我国葡萄灰霉病菌形态型和致病力分化
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摘要
我国葡萄(Vitis vinifera)种植范围广,葡萄灰霉病(grape gray mold)是危害葡萄生产的重要病害之一,其病原为灰葡萄孢(Botrytis cinerea),为了解我国不同葡萄种植气候区灰葡萄孢葡萄分离物的形态型变异和致病力分化情况,本研究对采集自不同葡萄种植气候区的143个灰葡萄孢菌株进行了形态型观察和生长速率测定,并采用菌丝块创伤接种法测定其对红地球(V.vinifera cv.'Redglobe')、马奶葡萄(V.vinifera cv.'Mare Nipple')和油菜(Brassica campestris)叶片的致病力。结果显示,供试菌株的形态型可以分为菌丝型和菌核型两类,其中40.56%为菌丝型,59.44%为菌核型,在菌丝型中有M1、M2、M3、M4、M5和M6等6个亚型,以M6亚型菌株最多,在菌核型中有S1、S2、S3、S4和S6等5个亚型,以S3亚型菌株最多;143个菌株的生长速率变化范围较大,从4.76mm/d到12.91 mm/d,聚类分析可以分为3个等级;所有菌对红地球和马奶均具有致病性,但是致病力分化严重,病斑面积分别在46.03~258.55 mm2和14.80~385.34mm2之间,聚类分析都可以分为4个等级;65个菌株中有62个灰葡萄孢菌株对油菜叶片具有致病性,致病半径在2.88~16.63 mm之间。菌株形态型、致病力与采集地、气候区、寄主来源的相关性分析表明,二者与采集地地理距离、寄主葡萄种类不具有明显相关性,但是与气候区来源具有一定相关性。菌株形态型、生长速率和致病力之间的相关性分析表明,灰葡萄孢对红地球、马奶和油菜的致病力两两之间具有较弱的正相关关系,菌株生长速率与对红地球的致病力具有不显著的弱负相关关系,与对马奶和油菜的致病力具有较显著的弱正相关关系,而灰葡萄孢生长速率和致病力与菌株形态型无明显相关性。本研究为进一步对灰葡萄孢新的形态型进行分子系统学分析,对灰葡萄孢致病力分化机理的研究提供了理论依据,为灰葡萄孢多样性研究和有效防控提供依据。
Grapevines(Vitis vinifera) are widely planted in China.Grape gray mold caused by Botrytis cinerea,is one of the most common diseases threating grape production.The aim of this work is to estimate the phenotypes and virulence variability of B.cinerea isolates from grapes of different viticulture climate zones in China.In this study,characters of 143 B.cinerea isolates,containing phenotype,growth rate and virulences to Red globe grape(V.vinifera cv.'Redglobe'),Mare nipple grape(V.vinifera cv.'Mare Nipple') and rape(Brassica campestris) leaves,were tested by applying wound inoculation with mycelial blocks.The results showed that there were two phenotypes,mycelial type and sclerotial type,accounting for 40.56% and 59.44%respectively.Six sub-mycelial-types containing M1,M2,M3,M4,M5,and M6,were found,and the number of M6 type isolates was more than that of other sub-mycelial-types.There were 5 sub-sclerotial-types containing S1,S2,S3,S4,and S6,and the number of S3 type isolates was more than that of other subsclerotial-types.The growth rate variation of 143 isolates was very high and ranged from 4.76 mm/d to 12.91 mm/d,and 3 levels were found by clustering analysis.All 143 isolates could successfully infect red globe grape and mare nipple grape,while virulence differentiation was very serious.The sizes of lesion area were46.03~258.55 mm~2 and 14.80~385.34 mm~2 respectively,which were divided into 4 levels by clustering analysis.62 of 65 isolates could successfully infect rape leaves,and the radius of the lesions was between2.88 mm and 16.63 mm.The correlation analysis between test data containing phenotype and virulence,and isolates' information containing collection regions,viticulture climate zones and grape hosts was performed.The results indicated that there was no significantly correlation between the two indexes and geographical distance and host grapes,while there was a certain correlation between phenotype and viticulture climate zone.The virulence also had some correlation with viticulture climate zone.The correlational analysis among phenotype,growth rate,and virulence were performed.The results showed that there was the slightly positive correlation among isolates' virulences to red globe grape,mare nipple grape and rape.While no significant negative correlation between growth rate and virulence to Red globe grape,and significant positive correlation between growth rate and virulence to Mare nipple grape and rape were found.In addition,phenotype was not obviously related to growth rate and virulence.This study lays the foundation for analysis of molecular phylogeny to the new phenotype and researching the mechanism of virulence differentiation.The results of this study also provide some helpful basis for studying B.cinerea diversity and controlling B.cinerea effectively.
