京郊快菜细菌性软腐病病原鉴定
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摘要
为明确京郊快菜软腐病的病原,对取自北京不同区县的发病快菜植株进行组织病原菌分离、培养和回接试验,在形态学观察的基础上,对病原菌株进行了生理生化测定、分子生物学鉴定和致病力分析。结果表明,在分离纯化得到的29株菌株中,17株被鉴定为胡萝卜软腐果胶杆菌巴西亚种,其余12株被鉴定为胡萝卜软腐果胶杆菌胡萝卜亚种;同时也识别到9株非典型Pcc菌株,表现为不能分解柠檬酸盐或不具备在37℃和7%Na Cl条件下生长的能力,可微弱利用D-阿拉伯糖醇、异麦芽酮糖醇和D-麦芽糖。致病力测试结果显示,仅2株Pcb菌株和1株Pcc菌株表现为中等致病力,其余菌株均表现为高等致病力。明确了引发北京地区快菜软腐病致病菌的种类为Pcb和Pcc,且致病力普遍较强,可为其防控提供理论依据。
To identify the pathogens causing bacterial soft rot on Chinese cabbage in Beijng,29 pathogenic strains were isolated and cultured from diseased samples,which were collected from several districts in Beijing.Morphology analyses,physiological and biochemical tests,molecular assay and pathogenicity test were performed to elucidate the causative bacterial pathogens. The results showed that the 17 strains were identified as Pectobacterium carotovorum subsp. brasiliensis( Pcb),and the remaining 12 strains were identified as P. carotovorum subsp. carotovorum( Pcc). Moreover,there were 9 atypical Pcc strains,which were unable to degrade citrate,or failed to grow at 37 ℃ and 7% Na Cl,or weakly utilized palatinose,D-maltose and D-arabitol. The results of the pathogenicity analysis showed that all the strains expressed high pathogenicity with the exception of 2 Pcb strains and 1 Pcc strain exhibiting medium pathogenicity. The bacterial soft rot disease of Brassica rapa subsp. pekinensis occurred in Beijing and was caused mainly by Pcb and Pcc with high virulence,which will provide the basis for the disease control.
To identify the pathogens causing bacterial soft rot on Chinese cabbage in Beijng,29 pathogenic strains were isolated and cultured from diseased samples,which were collected from several districts in Beijing.Morphology analyses,physiological and biochemical tests,molecular assay and pathogenicity test were performed to elucidate the causative bacterial pathogens. The results showed that the 17 strains were identified as Pectobacterium carotovorum subsp. brasiliensis( Pcb),and the remaining 12 strains were identified as P. carotovorum subsp. carotovorum( Pcc). Moreover,there were 9 atypical Pcc strains,which were unable to degrade citrate,or failed to grow at 37 ℃ and 7% Na Cl,or weakly utilized palatinose,D-maltose and D-arabitol. The results of the pathogenicity analysis showed that all the strains expressed high pathogenicity with the exception of 2 Pcb strains and 1 Pcc strain exhibiting medium pathogenicity. The bacterial soft rot disease of Brassica rapa subsp. pekinensis occurred in Beijing and was caused mainly by Pcb and Pcc with high virulence,which will provide the basis for the disease control.
引文
[1]Davidsson P R,Kariola T,Niemi O,et al.Pathogenicity of and plant immunity to soft rot pectobacteria[J].Front Plant Science,2013,4:191.
[2]Nabhan S,Wydra K,Linde M,et al.The use of two complementary DNA assays,AFLP and MLSA,for epidemic and phylogenetic studies of pectolytic enterobacterial strains with focus on the heterogeneous species Pectobacterium carotovorum[J].Plant Pathology,2012,61(3):498-508.
[3]Alvarado I C M,Michereff S J,Mariano R L R,et al.Characterization and variability of soft rot-causing bacteria in Chinese cabbage in north eastern Brazil[J].Journal of Plant Pathology,2011,93(1):173-181.
[4]Shim H,Myung I S,Hong S K,et al.Outbreak of soft rot disease caused by Pectobacterium carotovorum subsp.carotovorum of Chinese cabbage followed by climate change in highlands of Korea[C].Korea:Society for plant pathology in China Annual Academic Symposium,2011:218-221.
[5]Waleron M,Waleron K,Lojkowska E.First report of Pectobacterium carotovorum subsp.Brasiliense causing soft rot on potato and other vegetables in Poland[J].Plant Disease,2015,99(9):1271.
[6]胡娜娜,李超,王茜,等.北京地区油菜软腐病病原菌的鉴定[J].微生物学报,2015,55(10):1253-1263.
[7]田宇,马亚丽,何付新,等.北京地区芹菜细菌性软腐病菌鉴定及其致病力分析[J].植物病理学报,2016,46(4):433-442.
[8]晋知文,宋加伟,谢学文,等.芹菜细菌性软腐病病原的分离与鉴定[J].植物病理学报,2016,46(3):304-312.
[9]方中达.植病研究方法[M].北京:农业出版社,1979:165-211.
[10]Schaad N W.Laboratory guide for identification of plant pathogenic bacteria[M].St.Paul:APS Press,2001.
