我国长江流域和南方地区花生青枯菌遗传多样性分析
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摘要
为明确不同青枯菌的遗传多样性和其在花生植株上的致病力差异,采用国际上新的青枯菌演化型分类模式,对从我国长江流域和南方地区9个花生种植区分离的95株花生青枯菌Ralstonia solanacearum菌株进行遗传多样性分析,基于内源葡聚糖酶基因egl对青枯菌进行系统发育研究,并对供试青枯菌的致病力进行测定。结果表明,所有95株菌株均属于青枯菌演化型I型,即亚洲分支类型。在序列变种分类上,所检测的9个花生种植区中有8个种植区的花生青枯菌菌株属于序列变种14,仅有1个种植区(广西壮族自治区贺州市)的花生青枯菌菌株属于序列变种48,表明我国长江流域和南方地区花生青枯菌群体遗传多样性水平较低。青枯菌致病力测定结果表明,来自赣州市的菌株GZ-1、贺州市的菌株HZ-2和宜昌市的菌株YC接种到花生植株14 d后,花生的病情指数分别为43.8、75.0和87.5,而来自其它6个花生种植区的菌株接种花生后,其病情指数均为100.0,表明菌株GZ-1和HZ-2的致病力较弱,而其它7个花生种植区代表性菌株的致病力均较强。
To illuminate the genetic diversity and pathogenic variation of different Ralstonia solanacearum strains in peanut Arachis hypogaea, the new hierarchical classification scheme of the R. solanacearum species complex comprising of species, phylotypes, and sequevars was applied to analysis of genetic diversity of 95 strains of R. solanacearum from nine peanut growing areas in Yangtze River Valley and southern China. The results showed that all strains belonged to phylotype I(Asia type). The phylogenetic tree based on the nucleotide sequences of egl from R. solanacearum peanut strains and reference strains was generated to divide them into sequevars. R. solanacearum strains isolated from eight of nine areas belonged to sequevar 14, only the strains obtained from Hezhou, Guangxi belonged to sequevar 48. The results indicated that the level of genetic diversity of R. solanacearum strains from peanut was low in Yangtze River Valley and southern China. Fourteen days after infection, the disease index of GZ-1, HZ-2 and YC strains were 43.8, 75.0 and 87.5, respectively. The disease index of represent strains from the other six peanut growing areas were all 100.0. The results of pathogenic tests of R. solanacearum strains demonstrated that the pathogenicity of GZ-1 and HZ-2 strains were weak, while the other represent strains isolated from seven areas showed more virulent.
To illuminate the genetic diversity and pathogenic variation of different Ralstonia solanacearum strains in peanut Arachis hypogaea, the new hierarchical classification scheme of the R. solanacearum species complex comprising of species, phylotypes, and sequevars was applied to analysis of genetic diversity of 95 strains of R. solanacearum from nine peanut growing areas in Yangtze River Valley and southern China. The results showed that all strains belonged to phylotype I(Asia type). The phylogenetic tree based on the nucleotide sequences of egl from R. solanacearum peanut strains and reference strains was generated to divide them into sequevars. R. solanacearum strains isolated from eight of nine areas belonged to sequevar 14, only the strains obtained from Hezhou, Guangxi belonged to sequevar 48. The results indicated that the level of genetic diversity of R. solanacearum strains from peanut was low in Yangtze River Valley and southern China. Fourteen days after infection, the disease index of GZ-1, HZ-2 and YC strains were 43.8, 75.0 and 87.5, respectively. The disease index of represent strains from the other six peanut growing areas were all 100.0. The results of pathogenic tests of R. solanacearum strains demonstrated that the pathogenicity of GZ-1 and HZ-2 strains were weak, while the other represent strains isolated from seven areas showed more virulent.
