四川省草莓主产区灰葡萄孢菌的遗传多样性
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摘要
为明确四川省草莓灰葡萄孢Botrytis cinerea群体遗传结构及其多样性水平,采用ISSR分子标记技术对分离自四川省10个县(市)的195株灰葡萄孢菌进行了遗传多态性分析。结果表明,四川省灰葡萄孢菌多态性丰富,6条ISSR引物共产生了63个多态性位点,应用Popgene32软件计算四川省不同主产区域(除德阳广汉种群外)种群的Nei’s基因多样性指数(H)和Shannon信息指数(I)均达到了H>0.2、I>0.3的水平,表明四川省的灰葡萄孢菌具有丰富的遗传多样性;灰葡萄孢菌群体的遗传多样性(Ht)均值为0.2976,种群内遗传多样性(Hs=0.2458)远远高于种群间(Dst=0.0518)的遗传多样性;遗传分化系数(Gst)均值0.1742,基因流(Nm)均值2.3696,说明该地区灰葡萄孢菌种群间遗传分化不明显,群体内基因交流频繁。通过UPGMA法和Omishare Tools热图软件均可将10个采集点分为3个类群,来自绵阳江油的菌株单独构成一个类群,来自成都崇州和德阳广汉的菌株构成一个类群,其余的菌株构成另外一个类群;利用Structure2.3软件对195份灰葡萄孢菌进行群体结构分析,可将134份菌株划分成21个群,另外61个菌株被列为混合群体。
A total of 195 isolates of Botrytis cinerea was investigated through inter-simple sequence repeat(ISSR) markers with six ISSR primers to determine its genetic structure and diversity in Sichuan Province. The results showed that the pathogen was genetically polymorphic and generated 63 polymorphic loci(96.83%).By using Popgene32 software, the Nei's gene diversity index(H) and Shannon information index(I) were H>0.2 and I>0.3 respectively, showing rich genetic diversity of B. cinerea in Sichuan Province, except for population of Deyang in Guanghan region. The average genetic diversity(Ht) index was 0.2976. The diversity index of intrapopulations(Hs=0.2458) was much higher than that of interpopulations(Dst=0.0518). The average genetic discrepancy coefficient(Gst) and average gene flow(Nm) reached 0.1742 and 2.3696 respectively,indicating that there was no obviousness in genetic differentiation of B. cinerea inter populations, but gene exchange frequently occurred within populations. The isolates of B. cinerea from ten main strawberry-growing regions in Sichuan Province could be divided into three geographical groups based on UPGMA and Omishare Tools thermogram analysis. The isolates from Jiangyou were clustered in the first group, those from Chongzhou and Guanghan in second group, while the remainder in the third group. Population structure analysis by using software 2.3 showed that 195 isolates of B. cinerea were clustered into 21 groups composed of 134 isolates and the other 61 isolates were regarded as mixed group.
A total of 195 isolates of Botrytis cinerea was investigated through inter-simple sequence repeat(ISSR) markers with six ISSR primers to determine its genetic structure and diversity in Sichuan Province. The results showed that the pathogen was genetically polymorphic and generated 63 polymorphic loci(96.83%).By using Popgene32 software, the Nei's gene diversity index(H) and Shannon information index(I) were H>0.2 and I>0.3 respectively, showing rich genetic diversity of B. cinerea in Sichuan Province, except for population of Deyang in Guanghan region. The average genetic diversity(Ht) index was 0.2976. The diversity index of intrapopulations(Hs=0.2458) was much higher than that of interpopulations(Dst=0.0518). The average genetic discrepancy coefficient(Gst) and average gene flow(Nm) reached 0.1742 and 2.3696 respectively,indicating that there was no obviousness in genetic differentiation of B. cinerea inter populations, but gene exchange frequently occurred within populations. The isolates of B. cinerea from ten main strawberry-growing regions in Sichuan Province could be divided into three geographical groups based on UPGMA and Omishare Tools thermogram analysis. The isolates from Jiangyou were clustered in the first group, those from Chongzhou and Guanghan in second group, while the remainder in the third group. Population structure analysis by using software 2.3 showed that 195 isolates of B. cinerea were clustered into 21 groups composed of 134 isolates and the other 61 isolates were regarded as mixed group.
