台湾省苹果上牛眼果腐病菌的截获鉴定
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摘要
为明确从宁波空港口岸入境旅客携带苹果上截获的1株明孢盘菌属菌株60017的分类地位,通过形态学特征观察、内转录间隔区(ITS)和β-微管蛋白基因(β-tubulin)序列比对分析以及致病性测定,对该菌株进行了种类鉴定。结果表明:菌株60017在PDA培养基上的菌落为圆形,边缘整齐,呈乳白色,气生菌丝较少,可产生大量分生孢子;基于ITS和β-tubulin序列构建的系统发育树中,菌株60017和苹果牛眼果腐病菌Neofabraea kienholzii处于同一分支;菌株60017接种苹果后,在接种部位能引起褐腐症状,与原发病症状一致。结合形态学特征、致病性测定结果以及分子鉴定结果,最终将该菌株鉴定为苹果牛眼果腐病菌N. kienholzii。
To identify the fungal strain 60017 isolated from the apple imported from Taiwan Province of China, the morphological observation, sequence analysis and pathogenicity test were carried out. The morphological results showed that the colony of the strain 60017 on PDA medium was round, with neat edges, milky white, less aerial hyphae, and could produce a large number of conidia. Its rDNA ITS andβ-tubulin sequences were sequenced. Analysis of these sequences with the related sequences in NCBI showed that the strain 60017 clustered into Neofabraea kienholzii clade. The pathogenicity test showed that the strain 60017 could infect apple fruit with the symptoms of brown rot. These results indicated that the fungal strain 60017 was N. kienholzii.
To identify the fungal strain 60017 isolated from the apple imported from Taiwan Province of China, the morphological observation, sequence analysis and pathogenicity test were carried out. The morphological results showed that the colony of the strain 60017 on PDA medium was round, with neat edges, milky white, less aerial hyphae, and could produce a large number of conidia. Its rDNA ITS andβ-tubulin sequences were sequenced. Analysis of these sequences with the related sequences in NCBI showed that the strain 60017 clustered into Neofabraea kienholzii clade. The pathogenicity test showed that the strain 60017 could infect apple fruit with the symptoms of brown rot. These results indicated that the fungal strain 60017 was N. kienholzii.
引文
Aguilar CG,Mazzola M,Xiao CL.2018.Control of bull’s-eye rot of apple caused by Neofabraea perennans and Neofabraea kienholzii using pre-and postharvest fungicides.Plant Disease,102(5):905-910
Cameldi I,Neri F,Menghini M,Pirondi A,NanniI M,Collina M,Mari M.2017.Characterization of Neofabraea vagabunda isolates causing apple bull’s eye rot in Italy(Emilia-Romagna region).Plant Pathology,66(9):1432-1444
Chen C,Verkley GJM,Sun GY,Groenewald JZ,Crous PW.2016.Redefining common endophytes and plant pathogens in Neofabraea,Pezicula,and related genera.Fungal Biology,120(11):1291-1322
Cunnington JH.2004.Three Neofabraea species on pome fruit in Australia.Australasian Plant Pathology,33(3):453-454
China Tourism Academy.2017.Annual report of China outboard tourism development 2017.Beijing:Tourism Education Press(in Chinese)[中国旅游研究院.2017.中国出境旅游发展年度报告2017.北京:旅游教育出版社]
de Jong SN,Levesque CA,Verkley GJM,Abeln ECA,Rahe JE,Braun PG.2001.Phylogenetic relationships among Neofabraea species causing tree cankers and bull’s-eye rot of apple based on DNAsequencing of ITS nuclear rDNA mitochondrial rDNA,and theβ-tubulin gene.Mycological Research,105(6):658-669
Fang ZD.1998.Research methods of plant disease(3rd edition).Beijing:China Agriculture Press(in Chinese)[方中达.1998.植病研究方法(第3版).北京:中国农业出版社]