引文
Anuratha CS,Gnananickam SS.1990.Biological control of bacterial wilt caused by Pseudomonas solanacearum in India with agonistic bacteria.Plant and Soil,124(1):109-116
Buddenhagen I,Kelman A.1964.Biological and physiological aspects of bacterial wilt caused by Pseudomonas solanacearum.Annual Review of Phytopathology,2:203-230
Chen BY,Jiang HF,Liao BS,Ren XP.2007.Progress on groundnut genetic enhancement for bacterial wilt resistance.Chinese Agricultural Science Bulletin,32(8):369-372(in Chinese)[陈本银,姜慧芳,廖伯寿,任小平.2007.中国花生青枯病抗性遗传改良研究进展.中国农学通报,32(8):369-372]
Cook D,Barlow E,Sequeira L.1989.Genetic diversity of Pseudomonas solanacearum:detection of restriction fragment polymorphisms with DNA probes that specify virulence and hypersensitive response.Molecular Plant-Microbe Interactions,2(3):113-121
Cook D,Sequeira L.1994.Strain differentiation of Pseudomonas solanacearum by molecular genetic methods.//Hayward AC,Hartman GL.Bacterial wilt:the disease and its causative agent,Pseudomonas solanacearum.United Kingdom:CAB International,pp.77-93
Denny T.2006.Plant pathogenic Ralstonia species.//Gnanamanickam SS.Plant associated bacteria.Dordrecht,Netherlands:Springer Publishing,pp.573-644
Fegan M,Prior P.2005.How complex is the"Ralstonia solanacearum species complex"?//Allen C,Prior P,Hayward AC.Bacterial wilt disease and the Ralstonia solanacearum species complex.Saint Paul,Minnesota:APS Press,pp.449-462
Genin S,Denny TP.2012.Pathogenomics of the Ralstonia solanacearum species complex.Annual Review of Phytopathology,50:67-89
Hayward AC.1991.Biology and epidemiology of bacterial wilt caused by Pseudomonas solanacearum.Annual Review of Phytopathology,29:65-87
Hayward AC.1994.Systematics and phylogeny of Pseudomonas solanacearum and related bacteria.//Hayward AC,Hartman GL.Bacterial wilt:the disease and its causative agent,Pseudomonas solanacearum.United Kingdom:CAB International,pp.123-135
He LY,Sequeira L,Kelma A.1983.Characteristics of strains of Pseudomonas solanacearum from China.Plant Disease,67(12):1357-1361
Holbmok CC,Stalker HT.2003.Peanut breeding and genetic resources.//Janick J.Plant breeding reviews:volume 22.Hoboken:John Wiley and Sons Ltd,pp.297-356
Horita M,Tsuchiya K.2009.Current status and future prospects of the classification system for the bacterial wilt pathogen Ralstonia solanacearum species complex.Japanese Journal of Phytopathology,75(4):297-306
Hu BY,Wang JS,Hou XY,Sun DX,Yu HB.1981.Study on bacterial wilt in peanut.Journal of Peanut Science,(4):1-8(in Chinese)[胡宝珏,王家绍,侯绪友,孙栋修,于恒宝.1981.花生青枯病的研究.花生科技,(4):1-8]
Hua JY,Zhang CL,He LY.1984.Biotypes and physiological variations of Ralstonia solanacearum Smith from China.Journal of Plant Protection,11(1):43-50(in Chinese)[华静月,张长龄,何礼远.1984.我国植物青枯病菌的生化型和其他生理差异.植物保护学报,11(1):43-50]
Jiang HF.2006.Genetic diversity among genotypes with bacterial wilt resistance and molecular markers related to resistance in peanut(Arachis hypogaea L).Ph.D Thesis.Wuhan:Huazhong Agricultural University(in Chinese)[姜慧芳.2006.花生抗青枯病种质的遗传多样性及抗性分子标记.博士学位论文.武汉:华中农业大学]
Li WR,Duan NX.1987.Study on pathogenicity of bacterial wilt in peanut.Journal of Peanut Science,(4):1-4(in Chinese)[李文溶,段迺雄.1987.花生青枯菌致病性的研究.花生科技,(4):1-4]
