茉莉酸不同方式处理水稻对稻瘟病的防控效果及对水稻防御体系的影响
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摘要
【目的】分析外源茉莉酸喷雾处理水稻后对水稻防御系统的影响,为深入开展茉莉酸在稻瘟病菌与水稻互作中的作用机制研究提供参考依据。【方法】将茉莉酸以两种方式处理水稻:将100和400μmol/L茉莉酸分别预先喷雾于水稻叶片上,6 h后再接种稻瘟病菌株孢子;用100和400μmol/L茉莉酸制备稻瘟病菌株孢子悬浮液直接喷雾接种水稻,调查水稻稻瘟病发病症状及实时荧光定量PCR(qRT-PCR)检测水稻防御相关基因的表达。【结果】分别用100和400μmol/L茉莉酸预先喷雾水稻6 h后再接种稻瘟病菌株孢子的诱抗效果分别为16.02%和25.51%;而以100和400μmol/L茉莉酸制备稻瘟病菌株孢子悬浮液直接喷雾水稻的诱抗效果分别为21.82%和34.09%。qRT-PCR检测发现受侵染水稻防御相关基因均有不同程度的上调或下调表达。与接种前(0 h)相比,在茉莉酸预先喷雾水稻后再接种稻瘟病菌孢子的水稻和茉莉酸制备孢子悬浮液直接接种的水稻中病程相关基因PR1a在稻瘟病菌侵染水稻早期表达量有所上调、侵染后期表达量下调,病程相关基因PR10a的表达量上调;水杨酸途径相关基因EDS1和PAL在稻瘟病菌侵染水稻的整个进程中一直处于较低的表达水平;茉莉酸途径相关基因AOS2的表达量显著上调(P<0.05)。【结论】水稻病程相关基因及茉莉酸途径相关基因参与了水稻防御体系对外源茉莉酸的响应。
【Objective】In order to provide references for deeply studying functional mechanism of jasmonic acids during interaction of rice and Magnaporthe oryzae,the effects of exogenous jasmonic acid spray on defense system of infested rice were analyzed.【Method】Rice leaves were treated with two different treatment patterns of jasmonic acids. The first treatment:jasmonic acids with two concentrations of 100 and 400 μmol/L were sprayed on rice leaves respectively,and then inoculated with M. oryzae spores 6 h later. The second treatment was:rice leaves were directly inoculated with M. oryzae spores suspension prepared by 100 and 400 μmol/L jasmonic acids respectively. The symptoms of rice blast and expression of defense-related genes were analyzed using real-time fluorescence quantitative PCR.【Result】The induced resistance effects were 16.02% and 25.51% for the rice leaves that were pre-sprayed with 100 and 400 μmol/L jasmonic acids,and then inoculated with M. oryzae spores 6 h later. And induced resistance effects were 21.82% and 34.09% for rice leaves were inoculated with M. oryzae spores suspension prepared by 100 and 400 μmol/L jasmonic acids. The rice defense-related genes up-regulated or down-regulated to different degrees in infected rice leaves using real-time fluorescence quantitative PCR. Comparing with before inoculation(0 h),disease related gene PR1 a in the two treatments above appeared up-regulation in the early stage of M. oryzae infection and down-regulation in the late stage of infection,disease related gene PR10 a appeared up-regulation during the whole infection. Salicylic acid pathway related genes EDS1 and PAL showed low expression level and jasmonic acid pathway related gene AOS2 appeared significant up-regulation in infested leaves treated with two different patterns of jasmonic acids(P<0.05).【Conclusion】Rice disease related genes and jasmonic acid related genes involve in response of rice defense system to jasmonic acids.