[11]Mukherjee A,Cui Y,Ma W,et al.hex A of Erwinia carotovora ssp.carotovora strain Ecc71 negatively regulates production of Rpo S and rsm B RNA,a global regulator of extracellular proteins,plant virulence and the quorum-sensing signal,N-(3-oxohexanoyl)-L-homoserine lactone[J].Environmental Microbiology,2000,2(2):203-215.
[12]Zhu L,Xie H,Chen S,et al.Rapid isolation,identification and phylogenetic analysis of Pectobacterium carotovorum ssp.[J].Journal of Plant Pathology,2010,92(2):479-483.
[13]Darrasse A,Priou S,Kotoujansky A,et al.PCR and restriction fragment length polymorphism of a pel gene as a tool to identify Erwinia carotovora in relation to potato diseases[J].Applied and Environmental Microbiology,1994,60(5):1437-1443.
[14]Duarte V,de Boer S H,Ward L J,et al.Characterization of atypical Erwinia carotovora strains causing blackleg of potato in Brazil[J].Journal of Applied Microbiology,2004,96(3):535-545.
[15]Toth I K,Avrova A O,Hyman L J.Rapid identification and differentiation of the soft rot erwinias by 16S-23S intergenic transcribed spacer-PCR and restriction fragment length polymorphism analyses[J].Applied and Environmental Microbiology,2001,67(9):4070-4076.
[16]Kettani-Halabi M,Terta M,Amdan M,et al.An easy,simple inexpensive test for the specific detection of Pectobacterium carotovorum subsp.carotovorum based on sequence analysis of the pmr A gene[J].BMC Microbiology,2013,13(1):176.
[17]Terta M,El Karkouri A,Mhand R,et al.Occurrence of Pectobacterium carotovorum strains isolated from potato soft rot in Morocco[J].Cellular and Molecular Biology,2010,56(S1):OL1324-OL1333.
[18]Gardan L,Gouy C,Christen R,et al.Elevation of three subspecies of Pectobacterium carotovorum to species level:Pectobacterium atrosepticum sp.nov.,Pectobacterium betavasculorum sp.nov.and Pectobacterium wasabiae sp.nov[J].International Journal of Systematic and Evolutionary Microbiology,2003,53(Pt 2):381-391.
[19]Perombelon M C,van der Wolf J M.Methods for the detection and quantification of Erwinia carotovora subsp.atroseptica on potatoes suitable for commercial use:a laboratory manual[M].Invergowrie,Dundee,Scotland:Scottish Crop Research Institute,2002.
[20]Faquihi H,Terta M,Amdan M,et al.Phenotypic and genotypic diversity of Pectobacterium carotovorum subsp.carotovorum causing soft rot disease of potatoes in Morocco[J].European Journal of Plant Pathology,2015,143(4):801-811.
[21]Meng X,Chai A,Li B,et al.Emergence of bacterial soft rot in cucumber caused by Pectobacterium carotovorum subsp.brasiliense in China[J].Plant Disease,2017,101(2):279-287.
[22]Queiroz M F,Gama M A S,Mariano R L R,et al.First report of soft rot in kale caused by Pectobacterium carotovorum subsp.brasiliensis in Brazil[J].Plant Disease,2017,101:1542.
[23]Staley J T.The bacterial species dilemma and the genomic-phylogenetic species concept[J].Philosophical Transactions of the Royal Society of London.Series B,Biological Sciences,2006,361(1475):1899-1909.
[24]W9sten M M,Groisman E A.Molecular characterization of the Pmr A regulon[J].The Journal of Biological Chemistry,1999,274(38):27185-27190.
[25]Nabhan S,De Boer S H,Maiss E,et al.Taxonomic relatedness between Pectobacterium carotovorum subsp.carotovorum,Pectobacterium carotovorum subsp.odoriferum and Pectobacterium carotovorum subsp.brasiliense subsp.nov[J].Journal of Applied Microbiology,2012,113(4):904-913.
[26]Baghaee-Ravari S,Rahimian H,Shams-Bakhsh M,et al.Characterization of Pectobacterium species from Iran using biochemical and molecular methods[J].European Journal of Plant Pathology,2011,129(3):413-425.
[2]Nabhan S,Wydra K,Linde M,et al.The use of two complementary DNA assays,AFLP and MLSA,for epidemic and phylogenetic studies of pectolytic enterobacterial strains with focus on the heterogeneous species Pectobacterium carotovorum[J].Plant Pathology,2012,61(3):498-508.
[3]Alvarado I C M,Michereff S J,Mariano R L R,et al.Characterization and variability of soft rot-causing bacteria in Chinese cabbage in north eastern Brazil[J].Journal of Plant Pathology,2011,93(1):173-181.
[4]Shim H,Myung I S,Hong S K,et al.Outbreak of soft rot disease caused by Pectobacterium carotovorum subsp.carotovorum of Chinese cabbage followed by climate change in highlands of Korea[C].Korea:Society for plant pathology in China Annual Academic Symposium,2011:218-221.
