一种由山葵果胶杆菌引起的马铃薯细菌性软腐病
|
摘要
【目的】本文确定了引发青海省马铃薯(Solanum tuberosum L.)细菌性软腐病的病原。【方法】结合选择性CVP培养基上菌落形态观察、致病性测定、16S r DNA序列以及Pectobacterium spp.的序列特征分析,对从青海大通县马铃薯软腐病样中的病原菌进行生物学鉴定。【结果】鉴定出肠杆菌科果胶杆菌属山葵果胶杆菌(Pectobacterium wasabiae)能引起青海马铃薯细菌性软腐病。生理生化试验表明,该病原菌株不能在37℃生长,能使明胶液化,不具有耐盐性,对红霉素不敏感;过氧化氢酶反应为阳性,氧化酶、蔗糖试验还原反应、吲哚产生和磷酸酶活性试验均为阴性;可以利用纤维二糖和柠檬酸盐,不能利用D-麦芽糖、山梨醇、D-阿拉伯糖、α-甲基葡萄糖、乳糖、棉子糖和蜜二糖。【结论】研究结果可为青海马铃薯软腐病的综合防治提供科学依据。
【Objective】This present study aimed to identify the causal pathogen causing soft rot of potato( Solanum tuberosum L.) in Qinghai province.【Method】One strain isolated from infected potato tubers was characterized by morphology on CVP medium,pathogenicity test,analyses of 16 S rRNA sequence as well as Pectobacterium spp. sequence characteristics.【Result】This strain was identified as Pectobacterium wasabiae and belonged to Enterobacteriaceae,Pectobacterium. The physiological and biochemical tests showed that the bacterium could not grow at 37 ℃,caused gelatin liquefaction,had not the tolerance to salt in 5 % Na Cl solution and had no sensitivity to erythromycin; The bacterium gave a positive response to catalase but negative responses to oxidase,phosphatase activity,indole production and sucrose reduction reaction. The bacterium utilized some substances including citrate and cellobiose but could not utilize D-maltose,D-sorbitol,D-arabitol,α-methy lglucoside,lactose,raffinose,and melibiose.【Conclusion】These results would provide a scientific basis for the integrated control of potato soft rot disease.
【Objective】This present study aimed to identify the causal pathogen causing soft rot of potato( Solanum tuberosum L.) in Qinghai province.【Method】One strain isolated from infected potato tubers was characterized by morphology on CVP medium,pathogenicity test,analyses of 16 S rRNA sequence as well as Pectobacterium spp. sequence characteristics.【Result】This strain was identified as Pectobacterium wasabiae and belonged to Enterobacteriaceae,Pectobacterium. The physiological and biochemical tests showed that the bacterium could not grow at 37 ℃,caused gelatin liquefaction,had not the tolerance to salt in 5 % Na Cl solution and had no sensitivity to erythromycin; The bacterium gave a positive response to catalase but negative responses to oxidase,phosphatase activity,indole production and sucrose reduction reaction. The bacterium utilized some substances including citrate and cellobiose but could not utilize D-maltose,D-sorbitol,D-arabitol,α-methy lglucoside,lactose,raffinose,and melibiose.【Conclusion】These results would provide a scientific basis for the integrated control of potato soft rot disease.
引文
[1]农业部新闻办公室.以科技创新引领马铃薯主粮化发展[CP/OL].[2015-01-06]http//www.moa.gov.cn/zwllm/zwdt/201501/t20150106_4323476.htm.
[2]王舰,王芳.2013年青海省马铃薯产业发展现状与可持续发展[C].//中国马铃薯大会,黑龙江大兴安岭加格达奇区,2014:162-165.
[3]王金生.植物病原细菌学[M].北京:中国农业出版社,2000:196-201.
[4]Lund M B.Bacterial soft rot of potato in:in Plant Pathogens(Lovelock D W Ed)[M].New York:Academic Press,1979:24-29.
[5]Lapwood D H,Read T P,Spokes J.Methods for assessing the susceptibility of potato of different cultivars to rotting by Erwinias carotovora subspecies atroseptica and carotovora[J].Plant Pathol,1984,33(1):13-20.
[6]Cother J E,Powell V.Physiological and pathological characteristics of Erwinia chrysanthemi isolates from potato tuber[J].J Appl Microbiol,1983,54(1):37-43.