【Objective】In order to provide references for deeply studying functional mechanism of jasmonic acids during interaction of rice and Magnaporthe oryzae,the effects of exogenous jasmonic acid spray on defense system of infested rice were analyzed.【Method】Rice leaves were treated with two different treatment patterns of jasmonic acids. The first treatment:jasmonic acids with two concentrations of 100 and 400 μmol/L were sprayed on rice leaves respectively,and then inoculated with M. oryzae spores 6 h later. The second treatment was:rice leaves were directly inoculated with M. oryzae spores suspension prepared by 100 and 400 μmol/L jasmonic acids respectively. The symptoms of rice blast and expression of defense-related genes were analyzed using real-time fluorescence quantitative PCR.【Result】The induced resistance effects were 16.02% and 25.51% for the rice leaves that were pre-sprayed with 100 and 400 μmol/L jasmonic acids,and then inoculated with M. oryzae spores 6 h later. And induced resistance effects were 21.82% and 34.09% for rice leaves were inoculated with M. oryzae spores suspension prepared by 100 and 400 μmol/L jasmonic acids. The rice defense-related genes up-regulated or down-regulated to different degrees in infected rice leaves using real-time fluorescence quantitative PCR. Comparing with before inoculation(0 h),disease related gene PR1 a in the two treatments above appeared up-regulation in the early stage of M. oryzae infection and down-regulation in the late stage of infection,disease related gene PR10 a appeared up-regulation during the whole infection. Salicylic acid pathway related genes EDS1 and PAL showed low expression level and jasmonic acid pathway related gene AOS2 appeared significant up-regulation in infested leaves treated with two different patterns of jasmonic acids(P<0.05).【Conclusion】Rice disease related genes and jasmonic acid related genes involve in response of rice defense system to jasmonic acids.
引文
崔伟婵,李霜雯,严善春.2016.不同光照下外源茉莉酸类物质对兴安落叶松防御蛋白的影响[J].东北林业大学学报,44(9):78-81.[Cui W C,Li S W,Yan S C.2016.E-ffects of exogenous jasmonic acid and methyl jasmonate on the defensive proteins of larix gmelinii under different light conditions[J].Journal of Northeast Forestry University,44(9):78-81.]
李兆举,徐新娟,齐红志,王二歌,宋思艺,王晓桠,赵博文.2017.MeJA浸种对干旱胁迫下玉米种子萌发及幼苗生理特性的影响[J].河南农业科学,46(12):36-41.[Li ZJ,Xu X J,Qi H Z,Wang E G,Song S Y,Wang X Y,Zhao B W.2017.Effects of seed soaking with meja on seed germination and seeding physiological characteristics of maize under drought stress[J].Journal of Henan Agricultural Sciences,46(12):36-41.]
刘霞,唐设,窦志,李刚华,刘正辉,王绍华,丁承强,丁艳锋.2016.茉莉酸甲酯对武运粳24和宁粳3号灌浆早期高温胁迫生理特性的影响[J].中国水稻科学,30(3):291-303.[Liu X,Tang S,Dou Z,Li G H,Liu Z H,Wang SH,Ding C Q,Ding Y F.2016.Effects of MeJA on the physiological characteristics of Japonica rice Wuyunjing24 and Ning-jing 3 during early grain filling stage under heat stress[J].Chinese Journal of Rice Science,30(3):291-303.]
王云锋,李晓杰,刘立娜,刘林,王长秘,李晓疆,李成云,杨静.2018a.稻瘟病菌株形态发育对外源茉莉酸敏感程度分析[J].分子植物育种,16(12):4098-4105.[Wang YF,Li X J,Liu L N,Liu L,Wang C M,Li X J,Li C Y,Yang J.2018a.Sensibility analysis for morphological development of rice blast strain to exogenous jasmonic acid[J].Molecular Plant Breeding,16(12):4098-4105.]
王云锋,王长秘,李春琴,刘林,李晓杰,李晓疆,杨静.2018b.稻瘟病菌侵染时水稻防御体系对外源茉莉酸的响应分析[J].南方农业学报,49(7):1324-1331.[Wang Y F,Wang C M,Li C Q,Liu L,Li X J,Li X J,Yang J.2018b.Response of rice defense system to exogenous jamonisic acids during rice blast strain infecting rice[J].Journal of Southern Agriculture,49(7):1324-1331.]
吴国昭,谢丽君,宋圆圆,陈敏,曾任森.2009.外源信号物质诱导广东高州普通野生稻抗稻瘟病的生理生化机理[J].西北农业学报,18(3):254-258.[Wu G Z,Xie L J,Song Y Y,Chen M,Zeng R S.2009.The physiological and biochemical mechanisms of disease resistance of Oryza rufipogon Griff.Indigenous to Gaozhou,Guangdong against Pyricularia grisea induced by exogenously applied plant signal compounds[J].Acta Agriculturae Boreali-Occidentalis Sinica,18(3):254-258.]
