稻瘟病菌侵染时水稻防御体系对外源茉莉酸的响应分析
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摘要
【目的】探究特定时期稻瘟病菌与水稻互作对外源茉莉酸(JA)的响应,为揭示稻瘟病菌与水稻互作时水稻防御体系对外源JA响应的分子机制提供理论依据。【方法】以稻瘟病菌株95234I-1b和普通感病水稻品种丽江新团黑谷(LTH)为试验材料,通过外源JA以两种不同的方式(处理1,分别用100和400μmol/L JA处理水稻叶片,6 h后接种95234I-1b菌株;处理2,水稻叶片接种95234I-1b菌株72 h后,分别用100和400μmol/L JA处理水稻叶片)处理与稻瘟病菌互作过程中特定时期的水稻,调查水稻稻瘟病发病症状,并用即时聚合酶链锁反应(q RT-PCR)检测水稻防御相关基因的表达情况。【结果】外源JA的两种不同处理方式均能减轻受侵染水稻稻瘟病发病症状,并诱导水稻防御相关基因不同程度的上调表达。用100和400μmol/L JA喷雾水稻6 h后再接种稻瘟病菌株的诱抗效果分别为15.06%和25.63%;稻瘟病菌株孢子接种水稻72 h再喷雾100和400μmol/L JA的诱抗效果分别为36.60%和47.12%。与对照相比,JA抑制了水稻水杨酸(SA)途径相关基因的大幅上调表达。在一定JA浓度范围内,高浓度JA对SA途径相关基因表达抑制程度高于低浓度JA的抑制程度;高浓度JA诱导JA途径相关基因上调表达幅度高于低浓度JA的诱导程度;高浓度JA诱导PR1a上调表达幅度小于低浓度JA诱导的上调幅度,高浓度JA诱导PR10a上调表达幅度大于低浓度JA诱导的上调幅度。【结论】JA以两种不同的方式处理与稻瘟病菌互作过程中特定时期的水稻,均能诱导水稻病程相关基因和JA途径相关基因上调表达,抑制SA途径相关基因的大幅上调表达,表明水稻防御体系中的病程相关基因及JA途径的相关基因主要参与了水稻防御体系对外源JA的响应。
【Objective】Through studying response of interaction between rice blast strain and rice to exogenous jasmonic acids(JA)at special time points,the theoretical basis for studying molecular mechanism on response of rice defense system to exogenous JA during rice blast strain infecting rice was provided.【Method】In this study,rice blast strain95234 I-1 b and common susceptible rice variety LTH were used as materials. Rice leaves during rice blast strain infecting rice were treated with exogenous jasmonic acids through two different methods(treatment 1:rice leaves were pre-sprayed with 100 and 400 μmol/L JA and then the pre-treated leaves were inoculated with rice blast strain 95234 I-1 b after 6 h;treatment 2:rice leaves were inoculated with rice blast strain 95234 I-1 b,the inoculated leaves were sprayed with 100 and 400 μmol/L JA after 72 h). The symptom of rice blast were investigated and expression level of rice defense-related genes were detected by real-time polymerase chain reaction(q RT-PCR).【Result】The exogenous JA through two different methods could alleviate the symptoms and induce up-regulation of defense-related genes of infected rice. The induced effects of 100 and 400 μmol/L JA spraying rice and then inoculated with rice blast strain after 6 h were 15.06% and 25.63% respectively. The induced effects of rice leaves which inoculated with rice blast strain and were sprayed by 100 and 400μmol/L JA after 72 h were 36.60% and 47.12% respectively. Compared to control,JA inhibited up-regulation of salicylic acid(SA)-pathway related genes. And within a certain concentration of JA,high concentration of JA could inhibit SApathway related genes more than those of low concentration of JA. High concentration of JA could induce larger degree of up-regulation of JA-pathway genes than low concentration of JA. High concentration of JA could induce smaller degree of up-regulation of PR1 a than low concentration of JA,and high concentration of JA could induce larger degree of up-regulation than low concentration of JA.【Conclusion】Exogenous JA through two methods and interaction with rice blast strain at specific periods could induce up-regulation of pathogenesis-related genes and JA-pathway genes,and inhibit up-regulation of SA-pathway genes. Therefore,pathogenesis-related genes and JA-pathway genes mainly involve in response of rice defense system to exogenous JA.