[7]Stawiak M,van Beckhoven J R C M,Speksnijder A G C L,et al.Biochemical and genetical analysis reveal a new clade of biovar 3Dickeya spp.Strains isolated from potato in Europe[J].Eur J Plant Pathol,2009,125(2):245-261.
[8]Toth I K,van der Wolf J M,Saddler G,et al.Dickeya species:an emerging problem for potato production in Europe[J].Plant Pathol,2011,60(3):385-399.
[9]Choi O,Kim J.Pectobacterium carotovorum subsp.brasiliense causing soft rot on paprika in Korea[J].J Phytopathol,2013,161(2):125-127.
[10]Perombelon M C,Kelman A.Ecology of the soft rot Erwinias[J].Annual Review of Phytopathology,1980,18(1):361-387.
[11]王金生,韦忠民,方中达.马铃薯软腐细菌的鉴定[J].植物病理学报,1985,15(1):25-30.
[12]Goto M,Matsumoto K.Erwinia carotovora subsp.wasabiae subsp.nov.isolated from diseased rhizomes and fibrous roots of Japanese horseradish[J].Int J Syst Bacteriol,1987,37(2):130-135.
[13]Kim H S,Ma B,Perna N T,et al.Phylogeny and virulence of naturally occurring type III secretion system-deficient Pectobacterium strains[J].Appl Environ Microb,2009,75(13):4539-4549.
[14]Pitman A R,Wright P J,Gailbraith M D,et al.Biochemical and genetic diversity of pectolytic enterobacteria causing soft rot disease of potatoes in New Zealand[J].Australas Plant Path,2008,37(6):559-568.
[15]Pitman A R,Harrow S A,Visnovsky S B.Genetic characterization of Pectobacterium wasabiae causing soft rot disease of potato in New Zealand[J].Eur J Plant Pathol,2010,126(3):423-435.
[16]Baghaee-Ravari S,Rahimian H,Shams-Bakhsh M,et al.Characterization of Pectobacterium species from Iran using biochemical and molecular methods[J].Eur J Plant Pathol,2011,129(3):413-425.
[17]方中达.植病研究方法[M].北京:中国农业出版社,1998.
[18]Garrity G.Bergey’s Manual of Systematic Bacteriology[M].Volume 2:The Proteobacteria.Berlin:Springer Verlag,2005.
[19]Onkendi E M,Moleleki L N.Characterization of Pectobacterium carotovorum subsp.carotovorum and brasiliense from diseased potatoes in Kenya[J].Eur J Plant Pathol,2014,139(3):557-566.
[20]Nabhan S,Boer S H D,Maiss E,et al.Taxonomic relatedness between Pectobacterium carotovorum subsp.carotovorum,Pectobacterium carotovorum subsp.odoriferum and Pectobacterium carotovorum subsp.brasiliense subsp.nov[J].Appl Environ Microb,2012,113(4):904-913.
[21]Yap M N,Barak J D,Charkowski A O.Genomic diversity of Erwinia carotovora subsp.carotovora and its correlation with virulence[J].Appl Environ Microb,2004,70(5):3013-3023.
[22]Glasner J D,Marquez-Villavicencio M,Kim H S,et al.Niche-specificity and the variable fraction of the Pectobacterium pan-genome[J].Plant Microbe In,2008,21(12):1549-1560.
[23]Rantakari A,Virtaharju O,Vhhmiko S,et al.Type III secretion contributes to the pathogenesis of the soft rot pathogen Erwinia carotovora:Partial characterization of the hrp gene cluster[J].Mol Plant Microbe In,2001,14(8):962-968.
[24]Holeva M C,Bell K S,Hyman L J,et al.Use of a pooled transposon mutation grid to demonstrate roles in disease development for Erwinia carotovora subsp.atroseptica putative type III secreted effector(Dsp E/A)and helper(Hrp N)proteins[J].Mol Plant Microbe In,2004,17(9):943-950.
[2]王舰,王芳.2013年青海省马铃薯产业发展现状与可持续发展[C].//中国马铃薯大会,黑龙江大兴安岭加格达奇区,2014:162-165.
[3]王金生.植物病原细菌学[M].北京:中国农业出版社,2000:196-201.
[4]Lund M B.Bacterial soft rot of potato in:in Plant Pathogens(Lovelock D W Ed)[M].New York:Academic Press,1979:24-29.