Li YY,Feng J,Liu HL,Wang L,Hsiang T,Li XH,Huang JB.2016.Genetic diversity and pathogenicity of Ralstonia solanacearum causing tobacco bacterial wilt in China.Plant disease,100(7):1288-1296
Li YY,Liu HL,Zheng L,Huang JB,Li XH.2015.Research progress on genetic diversity of Ralstonia solanacearum in China.Journal of Anhui Agricultural Sciences,43(14):107-110,112(in Chinese)[黎妍妍,刘海龙,郑露,黄俊斌,李锡宏.2015.我国植物青枯菌遗传多样性研究进展.安徽农业科学,43(14):107-110,112]
Liao BS,Li D,Shan ZH,Lei Y,Tan YJ,Duan NX,Tang GY.1997.Influence of latent infection of Ralstonia solanaceaum on groundnut.Chinese Journal of Oil Crop Sciences,19(4):55-58(in Chinese)[廖伯寿,李栋,单志慧,雷永,谈宇俊,段乃雄,唐桂英.1997.青枯菌潜伏侵染对花生的影响.中国油料作物学报,19(4):55-58]
Liao BS,Li WR,Song DR.1986.Study on genetic of resistance to bacteria wilt in peanut.Chinese Journal of Oil Crop Sciences,8(3):1-8(in Chinese)[廖伯寿,李文溶,孙大容.1986.花生青枯病杭性遗传研究.中国油料作物学报,8(3):1-8]
Liao BS,Shan ZH,Lei Y,Sun DR.1999.Discussion on the relationship between latent infection by Ralstonia solanaceaum and genetic improvement of resistance to bacteria wilt in peanut.Peanut Science and Technology,(S1):112-115(in Chinese)[廖伯寿,单志慧,雷永,孙大容.1999.论青枯菌潜伏侵染与花生抗性遗传改良的关系.花生科技,(S1):112-115]
Liu Y,Wu DS,Liu QP,Zhang ST,Tang YM,Jiang GF,Li SL,Ding W.2017.The sequevar distribution of Ralstonia solanacearum in tobacco-growing zones of China is structured by elevation.European Journal of Plant Pathology,147(3):541-551
Liu YQ,Kanda Y,Yano K,Kiba A,Hikichi Y,Aino M,Kawaguchi A,Mizoguchi S,Nakaho K,Shiomi H,et al.2009.Molecular typing of Japanese strains of Ralstonia solanacearum in relation to the ability to induce a hypersensitive reaction in tobacco.Journal of General Plant Pathology,75(5):369-380
Pastrik KH,Maiss E.2000.Detection of Ralstonia solanacearum in potato tubers by polymerase chain reaction.Journal of Phytopathology,148(12):619-626
Prior P,Fegan M.2005.Recent developments in the phylogeny and classification of Ralstonia solanacearum.Acta Horticulturae,695:127-136
Poussier S,Prior P,Luisetti J,Hayward C,Fegan M.2000.Partial sequencing of the hrpB and endoglucanase genes confirms and expands the known diversity within the Ralstonia solanacearum species complex.Systematic and Applied Microbiology,23(4):479-486
Remenant B,Coupat-Goutaland B,Guidot A,Cellier G,Wicker E,Allen C,Fegan M,Pruvost O,Elbaz M,Calteau A,et al.2010.Genomes of three tomato pathogens within the Ralstonia solanacearum species complex reveal significant evolutionary divergence.BMC Genomics,11:379
Su T,Lu M,Zhou Q,Wang F,Wang GF,Xie GL.2010.Evaluation and identification of antagonistic bacteria against different biovars of Ralstonia solanacearum and their active metabolites.Journal of Plant Protection,37(5):431-435(in Chinese)[苏婷,路梅,周青,王芳王国芬,谢关林.2010.抗不同生化型青枯菌的生防菌筛选鉴定及其活性分析.植物保护学报,37(5):431-435]
Tan YJ,Liao BS.1990.Review on bacteria wilt in peanut in domestic and abroad.Chinese Journal of Oil Crop Sciences,12(4):87-90(in Chinese)[谈宇俊,廖伯寿.1990.国内外花生青枯病研究述评.中国油料作物学报,12(4):87-90]