肖宇龙,李湘民,余传元,雷建国,王智权,王晓玲,何虎,邱在辉,肖宇华.2018.江西省稻瘟病生理小种致病力测定及1组水稻优质新种质的稻瘟病抗性表现及应用评估[J].江西农业学报,30(4):9-13.[Xiao Y L,Li X M,Yu CY,Lei J G,Wang Z Q,Wang X L,He H,Qiu Z H,Xiao Y H.2018.Pathogenicity of physiological races of rice blast disease in Jiangxi Province,blast-resistant performance and application evaluation of a set of new highquality rice germplasms[J].Acta Agriculturae Jiangxi,30(4):9-13.]
许志刚.2002.普通植物病理学[M].第3版.北京:中国农业出版社:319-335.[Xu Z G.2002.General Plant Pathology[M].The 3rdEdition.Beijing:China Agriculture Press:319-335.]
张知晓,泽桑梓,户连荣,刘凌,季梅.2018.茉莉酸甲酯生物活性研究进展[J].河南农业科学,47(11):1-7.[Zhang ZX,Ze S Z,Hu L R,Liu L,Ji M.2018.Research advance in biological activities of methyl jasmonate[J].Journal of Henan Agricultural Sciences,47(11):1-7.]
邹志燕,王振中.2006.茉莉酸诱导水稻幼苗对稻瘟病抗性作用研究[J].植物病理学报,36(5):432-438.[Zou Z Y,Wang Z Z.2006.Effect of jasmonic acid on induced resistance of rice seedlings to Magnaporthe grisea[J].Acta Phytopathologica Sinica,36(5):432-438.]
Ahn I P,Kim S,Kang S,Suh S C,Lee Y H.2005.Rice defense mechanisms against Cochliobolus miyabeanus and Magnaporthe grisea are distinct[J].Phytopathology,95(11):1248-1255.
Alkooranee J T,Aledan T R,Ali A K,Lu G,Zhang X,Wu J,Fu C,Li M.2017.Detecting the hormonal pathways in oilseed rape behind induced systemic resistance by Trichoderma harzianum TH12 to Sclerotinia sclerotiorum[J].PLoS One,12(1):e0168850.
Balbi V,Devoto A.2008.Jasmonate signaling network in Arbidops thaliana:Crucial regulatory nodes and new physiological scenarios[J].New Phytologist,177(2):301-318.
Bostock R M.2005.Signal crosstalk and induced resistance:Straddling the line between cost and benefit[J].Annual Review of Phytopathology,43:545-580.
Creelman R A,John E M.1997.Oligosaccharins,brassinotides and jasmonates:Nontr-adition airegu lators of plant growth,development,and gene expression[J].Plant Cell,9:121l-1223.
Duan Z B,LüG Z,Shen C C,Li Q Y,Qin Z,Niu J S.2014.The role of jasmonic acid signalling in wheat(triticum aestivum,l.)powdery mildew resistance reaction[J].European Journal of Plant Pathology,140(1):169-183.
Gravot A,Deleu C,Wagner G,Lariagon C,Lugan R,Todd C,Wendehenne D,Delourme R,Bouchereau A,ManzanaresDauleux M J.2012.Arginase induction represses gall development during clubroot infection inarabidopsis[J].Plant&Cell Physiology,53(5):901-911.
Halim V A,Vess A,Scheel D,Rosahl S.2006.The role of salicylic acid and jasmonic acid in pathogen defence[J].Plant Biology,8(3):307-313.
Hua L,Wang D R,Tan L B,Fu Y C,Liu F X,Xiao L T,Zhu Z F,Fu Q,Sun X Y,Gu P,Cai H W,McCouch S R,Sun C Q.2015.LABA1,a domestication gene associated with long,barbed awns in wild rice(Article)[J].Plant Cell,27(7):1875-1888.