Grapevines(Vitis vinifera) are widely planted in China.Grape gray mold caused by Botrytis cinerea,is one of the most common diseases threating grape production.The aim of this work is to estimate the phenotypes and virulence variability of B.cinerea isolates from grapes of different viticulture climate zones in China.In this study,characters of 143 B.cinerea isolates,containing phenotype,growth rate and virulences to Red globe grape(V.vinifera cv.'Redglobe'),Mare nipple grape(V.vinifera cv.'Mare Nipple') and rape(Brassica campestris) leaves,were tested by applying wound inoculation with mycelial blocks.The results showed that there were two phenotypes,mycelial type and sclerotial type,accounting for 40.56% and 59.44%respectively.Six sub-mycelial-types containing M1,M2,M3,M4,M5,and M6,were found,and the number of M6 type isolates was more than that of other sub-mycelial-types.There were 5 sub-sclerotial-types containing S1,S2,S3,S4,and S6,and the number of S3 type isolates was more than that of other subsclerotial-types.The growth rate variation of 143 isolates was very high and ranged from 4.76 mm/d to 12.91 mm/d,and 3 levels were found by clustering analysis.All 143 isolates could successfully infect red globe grape and mare nipple grape,while virulence differentiation was very serious.The sizes of lesion area were46.03~258.55 mm~2 and 14.80~385.34 mm~2 respectively,which were divided into 4 levels by clustering analysis.62 of 65 isolates could successfully infect rape leaves,and the radius of the lesions was between2.88 mm and 16.63 mm.The correlation analysis between test data containing phenotype and virulence,and isolates' information containing collection regions,viticulture climate zones and grape hosts was performed.The results indicated that there was no significantly correlation between the two indexes and geographical distance and host grapes,while there was a certain correlation between phenotype and viticulture climate zone.The virulence also had some correlation with viticulture climate zone.The correlational analysis among phenotype,growth rate,and virulence were performed.The results showed that there was the slightly positive correlation among isolates' virulences to red globe grape,mare nipple grape and rape.While no significant negative correlation between growth rate and virulence to Red globe grape,and significant positive correlation between growth rate and virulence to Mare nipple grape and rape were found.In addition,phenotype was not obviously related to growth rate and virulence.This study lays the foundation for analysis of molecular phylogeny to the new phenotype and researching the mechanism of virulence differentiation.The results of this study also provide some helpful basis for studying B.cinerea diversity and controlling B.cinerea effectively.
引文
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董金皋.2007.农业植物病理学(第二版)[M].中国农业出版社,北京.(Dong J G.2007.Agricultural Plant Pathology(Second Edition)[M].China Agriculture Press,Beijing.)
范咏梅,陈林凤,郝敬喆,等.2010.新疆灰霉病菌多态性及其致病力分化分析[J].中国生态农业学报,18(3):548-555.(Fan Y M,Chen L F,Hao J Z,et al.2010.Polymorphism analysis of Botrytis cinerea and its pathogenicity differentiation in Xinjiang[J].Chinese Journal of Eco-Agriculture,18(3):548-555.)
高俊明,刘慧平,王建明,等.2003.山西省蔬菜灰霉病菌营养体亲和性及菌丝融合群的初步研究[J].植物病理学报33(2):132-134.(Gao J M,Liu H P,Wang J M,et al2003.Study on vegetative compatibility and anastomosis group in Botrytis cinerea Pers.[J].Acta Phytopathologica Sinica,33(2):132-134.)
贺普超.1999.葡萄学[M].中国农业出版社,北京.(He P C1999.Grapevine[M].China Agriculture Press,Beijing.)
黄燕,朱振东,段灿星,等.2014.灰葡萄孢蚕豆分离物的遗传多样性[J].中国农业科学,47(12):2335-2347.(Huang Y,Zhu Z D,Duan C X,et al.2014.Genetic diversity of Botrytis cinerea isolates from broad bean[J].Scientia Agricultura Sinica,47(12):2335-2347.)