Tang RH,Zhou HQ.2000.Resistance to bacterial wilt in some interspecific derivatives of hybrids between cultivated peanut and wild species.Chinese Journal of Oil Crop Sciences,22(3):61-65(in Chinese)[唐荣华,周汉群.2000.花生属栽野杂种后代抗青枯病研究.中国油料作物学报,22(3):61-65]
Wang L.2016.Pathogenicity and genetic diversity of Ralstonia solanacearum causing potato brown rot in China.Master Thesis.Wuhan:Huazhong Agricultural University(in Chinese)[王丽.2016.中国马铃薯青枯菌致病力和遗传多样性研究.硕士学位论文.武汉:华中农业大学]
Wang Y.2013.Study on the comparative advantage of Chinese groundnut in main producing areas.Ph.D Thesis.Nanjing:Nanjing Agricultural University(in Chinese)[王艳.2013.中国花生主产区比较优势研究.博士学位论文.南京:南京农业大学]
Wicker E,Grassart L,Coranson-Beaudu R,Mian D,Guilbaud C,Fegan M,Prior P.2007.Ralstonia solanacearum strains from Martinique(French West Indies)exhibiting a new pathogenic potential.Applied and Environmental Microbiology,73(21):6790-6801
Xue QY,Yin YN,Yang W,Heuer H,Prior P,Guo JH,Smalla K.2011.Genetic diversity of Ralstonia solanacearum strains from China assessed by PCR-based fingerprints to unravel host plant-and sitedependent distribution patterns.FEMS Microbiology Ecology,75(3):507-519
Yabuuchi E,Kosako Y,Yano I,Hotta H,Nishiuchi Y.1995.Transfer of two Burkholderia and an Alcaligenes species to Ralstonia gen.nov.:proposal of Ralstonia pickettii(Ralston,Palleroni and Douderoff 1973)comb.nov.,Ralstonia solanacearum(Smith1896)comb.nov.and Ralstonia eutropha(Davis 1969)comb.nov.Microbiology and Immunology,39(11):897-904
Yan LY,Huang JQ,Lei Y,Wang SY,Liao BS.2010.Identification and mutation of Ralstonia solanacearum Hongan isolate.Chinese Journal of Oil Crop Sciences,32(1):144-146(in Chinese)[晏立英,黄家权,雷永,王圣玉,廖伯寿.2010.花生青枯菌红安分离物的鉴定.中国油料作物学报,32(1):144-146]
Buddenhagen I,Kelman A.1964.Biological and physiological aspects of bacterial wilt caused by Pseudomonas solanacearum.Annual Review of Phytopathology,2:203-230
Chen BY,Jiang HF,Liao BS,Ren XP.2007.Progress on groundnut genetic enhancement for bacterial wilt resistance.Chinese Agricultural Science Bulletin,32(8):369-372(in Chinese)[陈本银,姜慧芳,廖伯寿,任小平.2007.中国花生青枯病抗性遗传改良研究进展.中国农学通报,32(8):369-372]
Cook D,Barlow E,Sequeira L.1989.Genetic diversity of Pseudomonas solanacearum:detection of restriction fragment polymorphisms with DNA probes that specify virulence and hypersensitive response.Molecular Plant-Microbe Interactions,2(3):113-121
Cook D,Sequeira L.1994.Strain differentiation of Pseudomonas solanacearum by molecular genetic methods.//Hayward AC,Hartman GL.Bacterial wilt:the disease and its causative agent,Pseudomonas solanacearum.United Kingdom:CAB International,pp.77-93
Denny T.2006.Plant pathogenic Ralstonia species.//Gnanamanickam SS.Plant associated bacteria.Dordrecht,Netherlands:Springer Publishing,pp.573-644
Fegan M,Prior P.2005.How complex is the"Ralstonia solanacearum species complex"?//Allen C,Prior P,Hayward AC.Bacterial wilt disease and the Ralstonia solanacearum species complex.Saint Paul,Minnesota:APS Press,pp.449-462
Genin S,Denny TP.2012.Pathogenomics of the Ralstonia solanacearum species complex.Annual Review of Phytopathology,50:67-89
Hayward AC.1991.Biology and epidemiology of bacterial wilt caused by Pseudomonas solanacearum.Annual Review of Phytopathology,29:65-87
