广东省鼎湖山国家级自然保护区球孢白僵菌遗传多样性研究
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摘要
【目的】了解广东省球孢白僵菌Beauveria bassiana的群体遗传结构、基因流、菌株分化、遗传变异和寄主类型,以及与生态环境的相关性,为筛选优良生产菌株提供后备种质资源。【方法】采用SSR分子标记对广东省鼎湖山国家级自然保护区(以下简称"DHS")采集的81株球孢白僵菌进行遗传结构和多样性分析。【结果】使用8对SSR引物扩增81个菌株产生多态位点数为58个,多态位点比率为100%,其中,引物Ba12多态位点数最多(10个)。通过Popgene 32软件包分析发现,DHS球孢白僵菌种群Nei’s基因多样性指数(h)为0.212 6,Shannon信息指数(I_s)为0.348 7,可见其遗传多样性水平较高,群体异质性较强。将群体分别按照寄主和土壤、不同寄主目划分为不同亚群进一步分析,发现土壤亚群遗传多样性水平(h=0.192 5,I_s=0.309 9)略高于寄主亚群(h=0.176 9,I_s=0.278 3),种群间遗传分化程度较弱(N_m=1.662 9,G_(st)=0.130 7);不同寄主目群体间的遗传分化较小且基因流明显。基于UPGMA聚类分析发现,各分支菌株的分布趋势与分离基质、不同寄主目均没有相关性。【结论】DHS球孢白僵菌遗传谱系与分离基质和寄主来源均无明显相关性,不同生态环境和寄主类型的影响共同维持了DHS球孢白僵菌种群内遗传变异的多样性。
【Objective】To understand the population genetic structure, gene flow, strain differentiation of Beauveria bassiana in Guangdong Province, analyze the correlation between genetic variation and host type,ecological environment, and provide a large number of reserved germplasm resources for screening and production of superior strains.【Method】The genetic variation and diversity of 81 B. bassiana isolates from Dinghu Mountain National Nature Reserve of Guangdong Province(DHS) were assessed using SSR markers.【Result】Eight SSR primers were amplified to produce 58 polymorphic points, the percentage of polymorphic loci was 100%, and the number of polymorphic points of primer Ba12 was the largest(10). Nei's genetic diversity index(h) was 0.212 6 and Shannon's information index(I_s) was 0.348 7 based on the analysis using the Popgene32 software package, which indicated the genetic diversity and population heterogeneity levels of B. bassiana were high in DHS. Eighty-one strains of B. bassiana were divided into different subpopulations according to host/soil and insect order respectively. The genetic diversity of soil subpopulation(h=0.192 5, I_s=0.309 9) was slightly higher than that of host subpopulation(h=0.176 9, I_s=0.278 3), suggesting a generally low genetic differentiation among subpopulations(N_m=1.662 9, G_(st)=0.130 7).Genetic differentiation among subpopulations in different insect order was low and was associated with strong gene flow. The distribution of isolates had no obvious correlation with isolation medium or insect orders based on UPGMA clustering.【Conclusion】The genetic lineage of B. bassiana in DHS is not correlated with isolation medium or insect orders. The diversification of B. bassiana population in DHS is maintained by different ecological environment and host type.
【Objective】To understand the population genetic structure, gene flow, strain differentiation of Beauveria bassiana in Guangdong Province, analyze the correlation between genetic variation and host type,ecological environment, and provide a large number of reserved germplasm resources for screening and production of superior strains.【Method】The genetic variation and diversity of 81 B. bassiana isolates from Dinghu Mountain National Nature Reserve of Guangdong Province(DHS) were assessed using SSR markers.【Result】Eight SSR primers were amplified to produce 58 polymorphic points, the percentage of polymorphic loci was 100%, and the number of polymorphic points of primer Ba12 was the largest(10). Nei's genetic diversity index(h) was 0.212 6 and Shannon's information index(I_s) was 0.348 7 based on the analysis using the Popgene32 software package, which indicated the genetic diversity and population heterogeneity levels of B. bassiana were high in DHS. Eighty-one strains of B. bassiana were divided into different subpopulations according to host/soil and insect order respectively. The genetic diversity of soil subpopulation(h=0.192 5, I_s=0.309 9) was slightly higher than that of host subpopulation(h=0.176 9, I_s=0.278 3), suggesting a generally low genetic differentiation among subpopulations(N_m=1.662 9, G_(st)=0.130 7).Genetic differentiation among subpopulations in different insect order was low and was associated with strong gene flow. The distribution of isolates had no obvious correlation with isolation medium or insect orders based on UPGMA clustering.【Conclusion】The genetic lineage of B. bassiana in DHS is not correlated with isolation medium or insect orders. The diversification of B. bassiana population in DHS is maintained by different ecological environment and host type.
