西北地区马铃薯疮痂病病原菌鉴定及其生物学特性
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摘要
为明确西北地区马铃薯疮痂病病原菌的种类和生物学特性,分别采用常规组织分离法和土壤混悬液分离法从宁夏、陕西和甘肃3个省区采集的29份疮痂病发病薯块和8份发病地块土壤中进行病原菌分离,并利用形态特征、生理生化特性和16S rDNA序列分析对病原菌进行鉴定。结果表明,从发病薯块和发病土壤中共分离到50株链霉菌Streptomyces spp.,通过回接法验证获得6株马铃薯疮痂病致病菌株。6株致病菌株的培养特性和形态特征差别较大;其中菌株G4-1、G9和SYN13不能以果糖和木糖为单一碳源,菌株SYNT3不能以棉子糖为单一碳源;除菌株NLG4-1外,其余5株菌株均能在络氨酸琼脂培养基上产生黑色素。经16S rDNA序列分析,菌株G4-1、G9与疮痂病链霉菌S. scabiei的相似率分别达99.47%和99.34%,菌株NLG4-1、SYNT3与S. enissocaesilis的相似率分别达97.90%和98.18%,菌株GBH2与加利利链霉菌S. galilaeus的相似率达99.93%,菌株SYN13与S. turgidiscabies的相似率达97.56%,表明西北地区马铃薯疮痂病病原菌至少存在4个种。
To clarify the species and biological characteristics of the pathogenic bacteria of potato scab disease in northwestern China, the pathogens were isolated from 29 samples of pathogenic potato chips and eight samples of pathogenic soil collected from Ningxia, Shaanxi and Gansu provinces with conventional tissue separation and soil suspension separation, and the pathogens were identified with biological, physiological and biochemical characteristics and 16S rDNA sequence analysis. The results showed that 50 strains of Streptomyces were isolated from the pathogenic potato and soil, and six were obtained by reattachment test. The cultural characteristics and morphological features of the six pathogenic strains were quite different. Among them, D-fructose and D-xylose could not be used as a single carbon source for strains G4-1, G9 and SYN13. Strain SYNT3 could not use raffinose as single carbon source.All the five strains except NLG4-1 could produce melanin in the Tyrosine-agar. According to 16S rDNA sequence analysis, the similarity rates of strains G4-1 and G9 with S. scabiei were 99.47% and 99.34%,respectively; the similarity rates of strains NLG4-1 and SYNT3 with S. enissocaesilis reached 97.90%and 98.18%, respectively; the similarity of the strain GBH2 with S. galilaeus was 99.93%, and the similarity of the strain SYN13 with S. turgidiscabies was 97.56%. It indicated that there were at least four species of pathogenic bacteria of potato scab disease in northwestern China.
To clarify the species and biological characteristics of the pathogenic bacteria of potato scab disease in northwestern China, the pathogens were isolated from 29 samples of pathogenic potato chips and eight samples of pathogenic soil collected from Ningxia, Shaanxi and Gansu provinces with conventional tissue separation and soil suspension separation, and the pathogens were identified with biological, physiological and biochemical characteristics and 16S rDNA sequence analysis. The results showed that 50 strains of Streptomyces were isolated from the pathogenic potato and soil, and six were obtained by reattachment test. The cultural characteristics and morphological features of the six pathogenic strains were quite different. Among them, D-fructose and D-xylose could not be used as a single carbon source for strains G4-1, G9 and SYN13. Strain SYNT3 could not use raffinose as single carbon source.All the five strains except NLG4-1 could produce melanin in the Tyrosine-agar. According to 16S rDNA sequence analysis, the similarity rates of strains G4-1 and G9 with S. scabiei were 99.47% and 99.34%,respectively; the similarity rates of strains NLG4-1 and SYNT3 with S. enissocaesilis reached 97.90%and 98.18%, respectively; the similarity of the strain GBH2 with S. galilaeus was 99.93%, and the similarity of the strain SYN13 with S. turgidiscabies was 97.56%. It indicated that there were at least four species of pathogenic bacteria of potato scab disease in northwestern China.