Hayward AC.1994.Systematics and phylogeny of Pseudomonas solanacearum and related bacteria.//Hayward AC,Hartman GL.Bacterial wilt:the disease and its causative agent,Pseudomonas solanacearum.United Kingdom:CAB International,pp.123-135
He LY,Sequeira L,Kelma A.1983.Characteristics of strains of Pseudomonas solanacearum from China.Plant Disease,67(12):1357-1361
Holbmok CC,Stalker HT.2003.Peanut breeding and genetic resources.//Janick J.Plant breeding reviews:volume 22.Hoboken:John Wiley and Sons Ltd,pp.297-356
Horita M,Tsuchiya K.2009.Current status and future prospects of the classification system for the bacterial wilt pathogen Ralstonia solanacearum species complex.Japanese Journal of Phytopathology,75(4):297-306
Hu BY,Wang JS,Hou XY,Sun DX,Yu HB.1981.Study on bacterial wilt in peanut.Journal of Peanut Science,(4):1-8(in Chinese)[胡宝珏,王家绍,侯绪友,孙栋修,于恒宝.1981.花生青枯病的研究.花生科技,(4):1-8]
Hua JY,Zhang CL,He LY.1984.Biotypes and physiological variations of Ralstonia solanacearum Smith from China.Journal of Plant Protection,11(1):43-50(in Chinese)[华静月,张长龄,何礼远.1984.我国植物青枯病菌的生化型和其他生理差异.植物保护学报,11(1):43-50]
Jiang HF.2006.Genetic diversity among genotypes with bacterial wilt resistance and molecular markers related to resistance in peanut(Arachis hypogaea L).Ph.D Thesis.Wuhan:Huazhong Agricultural University(in Chinese)[姜慧芳.2006.花生抗青枯病种质的遗传多样性及抗性分子标记.博士学位论文.武汉:华中农业大学]
Li WR,Duan NX.1987.Study on pathogenicity of bacterial wilt in peanut.Journal of Peanut Science,(4):1-4(in Chinese)[李文溶,段迺雄.1987.花生青枯菌致病性的研究.花生科技,(4):1-4]
Li YY,Feng J,Liu HL,Wang L,Hsiang T,Li XH,Huang JB.2016.Genetic diversity and pathogenicity of Ralstonia solanacearum causing tobacco bacterial wilt in China.Plant disease,100(7):1288-1296
Li YY,Liu HL,Zheng L,Huang JB,Li XH.2015.Research progress on genetic diversity of Ralstonia solanacearum in China.Journal of Anhui Agricultural Sciences,43(14):107-110,112(in Chinese)[黎妍妍,刘海龙,郑露,黄俊斌,李锡宏.2015.我国植物青枯菌遗传多样性研究进展.安徽农业科学,43(14):107-110,112]
Liao BS,Li D,Shan ZH,Lei Y,Tan YJ,Duan NX,Tang GY.1997.Influence of latent infection of Ralstonia solanaceaum on groundnut.Chinese Journal of Oil Crop Sciences,19(4):55-58(in Chinese)[廖伯寿,李栋,单志慧,雷永,谈宇俊,段乃雄,唐桂英.1997.青枯菌潜伏侵染对花生的影响.中国油料作物学报,19(4):55-58]
Liao BS,Li WR,Song DR.1986.Study on genetic of resistance to bacteria wilt in peanut.Chinese Journal of Oil Crop Sciences,8(3):1-8(in Chinese)[廖伯寿,李文溶,孙大容.1986.花生青枯病杭性遗传研究.中国油料作物学报,8(3):1-8]
Liao BS,Shan ZH,Lei Y,Sun DR.1999.Discussion on the relationship between latent infection by Ralstonia solanaceaum and genetic improvement of resistance to bacteria wilt in peanut.Peanut Science and Technology,(S1):112-115(in Chinese)[廖伯寿,单志慧,雷永,孙大容.1999.论青枯菌潜伏侵染与花生抗性遗传改良的关系.花生科技,(S1):112-115]
Liu Y,Wu DS,Liu QP,Zhang ST,Tang YM,Jiang GF,Li SL,Ding W.2017.The sequevar distribution of Ralstonia solanacearum in tobacco-growing zones of China is structured by elevation.European Journal of Plant Pathology,147(3):541-551
Liu YQ,Kanda Y,Yano K,Kiba A,Hikichi Y,Aino M,Kawaguchi A,Mizoguchi S,Nakaho K,Shiomi H,et al.2009.Molecular typing of Japanese strains of Ralstonia solanacearum in relation to the ability to induce a hypersensitive reaction in tobacco.Journal of General Plant Pathology,75(5):369-380
