10株葡萄源丝状真菌分离鉴定及其致病力分析
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摘要
采用组织分离法从京郊地区腐烂的葡萄果实及葡萄叶上分离到10株丝状真菌,其中从腐烂的葡萄果实中分离到3株丝状真菌,命名为814~#、815~#和880~#;从葡萄叶上分离到7株丝状真菌,命名为794~#、797~#、798~#、811~#、902~#、903~#和1397~#。结合形态学特征及真菌r DNA ITS区序列分析结果,菌株814~#为茎点霉(Phoma sp.),菌株815~#为刺孢曲霉(Aspergillus aculeatus),菌株880~#为灰葡萄孢(Botrytis cinerea),菌株794~#为日本曲霉(Aspergillus japonicus),菌株797~#为粉红单端孢(Trichothecium roseum),菌株798~#为大蒜盲种葡萄孢(Botrytis porri),菌株811~#为果生炭疽菌(Colletotrichum fructicola),菌株902~#为链格孢菌(Alternaria eichhorniae),菌株903~#为芬芳镰孢菌(Fusarium redolens)和菌株1397~#为脉孢菌(Neurospora terricola),其中A.aculeatus、A.japonicus、C.fructicola、A.eichhorniae、F.redolens和N.terricola未见在葡萄上分离到的报道。将10株丝状真菌分别接种到健康的葡萄、圣女果、苹果和梨果实上,B.cinerea、A.japonicus、A.aculeatus、T.roseum、B.porri、C.fructicola、A.eichhorniae和F.redolens均可使4种果实发病,其中B.cinerea、A.japonicus、B.porri和F.redolens致病力较强;而Phoma sp.和N.terricola只能使葡萄和圣女果发病。
Ten strains were isolated from infected grapes and grape leaves in the suburb of Beijing and analysis the pathogenicity of different pathogenic bacteria to four fruits. In order to provide reference for the study of interspecific relationship and disease control of fungi. Three strains 814~#,815~#and 880~#were isolated from infected grapes. Seven strains 794~#,797~#,798~#,811~#,902~#,903~#and 1397~#were isolated from infected grape leaves. The fungal pathogens 814~#,815~#,880~#,794~#,797~#,798~#,811~#,902~#,903~#and 1397~#were identified as Phoma sp.,Aspergillus aculeatus,Botrytis cinerea,Aspergillus japonicas,Trichothecium roseum,Botrytis porri,Colletotrichum fructicola,Alternaria eichhorniae,Fusarium redolens and Neurospora terricola by morphological characterization and rDNA ITS sequence analysis. In addition,A. aculeatus was firstly isolated from grapes and A. japonicas,C. fructicola,A. eichhorniae,F. redolens and N. terricola were firstly isolated from grape leaves. Reinoculated fruits such as grapes,cherry tomatoes,apples and pears showed that B. cinerea,A. japonicus,A. aculeatus,T. roseum,B. porri,C. fructicola,A. eichhorniae and F. redolens can cause all the fruits disease and B. cinerea,A. japonicus,B. porri and F. redolens showed strong pathogenicity. In contract,Phoma sp. and N. terricola can only cause grapes and cherry tomatoes disease.
Ten strains were isolated from infected grapes and grape leaves in the suburb of Beijing and analysis the pathogenicity of different pathogenic bacteria to four fruits. In order to provide reference for the study of interspecific relationship and disease control of fungi. Three strains 814~#,815~#and 880~#were isolated from infected grapes. Seven strains 794~#,797~#,798~#,811~#,902~#,903~#and 1397~#were isolated from infected grape leaves. The fungal pathogens 814~#,815~#,880~#,794~#,797~#,798~#,811~#,902~#,903~#and 1397~#were identified as Phoma sp.,Aspergillus aculeatus,Botrytis cinerea,Aspergillus japonicas,Trichothecium roseum,Botrytis porri,Colletotrichum fructicola,Alternaria eichhorniae,Fusarium redolens and Neurospora terricola by morphological characterization and rDNA ITS sequence analysis. In addition,A. aculeatus was firstly isolated from grapes and A. japonicas,C. fructicola,A. eichhorniae,F. redolens and N. terricola were firstly isolated from grape leaves. Reinoculated fruits such as grapes,cherry tomatoes,apples and pears showed that B. cinerea,A. japonicus,A. aculeatus,T. roseum,B. porri,C. fructicola,A. eichhorniae and F. redolens can cause all the fruits disease and B. cinerea,A. japonicus,B. porri and F. redolens showed strong pathogenicity. In contract,Phoma sp. and N. terricola can only cause grapes and cherry tomatoes disease.