引文
Asadollahi M,Fekete E,Karaffa L,Flipphi M,árnyasi M,Esmaeili M,Váczy KZ,Sándor E,2013.Comparison of Botrytis cinerea populations isolated from two open-field cultivated host plants.Microbiological Research,168(6):379-388
Duan HJ,Zhang CY,Li XH,Guo XM,Ma SY,2008.Assessment of genetic diversity in Fusarium oxysporum f.sp.niveum by RAPD,ISSR and AFLP analysis.Mycosystema,27(2):267-276(in Chinese)
Dwiedi A,Khim N,Reynes C,Ravel P,Ma L,Tichit M,Bourchier C,Kim S,Dourng D,Khean C,Chim P.Siv SFrutos R,Lek D,Mercereau-Puijalon O,Ariey F,Menard D,Cornillot E,2016.Plasmodium falciparum,parasite population structure and gene flow associated to anti-malarial drugs resistance in Cambodia.Malaria Journal,15(1):319
Esterio M,Muňoz G,Ramos C,CofréG,Estévez R,Salinas A,Auger J,2011.Characterization of Botrytis cinerea isolates present in Thompson seedless table grapes in the central valley of Chile.Plant Disease,95(6):683-690
Feketeé,Fekete E,Irinyi L,Karaffa L,árnyasi M,Asadollahi M,Sándo E,2012.Genetic diversity of a Botrytis cinerea cryptic species complex in Hungary.Microbiological Research,167(5):283-291
Feng HJ,Wen CY,Yin XM,Hong KQ,Zang R,2017.Population genetic diversity analysis of Chinese jujube branch canker pathogens(Botryosphaeria dothidea)in China using ISSR markers.Journal of Fruit Science,34(8):55-65(in Chinese)
Foumier E,Giraud T,Loiseau A,Vautrin D,Estoup A,Solignac M,Cornue JM,Brygoo Y,2002.Characterization of nine polymorphic microsatellite loci in the fungus Botrytis cinerea(Ascomycota).Molecular Ecology Notes,2(3):253-255
Fournier E,Gladieux P,Giraud T,2013.The‘Dr Jekyll and Mr Hyde fungus’:noble rot versus gray mold symptoms of Botrytis cinerea on grapes.Evolutionary Applications,6(6):960-969
Gao CZ,Yang HL,Huangxia YQ,Huang JB,Li GQ,Zheng L,2017.Occurrence of grey mould disease in greenhouse-grown strawberry and its correlations with epidemic factors in Hubei Province.Scientia Agricultura Sinica,50(9):1617-1623(in Chinese)
Gong GS,Xu Q,Zhang M,Yang JZ,Chen HB,Shen SA,Tang TF,2010.A simple method for isolation of monospores from pathogenic fungi.Corn Science,18(1):126-127(in Chinese)
Huang Y,Zhu ZD,Duan CX,Wu XF,DongFang Y,2014.Genetic diversity of Botrytis cinerea isolates from broad bean.Scientia Agricultura Sinica,47(12):2335-2347(in Chinese)
Ibrahim AAA,Muhammad AAG,William B,Elsayed EH,2017.Multilocus genetic techniques,RAPD-PCR and ISSR-PCRmarkers and polygalacturonase activity as tools for differentiation among Alternaria solani isolates on tomato fruits and relation to their pathogenicity in Egypt.Asian Journal of Plant Pathology,11(1):18-27
Kumari S,Tayal P,Sharma E,Kapoor R,2014.Analyses of genetic and pathogenic variability among Botrytis cinerea isolates.Microbiological Research,169(11):862-872
Latoore BA,Elfarr K,Ferrada EE,2015.Gray mold caused by Botrytis cinerea limits grape production in Chile.Ciencia E Investigación Agraria,42(3):305-330