Gariepy TD,Rahe JE,Levesque CA,Spotts RA,Sugar DL,Henriquez JL.2005.Neofabraea species associated with bull’s-eye rot and cankers of apple and pear in the Pacific Northwest.Canadian Journal of Plant Pathology,27(1):118-124
Henriquez JL,Sugar D,Spotts RA.2004.Etiology of bull’s eye rot of pear caused by Neofabraea spp.in Oregon,Washington,and California.Plant Disease,88(10):1134-1138
Jin YM,Wan MW,Wang WL,Kong LB,Pan MY,Zhang SJ,Gao C,Zhou Q,Liu J.2012.Analysis of quarantining and supervising goods carried by passengers entering at Shanghai International Airports.Acta Agriculturae Shanghai,28(3):26-31(in Chinese)[金亦民,万明伟,王雯丽,孔令斌,潘梅莹,张书俊,高超,周琦,刘俊.2012.上海空港口岸入境旅客携带物检疫监管情况分析.上海农业学报,28(3):26-31]
Johnston PR,Seifert KA,Stone JK,Rossman AY,MarvanováL.2014.Recommendations on generic names competing for use in Leotiomycetes(Ascomycota).IMA Fungus,5(1):91-120
Kingsnorth J,Perrine J,Berrie A,Saville R.2017.First report of Neofabraea kienholzii causing bull’s eye rot of apple in the UK.New Disease Reports,36:15
Lin HJ,Wang WF,Yi JP,Hu XN,He RR,Jiang X.2016.Development of TaqMan probes real-time fluorescence PCR method for the rapid detection of the causal fungal agents of bull’s-eye rot on apple.Plant Quarantine,30(2):55-62(in Chinese)[林惠娇,王卫芳,易建平,胡学难,何日荣,蒋湘.2016.TaqMan探针实时荧光PCR快速检测苹果牛眼果腐病菌方法的建立.植物检疫,30(2):55-62]
Lin SC,Wu YN,Xu L,Chen YR.2009.Quarantine supervision of plants and plant products carried by passengers from Thailand.Plant Quarantine,23(4):59-60(in Chinese)[林盛才,吴毓南,许玲,陈粤榕.2009.对泰国入境旅客携带植物及其产品的检疫监管.植物检疫,23(4):59-60]
Liu F.2010.Research on apple industry development of Hebei Province.Master Thesis.Baoding:Agricultural University of Hebei(in Chinese)[刘峰.2010.河北省苹果产业发展研究.硕士学位论文.保定:河北农业大学]
Michalecka M,Bryk H,Poniatowska A,Pu?awska J.2016.Identification of Neofabraea species causing bull’s eye rot of apple in Poland and their direct detection in apple fruit using multiplex PCR.Plant Pathology,65(4):643-654
Ministry of Agriculture.2010.Apple’s superior regional layout planning(2008-2015).Agriculture Engineering Technology(Agricultural Product Processing Industry),(3):16-17(in Chinese)[农业部.2010.苹果优势区域布局规划(2008-2015年).农业工程技术(农产品加工业),(3):16-17]
Office of Chief Scientist of National Apple Industry Technical System.2017.Trends and suggestions of apple industry in 2017.Fruit Growers’Friend,(5):1-3(in Chinese)[国家苹果产业技术体系首席科学家办公室.2017.2017年苹果产业发展趋势与建议.果农之友,(5):1-3]
Pe?icováK,Kola?ík M,HortováB,NovotnyD.2017.Diversity and identification of Neofabraea species causing bull’s eye rot in the Czech Republic.European Journal of Plant Pathology,147(3):683-693
Seifert KA.2013.Nomenclatural novelties:Neofabraea kienholzii.Index Fungorum,(28):1
Spotts RA,Seifert KA,Wallis KM,Sugar D,Xiao CL,Serdani M,Henriquez JL.2009.Description of Cryptosporiopsis kienholzii and species profiles of Neofabraea in major pome fruit growing districts in the Pacific Northwest USA.Mycological Research,113(11):1301-1311
Verkley GJM.1999.A monograph of the genus Pezicula and its anamorphs:studies in mycology no.44.Centraalbureau voor Schimmelultures,Baarn:The Netherlands,pp.1-180