引文
Archuleta JG,Easton GD.1981.The cause of deep-pitted scab of potatoes.American Journal of Potato Research,58(8):385-392
Bouchek-Mechiche K,Gardan L,Normand P,Jouan B.2000.DNA relatedness among strains of Streptomyces pathogenic to potato in France:description of three new species,S.europaeiscabiei sp.nov.and S.stelliscabiei sp.nov.associated with common scab,and S.reticuliscabiei sp.nov.associated with netted scab.International Journal of Systematic and Evolutionary Microbiology,50(1):91-99
Bukhalid RA,Chung SY,Loria R.1998.nec1,a gene conferring a necrogenic phenotype,is conserved in plant-pathogenic Streptomyces spp.,and linked to a transposase pseudogene.Molecular Plant-Microbe Interactions,11(10):960-967
Bukhalid RA,Takeuchi T,Labeda D,Loria R.2002.Horizontal transfer of the plant virulence gene,nec1,and flanking sequences among genetically distinct Streptomyces strains in the diastatochromogenes cluster.Applied and Environmental Microbiology,68(2):738-744
Doering-Saad C,Kampfer P,Manulis S,Kritaman G,Schneider J,Zakrzewska-Czerwinska J,Schrempf H,Barash I.1992.Diversity among Streptomyces strains causing potato scab.Applied and Environmental Microbiology,58(12):3932-3940
Fan CX.2012.The technology of prevention and control on potato scab.Rural Science and Technology,(12):41(in Chinese)[范彩霞.2012.马铃薯疮痂病防治技术.农村科技,(12):41]
Goyer C,Faucher E,Beaulieu C.1996.Streptomyces caviscabies sp.nov.,from deep-pitted lesions in potatoes in Quebec,Canada.International Journal of Systematic Bacteriology,46(3):635-639
Healy FG,Bukhalid RA,Loria R.1999.Characterization of an insertion sequence element associated with genetically diverse plant pathogenic Streptomyces spp.Journal of Bacteriology,181(5):1562-1568
Hiltunen LH,Weckman A,Ylhainen A,Rita H,Richter E,Tvalkonen JPT.2005.Responses of potato cultivars to the common scab pathogens,Streptomyces scabies and S.turgidiscabies.Annals of Applied Biology,146:395-403
Kang R,Wang SR.2013.Preliminary identification of the pathogen of potato scab in Gansu.Plant Protection,39(3):78-82(in Chinese)[康蓉,王生荣.2013.甘肃马铃薯疮痂病病原初步鉴定.植物保护,39(3):78-82]
Kers JA,Cameron KD,Joshi MV,Bukhalid RA,Morello JE,Wach MJ,Gibson DM,Loria R.2005.A large mobile pathogenicity island confers plant pathogenicity on Streptomyces species.Molecular Microbiology,55(4):1025-1033
Lambert DH,Loria R.1989.Streptomyces scabies sp.nov.,nom.rev.International Journal of Systematic Bacteriology,39(4):387-392
Li QQ,Li JP.2006.Effects of 5 fungicides in different dilute solution irrigation on controlling potato scab.Gansu Agricultural Science and Technology,(8):31-32(in Chinese)[李青青,李继平.2006.5种药剂不同稀释液浇灌防治脱毒马铃薯疮痂病效果初报.甘肃农业科技,(8):31-32]
Miyajima K,Tanaka F,Takeuchi T,Kuninaqa S.1998.Streptomyces turgidiscabies sp.nov.International Journal of Systematic Bacteriology,48(2):495-502
Wanner LA.2009.A patchwork of Streptomyces species isolated from potato common scab lesions in North America.American Journal of Potato Research,86(4):247-64