Lee M W,Qi M,Yang Y.2001.A novel jasmonic acid-inducible rice myb gene associates with fungal infection and host cell death[J].Molecular Plant-Microbe Interactions,14(4):527-535.
Liu S L,Wu J,Zhang P,Hasi G,Huang Y,Lu J,Zhang Y L.2016.Response of phytohormones and correlation of sar signal pathway genes to the different resistance levels of grapevine against Plasmopara viticola infection[J].Plant Physiology and Biochemistry,107:56-66.
Nalley L,Tsiboe F,Durandmorat A,Shew A,Thoma G.2016.Economic and environmental impact of rice blast pathogen(Magnaporthe oryzae)alleviation in the United States[J].PLoS One,11(12):e0167295.
Osés-Ruiz M,Sakulkoo W,Littlejohn G R,Martin-Urdiroz M,Talbot N J.2017.Two independent S-phase checkpoints regulate appressorium-mediated plant infection by the rice blast fungus Magnaporthe oryzae[J].Proceedings of the National Academy of Sciences of the United States of America,114(2):237-244.
Roy-Barman S,Chattoo B B.2005.Rice blast fungus sequenced[J].Currentence,89(6):930-931.
Satapathy L,Kumar D,Kumar M,Mukhopadhyay K.2018.Functional and DNA-protein binding studies of WRKYtranscription factors and their expression analysis in response to biotic and abiotic stress in wheat(Triticum aestivum L.)[J].Biotech,8(1):40.
Séverine L,Alexandre R S,Christine L,Jocelyne L,Nathalie M,Melanie J,Maria J D,Antoine G.2015.Both the jasmonic acid and the salicylic acid pathways contribute to resistance to the biotrophic clubroot agent plasmodiophora brassicae in arabidopsis[J].Plant&Cell Physiology,56(11):2158-2168.
Skamnioti P,Gurr S J.2009.Against the grain:Safeguarding rice from rice blast disease[J].Trends in Biotechnology,27(3):141-150.
Smith J L,Moraes C,Mescher M C.2009.Jasmonate-and salicylate-mediated plant defense responses to insect herbivores,pathogens and parasitic plants[J].Pest Management Science,65(5):497-503.
Yan X,Talbot N J.2016.Investigating the cell biology of plant infection by the rice blast fungus Magnaporthe oryzae[J].Current Opinion in Microbiology,34:147-153.
Yi S Y,Ken S,Moon J S,Lee S G,Kwon S Y.2014.The activated SA and JA signaling pathways have an influence on flg22-triggered oxidative burst and callose deposition[J].PLoS One,9(2):e88951.
Zhang W,Corwin J A,Copeland D,Feusier J,Eshbauqh R,Chen F,Atwell S,Kliebenstein D J.2017.Plastic transcriptomes stabilize immunity to pathogen diversity:The jasmonic acid and salicylic acid networks within the arabidopsis/Botrytis pathosystem[J].Plant Cell,29(11):2727-2752.
李兆举,徐新娟,齐红志,王二歌,宋思艺,王晓桠,赵博文.2017.MeJA浸种对干旱胁迫下玉米种子萌发及幼苗生理特性的影响[J].河南农业科学,46(12):36-41.[Li ZJ,Xu X J,Qi H Z,Wang E G,Song S Y,Wang X Y,Zhao B W.2017.Effects of seed soaking with meja on seed germination and seeding physiological characteristics of maize under drought stress[J].Journal of Henan Agricultural Sciences,46(12):36-41.]
刘霞,唐设,窦志,李刚华,刘正辉,王绍华,丁承强,丁艳锋.2016.茉莉酸甲酯对武运粳24和宁粳3号灌浆早期高温胁迫生理特性的影响[J].中国水稻科学,30(3):291-303.[Liu X,Tang S,Dou Z,Li G H,Liu Z H,Wang SH,Ding C Q,Ding Y F.2016.Effects of MeJA on the physiological characteristics of Japonica rice Wuyunjing24 and Ning-jing 3 during early grain filling stage under heat stress[J].Chinese Journal of Rice Science,30(3):291-303.]
