广西甘蔗白条病病原菌的分离鉴定
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摘要
为明确引起我国甘蔗检疫性细菌病害甘蔗白条病的病原菌及其症状特点,从广西来宾和崇左甘蔗主产区采集疑似白条病病害样本,采用组织分离法进行病原菌的分离,利用形态学与基因序列分析相结合的方法鉴定病原菌,柯赫氏法则回接验证病原菌的致病性。结果表明,引起广西甘蔗白条病的病原菌为白条黄单胞杆菌(Xanthomonas albilineans),16S-23S rRNA基因转录间隔区(ITS)基因序列分析以及白条黄单胞杆菌特异基因分析结果显示分离到的白条病病原菌与法国、巴西以及新西兰等国家的白条病病原菌同源性达到99%～100%。人工接种条件下,该病原菌还可侵染玉米并发生类似病症。本研究成功分离鉴定我国广西甘蔗白条病病原菌,为今后的致病机理研究和白条病抗病育种等方面奠定基础。
In order to clarify the symptoms and pathogens of the bacterial quarantine disease,sugarcane leaf scald disease,infected samples showing suspected symptoms were collected from Laibin and Chongzuo,main sugarcane growing areas in Guangxi Province. Pathogenic isolates were obtained and purified by tissue isolation method,identified based on morphological characteristics and gene sequencing analysis. Their pathogenicity was proved by the Koch's. The results showed that the causal agent of sugarcane leaf scald disease in Guangxi is Xanthomonas albilineans. 16 S-23 S ribosomal RNA intergenic spacer(ITS) and specific gene sequence analysis results showed that leaf scald pathogen isolated from Guangxi shared 99%-100% similarity with the pathogen isolated in France,Brazil and New Zealand. Under artificial inoculation conditions,the pathogen can also infect corn and develop similar disease symptoms. This study isolated and identified the pathogen of sugarcane leaf scald in Guangxi,China. The results lay a foundation for future research on disease mechanism of leaf scald and disease resistance breeding.
In order to clarify the symptoms and pathogens of the bacterial quarantine disease,sugarcane leaf scald disease,infected samples showing suspected symptoms were collected from Laibin and Chongzuo,main sugarcane growing areas in Guangxi Province. Pathogenic isolates were obtained and purified by tissue isolation method,identified based on morphological characteristics and gene sequencing analysis. Their pathogenicity was proved by the Koch's. The results showed that the causal agent of sugarcane leaf scald disease in Guangxi is Xanthomonas albilineans. 16 S-23 S ribosomal RNA intergenic spacer(ITS) and specific gene sequence analysis results showed that leaf scald pathogen isolated from Guangxi shared 99%-100% similarity with the pathogen isolated in France,Brazil and New Zealand. Under artificial inoculation conditions,the pathogen can also infect corn and develop similar disease symptoms. This study isolated and identified the pathogen of sugarcane leaf scald in Guangxi,China. The results lay a foundation for future research on disease mechanism of leaf scald and disease resistance breeding.
引文
[1]Rott P,Davis M J.Leaf Scald//Rott P,Bailey R A,Comstock J C,et al.A Guide to Sugarcane Diseases.Montpellier:La Librairie du Cirad,2000:38-44.
[2]Hoy J W,Grisham M P.Sugarcane leaf scald distribution,symptomatology,and effect on yield in Louisiana.Plant Disease,1994,78(11):1083-1087.
[3]Rott P,Soupa D,Brunet Y,et al.Leaf scald(Xanthomonas albilineans)incidence and its effect on yield in seven sugarcane cultivars in Guadeloupe.Plant Pathology,1995,44(6):1075-1084.
[4]Li Y R,Yang L T.Research and development priorities for sugar industry of China:recent research highlights.Sugar Tech,2015,17(1):9-12.
[5]Zhang R Y,Shan H L,Li W F,et al.First report of sugarcane leaf scald caused by Xanthomonas albilineans(Ashby)Dowson in the province of Guangxi,China.Plant Disease,2017,101(8):1541.
[6]刘刚.《中华人民共和国进境植物检疫性有害生物名录》发布农药市场信息,2007(13):40-41.
[7]Davis M J,Rott P,Baudin P,et al.Evaluation of selective media and immunoassays for detection of Xanthomonas albilineans,causal agent of sugarcane leaf scald disease Plant Disease,1994,78(1):78-82.
[8]Pan Y B,Grisham M P,Burner D M,et al.Development of polymerase chain reaction primers highly specific for Xanthomonas albilineans,the causal bacterium of sugarcane leaf scald disease.Plant Disease,1999,83(3):218-222.
[9]Davis M J,Rott P,Monge G A.Multiplex polymerase chain reaction(PCR)for diagnosis of leaf scald and ratoon stunting diseases.Sugary Azucar,1997,92(6):26.
[10]Koike H.Aluminum-cap method for testing sugarcane varieties against leaf scald disease.Phytopathology,1965,55:317-319.
[11]Ricaud C,Ryan C C.Leaf scald//Ricaud C,Egan B T,Gillaspie A G J,et al.Diseases of sugarcane major diseases.Amsterdam,The Netherlands:Elsevier Publishing Co.,1989:39-58.
[12]Daugrois J H,Boisne-Noc B R,Champoiseau B P,et al.The revisited infection cycle of Xanthomonas albilineans,the causal agent of leaf scald of sugarcane.Functional Plant Science and Biotechnology,2012,6(2):91-97.
[13]李文凤,单红丽,张荣跃,等.广西蔗区检测发现由白条黄单菌引起的检疫性病害甘蔗白条病//中国植物病理学会2017年学术年会论文集.北京:中国植物病理学会,2017:281.