引文
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[8]GARRIDO-JURADO I,MARQUEZ M,ORTIZ-UR-QUIZA A,et al.Genetic analyses place most Spanish isolates of Beauveria bassiana in a molecular group with word-wide distribution[J].BMC Microbiol,2011,11(1):84.
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[10]WANG C,SHAH F A,PATEL N,et al.Molecular in-vestigation on strain genetic relatedness and population structure of Beauveria bassiana[J].Environ Microbiol,2003,5(10):908-915.
[11]胡晓磊,何玲敏,陈雪,等.中国南方球孢白僵菌的遗传多样性和种群遗传结构[J].中国生物防治学报,2013,29(1):31-41.
[12]何玲敏,胡晓磊,陈雪,等.中国北方球孢白僵菌的遗传多样性和种群遗传结构[J].应用生态学报,2012,23(11):3087-3095.
[13]MEYLING N V,LUBECK M,BUCKLEY E P,et al.Community composition,host range and genetic structure of the fungal entomopathogen Beauveria in adjoining agricultural and seminatural habitats[J].Mol Ecol,2009,18(6):1282-1293.
[14]MEYLING N V,PILZ C,KELLER S,et al.Diversity of Beauveria spp.isolates from pollen beetles Meligethes aeneus in Switzerland[J].J Invert Pathol,2012,109(1):76-82.
[15]王滨,樊美珍,李增智.球孢白僵菌选择性培养基的筛选[J].安徽农业大学学报,2000,27(1):23-28.
[16]蔡悦.球孢白僵菌的繁殖与自然群体交配型关系的研究[D].合肥:安徽农业大学,2012.
[17]蒲顺昌,秦丽,陈名君,等.马尾松林中球孢白僵菌寄主转移和专化性的SSR标记分析[J].菌物学报,2013,32(4):698-709.
[18]盖红梅,任民.SSR数据处理宏程序DataTrans 1.0[J/OL].分子植物育种(网络版),2011,9(48):1359-1365.http://mpb.chinese.sophiapublisher.com.doi:10.5376/mpb.cn.2011.09.0048
[19]TAKAHATA N,NEI M.Fst and Gst Statistics in the finite island model[J].Genetics,1984,107(3):501-504.
[20]张正坤,孟鑫睿,张佳诗,等.吉林省球孢白僵菌遗传多样性与亚洲玉米螟化性相关性分析[J].中国生物防治学报,2015,31(6):836-844.
[21]AMOS W,HARWOOD J.Factors affecting levels of genetic diversity in natural populations[J].Philos Transe RSoc B:Biol Sci,1998,353(1366):177-186.
[22]李旻,王四宝,樊美珍,等.森林生态系中球孢白僵菌遗传多样性的ISSR分析[J].遗传,2006,28(8):977-983.
[23]陈雪娇,李云飞,孙娟娟,等.利用ISSR标记分析红脂大小蠹微生境中球孢白僵菌遗传多样性[J].菌物学报,2014,33(5):1015-1024.
[24]BIDOCHKA M,MENZIES F,KAMP A.Genetic groups of the insect-pathogenic fungus Beauveria bassiana are associated with habitat and thermal growth preferences[J].Arch Microbiol,2002,178(6):531-537.
[25]LUAN F,ZHANG S,CAI Y,et al.Identification of the molecular origin and development of a panzootic caused by Beauveria bassiana in praying mantis populations in eastern China[J].J Invert Pathol,2011,108:98-105.
[2]MEYLING N V,EILENBERG J.Ecology of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae in temperate agroecosystems:Potential for conservation biological control[J].Biol Control,2007,43(2):145-155.
[3]李增智.我国利用真菌防治害虫的历史、进展及现状[J].中国生物防治学报,2015,31(5):699-711.
[4]RODRIGUEZ-GONZáLEZá,MAYO S,GONZáLEZ-LóPEZó,et al.Inhibitory activity of Beauveria bassiana and Trichoderma spp.on the insect pests Xylotrechus arvicola(Coleoptera:Cerambycidae)and Acanthoscelides obtectus(Coleoptera:Chrisomelidae:Bruchinae)[J].Environ Monit Assess,2017,189(12):1-8.