【Objective】Through studying response of interaction between rice blast strain and rice to exogenous jasmonic acids(JA)at special time points,the theoretical basis for studying molecular mechanism on response of rice defense system to exogenous JA during rice blast strain infecting rice was provided.【Method】In this study,rice blast strain95234 I-1 b and common susceptible rice variety LTH were used as materials. Rice leaves during rice blast strain infecting rice were treated with exogenous jasmonic acids through two different methods(treatment 1:rice leaves were pre-sprayed with 100 and 400 μmol/L JA and then the pre-treated leaves were inoculated with rice blast strain 95234 I-1 b after 6 h;treatment 2:rice leaves were inoculated with rice blast strain 95234 I-1 b,the inoculated leaves were sprayed with 100 and 400 μmol/L JA after 72 h). The symptom of rice blast were investigated and expression level of rice defense-related genes were detected by real-time polymerase chain reaction(q RT-PCR).【Result】The exogenous JA through two different methods could alleviate the symptoms and induce up-regulation of defense-related genes of infected rice. The induced effects of 100 and 400 μmol/L JA spraying rice and then inoculated with rice blast strain after 6 h were 15.06% and 25.63% respectively. The induced effects of rice leaves which inoculated with rice blast strain and were sprayed by 100 and 400μmol/L JA after 72 h were 36.60% and 47.12% respectively. Compared to control,JA inhibited up-regulation of salicylic acid(SA)-pathway related genes. And within a certain concentration of JA,high concentration of JA could inhibit SApathway related genes more than those of low concentration of JA. High concentration of JA could induce larger degree of up-regulation of JA-pathway genes than low concentration of JA. High concentration of JA could induce smaller degree of up-regulation of PR1 a than low concentration of JA,and high concentration of JA could induce larger degree of up-regulation than low concentration of JA.【Conclusion】Exogenous JA through two methods and interaction with rice blast strain at specific periods could induce up-regulation of pathogenesis-related genes and JA-pathway genes,and inhibit up-regulation of SA-pathway genes. Therefore,pathogenesis-related genes and JA-pathway genes mainly involve in response of rice defense system to exogenous JA.
引文
宾金华,姜胜,黄胜琴,潘瑞炽.2000.茉莉酸甲酯诱导烟草幼苗抗炭疽病与PAL活性及细胞壁物质的关系[J].植物生理学报,26(1):1-6.[Bin J H,Jiang S,Huang S Q,Pan R C.2000.The relationship between methyl jasmonate induced arthracnose resistance of tobacco seedlings and phenylalanine ammomaly reactivity and cell wall substances[J].Acta Phytophysiologica Sinica,26(1):1-6.]
王云锋,王春梅,王长秘,张娅玲,刘林,李成云,杨静.2017.外源水杨酸对稻瘟病菌效应蛋白BAS4过表达菌株耐受性的影响[J].南方农业学报,48(12):2169-2175.[Wang Y F,Wang C M,Wang C M,Zhang Y L,Liu L,Li C Y,Yang J.2017.Effects of exogenous salicylic acid on tolerance of effector protein BAS4 overexpression strain of Magnaporthe oryzae[J].Journal of Southern Agriculture,48(12):2169-2175]
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 andbiochemical mechanisms of disease resistance of Oryza rufipogon Griff.indigenous to Gaozhou,Guangdong against Pyricularia grisea induced by exogenously applied plant signa[lJ].Acta Agriculturae Boreali-occidentalis Sinica,18(3):254-258.]
向妙莲,何永明,付永琦,曾晓春,黄俊宝,黄友明.2013.茉莉酸甲酯对水稻白叶枯病的诱导抗性及相关防御酶活性的影响[J].植物保护学报,40(2):97-101.[Xiang M L,He Y M,Fu Y Q,Zeng X C,Huang J B,Huang Y M.2013.Effect of methyl jasmonate on induced resistance of rice seedlings against bacterial leaf blight and activities of related defense enzymes[J].Acta Phytophylacica Sinica,40(2):97-101.]
