大丽轮枝菌侵染拟南芥的过程及诱导的防御反应研究
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摘要
为进一步了解大丽轮枝菌的致病机制,本研究以绿色荧光蛋白标记的大丽轮枝菌菌系为材料,研究其对拟南芥的侵染过程和诱导拟南芥产生的防御反应。结果表明,大丽轮枝菌能侵染拟南芥,接菌6h后,分生孢子吸附在根表面,2~3 d到达皮层,9 d到达维管束,随后迅速扩展,接菌后11 d完成系统侵染,产生黄萎病症状。大丽轮枝菌在侵染过程中,能诱导拟南芥胼胝质、活性氧的产生和防御酶活性升高,植物产生的防御反应特征明显。本研究结果为大丽轮枝菌致病机理研究提供了理论依据。
In order to further understand the pathogenic mechanism of Verticillium dahliae,this study used V. dahliae strain with green fluorescent protein as materials,the infection process and the defense response induced in Arabidopsis thaliana was studied. The results showed that V. dahliae could infect A. thaliana,colonized in root surface at 6 h after inoculation,reached cortex at 2 ~ 3 d,arrived in vascular tissues at 9 d,then expanded rapidly,finished colonization at11 d after inoculation,caused symptoms of Verticillium wilt. During the infection process,V. dahliae could induce production of callose and H_2O_2,and increase the activity of the defense-related enzymes in the A. thaliana. The characteristics of defense response in plants are obvious. The results of this study provide a theoretical basis for the research on the pathogenic mechanism of V. dahliae.
In order to further understand the pathogenic mechanism of Verticillium dahliae,this study used V. dahliae strain with green fluorescent protein as materials,the infection process and the defense response induced in Arabidopsis thaliana was studied. The results showed that V. dahliae could infect A. thaliana,colonized in root surface at 6 h after inoculation,reached cortex at 2 ~ 3 d,arrived in vascular tissues at 9 d,then expanded rapidly,finished colonization at11 d after inoculation,caused symptoms of Verticillium wilt. During the infection process,V. dahliae could induce production of callose and H_2O_2,and increase the activity of the defense-related enzymes in the A. thaliana. The characteristics of defense response in plants are obvious. The results of this study provide a theoretical basis for the research on the pathogenic mechanism of V. dahliae.
引文
[1]Fradin E F,Thomma B P.Physiology and molecular aspects of verticillium wilt diseases caused by V.dahliae and V.albo-atrum[J].Molecular Plant Pathology,2006,7(2):71-86
[2]Veronese P,Narasimhan M L,Stevenson R A,Zhu J K,Weller S C,Subbarao K V,Bressan R A.Identification of a locus controlling Verticillium disease symptom response in Arabidopsis thaliana[J].Plant Journal,2003,35(5):574-587
[3]Newman M A,Sundelin T,Nielsen J T,Erbs G.MAMP(microbeassociated molecular pattern)triggered immunity in plants[J].Frontiers in Plant Science,2013,4(2):139
[4]Shi X,Han X,Lu T G.Callose synthesis during reproductive development in monocotyledonous and dicotyledonous plants[J].Plant Signaling&Behavior,2016,11(2):e1062196
[5]Alvarez M E,Pennell R I,Meijer P J,Ishikawa A,Dixon R A,Lamb C.Reactive oxygen intermeoiates mediate a systemic signal network in the establishment of plant immunity[J].Cell,1998,92(6):773-784
[6]薛鑫,张芊,吴金霞.植物体内活性氧的研究及其在植物抗逆方面的应用[J].生物技术通报,2013(10):6-11