Xing YY,LüDQ,Wei Q,Wan SM,Dong XZ,Qiu CL,Jin GH.2016.Species and pathogenicity identification of Streptomyces species causing potato common scab in part of Heilongjiang Province.Plant Protection,42(1):26-32,50(in Chinese)[邢莹莹,吕典秋,魏琪,万书明,董学志,邱彩玲,金光辉.2016.黑龙江省部分地区马铃薯疮痂病病菌种类及致病性鉴定.植物保护,42(1):26-32,50]
Yang MP,Wang RX,Du WF,Gong C,Yu DC,Zhao WQ,Song SW,Zhang HJ.2018.The pathogenic Streptomyces species causing potato scab in Yunnan Province.Acta Phytopathologica Sinica,48(4):445-454(in Chinese)[杨梦平,王瑞仙,杜魏甫,巩晨,于德才,赵伟全,宋双伟,张红骥.2018.云南省马铃薯疮痂病致病链霉菌种类组成研究.植物病理学报,48(4):445-454]
Zhang HY,Guo FL,Xu HM,Zhao WQ,Liu DQ.2014.Identification of the pathogen of potato scab in Zhangbei areas of Hebei.Science Agricultura Jiangsu,42(10):131-134(in Chinese)[张海颖,郭凤柳,许华民,赵伟全,刘大群.2014.河北省张北地区马铃薯疮痂病的病菌鉴定.江苏农业科学,42:(10):131-134]
Zhang JP,Yi YH,Yan RP,Ha S,Lin TR,Cheng YC,Hu B.2013.Occurrence and control approach of potato common scab(Streptomyces spp.)in Inner Mongolia.Chinese Potato Journal,27(1):56-59(in Chinese)[张建平,尹玉和,闫任沛,哈斯,林团荣,程玉臣,胡冰.2013.内蒙古马铃薯疮痂病发生与防治途径.中国马铃薯,27(1):56-59]
Zhang M,Zhao WQ,Yu XM,Yang WX,Zhang T,Liu DQ.2009.Genetic diversity of potato scab pathogens in China based on 16SrDNA sequences.Scientia Agricultura Sinica,42(2):499-504(in Chinese)[张萌,赵伟全,于秀梅,杨文香,张汀,刘大群.2009.中国马铃薯疮痂病病原菌16S rDNA的遗传多样性分析.中国农业科学,42(2):499-504]
Zhao WQ,Yang WX,Li YN,Liu DQ,Meng QF,Zhang T.2006.Characterization and identification on the pathogen of potato scab in China.Science Agricultura Sinica,39(2):313-318(in Chinese)[赵伟全,杨文香,李亚宁,刘大群,孟庆芳,张汀.2006.中国马铃薯疮痂病菌的鉴定.中国农业科学,39(2):313-318]
Bouchek-Mechiche K,Gardan L,Normand P,Jouan B.2000.DNA relatedness among strains of Streptomyces pathogenic to potato in France:description of three new species,S.europaeiscabiei sp.nov.and S.stelliscabiei sp.nov.associated with common scab,and S.reticuliscabiei sp.nov.associated with netted scab.International Journal of Systematic and Evolutionary Microbiology,50(1):91-99
Bukhalid RA,Chung SY,Loria R.1998.nec1,a gene conferring a necrogenic phenotype,is conserved in plant-pathogenic Streptomyces spp.,and linked to a transposase pseudogene.Molecular Plant-Microbe Interactions,11(10):960-967
Bukhalid RA,Takeuchi T,Labeda D,Loria R.2002.Horizontal transfer of the plant virulence gene,nec1,and flanking sequences among genetically distinct Streptomyces strains in the diastatochromogenes cluster.Applied and Environmental Microbiology,68(2):738-744
Doering-Saad C,Kampfer P,Manulis S,Kritaman G,Schneider J,Zakrzewska-Czerwinska J,Schrempf H,Barash I.1992.Diversity among Streptomyces strains causing potato scab.Applied and Environmental Microbiology,58(12):3932-3940
Fan CX.2012.The technology of prevention and control on potato scab.Rural Science and Technology,(12):41(in Chinese)[范彩霞.2012.马铃薯疮痂病防治技术.农村科技,(12):41]
Goyer C,Faucher E,Beaulieu C.1996.Streptomyces caviscabies sp.nov.,from deep-pitted lesions in potatoes in Quebec,Canada.International Journal of Systematic Bacteriology,46(3):635-639