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杨庆.2017.水稻抗稻瘟病基因与育种研究进展[J].北方水稻,47(2):47-50.[Yang Q.2017.Review of reserches on gene and breeding of blast resistance on rice[J].Northern Rice,47(2):47-50.]
杨艳丽.2010.外源茉莉酸影响马铃薯对晚疫病抗性的初步研究[D].长沙:湖南农业大学.[Yang Y L.2010.Preliminary study of exogenous jasmonic acid effects on potato resistance to late blight[D].Changsha:Hunan Agricultural University.]
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 ZY,Wang Z Z.2006.Effect of jasmonic acid on induced resistance of rice seedlings to Magnaportheg risea[J].Acta Phytopathologica Sinica,36(5):432-438.]
Blechert S,Brodschelm W,Holder S,Kammerer L,Kutchan TM,Mueller M J,Xia Z Q,Zenk M H.1995.The octadecanoic pathway signal molecules for the regulation of secondary pathways[J].Proceedings of the National Academy of Science,92(10):4099-4105.
Blée E.2002.Impact of phyto-oxylipins in plant defense[J].Trends in Plant Science,7(7):315-322.
Browse J.2009.Jasmonate passes muster:A receptor and targets for the defense hormone[J].Annual Review of Plant Biology,60(1):183-205.
Creelman R A,Mullet J E.1995.Jasmonic acid distribution and action in plants:Regulation during development and response to biotic and abiotic stress[J].National Academy of Science,92:4114-4119.
Dohondt S,Geoffroy P,Stelmach B A,Legrand M,Heiz T.2000.Soluble phospholipase A2 activity is induced before oxylipin accumulation in tobacco mosaic virus-infected tobacco leaves and is contributed by patatin-like enzymes[J].Plant Journal,23(4):431-440.
Donald F C,Ahnya M R.1995.Age-Dependent effects of jasmonic acid treatment and wind exposure on foliar oxidase activity and insect Resistance in tomato[J].Journal of Chemical Ecology,25(2):271-281.
Durrant W E,Dong X.2004.Systematic acquired resistance[J].Annual Reviews of Phytopathol,42(4):185-209.
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 in Arabidopsis[J].Plant and Cell Physiology,53(5):901-911.
He Y Q,Zhang,H H,Sun Z T,Li J M,Hong G J,Zhu Q S,Zhu X B,Mac Farlane S,Yan F B,Chen J P.2017.Jasmonic acid-mediated defense suppresses brassinosteroid-mediated susceptibility to Rice black streaked dwarf virus infection in rice[J].New Phytologist,214(1):388-399.
Lee Y H,Hong J K.2014.Differential defence responses of susceptible and resistant kimchi cabbage cultivars to anthracnose,black spot and black rot diseases[J].Plant Pathology,64(2):406-415.
LemariéS,Robert-Seilaniantz A,Lariagon C,Lemoine J,Marnet N,Jubault M,Manzanares-Dauleux M J,Gravot A.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 Physiol,56(11):2158-2168.
Mei C,Qi M,Sheng G,Yang Y.2006.Inducible overexpression of a rice allene oxide synthase gene increases the endogenous jasmonic acid level,PR gene expression,and host resistance to fungal infection[J].Molecular Plant Microbe Interactions,19(10):1127-1137.
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].PLo S One,11(12):e0167295.
Oka K,Kobayashi M,Mitsuhara I,Seo S.2013.Jasmonic acid negatively regulates resistance to Tobacco mosaic virus in tobacco[J].Plant&Cell Physiology,54(12):1999-2010.
Seo S,Seto H,Yamakawa H,Ohashi Y.2001.Transient accumulation of jasmonic acid during the synchronized hypersensitive cell death in tobacco mosaic virus-infected tobacco leaves[J].Molecular Plant Microbe Interactions,14(2):261-264.
Thaler J S.1999.Jasmonate-inducible plant defences cause increased parasitism of herbivores[J].Nature,399:686-688.
Walters D,Cowley T,Mitchell A.2002.Methyl jasmonate alters polyamine metabolism and induces systemic protection against powdery mildew infection in barley seedlings[J].Journal of Experimental Botany,53(369):747-756.