[7]张琳,崔红米,王建军,侯喜林,李英.镉胁迫对不结球白菜Vc合成L-半乳糖途径基因表达及抗氧化系统的影响[J].植物生理学报,2015,51(7):1099-1108
[8]Mayer A M.Polyphenol oxidases in plants and fungi:goingplaces?A Review[J].Phytochemistry,2007,38(5):2318-2331
[9]Kachroo A,He Z H,Rajesh P,Zhu Q,Zhong J P,Li D B,Ronald P,Lamb C,Chattoo B B.Induction of H2O2in transgenic rice leads to cell death and enhanced resistance to both bacterial and fungal pathogens[J].Transgenic Research,2003,12(5):577-586
[10]胡慧兰.转GFP基因大丽轮枝菌的构建及侵染棉花过程的研究[D].南京:南京农业大学,2012
[11]王晓楠.绿色荧光蛋白标记的大丽轮枝菌对棉花叶片和单子叶植物的侵染研究[D].南京:南京农业大学,2014
[12]赵凤轩.绿色荧光蛋白标记的大丽轮枝菌的获得及其在棉花中侵染过程研究[D].北京:中国农业科学院,2010
[13]陆燕,李澄,陈志德,杨宜红,唐灿明.解淀粉芽孢杆菌41B-1对花生白绢病的生防效果[J].中国作物油料学报,2016,38(4):487-494
[14]杨宜红,陆燕,吴颖静,冯自力,唐灿明.棉花品种黄萎病抗性与免疫反应的关系[J].核农学报,2017,31(3):588-596
[15]Krikun J,Bernier C C.Morphology of microsclerotia of Verticillium dahlia in roots of gramineousplants[J].European Journal of Plant Pathology,1990,12(4):439-441
[16]Nagtzaam M P M,Termorshuizen A J,Bollen G J.The relationship between soil inoculum density and plant infection as a basis for a quantitative bioassay of Verticillium dahlia[J].European Journal of Plant Pathology,1997,103(1):597-605
[17]Eynck C,Koopmann B,Grunewaldtstoecker G,Karlovsky P,Tiedemann A V.Differential interactions of Verticillium longisporum and V.dahliae with Brassica napus detected with molecular and histological techniques[J].European Journal of Plant Pathology,2007,118(3):259-274
[18]Zhou L,Hu Q,Johansson A,Dixelius C.Venicillium longisporum and V.dahliae infection and disease in Brassica napus[J].Plant Physiology,2006,55(1):137-144
[19]Garber R H,Houston B R.Penetration and development of Verticillium albo-atrum in the cotton plant[J].Phytopathology,1966,56(10):1121-1126
[20]Bowers J H,Nameth S T,Riedel R M,Rowe R C.Infection and colonization of potato roots by Verticilliumdahliae as affected by Pratylenchus penetrans and P.crenatus[J].Phytopathology,1996,86(6):614-621
[21]Vallad G E,Subbarao K V.Colonization of resistant and susceptible lettuce cultivars by a green fluorescent protein-tagged isolate of Verticilliumdahlia[J].Phytopathology,2008,98(8):871-885
[22]Eckert M,Maguire K,Urban M,Foster S,Fitt B,Lucas J,Hammond-Kosack K.Agrobacterium tumefaciensmediated transformation of Leptosphaeria spp.and Oculimacula spp.with the reef coral gene DsRed and the jellyfish gene gfp[J].FEMS Microbiology Letters,2005,253(1):67-74
[23]张超.加工番茄抗黄萎病基因qtl定位及品种(系)遗传多样性分析[D].石河子:石河子大学,2011
[24]姚技强,林柏青,肖蕴华.茄子黄萎病抗病材料的组织病理学研究[J].植物病理学报,1996,26(2):159-163
[25]王建美,田呈明.黄栌枯萎病病原菌及组织病理学研究[C]//中国植物病理学会学术年会.北京:西北农林科技大学出版社,2007:36-37
[26]王妍.大丽轮枝菌对黄栌植株的侵染特点及其定量检测研究[D].北京:北京林业大学,2012
[27]Zavaliev R,Ueki S,Epel B L,Citovsky V.Biology of callose(β-1,3-glucan)turnover at plasmodesmata[J].Protoplasma,2011,248(1):117-130
[28]Xie B,Wang X,Zhu M,Zhang Z,Hong Z.Cal S7 encodes a callose synthase responsible for callose deposition in the phloem[J].Plant Journal,2011,65(1):1-14
[29]Wan L,Zha W,Cheng X,Liu C,Lv L,Liu C,Wang Z,Du B,Chen R,Zhu L,He G.A riceβ-1,3-glucanase gene Osg1 is required for callose degradation in pollen development[J].Planta,2011,233(2):309-323