[5]Lapwood D H,Read T P,Spokes J.Methods for assessing the susceptibility of potato of different cultivars to rotting by Erwinias carotovora subspecies atroseptica and carotovora[J].Plant Pathol,1984,33(1):13-20.
[6]Cother J E,Powell V.Physiological and pathological characteristics of Erwinia chrysanthemi isolates from potato tuber[J].J Appl Microbiol,1983,54(1):37-43.
[7]Stawiak M,van Beckhoven J R C M,Speksnijder A G C L,et al.Biochemical and genetical analysis reveal a new clade of biovar 3Dickeya spp.Strains isolated from potato in Europe[J].Eur J Plant Pathol,2009,125(2):245-261.
[8]Toth I K,van der Wolf J M,Saddler G,et al.Dickeya species:an emerging problem for potato production in Europe[J].Plant Pathol,2011,60(3):385-399.
[9]Choi O,Kim J.Pectobacterium carotovorum subsp.brasiliense causing soft rot on paprika in Korea[J].J Phytopathol,2013,161(2):125-127.
[10]Perombelon M C,Kelman A.Ecology of the soft rot Erwinias[J].Annual Review of Phytopathology,1980,18(1):361-387.
[11]王金生,韦忠民,方中达.马铃薯软腐细菌的鉴定[J].植物病理学报,1985,15(1):25-30.
[12]Goto M,Matsumoto K.Erwinia carotovora subsp.wasabiae subsp.nov.isolated from diseased rhizomes and fibrous roots of Japanese horseradish[J].Int J Syst Bacteriol,1987,37(2):130-135.
[13]Kim H S,Ma B,Perna N T,et al.Phylogeny and virulence of naturally occurring type III secretion system-deficient Pectobacterium strains[J].Appl Environ Microb,2009,75(13):4539-4549.
[14]Pitman A R,Wright P J,Gailbraith M D,et al.Biochemical and genetic diversity of pectolytic enterobacteria causing soft rot disease of potatoes in New Zealand[J].Australas Plant Path,2008,37(6):559-568.
[15]Pitman A R,Harrow S A,Visnovsky S B.Genetic characterization of Pectobacterium wasabiae causing soft rot disease of potato in New Zealand[J].Eur J Plant Pathol,2010,126(3):423-435.
[16]Baghaee-Ravari S,Rahimian H,Shams-Bakhsh M,et al.Characterization of Pectobacterium species from Iran using biochemical and molecular methods[J].Eur J Plant Pathol,2011,129(3):413-425.
[17]方中达.植病研究方法[M].北京:中国农业出版社,1998.
[18]Garrity G.Bergey’s Manual of Systematic Bacteriology[M].Volume 2:The Proteobacteria.Berlin:Springer Verlag,2005.
[19]Onkendi E M,Moleleki L N.Characterization of Pectobacterium carotovorum subsp.carotovorum and brasiliense from diseased potatoes in Kenya[J].Eur J Plant Pathol,2014,139(3):557-566.
[20]Nabhan S,Boer S H D,Maiss E,et al.Taxonomic relatedness between Pectobacterium carotovorum subsp.carotovorum,Pectobacterium carotovorum subsp.odoriferum and Pectobacterium carotovorum subsp.brasiliense subsp.nov[J].Appl Environ Microb,2012,113(4):904-913.
[21]Yap M N,Barak J D,Charkowski A O.Genomic diversity of Erwinia carotovora subsp.carotovora and its correlation with virulence[J].Appl Environ Microb,2004,70(5):3013-3023.
[22]Glasner J D,Marquez-Villavicencio M,Kim H S,et al.Niche-specificity and the variable fraction of the Pectobacterium pan-genome[J].Plant Microbe In,2008,21(12):1549-1560.
[23]Rantakari A,Virtaharju O,Vhhmiko S,et al.Type III secretion contributes to the pathogenesis of the soft rot pathogen Erwinia carotovora:Partial characterization of the hrp gene cluster[J].Mol Plant Microbe In,2001,14(8):962-968.
[24]Holeva M C,Bell K S,Hyman L J,et al.Use of a pooled transposon mutation grid to demonstrate roles in disease development for Erwinia carotovora subsp.atroseptica putative type III secreted effector(Dsp E/A)and helper(Hrp N)proteins[J].Mol Plant Microbe In,2004,17(9):943-950.