[5]Waleron M,Waleron K,Lojkowska E.First report of Pectobacterium carotovorum subsp.Brasiliense causing soft rot on potato and other vegetables in Poland[J].Plant Disease,2015,99(9):1271.
[6]胡娜娜,李超,王茜,等.北京地区油菜软腐病病原菌的鉴定[J].微生物学报,2015,55(10):1253-1263.
[7]田宇,马亚丽,何付新,等.北京地区芹菜细菌性软腐病菌鉴定及其致病力分析[J].植物病理学报,2016,46(4):433-442.
[8]晋知文,宋加伟,谢学文,等.芹菜细菌性软腐病病原的分离与鉴定[J].植物病理学报,2016,46(3):304-312.
[9]方中达.植病研究方法[M].北京:农业出版社,1979:165-211.
[10]Schaad N W.Laboratory guide for identification of plant pathogenic bacteria[M].St.Paul:APS Press,2001.
[11]Mukherjee A,Cui Y,Ma W,et al.hex A of Erwinia carotovora ssp.carotovora strain Ecc71 negatively regulates production of Rpo S and rsm B RNA,a global regulator of extracellular proteins,plant virulence and the quorum-sensing signal,N-(3-oxohexanoyl)-L-homoserine lactone[J].Environmental Microbiology,2000,2(2):203-215.
[12]Zhu L,Xie H,Chen S,et al.Rapid isolation,identification and phylogenetic analysis of Pectobacterium carotovorum ssp.[J].Journal of Plant Pathology,2010,92(2):479-483.
[13]Darrasse A,Priou S,Kotoujansky A,et al.PCR and restriction fragment length polymorphism of a pel gene as a tool to identify Erwinia carotovora in relation to potato diseases[J].Applied and Environmental Microbiology,1994,60(5):1437-1443.
[14]Duarte V,de Boer S H,Ward L J,et al.Characterization of atypical Erwinia carotovora strains causing blackleg of potato in Brazil[J].Journal of Applied Microbiology,2004,96(3):535-545.
[15]Toth I K,Avrova A O,Hyman L J.Rapid identification and differentiation of the soft rot erwinias by 16S-23S intergenic transcribed spacer-PCR and restriction fragment length polymorphism analyses[J].Applied and Environmental Microbiology,2001,67(9):4070-4076.
[16]Kettani-Halabi M,Terta M,Amdan M,et al.An easy,simple inexpensive test for the specific detection of Pectobacterium carotovorum subsp.carotovorum based on sequence analysis of the pmr A gene[J].BMC Microbiology,2013,13(1):176.
[17]Terta M,El Karkouri A,Mhand R,et al.Occurrence of Pectobacterium carotovorum strains isolated from potato soft rot in Morocco[J].Cellular and Molecular Biology,2010,56(S1):OL1324-OL1333.
[18]Gardan L,Gouy C,Christen R,et al.Elevation of three subspecies of Pectobacterium carotovorum to species level:Pectobacterium atrosepticum sp.nov.,Pectobacterium betavasculorum sp.nov.and Pectobacterium wasabiae sp.nov[J].International Journal of Systematic and Evolutionary Microbiology,2003,53(Pt 2):381-391.
[19]Perombelon M C,van der Wolf J M.Methods for the detection and quantification of Erwinia carotovora subsp.atroseptica on potatoes suitable for commercial use:a laboratory manual[M].Invergowrie,Dundee,Scotland:Scottish Crop Research Institute,2002.
[20]Faquihi H,Terta M,Amdan M,et al.Phenotypic and genotypic diversity of Pectobacterium carotovorum subsp.carotovorum causing soft rot disease of potatoes in Morocco[J].European Journal of Plant Pathology,2015,143(4):801-811.
[21]Meng X,Chai A,Li B,et al.Emergence of bacterial soft rot in cucumber caused by Pectobacterium carotovorum subsp.brasiliense in China[J].Plant Disease,2017,101(2):279-287.
[22]Queiroz M F,Gama M A S,Mariano R L R,et al.First report of soft rot in kale caused by Pectobacterium carotovorum subsp.brasiliensis in Brazil[J].Plant Disease,2017,101:1542.
[23]Staley J T.The bacterial species dilemma and the genomic-phylogenetic species concept[J].Philosophical Transactions of the Royal Society of London.Series B,Biological Sciences,2006,361(1475):1899-1909.
[24]W9sten M M,Groisman E A.Molecular characterization of the Pmr A regulon[J].The Journal of Biological Chemistry,1999,274(38):27185-27190.
[25]Nabhan S,De Boer S H,Maiss E,et al.Taxonomic relatedness between Pectobacterium carotovorum subsp.carotovorum,Pectobacterium carotovorum subsp.odoriferum and Pectobacterium carotovorum subsp.brasiliense subsp.nov[J].Journal of Applied Microbiology,2012,113(4):904-913.
[26]Baghaee-Ravari S,Rahimian H,Shams-Bakhsh M,et al.Characterization of Pectobacterium species from Iran using biochemical and molecular methods[J].European Journal of Plant Pathology,2011,129(3):413-425.