菜豆普通细菌性疫病病原菌鉴定

doi:10.3864/j.issn.0578-1752.2012.13.006

摘要/Abstract

摘要： 【目的】明确中国北方地区菜豆普通细菌性疫病病原菌。【方法】应用菌落特征、油菜黄单胞菌菜豆变种诊断试剂检测、致病性测定、16S rDNA和16S-23S rDNA ITS序列分析、特异PCR检测、脂肪酸分析及生理生化反应特征对病原菌分离物进行鉴定。【结果】从病害样品和菜豆种子样品中均分离到类似黄单胞杆菌的细菌分离物，选取25个代表分离物进行致病性测定，有24个分离物在菜豆品种“英国红”上导致典型菜豆普通细菌性疫病症状。结合16S rDNA及16S-23S rDNA ITS序列比对分析、特异性PCR检测及生理生化反应的结果，24个分离物中7株被鉴定为地毯草黄单胞菌菜豆变种，17株为褐色黄单胞菌褐色亚种。【结论】中国北方地区发生的菜豆普通细菌性疫病由地毯草黄单胞菌菜豆变种或（和）褐色黄单胞菌褐色亚种引起；褐色黄单胞菌褐色亚种在中国属首次报道。

关键词: 菜豆, 菜豆普通细菌性疫病, 地毯草黄单胞菌菜豆变种, 褐色黄单胞菌褐色亚种

Abstract: 【Objective】The objective of this study is to identify the pathogens that cause common bacterial blight on common bean in north China.【Method】 The causal agents were identified by colony characterization, detection using Xanthomonas campestris pv. phaseoli diagonsis kit (ADGEN EXPRESSTM), pathogenicity test, sequencing 16 rDNA and 16S-23S rDNA intergenic spacer (ITS) regions, PCR detection using specific primers for X. axonopodis pv. phaseoli and X. fuscans subsp. fuscans, fatty acid methyl esters (FAME) analysis and biochemical characterization tests.【Result】Bacterial isolates of Xanthmonads-like were isolated from all diseased plant and seed samples. Twenty-five representative isolates were selected for pathogenicity test, and 24 tested isolates casused typical symptoms of common bacterial blight on the inoculated plants of common bean cutivar ‘Yingguohong’. Based on pathenogenicity test, and combined with the results of diagonsis kit and specific PCR detection, biochemical and molecular characterization, 7 isoaltes were identifed as X. axonopodis pv. phaseoli, and 17 isolates were identified as X. fuscans subsp. fuscans.【Conclusion】 The common bacterial blight on common bean in north China was incited by X. axonopodis pv. phaseoli or/and X. fuscans subsp. fuscans. It was the first report of X. fuscans subsp. fuscans in China.

Key words: common bean, common bacterial blight, Xanthomonas axonopodis pv. phaseoli, X. fuscans subsp. fuscans

陈泓宇, 徐新新, 段灿星, 王述民, 朱振东. 菜豆普通细菌性疫病病原菌鉴定[J]. 中国农业科学, 2012, 45(13): 2618-2627.

CHEN Hong-Yu, XU Xin-Xin, DUAN Can-Xing, WANG Shu-Min, ZHU Zhen-Dong. Identification of Pathogens Causing Common Bacterial Blight on Common Bean (Phaseolus vulgaris L.)[J]. Scientia Agricultura Sinica, 2012, 45(13): 2618-2627.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2012.13.006

https://www.chinaagrisci.com/CN/Y2012/V45/I13/2618

参考文献

[1]Broughton W J, Hernández G, Blair M, Beebe S, Gepts P, Vanderleyden J. Beans (Phaseolus spp.) -model food legumes. Plant and Soil, 2003, 252(1): 55-128.

[2]冯佰利, 鱼欢, 高小丽, 高金峰, 柴岩. 中国芸豆品牌发展战略研究. 中国农学通报, 2005, 21(6): 454-457.

Feng B L, Yu H, Gao X L, Gao J F, Chai Y. Study on brand agriculture of kidney bean in China. Chinese Agricultural Science Bulletin, 2005, 21(6): 454-457. (in Chinese)

[3]Zapata M, Beaver J S, Porch T G. Dominant gene for common bean resistance to common bacterial blight caused by Xanthomonas axonopodis pv. phaseoli. Euphytica, 2011, 179(3): 373-382.

[4]Biljana T, Svetlana M, Emil R, Ivana P, Miloš S. Susceptibility of bean genotypes to Xanthomonas campestris pv. phaseoli in greenhouse conditions. Pesticides and Phytomedicine, 2008, 23: 167-173.

[5]Mkandawire A B C, Mabagala R B, Guzmán P, Gepts P, Gilbertson R L. Genetic diversity and pathogenic variation of common blight bacteria (Xanthomonas campestris pv. phaseoli and X. campestris pv. phaseoli var. fuscans) suggests pathogen coevolution with the common bean. Phytopathology, 2004, 94(6): 593-603.

[6]Vauterin L, Hoste B, Kersters K, Swings J. Reclassification of Xanthomonas. International Journal of Systematic and Evolutionary Microbiology, 1995, 45(3): 472-489.