[14]韦金菊,魏春燕,宋修鹏,等.广西北海蔗区甘蔗白条病发生情况调查.南方农业学报,2018,49(2):264-270.
[15]Autrey L J C,Saumtally S,Dookun A.Studies on variation in the leaf scald pathogen,Xanthomonas albilineans[EB/OL].[2018-10-23].http://www.beep.ird.fr/cgi-bin/library.cgi?e=d-01000-00---off-0staffpu1--00-1----0-10-0---0---0direct-10---4-------0-1l--11-fr-50---20-about---01-2-1-00-0-0-11-1-0utf Zz-8-00&a=d&c=staffpu1&cl=CL3.1.32&d=HASH0121109142afe0df33d2e645s1424.
[16]Daugrois J H,Dumont V,Champoiseau P,et al.Aerial contamination of sugarcane in Guadeloupe by two strains of Xanthomonas albilineans.European Journal of Plant Pathology,2003,109(5):445-458.
[17]Champoiseau P,Daugrois J H,Girard J C,et al.Variation in albicidin biosynthesis genes and in pathogenicity of Xanthomonas albilineans,the sugarcane leaf scald pathogen.Phytopathology,2006,96(1):33-45.
[18]Huertalara M,Rojasmartinez R I,Bautistacalles J,et al.Genetic and pathogenic diversity of Xanthomonas albilineans Ashby(Dowson),in Mexico.Research Journal of Biological Sciences,2009,4(3):312-319.
[19]Rott P,Abel M,Soupa D,et al.Population dynamics of Xanthomonas albilineans in sugarcane plants as determined with an antibiotic resistant mutant.Plant Disease,1994,78(3):241-247.
[2]Hoy J W,Grisham M P.Sugarcane leaf scald distribution,symptomatology,and effect on yield in Louisiana.Plant Disease,1994,78(11):1083-1087.
[3]Rott P,Soupa D,Brunet Y,et al.Leaf scald(Xanthomonas albilineans)incidence and its effect on yield in seven sugarcane cultivars in Guadeloupe.Plant Pathology,1995,44(6):1075-1084.
[4]Li Y R,Yang L T.Research and development priorities for sugar industry of China:recent research highlights.Sugar Tech,2015,17(1):9-12.
[5]Zhang R Y,Shan H L,Li W F,et al.First report of sugarcane leaf scald caused by Xanthomonas albilineans(Ashby)Dowson in the province of Guangxi,China.Plant Disease,2017,101(8):1541.
[6]刘刚.《中华人民共和国进境植物检疫性有害生物名录》发布农药市场信息,2007(13):40-41.
[7]Davis M J,Rott P,Baudin P,et al.Evaluation of selective media and immunoassays for detection of Xanthomonas albilineans,causal agent of sugarcane leaf scald disease Plant Disease,1994,78(1):78-82.
[8]Pan Y B,Grisham M P,Burner D M,et al.Development of polymerase chain reaction primers highly specific for Xanthomonas albilineans,the causal bacterium of sugarcane leaf scald disease.Plant Disease,1999,83(3):218-222.
[9]Davis M J,Rott P,Monge G A.Multiplex polymerase chain reaction(PCR)for diagnosis of leaf scald and ratoon stunting diseases.Sugary Azucar,1997,92(6):26.
[10]Koike H.Aluminum-cap method for testing sugarcane varieties against leaf scald disease.Phytopathology,1965,55:317-319.
[11]Ricaud C,Ryan C C.Leaf scald//Ricaud C,Egan B T,Gillaspie A G J,et al.Diseases of sugarcane major diseases.Amsterdam,The Netherlands:Elsevier Publishing Co.,1989:39-58.
[12]Daugrois J H,Boisne-Noc B R,Champoiseau B P,et al.The revisited infection cycle of Xanthomonas albilineans,the causal agent of leaf scald of sugarcane.Functional Plant Science and Biotechnology,2012,6(2):91-97.
[13]李文凤,单红丽,张荣跃,等.广西蔗区检测发现由白条黄单菌引起的检疫性病害甘蔗白条病//中国植物病理学会2017年学术年会论文集.北京:中国植物病理学会,2017:281.
[14]韦金菊,魏春燕,宋修鹏,等.广西北海蔗区甘蔗白条病发生情况调查.南方农业学报,2018,49(2):264-270.
[15]Autrey L J C,Saumtally S,Dookun A.Studies on variation in the leaf scald pathogen,Xanthomonas albilineans[EB/OL].[2018-10-23].http://www.beep.ird.fr/cgi-bin/library.cgi?e=d-01000-00---off-0staffpu1--00-1----0-10-0---0---0direct-10---4-------0-1l--11-fr-50---20-about---01-2-1-00-0-0-11-1-0utf Zz-8-00&a=d&c=staffpu1&cl=CL3.1.32&d=HASH0121109142afe0df33d2e645s1424.
[16]Daugrois J H,Dumont V,Champoiseau P,et al.Aerial contamination of sugarcane in Guadeloupe by two strains of Xanthomonas albilineans.European Journal of Plant Pathology,2003,109(5):445-458.
[17]Champoiseau P,Daugrois J H,Girard J C,et al.Variation in albicidin biosynthesis genes and in pathogenicity of Xanthomonas albilineans,the sugarcane leaf scald pathogen.Phytopathology,2006,96(1):33-45.
[18]Huertalara M,Rojasmartinez R I,Bautistacalles J,et al.Genetic and pathogenic diversity of Xanthomonas albilineans Ashby(Dowson),in Mexico.Research Journal of Biological Sciences,2009,4(3):312-319.
[19]Rott P,Abel M,Soupa D,et al.Population dynamics of Xanthomonas albilineans in sugarcane plants as determined with an antibiotic resistant mutant.Plant Disease,1994,78(3):241-247.