Pastrik KH,Maiss E.2000.Detection of Ralstonia solanacearum in potato tubers by polymerase chain reaction.Journal of Phytopathology,148(12):619-626
Prior P,Fegan M.2005.Recent developments in the phylogeny and classification of Ralstonia solanacearum.Acta Horticulturae,695:127-136
Poussier S,Prior P,Luisetti J,Hayward C,Fegan M.2000.Partial sequencing of the hrpB and endoglucanase genes confirms and expands the known diversity within the Ralstonia solanacearum species complex.Systematic and Applied Microbiology,23(4):479-486
Remenant B,Coupat-Goutaland B,Guidot A,Cellier G,Wicker E,Allen C,Fegan M,Pruvost O,Elbaz M,Calteau A,et al.2010.Genomes of three tomato pathogens within the Ralstonia solanacearum species complex reveal significant evolutionary divergence.BMC Genomics,11:379
Su T,Lu M,Zhou Q,Wang F,Wang GF,Xie GL.2010.Evaluation and identification of antagonistic bacteria against different biovars of Ralstonia solanacearum and their active metabolites.Journal of Plant Protection,37(5):431-435(in Chinese)[苏婷,路梅,周青,王芳王国芬,谢关林.2010.抗不同生化型青枯菌的生防菌筛选鉴定及其活性分析.植物保护学报,37(5):431-435]
Tan YJ,Liao BS.1990.Review on bacteria wilt in peanut in domestic and abroad.Chinese Journal of Oil Crop Sciences,12(4):87-90(in Chinese)[谈宇俊,廖伯寿.1990.国内外花生青枯病研究述评.中国油料作物学报,12(4):87-90]
Tang RH,Zhou HQ.2000.Resistance to bacterial wilt in some interspecific derivatives of hybrids between cultivated peanut and wild species.Chinese Journal of Oil Crop Sciences,22(3):61-65(in Chinese)[唐荣华,周汉群.2000.花生属栽野杂种后代抗青枯病研究.中国油料作物学报,22(3):61-65]
Wang L.2016.Pathogenicity and genetic diversity of Ralstonia solanacearum causing potato brown rot in China.Master Thesis.Wuhan:Huazhong Agricultural University(in Chinese)[王丽.2016.中国马铃薯青枯菌致病力和遗传多样性研究.硕士学位论文.武汉:华中农业大学]
Wang Y.2013.Study on the comparative advantage of Chinese groundnut in main producing areas.Ph.D Thesis.Nanjing:Nanjing Agricultural University(in Chinese)[王艳.2013.中国花生主产区比较优势研究.博士学位论文.南京:南京农业大学]
Wicker E,Grassart L,Coranson-Beaudu R,Mian D,Guilbaud C,Fegan M,Prior P.2007.Ralstonia solanacearum strains from Martinique(French West Indies)exhibiting a new pathogenic potential.Applied and Environmental Microbiology,73(21):6790-6801
Xue QY,Yin YN,Yang W,Heuer H,Prior P,Guo JH,Smalla K.2011.Genetic diversity of Ralstonia solanacearum strains from China assessed by PCR-based fingerprints to unravel host plant-and sitedependent distribution patterns.FEMS Microbiology Ecology,75(3):507-519
Yabuuchi E,Kosako Y,Yano I,Hotta H,Nishiuchi Y.1995.Transfer of two Burkholderia and an Alcaligenes species to Ralstonia gen.nov.:proposal of Ralstonia pickettii(Ralston,Palleroni and Douderoff 1973)comb.nov.,Ralstonia solanacearum(Smith1896)comb.nov.and Ralstonia eutropha(Davis 1969)comb.nov.Microbiology and Immunology,39(11):897-904
Yan LY,Huang JQ,Lei Y,Wang SY,Liao BS.2010.Identification and mutation of Ralstonia solanacearum Hongan isolate.Chinese Journal of Oil Crop Sciences,32(1):144-146(in Chinese)[晏立英,黄家权,雷永,王圣玉,廖伯寿.2010.花生青枯菌红安分离物的鉴定.中国油料作物学报,32(1):144-146]