[7]Schaad N W, Postnikova E, Lacy G H, Sechler A, Agarkova I, Stromberg P E, Stromberg V K, Vidaver A K. Reclassification of Xanthomonas campestris pv. citri (ex Hasse 1915) Dye 1978 forms A, B/C/D, and E as X. smithii subsp. citri (ex Hasse) sp. nov. nom. rev. comb. nov., X. fuscans subsp. aurantifolii (ex Gabriel 1989) sp. nov. nom. rev. comb. nov., and X. alfalfae subsp. citrumelo (ex Riker and Jones) Gabriel et al., 1989 sp. nov. nom. rev. comb. nov.; X. campestris pv. malvacearum (ex Smith 1901) Dye 1978 as X. smithii subsp. smithii nov. comb. nov. nom. nov.; X. campestris pv. alfalfae (ex Riker and Jones, 1935) Dye 1978 as X. alfalfae subsp. alfalfae (ex Riker et al., 1935) sp. nov. nom. rev.; and “var. fuscans” of X. campestris pv. phaseoli (ex Smith, 1987) Dye 1978 as X. fuscans subsp. fuscans sp. nov. Systematic and Applied Microbiology, 2005, 28: 494-518.

[8]Alavi S M, Sanjari S, Durand F, Brin C, Manceau C, Poussier S. Assessment of the genetic diversity of Xanthomonas axonopodis pv. phaseoli and Xanthomonas fuscans subsp. fuscans as a basis to identify putative pathogenicity genes and a type III secretion system of the SPI-1 family by multiple suppression subtractive hybridizations. Applied and Environmental Microbiology, 2008, 74(10): 3295-3301.

[9]Mutlu N, Vidaver A K, Coyne D P, Steadman J R, Lambrecht P A, Reiser J. Differential pathogenicity of Xanthomonas campestris pv. phaseoli and X. fuscans subsp. fuscans strains on bean genotypes with common blight resistance. Plant Disease, 2008, 92(4): 546-554.

[10]Opio A F, Allen D J, Teri J M. Pathogenic variation in Xanthomonas campestris pv. phaseoli, the causal agent of common bacterial blight in Phaseolus beans. Plant Pathology, 1996, 45(6): 1126-1133.

[11]Goodwin P H, Sopher C R. Brown pigmentation of Xanthomonas campestris pv. phaseoli associated with homogentisic acid. Canadian Journal of Microbiology, 1994, 40(1): 28-34.

[12]Chan J W Y F, Goodwin P H. Differentiation of Xanthomonas campestris pv. phaseoli from Xanthomonas campestris pv. phaseoli var. fuscans by PFGE and RFLP. European Journal of Plant Pathology, 1999, 105(9): 867-878.

[13]López R, Asensio C, Gilbertson R L. Phenotypic and genetic diversity in strains of common blight bacteria (Xanthomonas campestris pv. phaseoli and X. campestris pv. phaseoli var. fuscans) in a secondary center of diversity of the common bean host suggests multiple introduction events. Phytopathology, 2006, 96: 1204-1213.

[14]Mahuku G S, Jara C, Henriquez M A, Castellanos G, Cuasquer J. Genotypic characterization of the common bean bacterial blight pathogens Xanthomonas axonopodis pv. phaseoli and Xanthomonas axonopodis pv. phaseoli var. fuscans by rep-PCR and PCR-RFLP of the ribosomal genes. Journal of Phytopathology, 2006, 154(1): 35-44.

[15]Duncan R W, Singh S P, Gilbertson R L. Interaction of common bacterial blight bacteria with disease resistance quantitative trait loci in common bean. Phytopathology, 2011, 101(4): 425-435.

[16]郑继法, 沈东玲, 朱汉城. 菜豆细菌性疫病病原细菌的鉴定. 山东农业科学, 1989(2): 12-14.

Zheng J F, Shen D L, Zhu H C. On the pathogen identification of bean bacterial blight. Shandong Agricultural Sciences, 1989(2): 12-14. (in Chinese)

[17]金潜, 孙庆辉, 王钦英. 菜豆细菌性疫病病原细菌的鉴定及药剂敏感性测定. 新疆农垦科技, 1991(4): 21-24.

Jin Q, Sun Q H, Wang Q Y. Identification of the pathogen causing bean bacterial blight and its sensitivity determination to fungicides. Xinjiang Farmland Reclamation Science and Technology, 1991(4): 21-24. (in Chinese)

[18]Sheppard J W, Kurowski C, Remeeus P M. International Rules for Seed Testing, 7-021: Detection of Xanthomonas axonopodis pv. phaseoli and Xanthomonas axonopodis pv. phaseoli var. fuscans on Phaseolus vulgaris. Bassersdorf, Switzerland: International Seed Testing Association (ISTA), 2007.

[19]Goszczynska T, Serfontein J J, Serfontein S. Introduction to Practical Phytobacteriology. Switzerland: The Swiss Agency for Development and Cooperation (SDC), 2000.

[20]Fanelli V, Cariddi C, Finetti-Sialer M. Selective detection of Pseudomonas syringae pv. tomato using dot blot hybridization and real-time PCR. Plant Pathology, 2007, 56(4): 683-691.