Healy FG,Bukhalid RA,Loria R.1999.Characterization of an insertion sequence element associated with genetically diverse plant pathogenic Streptomyces spp.Journal of Bacteriology,181(5):1562-1568
Hiltunen LH,Weckman A,Ylhainen A,Rita H,Richter E,Tvalkonen JPT.2005.Responses of potato cultivars to the common scab pathogens,Streptomyces scabies and S.turgidiscabies.Annals of Applied Biology,146:395-403
Kang R,Wang SR.2013.Preliminary identification of the pathogen of potato scab in Gansu.Plant Protection,39(3):78-82(in Chinese)[康蓉,王生荣.2013.甘肃马铃薯疮痂病病原初步鉴定.植物保护,39(3):78-82]
Kers JA,Cameron KD,Joshi MV,Bukhalid RA,Morello JE,Wach MJ,Gibson DM,Loria R.2005.A large mobile pathogenicity island confers plant pathogenicity on Streptomyces species.Molecular Microbiology,55(4):1025-1033
Lambert DH,Loria R.1989.Streptomyces scabies sp.nov.,nom.rev.International Journal of Systematic Bacteriology,39(4):387-392
Li QQ,Li JP.2006.Effects of 5 fungicides in different dilute solution irrigation on controlling potato scab.Gansu Agricultural Science and Technology,(8):31-32(in Chinese)[李青青,李继平.2006.5种药剂不同稀释液浇灌防治脱毒马铃薯疮痂病效果初报.甘肃农业科技,(8):31-32]
Miyajima K,Tanaka F,Takeuchi T,Kuninaqa S.1998.Streptomyces turgidiscabies sp.nov.International Journal of Systematic Bacteriology,48(2):495-502
Wanner LA.2009.A patchwork of Streptomyces species isolated from potato common scab lesions in North America.American Journal of Potato Research,86(4):247-64
Xing YY,LüDQ,Wei Q,Wan SM,Dong XZ,Qiu CL,Jin GH.2016.Species and pathogenicity identification of Streptomyces species causing potato common scab in part of Heilongjiang Province.Plant Protection,42(1):26-32,50(in Chinese)[邢莹莹,吕典秋,魏琪,万书明,董学志,邱彩玲,金光辉.2016.黑龙江省部分地区马铃薯疮痂病病菌种类及致病性鉴定.植物保护,42(1):26-32,50]
Yang MP,Wang RX,Du WF,Gong C,Yu DC,Zhao WQ,Song SW,Zhang HJ.2018.The pathogenic Streptomyces species causing potato scab in Yunnan Province.Acta Phytopathologica Sinica,48(4):445-454(in Chinese)[杨梦平,王瑞仙,杜魏甫,巩晨,于德才,赵伟全,宋双伟,张红骥.2018.云南省马铃薯疮痂病致病链霉菌种类组成研究.植物病理学报,48(4):445-454]
Zhang HY,Guo FL,Xu HM,Zhao WQ,Liu DQ.2014.Identification of the pathogen of potato scab in Zhangbei areas of Hebei.Science Agricultura Jiangsu,42(10):131-134(in Chinese)[张海颖,郭凤柳,许华民,赵伟全,刘大群.2014.河北省张北地区马铃薯疮痂病的病菌鉴定.江苏农业科学,42:(10):131-134]
Zhang JP,Yi YH,Yan RP,Ha S,Lin TR,Cheng YC,Hu B.2013.Occurrence and control approach of potato common scab(Streptomyces spp.)in Inner Mongolia.Chinese Potato Journal,27(1):56-59(in Chinese)[张建平,尹玉和,闫任沛,哈斯,林团荣,程玉臣,胡冰.2013.内蒙古马铃薯疮痂病发生与防治途径.中国马铃薯,27(1):56-59]
Zhang M,Zhao WQ,Yu XM,Yang WX,Zhang T,Liu DQ.2009.Genetic diversity of potato scab pathogens in China based on 16SrDNA sequences.Scientia Agricultura Sinica,42(2):499-504(in Chinese)[张萌,赵伟全,于秀梅,杨文香,张汀,刘大群.2009.中国马铃薯疮痂病病原菌16S rDNA的遗传多样性分析.中国农业科学,42(2):499-504]
Zhao WQ,Yang WX,Li YN,Liu DQ,Meng QF,Zhang T.2006.Characterization and identification on the pathogen of potato scab in China.Science Agricultura Sinica,39(2):313-318(in Chinese)[赵伟全,杨文香,李亚宁,刘大群,孟庆芳,张汀.2006.中国马铃薯疮痂病菌的鉴定.中国农业科学,39(2):313-318]