张佳环;李娟;高洁;

• 1:吉林农业大学农学院

摘要(Abstract)：

[目的]明确hrpZPsg12基因对大豆细菌性斑点病菌致病性的影响。[方法]采用PCR方法从大豆细菌性斑点病菌中克隆hrpZPsg12基因,利用具有自杀特性的敲除质粒pKNOCK-Cm和具有功能互补作用的粘粒pUFR034,构建了hrpZPsg12基因的突变载体pKNOCK477-7和互补载体pUFR1026-68,并筛选出hrpZPsg12基因的突变体477-1及其功能互补子1026-5。进一步将野生型Psg12、突变体477-1和互补子反应斑。但是,反应斑的大小有差异,Psg12菌株接种的病斑较大,477-1的较小,互补子1026-5的接近野生型菌株Psg12。病斑中细菌繁殖量分析表明,野生型菌株Psg12繁殖量最高,突变体477-1的最低,互补子1026-5的接近野生型菌株Psg12的繁殖量。[结论]hrpZ-Psg12基因能够增强大豆细菌性斑点病菌对大豆的致病性,并且能够在烟草上产生过敏性反应。

关键词(KeyWords)： 大豆细菌性斑点病菌;hrpZPsg12基因;突变体;致病性

Abstract:

Keywords:

基金项目(Foundation): 吉林农业大学科研启动基金资助项目“大豆细菌性斑点病菌HrpZPsgl2蛋白功能的研究”(384);; 农业部“十一五”转基因生物新品种培育重大专项“抗病虫转基因大豆新品种培育”(2008ZX08004-004)

作者(Author): 张佳环;李娟;高洁;

Email:

DOI: 10.13989/j.cnki.0517-6611.2011.15.174

参考文献(References)：

• [1]刘亚光,马超,冯蕾.几种化学诱抗剂对大豆细菌性斑点病诱导抗性的研究[J].中国油料作物学报,2008,30(1):116-118.
• [2]姚红梅.上海地区大豆细菌性疫病病原鉴定及病害的发生危害和防治对策[D].杭州:浙江大学,2005.
• [3]MA W.Role of pseudomonas syringae type III secreted effectors in bacterialblight of soybeans[P/OL].[2009-01-01].http://www.reeis.usda.gov/web/crisprojectpages/212169.html.
• [4]KONG H,BLACKWOOD C,BUYER J S,et al.The genetic characterizationof Pseudomonas syringae pv.tagetis based on the 16S–23S rDNA inter-genic spacer regions[J].Biological Control,2005,32(3):356-362.
• [5]韩雪.大豆细菌斑点病的发生原因及防治[J].现代化农业,2006(1):14.
• [6]张佳环.大豆细菌性斑点病菌生理小种研究[D].长春:吉林农业大学,2003.
• [7]张佳环,高洁,袁美丽,等.大豆细菌性斑点病菌生理小种分布的研究[J].吉林农业大学学报,2003,25(1):24-26.
• [8]PARTRIDGE J E.Bacterial Blight of Soybeans[P/OL].http://nu-dis-tance.unl.edu/homer/disease/agron/soybean/sobacblt.html.
• [9]张淑珍,徐鹏飞,雷虹,等.黑龙江省大豆细菌性斑点病菌生理小种鉴定及抗性资源筛选[J].大豆科学,2006,25(2):141-144.
• [10]张佳环,高洁,许庆国,等.大豆品种对大豆细菌性斑点病的抗性鉴定[J].大豆科学,2000,19(2):180-183.
• [11]STEFANI E,CAFFIER D,FIORE N.The economic impact of the bacterialblight of soybean under European agroclimatic conditions[J].Journal ofPlant Pathology,1998,80(3):211-218.
• [12]FARHATULLAH,ROBIN W G,RAZIUDDIN,et al.Mapping gene for bac-terial blight(Rpg4 locus)in soybean[J].Pak J Bot,2010,42(3):2145-2149.
• [13]BRIAN H K,ADELA R R,JOANNE E M,et al.Identification of Harpinsin Pseudomonas syringae pv.tomato DC3000,which are functionally simi-lar to HrpK1 in promoting translocation of type III secretion system effec-tors[J].Journal of Bacteriology,2007,189(22):8059-8072.
• [14]CUNNAC S,LINDEBERG M,COLLMER A.Pseudomonas syringae typeIII secretion system effectors:repertoires in search of functions[J].Cur-rent Opinion in Microbiology,2009,12:53-60.
• [15]王玉全,李祥羽,徐鹏飞,等.黑龙江省大豆细菌性斑点病病原菌的分离鉴定及病害分布研究[J].黑龙江农业科学,2006(6):34-35.
• [16]姜兆远,邹晓威,高洁,等.大豆细菌性斑点病菌harpin编码基因的克隆与表达[J].微生物学报,2009,49(10):1403-1407.
• [17]张佳环.水稻黄单胞菌致病相关基因hpa2的研究[D].南京:南京农业大学,2008.
• [18]ZHANG J H,WANG X Y,ZHANG Y,et al.A conserved Hpa2 protein haslytic activity against the bacterial cell wall in phytopathogenic Xan-thomonas oryzae[J].Applied Microbiology Biotechnology,2008,79:605-616.
• [19]SASCHA D B,JANINE H,ANNETTE W,et al.In vitro Antibiosis byPseudomonas syringae Pss22d,Acting Against the Bacterial Blight Patho-gent of Soybean Plants,Does Not Influence In planta Biocontrol[J].Jour-nal of Phytopathology,2010,158(4):288-295.
• [20]王钧.诱导植物产生过敏反应的病原蛋白harpin和elicitin[J].植物生理学通讯,1995,31(6):451-454.
• [21]孙建和,严亚贤,陆承平.大肠杆菌vpr基因突变株的蛋白表达及功能研究[J].中国农业科学,2006,39(1):176-180.
• [22]CHO H J,PARK Y J,NOH T H,et al.Molecular analysis of the hrp genecluster in Xanthomonas oryzae pathovar oryzae KACC10859[J].MicrobialPathogenesis,2008,44(6):473-483.
• [23]ZHU W,MAGBANUA M M,WHITE F F.Identification of two novel hrp-associated genes in the hrp gene cluster of Xanthomonas oryzae pv.oryzae[J].Journal of Bacteriology,2000,182(7):1844-1853.
• [24]ALFANO J R,BAUER D W,MILOS T M,et al.Analysis of the role of thePseudomonas syringae pv.syringae HrpZ harpin in elicitation of the hy-persensitive response in tobacco using functionally non-polar hrpZ deletionmutations,truncated HrpZ fragments,and hrmA mutations[J].MolecularMicrobiology,1996,19(4):715-728.
• [25]ARLAT M,VAN G F,HUET J C,et al.PopA1,a protein which induces ahypersensitive-like response on specific Petunia genotypes,is secreted viathe Hrp pathway of Pseudomonas solanacearum[J].The EMBO Journal,1994,13(3):543-553.
• [26]CHARKOWSKI A O,ALFANO J R,PRESTON G,et al.The Pseudomonassyringae pv.tomato HrpW protein has domains similar to harpins and pec-tate lyases and can elicit the plant hypersensitive response and bind topectate[J].Journal of Bacteriology,1998,180(19):5211-5217.
• [27]WEI Z M,LABY R J,ZUMOFF C H,et al.Harpin,elicitor of the hyper-sensitive response produced by the plant pathogen Erwinia amylovora[J].Science,1992,257:85-87.
• [28]孟德,李志峰,李真和.内蒙古、黑龙江大豆生产技术的调查与分析[J].内蒙古农业科技,2008(3):82-84.
• [29]王帅,沈火林,吴晓霞,等.芹菜黄色突变体的遗传及生长表现研究[J].华北农学报,2010(S2):119-122.
• [30]盛积贵.黄淮海平原夏大豆超高产栽培技术[J].内蒙古农业科技,2008(3):114-115.
• [31]王益军,苗楠,施亚婷,等.一份玉米显性矮杆突变体的遗传分析[J].华北农学报,2010(5):90-93.
• [32]李雪巍,王小平,朱铁栓,等.赤峰市大豆与荞麦种植气候区划[J].内蒙古农业科技,2008(4):79,81.
• [33]齐春艳,梁正伟,杨福,等.水稻耐碱突变体ACR78在苏打盐碱胁迫下的生理响应[J].华北农学报,2009(1):20-25.