agrilus mali Matsumura. Acta Phytophy. Sin., 44(4): 559-566. [房冰, 段辛乐, 张彦龙, 杨忠岐, 陈茂华, 2017. 苹果小吉丁虫微卫星开发及其种群遗传结构分析. 植物保护学报, 44(4): 559-566]
Gao LN, Wang LX, Yang YQ, Gao GY, 1977. Preliminary investigation of Scythropus yasumatsui. For. Sci. Technol., (8): 18. [高犁牛, 王来喜, 杨有乾, 高冠玉, 1977. 枣飞象的初步调查研究. 林业科技通讯, (8): 18]
Glaubitz JC, 2004. Convert: a user-friendly program to reformat diploid genotypic data for commonly used population genetic software packages. Mol. Ecol. Notes, 4(2): 309-310.
Guzmán N, Contrerasdíaz H, Lanteri A, Juan C, Confalonieri V, 2013. Isolation and characterization of microsatellite loci in the fruit tree weevil Naupactus xanthographus (Coleoptera: Curculionidae): cross-amplification in related species of the Naupactus-Pantomorus complex. J. Genet., 89: e23-e27.
Hong B, Zhang F, Li YM, Zhang SL, Chen ZJ, 2017. Spatial distribution of Scythropus yasumatsui Kono et Morimoto adults in jujube orchard of Northern Shaanxi. Plant Prot., 43(6): 113-117, 122. [洪波, 张锋, 李英梅, 张淑莲, 陈志杰, 2017. 食芽象甲成虫在陕北枣园的空间分布格局. 植物保护, 43(6): 113-117, 122]
Hu WP, Liang TK, 2008. Forecast and control of Scythropus yasumatsui. Shanxi Agric., (1): 41. [胡维平, 梁廷康, 2008. 枣飞象的预测预报及防治. 山西农业, (1): 41]
Huang WZ, Li DC, 1993. Spatial distribution pattern of overwintering larvae of Scythropus yasumatsui. Entomol. Knowl., 30(6): 348-350. [黄维正, 李东成, 1993. 枣飞象越冬幼虫空间分布格局的研究. 昆虫知识, 30(6): 348-350]
Jarne P, Lagoda PJL, 1996. Microsatellites, from molecules to populations and back. Trends Ecol. Evol., 11(10): 424-429.
Jia ZB, Liu QX, Liu HY, 1991. Preliminary observation on life habit and control of Scythropus yasumatsui. J. Gansu For. Sci. Technol., (2): 49-50. [贾增波, 刘启雄, 刘鸿源, 1991.枣芽象甲生活习性初步观察及其防治. 甘肃林业科技, (2): 49-50]
Kirk H, Dorn S, Mazzi D, 2013. Molecular genetics and genomics generate new insights into invertebrate pest invasions. Evol. Appl., 6(5): 842-856.
Li MM, Li BL, Wu JX, Xu XL, 2018. Screening of microsatellite loci and the genetic diversity analysis of Mythimna separata (Lepidoptera: Noctuidae) populations in China. Acta Entomol. Sin., 61(6): 712-719. [李梅梅, 李伯辽, 仵均祥, 许向利, 2018. 我国粘虫种群的微卫星位点筛选及遗传多样性分析. 昆虫学报, 61(6): 712-719]
Li WA, Zhao P, Wang PX, 2007. The disease and pests of jujube and their control strategies in Shaanxi Province. J. Northwest For. Univ., 22(5): 120-123. [李文爱, 赵鹏, 王培新, 2007. 陕西枣树主要病虫害及防治对策研究. 西北林学院学报, 22(5): 120-123]
Li WF, Yang L, Tang K, Zeng L, Liang GW, 2007. Microsatellite polymorphism of Bactrocera dorsalis (Hendel) populations in China. Acta Entomol. Sin., 50(12): 1255-1262. [李伟丰, 杨朗, 唐侃, 曾玲, 梁广文, 2007. 中国桔小实蝇种群的微卫星多态性分析. 昆虫学报, 50(12): 1255-1262]
Liu GS, 1991. The life habit and control of Scythropus yasumatsui. Shanxi For. Sci. Technol., (3): 49. [刘光生, 1991. 枣飞象生活习性及防治. 山西林业科技, (3): 49]
Men QL, Chen MH, Zhang YL, Feng JN, 2012. Amplifying stability and genetic diversity of microsatellite loci in codling moth Cydia pomonella (Lepidoptera: Tortricidae) populations in China. Acta Phytophy. Sin., 39(4): 341-346. [门秋雷, 陈茂华, 张雅林, 冯纪年, 2012. 中国疫区内苹果蠹蛾微卫星位点的扩增稳定性及遗传多样性. 植物保护学报, 39(4): 341-346]
Miller MP, 1997. Tools for Population Genetic Analysis (TFPGA), Version 1.3: a Windows? program for the analysis of allozyme and molecular population genetic data. Department of Biological Sciences, Northern Arizona University, Flagstaff, AZ.
Nei M, 1978. Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics, 89(3): 583-590.
Powell W, Machray GC, Provan J, 1996. Polymorphism revealed by simple sequence repeats. Trends Plant Sci., 1(7): 215-222.
Ren Q, Cao LF, Zhao HX, Wang RS, Cheng S, Luo WH, Cao L, Ji CH, 2018. Analysis of genetic diversity of main Apis cerana populations in China. J. Henan Agric. Univ., 52(1): 91-95. [任勤, 曹联飞, 赵红霞, 王瑞生, 程尚, 罗文华, 曹兰, 姬聪慧, 2018. 中国主要东方蜜蜂种群的遗传多样性分析. 河南农业大学学报, 52(1): 91-95]
Rousset F, 2008. GENEPOP 007: a complete reimplementation of the GENEPOP software for Windows and Linux. Mol. Ecol. Resour., 8(1): 103-106.
Schuelke M, 2000. An economic method for the fluorescent labeling of PCR fragments. Nat. Biotechnol., 18(2): 233-234.
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