Li TD,Peng SW,Wang ZH,Lu GD,2010.Genetic diversity analysis of Rhizoctonia solani populations from Fujian using inter-simple sequence repeats(ISSR)technique.Acta Phytopathologica Sinica,40(2):186-194(in Chinese)
Li TG,Zhang L,Zhou SY,Li M,Li BD,2013.The study on species and genetic diversity of Botrytis cinerea populations in Shandong province.Chinese Agricultural Science Bulletin,29(10):137-143(in Chinese)
Montenegro J,Braun J,Fischer-Onaca O,Meier W,Matile S,2011.Synthetic polyion-counterion transport systems in polymersomes and gels.Organic&Biomolecular Chemistry,9(19):6623-6628
Moyano C,Alfonso C,Gallego J,Raposo R,Melgarejo P,2003.Comparison of RAPD and AFLP marker analysis as a means to study the genetic structure of Botrytis cinerea populations.European Journal of Plant Pathology,109(5):515-522
Rasiukevi?iūt?N,Rugienius R,?iksnianiene JB,2018.Genetic diversity of Botrytis cinerea from strawberry in Lithuania.Zemdirbyste-Agriculture,105(3):265-270
Rasiukevi?iūt?N,Valiuskaite A,Usellis N,Luk?ien?Z,2015.The effect of alternative plant protection measures on grey mould spread,strawberry growth and productivity.Sodininkyst?Ir Dar?ininkyst?,34:3-4
Wakeham A,Langton A,Adam SS,Kennedy R,2016.Interface of the environment and occurrence of Botrytis cinerea,in pre-symptomatic tomato crops.Crop Protection,90:27-33
Walker AS,Fournier E,2013.Habitat and host-specific differentiation in the multihost fungal pathogen Botryotinia fuckeliana and evidence for fungicide selection in populations.Modern Fungicides and Antifungal Compounds,7:167-172
Xu Z,Ali Z,Hou X,Li H,Yi JX,Abbasi PA,2013.Characterization of Chinese eggplant isolates of the fungal pathogen Verticillium dahliae from different geographic origins.Genetics and Molecular Research,12(1):183-195
Yamada K,Sonoda R,Ishikawa K,2016.Population genetic structure of QoI-resistant Pestalotiopsis longiseta isolates causing tea gray blight.Plant Disease,100:1686-1691
Yin DF,2015.The monitoring on fungicide resistance and the research on resistance mechanism of Botrytis cinerea from strawberry in Zhejiang Province.Master Thesis,Zhejiang University,Hangzhou.1-80(in Chinese)
Zhang J,2010.Studies on taxonomy of Botrytis species in Hubei Province and diversity of B.cinerea.PhDDissertation,Huazhong Agricultural University,Wuhan.5-10(in Chinese)
Zhang ZH,Gao XY,Du GD,Li H,2007.The trend of strawberryproduction:labour-saving cultivation.Chinese Agricultural Science Bulletin,23(10):101-103(in Chinese)
段会军,张彩英,李喜焕,郭小敏,马峙英,2008.基于RAPD、ISSR和AFLP对西瓜枯萎病菌遗传多样性的评价.菌物学报,27(2):267-276
冯慧静,文才艺,尹新明,洪坤奇,臧睿,2017.中国枣树干腐病菌(Botryosphaeria dothidea)群体遗传多样性的ISSR分析.果树学报,34(8):55-65
高翠珠,杨红玲,黄夏宇骐,黄俊斌,李国庆,郑露,2017.湖北省设施草莓灰霉病发生规律及流行因子分析.中国农业科学,50(9):1617-1623
龚国淑,徐琴,张敏,杨继芝,陈华保,申世安,唐太飞,2010.一种简便的病原真菌单孢分离方法研究.玉米科学,18(1):126-127
黄燕,朱振东,段灿星,武小菲,东方阳,2014.灰葡萄孢蚕豆分离物的遗传多样性.中国农业科学,47(12):2335-2347
李挺丹,彭世文,王宗华,鲁国东,2010.应用ISSR-PCR分析福建水稻纹枯菌的遗传多样性.植物病理学报,40(2):186-194
李廷刚,张路,周善跃,李敏,李宝笃,2013.山东省番茄灰霉病菌种类与遗传多样性研究.中国农学通报,29(10):137-143
尹大芳,2015.浙江省草莓灰霉病菌抗药性检测及抗性机制的研究.浙江大学硕士论文,杭州.1-80