[5]李鸿文,冯明光.球孢白僵菌不同菌株分生孢子的耐热能力[J].浙江大学学报(农业与生命科学版),2008,34(2):158-162.
[6]ST LEGER R J,ALLEE L L,MAY B,et al.World-wide distribution of genetic variation among isolates of Beauveria spp.[J].Mycol Res,1992,96(12):1007-1015.
[7]FERNANDES E K K,MORAES A M L,PACHECO RS,et al.Genetic diversity among Brazilian isolates of Beauveria bassiana:Comparisons with non-Brazilian isolates and other Beauveria species[J].J Appl Microbiol,2009,107(3):760-774.
[8]GARRIDO-JURADO I,MARQUEZ M,ORTIZ-UR-QUIZA A,et al.Genetic analyses place most Spanish isolates of Beauveria bassiana in a molecular group with word-wide distribution[J].BMC Microbiol,2011,11(1):84.
[9]PRABHUKARTHIKEYAN S R,KEERTHANA U,ARCHANA S,et al.Analysis of genetic diversity among different isolates of Beauveria bassiana by RAPDPCR[J/OL].J Biol Control,2017,31(1).https://doi.org/10.18311/jbc/2017/15581.
[10]WANG C,SHAH F A,PATEL N,et al.Molecular in-vestigation on strain genetic relatedness and population structure of Beauveria bassiana[J].Environ Microbiol,2003,5(10):908-915.
[11]胡晓磊,何玲敏,陈雪,等.中国南方球孢白僵菌的遗传多样性和种群遗传结构[J].中国生物防治学报,2013,29(1):31-41.
[12]何玲敏,胡晓磊,陈雪,等.中国北方球孢白僵菌的遗传多样性和种群遗传结构[J].应用生态学报,2012,23(11):3087-3095.
[13]MEYLING N V,LUBECK M,BUCKLEY E P,et al.Community composition,host range and genetic structure of the fungal entomopathogen Beauveria in adjoining agricultural and seminatural habitats[J].Mol Ecol,2009,18(6):1282-1293.
[14]MEYLING N V,PILZ C,KELLER S,et al.Diversity of Beauveria spp.isolates from pollen beetles Meligethes aeneus in Switzerland[J].J Invert Pathol,2012,109(1):76-82.
[15]王滨,樊美珍,李增智.球孢白僵菌选择性培养基的筛选[J].安徽农业大学学报,2000,27(1):23-28.
[16]蔡悦.球孢白僵菌的繁殖与自然群体交配型关系的研究[D].合肥:安徽农业大学,2012.
[17]蒲顺昌,秦丽,陈名君,等.马尾松林中球孢白僵菌寄主转移和专化性的SSR标记分析[J].菌物学报,2013,32(4):698-709.
[18]盖红梅,任民.SSR数据处理宏程序DataTrans 1.0[J/OL].分子植物育种(网络版),2011,9(48):1359-1365.http://mpb.chinese.sophiapublisher.com.doi:10.5376/mpb.cn.2011.09.0048
[19]TAKAHATA N,NEI M.Fst and Gst Statistics in the finite island model[J].Genetics,1984,107(3):501-504.
[20]张正坤,孟鑫睿,张佳诗,等.吉林省球孢白僵菌遗传多样性与亚洲玉米螟化性相关性分析[J].中国生物防治学报,2015,31(6):836-844.
[21]AMOS W,HARWOOD J.Factors affecting levels of genetic diversity in natural populations[J].Philos Transe RSoc B:Biol Sci,1998,353(1366):177-186.
[22]李旻,王四宝,樊美珍,等.森林生态系中球孢白僵菌遗传多样性的ISSR分析[J].遗传,2006,28(8):977-983.
[23]陈雪娇,李云飞,孙娟娟,等.利用ISSR标记分析红脂大小蠹微生境中球孢白僵菌遗传多样性[J].菌物学报,2014,33(5):1015-1024.
[24]BIDOCHKA M,MENZIES F,KAMP A.Genetic groups of the insect-pathogenic fungus Beauveria bassiana are associated with habitat and thermal growth preferences[J].Arch Microbiol,2002,178(6):531-537.
[25]LUAN F,ZHANG S,CAI Y,et al.Identification of the molecular origin and development of a panzootic caused by Beauveria bassiana in praying mantis populations in eastern China[J].J Invert Pathol,2011,108:98-105.