王云锋,李晓杰,刘立娜,刘林,王长秘,李晓疆,李成云,杨静.2018a.稻瘟病菌株形态发育对外源茉莉酸敏感程度分析[J].分子植物育种,16(12):4098-4105.[Wang YF,Li X J,Liu L N,Liu L,Wang C M,Li X J,Li C Y,Yang J.2018a.Sensibility analysis for morphological development of rice blast strain to exogenous jasmonic acid[J].Molecular Plant Breeding,16(12):4098-4105.]
王云锋,王长秘,李春琴,刘林,李晓杰,李晓疆,杨静.2018b.稻瘟病菌侵染时水稻防御体系对外源茉莉酸的响应分析[J].南方农业学报,49(7):1324-1331.[Wang Y F,Wang C M,Li C Q,Liu L,Li X J,Li X J,Yang J.2018b.Response of rice defense system to exogenous jamonisic acids during rice blast strain infecting rice[J].Journal of Southern Agriculture,49(7):1324-1331.]
吴国昭,谢丽君,宋圆圆,陈敏,曾任森.2009.外源信号物质诱导广东高州普通野生稻抗稻瘟病的生理生化机理[J].西北农业学报,18(3):254-258.[Wu G Z,Xie L J,Song Y Y,Chen M,Zeng R S.2009.The physiological and biochemical mechanisms of disease resistance of Oryza rufipogon Griff.Indigenous to Gaozhou,Guangdong against Pyricularia grisea induced by exogenously applied plant signal compounds[J].Acta Agriculturae Boreali-Occidentalis Sinica,18(3):254-258.]
肖宇龙,李湘民,余传元,雷建国,王智权,王晓玲,何虎,邱在辉,肖宇华.2018.江西省稻瘟病生理小种致病力测定及1组水稻优质新种质的稻瘟病抗性表现及应用评估[J].江西农业学报,30(4):9-13.[Xiao Y L,Li X M,Yu CY,Lei J G,Wang Z Q,Wang X L,He H,Qiu Z H,Xiao Y H.2018.Pathogenicity of physiological races of rice blast disease in Jiangxi Province,blast-resistant performance and application evaluation of a set of new highquality rice germplasms[J].Acta Agriculturae Jiangxi,30(4):9-13.]
许志刚.2002.普通植物病理学[M].第3版.北京:中国农业出版社:319-335.[Xu Z G.2002.General Plant Pathology[M].The 3rdEdition.Beijing:China Agriculture Press:319-335.]
张知晓,泽桑梓,户连荣,刘凌,季梅.2018.茉莉酸甲酯生物活性研究进展[J].河南农业科学,47(11):1-7.[Zhang ZX,Ze S Z,Hu L R,Liu L,Ji M.2018.Research advance in biological activities of methyl jasmonate[J].Journal of Henan Agricultural Sciences,47(11):1-7.]
邹志燕,王振中.2006.茉莉酸诱导水稻幼苗对稻瘟病抗性作用研究[J].植物病理学报,36(5):432-438.[Zou Z Y,Wang Z Z.2006.Effect of jasmonic acid on induced resistance of rice seedlings to Magnaporthe grisea[J].Acta Phytopathologica Sinica,36(5):432-438.]
Ahn I P,Kim S,Kang S,Suh S C,Lee Y H.2005.Rice defense mechanisms against Cochliobolus miyabeanus and Magnaporthe grisea are distinct[J].Phytopathology,95(11):1248-1255.
Alkooranee J T,Aledan T R,Ali A K,Lu G,Zhang X,Wu J,Fu C,Li M.2017.Detecting the hormonal pathways in oilseed rape behind induced systemic resistance by Trichoderma harzianum TH12 to Sclerotinia sclerotiorum[J].PLoS One,12(1):e0168850.
Balbi V,Devoto A.2008.Jasmonate signaling network in Arbidops thaliana:Crucial regulatory nodes and new physiological scenarios[J].New Phytologist,177(2):301-318.
Bostock R M.2005.Signal crosstalk and induced resistance:Straddling the line between cost and benefit[J].Annual Review of Phytopathology,43:545-580.
Creelman R A,John E M.1997.Oligosaccharins,brassinotides and jasmonates:Nontr-adition airegu lators of plant growth,development,and gene expression[J].Plant Cell,9:121l-1223.