张静,2010.湖北省灰霉病病菌区系和灰葡萄孢菌多样性研究.华中农业大学博士论文,武汉.5-10
张志宏,高秀岩,杜国栋,李贺,2007.草莓生产的发展趋势-省力化栽培.中国农学通报,23(10):101-103
Duan HJ,Zhang CY,Li XH,Guo XM,Ma SY,2008.Assessment of genetic diversity in Fusarium oxysporum f.sp.niveum by RAPD,ISSR and AFLP analysis.Mycosystema,27(2):267-276(in Chinese)
Dwiedi A,Khim N,Reynes C,Ravel P,Ma L,Tichit M,Bourchier C,Kim S,Dourng D,Khean C,Chim P.Siv SFrutos R,Lek D,Mercereau-Puijalon O,Ariey F,Menard D,Cornillot E,2016.Plasmodium falciparum,parasite population structure and gene flow associated to anti-malarial drugs resistance in Cambodia.Malaria Journal,15(1):319
Esterio M,Muňoz G,Ramos C,CofréG,Estévez R,Salinas A,Auger J,2011.Characterization of Botrytis cinerea isolates present in Thompson seedless table grapes in the central valley of Chile.Plant Disease,95(6):683-690
Feketeé,Fekete E,Irinyi L,Karaffa L,árnyasi M,Asadollahi M,Sándo E,2012.Genetic diversity of a Botrytis cinerea cryptic species complex in Hungary.Microbiological Research,167(5):283-291
Feng HJ,Wen CY,Yin XM,Hong KQ,Zang R,2017.Population genetic diversity analysis of Chinese jujube branch canker pathogens(Botryosphaeria dothidea)in China using ISSR markers.Journal of Fruit Science,34(8):55-65(in Chinese)
Foumier E,Giraud T,Loiseau A,Vautrin D,Estoup A,Solignac M,Cornue JM,Brygoo Y,2002.Characterization of nine polymorphic microsatellite loci in the fungus Botrytis cinerea(Ascomycota).Molecular Ecology Notes,2(3):253-255
Fournier E,Gladieux P,Giraud T,2013.The‘Dr Jekyll and Mr Hyde fungus’:noble rot versus gray mold symptoms of Botrytis cinerea on grapes.Evolutionary Applications,6(6):960-969
Gao CZ,Yang HL,Huangxia YQ,Huang JB,Li GQ,Zheng L,2017.Occurrence of grey mould disease in greenhouse-grown strawberry and its correlations with epidemic factors in Hubei Province.Scientia Agricultura Sinica,50(9):1617-1623(in Chinese)
Gong GS,Xu Q,Zhang M,Yang JZ,Chen HB,Shen SA,Tang TF,2010.A simple method for isolation of monospores from pathogenic fungi.Corn Science,18(1):126-127(in Chinese)
Huang Y,Zhu ZD,Duan CX,Wu XF,DongFang Y,2014.Genetic diversity of Botrytis cinerea isolates from broad bean.Scientia Agricultura Sinica,47(12):2335-2347(in Chinese)
Ibrahim AAA,Muhammad AAG,William B,Elsayed EH,2017.Multilocus genetic techniques,RAPD-PCR and ISSR-PCRmarkers and polygalacturonase activity as tools for differentiation among Alternaria solani isolates on tomato fruits and relation to their pathogenicity in Egypt.Asian Journal of Plant Pathology,11(1):18-27
Kumari S,Tayal P,Sharma E,Kapoor R,2014.Analyses of genetic and pathogenic variability among Botrytis cinerea isolates.Microbiological Research,169(11):862-872
Latoore BA,Elfarr K,Ferrada EE,2015.Gray mold caused by Botrytis cinerea limits grape production in Chile.Ciencia E Investigación Agraria,42(3):305-330
Li TD,Peng SW,Wang ZH,Lu GD,2010.Genetic diversity analysis of Rhizoctonia solani populations from Fujian using inter-simple sequence repeats(ISSR)technique.Acta Phytopathologica Sinica,40(2):186-194(in Chinese)