[30]Chen X Y,Kim J Y.Callose synthesis in higher plants[J].Plant Signaling&Behavior,2009,4(6):489-492
[31]杨和民,郑重,Leach J E.水稻受稻瘟菌侵染后发病初期的细胞学反应[J].实验生物学报,2004,37(5):344-350
[32]Orozeo-Cárdenas M,Ryan C A.Hydrogen peroxide is generated systemically in plant leaves by wounding and systemin via the octadecanoid pathway[J].Proceedings of National Academy of Sciences of the United States of America,1999,96(11):6553-6557
[33]Huckelhoven R,Fodor J,Preis C,Kogel K H.Hypersensitive cell death and papilla formation in barley attacked by the powdery mildew fungus are associated with hydrogen peroxide but not with salicylic acid accumulation[J].Plant Physiology,1999,119(4):1251-1260
[34]Dang J L,Dietrich R A,Richberg M H.Death don’t have no mercy:cell death program in plant-microbe interactions[J].Plant Cell,1996,8(10):1793-1807
[35]田丽波,杨衍,商桑,司龙亭.不同苦瓜品系的抗白粉病能力及其与防御酶活性的相关性[J].沈阳农业大学学报,2015,46(3):284-291
[36]李淑菊,马德华,庞金安,霍振荣.黄瓜感染黑星病菌后的生理变化及抗病性的产生[J].华北农学报,2003,18(3):74-77
[37]林陈强,李占飞,张慧,林戎斌,林新坚,陈济琛.枯草芽孢杆菌CS16诱导香蕉抗病性相关防御酶系的研究[J].福建农业学报,2013,28(6):570-574
[2]Veronese P,Narasimhan M L,Stevenson R A,Zhu J K,Weller S C,Subbarao K V,Bressan R A.Identification of a locus controlling Verticillium disease symptom response in Arabidopsis thaliana[J].Plant Journal,2003,35(5):574-587
[3]Newman M A,Sundelin T,Nielsen J T,Erbs G.MAMP(microbeassociated molecular pattern)triggered immunity in plants[J].Frontiers in Plant Science,2013,4(2):139
[4]Shi X,Han X,Lu T G.Callose synthesis during reproductive development in monocotyledonous and dicotyledonous plants[J].Plant Signaling&Behavior,2016,11(2):e1062196
[5]Alvarez M E,Pennell R I,Meijer P J,Ishikawa A,Dixon R A,Lamb C.Reactive oxygen intermeoiates mediate a systemic signal network in the establishment of plant immunity[J].Cell,1998,92(6):773-784
[6]薛鑫,张芊,吴金霞.植物体内活性氧的研究及其在植物抗逆方面的应用[J].生物技术通报,2013(10):6-11
[7]张琳,崔红米,王建军,侯喜林,李英.镉胁迫对不结球白菜Vc合成L-半乳糖途径基因表达及抗氧化系统的影响[J].植物生理学报,2015,51(7):1099-1108
[8]Mayer A M.Polyphenol oxidases in plants and fungi:goingplaces?A Review[J].Phytochemistry,2007,38(5):2318-2331
[9]Kachroo A,He Z H,Rajesh P,Zhu Q,Zhong J P,Li D B,Ronald P,Lamb C,Chattoo B B.Induction of H2O2in transgenic rice leads to cell death and enhanced resistance to both bacterial and fungal pathogens[J].Transgenic Research,2003,12(5):577-586
[10]胡慧兰.转GFP基因大丽轮枝菌的构建及侵染棉花过程的研究[D].南京:南京农业大学,2012
[11]王晓楠.绿色荧光蛋白标记的大丽轮枝菌对棉花叶片和单子叶植物的侵染研究[D].南京:南京农业大学,2014
[12]赵凤轩.绿色荧光蛋白标记的大丽轮枝菌的获得及其在棉花中侵染过程研究[D].北京:中国农业科学院,2010
[13]陆燕,李澄,陈志德,杨宜红,唐灿明.解淀粉芽孢杆菌41B-1对花生白绢病的生防效果[J].中国作物油料学报,2016,38(4):487-494
[14]杨宜红,陆燕,吴颖静,冯自力,唐灿明.棉花品种黄萎病抗性与免疫反应的关系[J].核农学报,2017,31(3):588-596
[15]Krikun J,Bernier C C.Morphology of microsclerotia of Verticillium dahlia in roots of gramineousplants[J].European Journal of Plant Pathology,1990,12(4):439-441
[16]Nagtzaam M P M,Termorshuizen A J,Bollen G J.The relationship between soil inoculum density and plant infection as a basis for a quantitative bioassay of Verticillium dahlia[J].European Journal of Plant Pathology,1997,103(1):597-605