引文
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[2]中国统计局.中国农业统计年鉴[M].北京:中国统计出版社,1999:3-100.
[3]苏梅.葡萄灰霉病发生规律与综合防治技术[J].山西果树,2013(3):31.SU M.Occurrence regularity and integrated control techniques of Botrytis cinerea[J].Shanxi Fruits,2013(3):31.
[4]ABRUNHOSA L,PATERSON R R,KOZAKIEWICZ Z,et al.Mycotoxin production from fungi isolated from grapes[J].Letters in Applied Microbiology,2001,32(4):240-242.
[5]侯平军,崔永辉.葡萄穗轴褐枯病的发生与防治[J].河南农业,2016(22):36.HOU P J,CUI Y H.The occurrence and control of brown rot of grape spike stalk[J].Agriculture of Henan,2016(22):36.
[6]CAPPA F,COCCONCELLI P S.Identification of fungi from dairy products by means of 18S rRNA analysis[J].International Journal of Food Microbiology,2001,69(1/2):157-160.
[7]杨济徽,李鹏,张龙,等.海南龙眼红粉病的病原鉴定[J].果树学报,2016,33(8):993-999.YANG J H,LI P,ZHANG L,et al.Identification of the pink disease for longan fruit in Hainan province[J].Journal of Fruit Science,2016,33(8):993-999.
[8]王友升,郭晓敏,姚子鹏,等.1株桃果实采后病原真菌的鉴定以及碳源代谢指纹图谱分析[J].北京工商大学学报(自然科学版),2011,29(1):47-53.WANG Y S,GUO X M,YAO Z P,et al.Identification and carbon metabolic fingerprinting analysis of a pathogen isolated from postharvest peach fruit[J].Journal of Beijing Technology and Business University(Natural Science Edition),2011,29(1):47-53.
[9]任向峰,姚婷,张萌,等.4株水果采后链格孢属真菌的r DNA ITS区序列与碳源代谢指纹图谱差异性分析[J].食品科学技术学报,2018,36(1):72-78.REN X F,YAO T,ZHANG M,et al.Comparasion of r DNA ITS sequence and carbon metabolic fingerprinting of Alternaria nees isolated from four fruits[J].Journal of Food Science and Technology,2018,36(1):72-78.
[10]姚婷,陈其葳,张燕,等.3株果实采后葡萄孢属真菌ITS区r DNA序列与碳源代谢指纹图谱分析[J].食品科学技术学报,2017,35(4):49-55.YAO T,CHEN Q W,ZHANG Y,et al.Analysis of r DNA ITS and carbon metabolic fingerprinting of Botrytis nees isolated from three fruits[J].Journal of Food Science and Technology,2017,35(4):49-55.
[11]王祥红,汤志宏,李静.曲霉菌的分离及其形态观察[J].生物学通报,2013,48(7):54-55.WANG X H,TANG Z H,LI J.Isolation and morphological observation of Aspergillus[J].Bulletin of Biology,2013,48(7):54-55.
[12]OH S Y,NAM K W,YOON D H.Identification of Acremonium acutatum and Trichothecium roseum isolated from grape with white stain symptom in Korea[J].Mycobiology,2014,42(3):269-273.
[13]魏景超.真菌鉴定手册[M].上海:上海科学技术出版社,1979:566-614.
[14]SERRA R,LOURENO A,ALPIO P,et al.Influence of the region of origin on the mycobiota of grapes with emphasis on Aspergillus and Penicillium species[J].Mycological Research,2006,110(8):971-978.
[15]HAMID M I,HUSSAIN M,GHAZANFAR M U,et al.Trichothecium roseum causes fruit rot of tomato,orange,and apple in Pakistan[J].Plant Disease,2014,98(9):1271.
[16]ALLEWELDT G,POSSINGHAM J V.Progress in grapevine breeding[J].Theoretical and Applied Genetics,1988,75(5):669-673.
[17]FU D D,WANG W,QIN R F,et al.Colletotrichum fructicola,first record of bitter rot of apple in China[J].Mycotaxon,2013,126(1):23-30.
[18]刘志.环渤海湾地区苹果再植病病原真菌分离鉴定及生防木霉菌株的筛选[D].泰安:山东农业大学,2013.
[19]LI H N,JIANG J J,HONG N,et al.First report of Colletotrichum fructicola causing bitter rot of pear(Pyrus bretschneideri)in China[J].Plant Disease,2013,97(7):1000.