Wang WF,Hu J,Zhang YY,Li XF,Chen MK,Fang JF.2010.The first and repetitious interception in Guangdong Port of bull’s eye rot on apple from US.Plant Quarantine,24(1):35-37(in Chinese)[王卫芳,胡佳,张永瑜,李新芳,陈茂坤,方剑锋.2010.广东口岸首次截获美国苹果牛眼果腐病.植物检疫,24(1):35-37]
Wenneker M,Pham KTK,Boekhoudt LC,de Boer FA,van Leeuwen PJ,Hollinger TC,Thomma BPHJ.2017.First report of Neofabraea kienholzii causing bull’s eye rot on pear(Pyrus communis)in the Netherlands.Plant Disease,101(4):634
White TJ,Bruns T,Lee S,Taylor J.1990.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics.//Innis MA,Gelfand DH,Sninsky JJ,White TJ.PCR protocols:a guide to methods and applications.San Diego,California:Academic Press,pp.315-322
Zhang LX,Hu PL,Li W,Yu ZX,Jiao BB,Yi JP,Yu C,Yang CY.2016.Identification of Neofabraea kienholzii on imported Malus pumila.Plant Quarantine,30(1):36-39(in Chinese)[张露茜,胡培龙,李薇,于子翔,焦彬彬,易建平,于翠,杨翠云.2016.进境苹果果实中苹果牛眼果腐病菌的检疫鉴定.植物检疫,30(1):36-39]
Zhang YJ,Luo JF,Liu P,Liu YT,Liao F.2012.Identification of Neofabraea alba from Chile apple.Plant Quarantine,26(2):47-49(in Chinese)[张裕君,罗加凤,刘鹏,刘跃庭,廖芳.2012.智利苹果中牛眼果腐病菌的分离鉴定研究.植物检疫,26(2):47-49]
Cameldi I,Neri F,Menghini M,Pirondi A,NanniI M,Collina M,Mari M.2017.Characterization of Neofabraea vagabunda isolates causing apple bull’s eye rot in Italy(Emilia-Romagna region).Plant Pathology,66(9):1432-1444
Chen C,Verkley GJM,Sun GY,Groenewald JZ,Crous PW.2016.Redefining common endophytes and plant pathogens in Neofabraea,Pezicula,and related genera.Fungal Biology,120(11):1291-1322
Cunnington JH.2004.Three Neofabraea species on pome fruit in Australia.Australasian Plant Pathology,33(3):453-454
China Tourism Academy.2017.Annual report of China outboard tourism development 2017.Beijing:Tourism Education Press(in Chinese)[中国旅游研究院.2017.中国出境旅游发展年度报告2017.北京:旅游教育出版社]
de Jong SN,Levesque CA,Verkley GJM,Abeln ECA,Rahe JE,Braun PG.2001.Phylogenetic relationships among Neofabraea species causing tree cankers and bull’s-eye rot of apple based on DNAsequencing of ITS nuclear rDNA mitochondrial rDNA,and theβ-tubulin gene.Mycological Research,105(6):658-669
Fang ZD.1998.Research methods of plant disease(3rd edition).Beijing:China Agriculture Press(in Chinese)[方中达.1998.植病研究方法(第3版).北京:中国农业出版社]
Gariepy TD,Rahe JE,Levesque CA,Spotts RA,Sugar DL,Henriquez JL.2005.Neofabraea species associated with bull’s-eye rot and cankers of apple and pear in the Pacific Northwest.Canadian Journal of Plant Pathology,27(1):118-124
Henriquez JL,Sugar D,Spotts RA.2004.Etiology of bull’s eye rot of pear caused by Neofabraea spp.in Oregon,Washington,and California.Plant Disease,88(10):1134-1138
Jin YM,Wan MW,Wang WL,Kong LB,Pan MY,Zhang SJ,Gao C,Zhou Q,Liu J.2012.Analysis of quarantining and supervising goods carried by passengers entering at Shanghai International Airports.Acta Agriculturae Shanghai,28(3):26-31(in Chinese)[金亦民,万明伟,王雯丽,孔令斌,潘梅莹,张书俊,高超,周琦,刘俊.2012.上海空港口岸入境旅客携带物检疫监管情况分析.上海农业学报,28(3):26-31]
Johnston PR,Seifert KA,Stone JK,Rossman AY,MarvanováL.2014.Recommendations on generic names competing for use in Leotiomycetes(Ascomycota).IMA Fungus,5(1):91-120
Kingsnorth J,Perrine J,Berrie A,Saville R.2017.First report of Neofabraea kienholzii causing bull’s eye rot of apple in the UK.New Disease Reports,36:15