[21]朱伟杰, 王楠, 郁雪平, 王伟. 生防菌Pseudomonas fluorescens 2P24对甜瓜根围土壤微生物的影响. 中国农业科学, 2010, 43(7): 1389-1396.

Zhu W J, Wang N, Yu X P, Wang W. Effects of the biocontrol agent Pseudomonas fluorescens 2P24 on microbial community diversity in the melon rhizosphere. Scientia Agricultura Sinica, 2010, 43(7): 1389-1396. (in Chinese)

[22]Li X, de Boer S H. Selection of polymerase chain reaction primers from an RNA intergenic spacer region for specific detection of Clavibacter michiganensis subsp. spedonicus. Phytopathology, 1995, 85(8): 837-842.

[23]Khoodoo M H R, Sahin F, Donmez M F, Fakim Y J. Molecular characterisation of Xanthomonas strains isolated from aroids in Mauritius. Systematic and Applied Microbiology, 2005, 28(4): 366-380.

[24]Toth I K, Hyman L J, Taylor R, Birch P R J. PCR-based detection of Xanthomonas campestris pv. phaseoli var. fuscans in plant material and its differentiation from X. c. pv. phaseoli. Journal of Applied Microbiology, 1998, 85: 327-336.

[25]Audy P, Braat C E, Saindon G, Huang H C, Laroche A. A rapid and sensitive PCR-based assay for concurrent detection of bacteria causing common and halo blights in bean seed. Phytopathology, 1996, 86(4): 361-366.

[26]Zanatta Z G C N, Moura A B, Maia L C, Santos A S. Bioassay for selection of biocontroller bacteria against bean common blight (Xanthomonas axonopodis pv. phaseoli). Brazilian Journal of Microbiology, 2007, 38(3): 511-515.

[27]邱文, 徐福寿, 谢关林, 徐丽慧, 怀雁, 李斌, 余山红, 钱军. 引起梨花枯和芽枯的Pseudomonas syringae pv. syringae 病原细菌鉴定. 中国农业科学, 2008, 41(9): 2657-2662.

Qiu W, Xu F S, Xie G L, Xu L L, Huai Y, Li B, Yu S H, Qian J. Identification of Pseudomonas syringae pv. syringae causing pear blossom and bud blast in China. Scientia Agricultura Sinica, 2008, 41(9): 2657-2662. (in Chinese)

[28]Weller D M, Saettler A W. Evaluation of seedborne Xanthomonas phaseoli and Xanthomonas phaseoli var. fuscans as primary inocula in bean blights. Phytopathology, 1980, 70: 148-152.

[29]Mabagala R B. The effect of populations of Xanthomonas campestris pv. phaseoli in bean reproductive tissues on seed infection of resistant and susceptible bean genotypes. European Journal of Plant Pathology, 1997, 103(2): 175-181.

[30]Darrasse A, Bureau C, Samson R, Morris C E, Jacques M A. Contamination of bean seeds by Xanthomonas axonopodis pv. phaseoli associated with low bacterial densities in the phyllosphere under field and greenhouse conditions. European Journal of Plant Pathology, 2007, 119(2): 203-215.

[31]方仲达, 任欣正. 我国植物细菌病害名录补志. 南京农学院学报, 1981(3): 29-32.

Fang Z D, Ren X Z. Additional notes on the list of bacterial plant pathogens. Journal of Nanjing Agricultural College, 1981(3): 29-32. (in Chinese)

[32]方仲达, 任欣正. 中国植物病原细菌名录. 南京农业大学学报, 1992, 15(4): 1-6.

Fang Z D, Ren X Z. Notes on list of bacterial plant pathogens in China. Journal of Nanjing Agricultural University, 1992, 15(4): 1-6. (in Chinese)

[33]王晓鸣. 菜豆病害和菜豆资源的抗病性概述//刘仪. 植物病害研究与防治. 北京: 中国农业科技出版社, 1998: 401-403.

Wang X M. An overview of common bean diseases and disease resistance resources//Liu Y. Studies and Control of Plant Diseases. Beijing: China Agricultural Science and Technology Press, 1998: 401-403. (in Chinese)

[34]Kwak M, Gepts P. Structure of genetic diversity in the two major gene pools of common bean (Phaseolus vulgaris L., Fabaceae). Theoretical and Applied Genetics, 2009, 118(5): 979-992.

[35]Zhang X, Blair M W, Wang S. Genetic diversity of Chinese common bean (Phaseolus vulgaris L.) landraces assessed with simple sequence repeat markers. Theoretical and Applied Genetics, 2008, 117(4): 629-640.

[36]张晓艳, 王坤, Blair M W, 王述民. 中国普通菜豆形态性状分析及分类. 植物遗传资源学报, 2007, 8(4): 406-410.

Zhang X Y, Wang K, Blair M W, Wang S M. Phenotypic diversity analysis and classification of common bean (Phaseolus vulgaris L.) landraces in China. Journal of Plant Genetic Resources, 2007, 8(4): 406-410. (in Chinese)