寇宏达,张艳杰,乔丹娜,等.2013.灰葡萄孢霉对不同葡萄品种致病力差异的检测和分析[J].西北农业学报,22(8)138-143.(Kou H D,Zhang Y J,Qiao D N,et al.2013.Detection and analysis of pathogenicity difference of Botrytis cinerea infecting different grape varieties[J].Acta Agriculturae Boreali-occidentalis Sinica,22(8):138-143.)
雷百战,李国英,石在强.2004.葡萄灰霉病病原鉴定和生物学特性研究[J].石河子大学学报(自然科学版),22(增刊):145-149.(Lei B Z,Li G Y,Shi Z Q.2004.Identification and biological characteristics of the pathogen of Botrytis cinerea on grape[J].Journal of Shihezi University(Natural Science),22(Supplement):145-149.)
李娜.2015.湖北省保护地番茄/草莓灰霉病菌多样性研究[D].硕士学位论文,华中农业大学,导师:李国庆.(Li N.2015.Studies on diversity of Botrytis cinerea isolated from tomato and strawberry[D].Thesis for M.S.,Huazhong Agricultural University,Supervisor:Li G Q.)
李廷刚,李敏,李宝笃.2013.山东省番茄灰霉病病菌致病力分化研究[J].山东农业科学,45(3):97-99,106.(Li T G,Li M,Li B D.2013.Study on pathogenicity differentiation of Botrytis cinerea in Shandong province[J].Shandong Agricultural Sciences,45(3):97-99,106.)
李喜玲.2008.灰葡萄孢致病力分化及与胞壁降解酶活性的关系研究[D].硕士学位论文,安徽农业大学,导师:高智谋.(Li X L.2008.On pathogenicity differentiation of Botrytis cinerea and relationship between pathogenicity and cell-wall-degrading enzyme activities[D].Thesis for M.S.,Anhui Agricultural University,Supervisor:Gao ZM.)
张从宇,张巨全,胡能兵,等.2003.番茄灰霉病菌寄主范围及致病力分化研究[J].安徽技术师范学院学报,17(3):239-242.(Zhang C Y,Zhang J Q,Hu N B,et al.2003.Study on host range and pathogenicity differentiation of Botrytis cinerea[J].Journal of Anhui Technical Teachers College,17(3):239-242.)
张佳.2015.中国草莓主产区灰霉病菌的多样性研究[D].博士学位论文,中国农业大学,导师:张国珍.(Zhang J.2015.Diversity of pathogens of gray mold from the main strawberry production regions in China[D].Thesis for ph.D.,China Agricultural University,Supervisor:Zhang GZ.)
张静.2010.湖北省灰霉病病菌区系和灰葡萄孢菌多样性研究[D].博士学位论文,华中农业大学,导师:李国庆.(Zhang J.2010.Studies on taxonomy of Botrytis species in Hubei province and diversity of B.cinerea[D].Thesis for ph.D.,Huazhong Agricultural University,Supervisor:Li G Q.)
张玮,乔广行,黄金宝,等.2013.中国葡萄灰霉病菌对嘧霉胺的抗药性检测[J].中国农业科学,46(6):1208-1212.(Zhang W,Qiao G H,Huang J B,et al.2013.Evaluation on resistance of grape gray mold pathogen Botrytis cinerea to pyrimethanil in China[J].Scientia Agricultura Sinica,46(6):1208-1212.)
周梓良.2015.湖北省灰霉病菌致病力分化及菌株HBstr-470中真菌病毒的研究[D].硕士学位论文,华中农业大学,导师:李国庆.(Zhou Z L.2015.Pathogenicity differentiation of Botrytis cinerea in Hubei province and the mycoviruses present in strain HBstr-470[D].Thesis for M.S.,Huazhong Agricultural University,Supervisor:LG Q.)
Cotoras M,Silva E.2005.Differences in the initial events of infection of Botrytis cinerea strains isolated from tomato and grape[J].Mycologia,97:485-492.
Dean R,Van Kan J A L,Pretorius Z A,et al.2012.The top 10fungal pathogens in molecular plant pathology[J].Molecular Plant Pathology,13(4):414-430.
Derckel J P,Baillieul F,Manteau S,et al.1999.Differential induction of grapevine defenses by two strains of Botrytis cinerea[J].Phytopathology,89(3):197-203.
Diolez A,Marches F,Fortini D,et al.1995.Boty,a long-terminal-repeat retroelement in the phytopathogenic fungus Botrytis cinerea[J].Applied and Environmental Microbiology,61(1):103-108.
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