[17]Eynck C,Koopmann B,Grunewaldtstoecker G,Karlovsky P,Tiedemann A V.Differential interactions of Verticillium longisporum and V.dahliae with Brassica napus detected with molecular and histological techniques[J].European Journal of Plant Pathology,2007,118(3):259-274
[18]Zhou L,Hu Q,Johansson A,Dixelius C.Venicillium longisporum and V.dahliae infection and disease in Brassica napus[J].Plant Physiology,2006,55(1):137-144
[19]Garber R H,Houston B R.Penetration and development of Verticillium albo-atrum in the cotton plant[J].Phytopathology,1966,56(10):1121-1126
[20]Bowers J H,Nameth S T,Riedel R M,Rowe R C.Infection and colonization of potato roots by Verticilliumdahliae as affected by Pratylenchus penetrans and P.crenatus[J].Phytopathology,1996,86(6):614-621
[21]Vallad G E,Subbarao K V.Colonization of resistant and susceptible lettuce cultivars by a green fluorescent protein-tagged isolate of Verticilliumdahlia[J].Phytopathology,2008,98(8):871-885
[22]Eckert M,Maguire K,Urban M,Foster S,Fitt B,Lucas J,Hammond-Kosack K.Agrobacterium tumefaciensmediated transformation of Leptosphaeria spp.and Oculimacula spp.with the reef coral gene DsRed and the jellyfish gene gfp[J].FEMS Microbiology Letters,2005,253(1):67-74
[23]张超.加工番茄抗黄萎病基因qtl定位及品种(系)遗传多样性分析[D].石河子:石河子大学,2011
[24]姚技强,林柏青,肖蕴华.茄子黄萎病抗病材料的组织病理学研究[J].植物病理学报,1996,26(2):159-163
[25]王建美,田呈明.黄栌枯萎病病原菌及组织病理学研究[C]//中国植物病理学会学术年会.北京:西北农林科技大学出版社,2007:36-37
[26]王妍.大丽轮枝菌对黄栌植株的侵染特点及其定量检测研究[D].北京:北京林业大学,2012
[27]Zavaliev R,Ueki S,Epel B L,Citovsky V.Biology of callose(β-1,3-glucan)turnover at plasmodesmata[J].Protoplasma,2011,248(1):117-130
[28]Xie B,Wang X,Zhu M,Zhang Z,Hong Z.Cal S7 encodes a callose synthase responsible for callose deposition in the phloem[J].Plant Journal,2011,65(1):1-14
[29]Wan L,Zha W,Cheng X,Liu C,Lv L,Liu C,Wang Z,Du B,Chen R,Zhu L,He G.A riceβ-1,3-glucanase gene Osg1 is required for callose degradation in pollen development[J].Planta,2011,233(2):309-323
[30]Chen X Y,Kim J Y.Callose synthesis in higher plants[J].Plant Signaling&Behavior,2009,4(6):489-492
[31]杨和民,郑重,Leach J E.水稻受稻瘟菌侵染后发病初期的细胞学反应[J].实验生物学报,2004,37(5):344-350
[32]Orozeo-Cárdenas M,Ryan C A.Hydrogen peroxide is generated systemically in plant leaves by wounding and systemin via the octadecanoid pathway[J].Proceedings of National Academy of Sciences of the United States of America,1999,96(11):6553-6557
[33]Huckelhoven R,Fodor J,Preis C,Kogel K H.Hypersensitive cell death and papilla formation in barley attacked by the powdery mildew fungus are associated with hydrogen peroxide but not with salicylic acid accumulation[J].Plant Physiology,1999,119(4):1251-1260
[34]Dang J L,Dietrich R A,Richberg M H.Death don’t have no mercy:cell death program in plant-microbe interactions[J].Plant Cell,1996,8(10):1793-1807
[35]田丽波,杨衍,商桑,司龙亭.不同苦瓜品系的抗白粉病能力及其与防御酶活性的相关性[J].沈阳农业大学学报,2015,46(3):284-291
[36]李淑菊,马德华,庞金安,霍振荣.黄瓜感染黑星病菌后的生理变化及抗病性的产生[J].华北农学报,2003,18(3):74-77
[37]林陈强,李占飞,张慧,林戎斌,林新坚,陈济琛.枯草芽孢杆菌CS16诱导香蕉抗病性相关防御酶系的研究[J].福建农业学报,2013,28(6):570-574