Li TG,Zhang L,Zhou SY,Li M,Li BD,2013.The study on species and genetic diversity of Botrytis cinerea populations in Shandong province.Chinese Agricultural Science Bulletin,29(10):137-143(in Chinese)
Montenegro J,Braun J,Fischer-Onaca O,Meier W,Matile S,2011.Synthetic polyion-counterion transport systems in polymersomes and gels.Organic&Biomolecular Chemistry,9(19):6623-6628
Moyano C,Alfonso C,Gallego J,Raposo R,Melgarejo P,2003.Comparison of RAPD and AFLP marker analysis as a means to study the genetic structure of Botrytis cinerea populations.European Journal of Plant Pathology,109(5):515-522
Rasiukevi?iūt?N,Rugienius R,?iksnianiene JB,2018.Genetic diversity of Botrytis cinerea from strawberry in Lithuania.Zemdirbyste-Agriculture,105(3):265-270
Rasiukevi?iūt?N,Valiuskaite A,Usellis N,Luk?ien?Z,2015.The effect of alternative plant protection measures on grey mould spread,strawberry growth and productivity.Sodininkyst?Ir Dar?ininkyst?,34:3-4
Wakeham A,Langton A,Adam SS,Kennedy R,2016.Interface of the environment and occurrence of Botrytis cinerea,in pre-symptomatic tomato crops.Crop Protection,90:27-33
Walker AS,Fournier E,2013.Habitat and host-specific differentiation in the multihost fungal pathogen Botryotinia fuckeliana and evidence for fungicide selection in populations.Modern Fungicides and Antifungal Compounds,7:167-172
Xu Z,Ali Z,Hou X,Li H,Yi JX,Abbasi PA,2013.Characterization of Chinese eggplant isolates of the fungal pathogen Verticillium dahliae from different geographic origins.Genetics and Molecular Research,12(1):183-195
Yamada K,Sonoda R,Ishikawa K,2016.Population genetic structure of QoI-resistant Pestalotiopsis longiseta isolates causing tea gray blight.Plant Disease,100:1686-1691
Yin DF,2015.The monitoring on fungicide resistance and the research on resistance mechanism of Botrytis cinerea from strawberry in Zhejiang Province.Master Thesis,Zhejiang University,Hangzhou.1-80(in Chinese)
Zhang J,2010.Studies on taxonomy of Botrytis species in Hubei Province and diversity of B.cinerea.PhDDissertation,Huazhong Agricultural University,Wuhan.5-10(in Chinese)
Zhang ZH,Gao XY,Du GD,Li H,2007.The trend of strawberryproduction:labour-saving cultivation.Chinese Agricultural Science Bulletin,23(10):101-103(in Chinese)
段会军,张彩英,李喜焕,郭小敏,马峙英,2008.基于RAPD、ISSR和AFLP对西瓜枯萎病菌遗传多样性的评价.菌物学报,27(2):267-276
冯慧静,文才艺,尹新明,洪坤奇,臧睿,2017.中国枣树干腐病菌(Botryosphaeria dothidea)群体遗传多样性的ISSR分析.果树学报,34(8):55-65
高翠珠,杨红玲,黄夏宇骐,黄俊斌,李国庆,郑露,2017.湖北省设施草莓灰霉病发生规律及流行因子分析.中国农业科学,50(9):1617-1623
龚国淑,徐琴,张敏,杨继芝,陈华保,申世安,唐太飞,2010.一种简便的病原真菌单孢分离方法研究.玉米科学,18(1):126-127
黄燕,朱振东,段灿星,武小菲,东方阳,2014.灰葡萄孢蚕豆分离物的遗传多样性.中国农业科学,47(12):2335-2347
李挺丹,彭世文,王宗华,鲁国东,2010.应用ISSR-PCR分析福建水稻纹枯菌的遗传多样性.植物病理学报,40(2):186-194
李廷刚,张路,周善跃,李敏,李宝笃,2013.山东省番茄灰霉病菌种类与遗传多样性研究.中国农学通报,29(10):137-143
尹大芳,2015.浙江省草莓灰霉病菌抗药性检测及抗性机制的研究.浙江大学硕士论文,杭州.1-80
张静,2010.湖北省灰霉病病菌区系和灰葡萄孢菌多样性研究.华中农业大学博士论文,武汉.5-10
张志宏,高秀岩,杜国栋,李贺,2007.草莓生产的发展趋势-省力化栽培.中国农学通报,23(10):101-103