Lin HJ,Wang WF,Yi JP,Hu XN,He RR,Jiang X.2016.Development of TaqMan probes real-time fluorescence PCR method for the rapid detection of the causal fungal agents of bull’s-eye rot on apple.Plant Quarantine,30(2):55-62(in Chinese)[林惠娇,王卫芳,易建平,胡学难,何日荣,蒋湘.2016.TaqMan探针实时荧光PCR快速检测苹果牛眼果腐病菌方法的建立.植物检疫,30(2):55-62]
Lin SC,Wu YN,Xu L,Chen YR.2009.Quarantine supervision of plants and plant products carried by passengers from Thailand.Plant Quarantine,23(4):59-60(in Chinese)[林盛才,吴毓南,许玲,陈粤榕.2009.对泰国入境旅客携带植物及其产品的检疫监管.植物检疫,23(4):59-60]
Liu F.2010.Research on apple industry development of Hebei Province.Master Thesis.Baoding:Agricultural University of Hebei(in Chinese)[刘峰.2010.河北省苹果产业发展研究.硕士学位论文.保定:河北农业大学]
Michalecka M,Bryk H,Poniatowska A,Pu?awska J.2016.Identification of Neofabraea species causing bull’s eye rot of apple in Poland and their direct detection in apple fruit using multiplex PCR.Plant Pathology,65(4):643-654
Ministry of Agriculture.2010.Apple’s superior regional layout planning(2008-2015).Agriculture Engineering Technology(Agricultural Product Processing Industry),(3):16-17(in Chinese)[农业部.2010.苹果优势区域布局规划(2008-2015年).农业工程技术(农产品加工业),(3):16-17]
Office of Chief Scientist of National Apple Industry Technical System.2017.Trends and suggestions of apple industry in 2017.Fruit Growers’Friend,(5):1-3(in Chinese)[国家苹果产业技术体系首席科学家办公室.2017.2017年苹果产业发展趋势与建议.果农之友,(5):1-3]
Pe?icováK,Kola?ík M,HortováB,NovotnyD.2017.Diversity and identification of Neofabraea species causing bull’s eye rot in the Czech Republic.European Journal of Plant Pathology,147(3):683-693
Seifert KA.2013.Nomenclatural novelties:Neofabraea kienholzii.Index Fungorum,(28):1
Spotts RA,Seifert KA,Wallis KM,Sugar D,Xiao CL,Serdani M,Henriquez JL.2009.Description of Cryptosporiopsis kienholzii and species profiles of Neofabraea in major pome fruit growing districts in the Pacific Northwest USA.Mycological Research,113(11):1301-1311
Verkley GJM.1999.A monograph of the genus Pezicula and its anamorphs:studies in mycology no.44.Centraalbureau voor Schimmelultures,Baarn:The Netherlands,pp.1-180
Wang WF,Hu J,Zhang YY,Li XF,Chen MK,Fang JF.2010.The first and repetitious interception in Guangdong Port of bull’s eye rot on apple from US.Plant Quarantine,24(1):35-37(in Chinese)[王卫芳,胡佳,张永瑜,李新芳,陈茂坤,方剑锋.2010.广东口岸首次截获美国苹果牛眼果腐病.植物检疫,24(1):35-37]
Wenneker M,Pham KTK,Boekhoudt LC,de Boer FA,van Leeuwen PJ,Hollinger TC,Thomma BPHJ.2017.First report of Neofabraea kienholzii causing bull’s eye rot on pear(Pyrus communis)in the Netherlands.Plant Disease,101(4):634
White TJ,Bruns T,Lee S,Taylor J.1990.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics.//Innis MA,Gelfand DH,Sninsky JJ,White TJ.PCR protocols:a guide to methods and applications.San Diego,California:Academic Press,pp.315-322
Zhang LX,Hu PL,Li W,Yu ZX,Jiao BB,Yi JP,Yu C,Yang CY.2016.Identification of Neofabraea kienholzii on imported Malus pumila.Plant Quarantine,30(1):36-39(in Chinese)[张露茜,胡培龙,李薇,于子翔,焦彬彬,易建平,于翠,杨翠云.2016.进境苹果果实中苹果牛眼果腐病菌的检疫鉴定.植物检疫,30(1):36-39]
Zhang YJ,Luo JF,Liu P,Liu YT,Liao F.2012.Identification of Neofabraea alba from Chile apple.Plant Quarantine,26(2):47-49(in Chinese)[张裕君,罗加凤,刘鹏,刘跃庭,廖芳.2012.智利苹果中牛眼果腐病菌的分离鉴定研究.植物检疫,26(2):47-49]