Duan Z B,LüG Z,Shen C C,Li Q Y,Qin Z,Niu J S.2014.The role of jasmonic acid signalling in wheat(triticum aestivum,l.)powdery mildew resistance reaction[J].European Journal of Plant Pathology,140(1):169-183.
Gravot A,Deleu C,Wagner G,Lariagon C,Lugan R,Todd C,Wendehenne D,Delourme R,Bouchereau A,ManzanaresDauleux M J.2012.Arginase induction represses gall development during clubroot infection inarabidopsis[J].Plant&Cell Physiology,53(5):901-911.
Halim V A,Vess A,Scheel D,Rosahl S.2006.The role of salicylic acid and jasmonic acid in pathogen defence[J].Plant Biology,8(3):307-313.
Hua L,Wang D R,Tan L B,Fu Y C,Liu F X,Xiao L T,Zhu Z F,Fu Q,Sun X Y,Gu P,Cai H W,McCouch S R,Sun C Q.2015.LABA1,a domestication gene associated with long,barbed awns in wild rice(Article)[J].Plant Cell,27(7):1875-1888.
Lee M W,Qi M,Yang Y.2001.A novel jasmonic acid-inducible rice myb gene associates with fungal infection and host cell death[J].Molecular Plant-Microbe Interactions,14(4):527-535.
Liu S L,Wu J,Zhang P,Hasi G,Huang Y,Lu J,Zhang Y L.2016.Response of phytohormones and correlation of sar signal pathway genes to the different resistance levels of grapevine against Plasmopara viticola infection[J].Plant Physiology and Biochemistry,107:56-66.
Nalley L,Tsiboe F,Durandmorat A,Shew A,Thoma G.2016.Economic and environmental impact of rice blast pathogen(Magnaporthe oryzae)alleviation in the United States[J].PLoS One,11(12):e0167295.
Osés-Ruiz M,Sakulkoo W,Littlejohn G R,Martin-Urdiroz M,Talbot N J.2017.Two independent S-phase checkpoints regulate appressorium-mediated plant infection by the rice blast fungus Magnaporthe oryzae[J].Proceedings of the National Academy of Sciences of the United States of America,114(2):237-244.
Roy-Barman S,Chattoo B B.2005.Rice blast fungus sequenced[J].Currentence,89(6):930-931.
Satapathy L,Kumar D,Kumar M,Mukhopadhyay K.2018.Functional and DNA-protein binding studies of WRKYtranscription factors and their expression analysis in response to biotic and abiotic stress in wheat(Triticum aestivum L.)[J].Biotech,8(1):40.
Séverine L,Alexandre R S,Christine L,Jocelyne L,Nathalie M,Melanie J,Maria J D,Antoine G.2015.Both the jasmonic acid and the salicylic acid pathways contribute to resistance to the biotrophic clubroot agent plasmodiophora brassicae in arabidopsis[J].Plant&Cell Physiology,56(11):2158-2168.
Skamnioti P,Gurr S J.2009.Against the grain:Safeguarding rice from rice blast disease[J].Trends in Biotechnology,27(3):141-150.
Smith J L,Moraes C,Mescher M C.2009.Jasmonate-and salicylate-mediated plant defense responses to insect herbivores,pathogens and parasitic plants[J].Pest Management Science,65(5):497-503.
Yan X,Talbot N J.2016.Investigating the cell biology of plant infection by the rice blast fungus Magnaporthe oryzae[J].Current Opinion in Microbiology,34:147-153.
Yi S Y,Ken S,Moon J S,Lee S G,Kwon S Y.2014.The activated SA and JA signaling pathways have an influence on flg22-triggered oxidative burst and callose deposition[J].PLoS One,9(2):e88951.
Zhang W,Corwin J A,Copeland D,Feusier J,Eshbauqh R,Chen F,Atwell S,Kliebenstein D J.2017.Plastic transcriptomes stabilize immunity to pathogen diversity:The jasmonic acid and salicylic acid networks within the arabidopsis/Botrytis pathosystem[J].Plant Cell,29(11):2727-2752.
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