Zhang S,Li X,Sun Z,Shao S,Hu L,Ye M,Zhou Y,Xia X,Yu J,Shi K.2015.Antagonism between phytohormone signalling underliesthe variation in disease susceptibility of tomato plants under elevated CO[2J].Journal of Experimental Botany,66(7):1951-1963.
Zhu F,Xi D H,Yuan S,Xu F,Zhang D W,Lin H H.2014.Salicylic acid and jasmonic acid are essential for systemic resistance against Tobacco mosaic virus in nicotiana benthamiana[J].Molecular Plant Microbe Interactions,27(6):567-577.
王云锋,王春梅,王长秘,张娅玲,刘林,李成云,杨静.2017.外源水杨酸对稻瘟病菌效应蛋白BAS4过表达菌株耐受性的影响[J].南方农业学报,48(12):2169-2175.[Wang Y F,Wang C M,Wang C M,Zhang Y L,Liu L,Li C Y,Yang J.2017.Effects of exogenous salicylic acid on tolerance of effector protein BAS4 overexpression strain of Magnaporthe oryzae[J].Journal of Southern Agriculture,48(12):2169-2175]
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 andbiochemical mechanisms of disease resistance of Oryza rufipogon Griff.indigenous to Gaozhou,Guangdong against Pyricularia grisea induced by exogenously applied plant signa[lJ].Acta Agriculturae Boreali-occidentalis Sinica,18(3):254-258.]
向妙莲,何永明,付永琦,曾晓春,黄俊宝,黄友明.2013.茉莉酸甲酯对水稻白叶枯病的诱导抗性及相关防御酶活性的影响[J].植物保护学报,40(2):97-101.[Xiang M L,He Y M,Fu Y Q,Zeng X C,Huang J B,Huang Y M.2013.Effect of methyl jasmonate on induced resistance of rice seedlings against bacterial leaf blight and activities of related defense enzymes[J].Acta Phytophylacica Sinica,40(2):97-101.]
许志刚.2002.普通植物病理学[M].第3版.北京:中国农业出版社:319-322.[Xu Z G.2002.General Plant Pathology[M].The 3ndEdition.Beijing:China Agriculture Press:319-322.]
杨庆.2017.水稻抗稻瘟病基因与育种研究进展[J].北方水稻,47(2):47-50.[Yang Q.2017.Review of reserches on gene and breeding of blast resistance on rice[J].Northern Rice,47(2):47-50.]
杨艳丽.2010.外源茉莉酸影响马铃薯对晚疫病抗性的初步研究[D].长沙:湖南农业大学.[Yang Y L.2010.Preliminary study of exogenous jasmonic acid effects on potato resistance to late blight[D].Changsha:Hunan Agricultural University.]
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 ZY,Wang Z Z.2006.Effect of jasmonic acid on induced resistance of rice seedlings to Magnaportheg risea[J].Acta Phytopathologica Sinica,36(5):432-438.]
Blechert S,Brodschelm W,Holder S,Kammerer L,Kutchan TM,Mueller M J,Xia Z Q,Zenk M H.1995.The octadecanoic pathway signal molecules for the regulation of secondary pathways[J].Proceedings of the National Academy of Science,92(10):4099-4105.
Blée E.2002.Impact of phyto-oxylipins in plant defense[J].Trends in Plant Science,7(7):315-322.
Browse J.2009.Jasmonate passes muster:A receptor and targets for the defense hormone[J].Annual Review of Plant Biology,60(1):183-205.
Creelman R A,Mullet J E.1995.Jasmonic acid distribution and action in plants:Regulation during development and response to biotic and abiotic stress[J].National Academy of Science,92:4114-4119.
Dohondt S,Geoffroy P,Stelmach B A,Legrand M,Heiz T.2000.Soluble phospholipase A2 activity is induced before oxylipin accumulation in tobacco mosaic virus-infected tobacco leaves and is contributed by patatin-like enzymes[J].Plant Journal,23(4):431-440.
Donald F C,Ahnya M R.1995.Age-Dependent effects of jasmonic acid treatment and wind exposure on foliar oxidase activity and insect Resistance in tomato[J].Journal of Chemical Ecology,25(2):271-281.
Durrant W E,Dong X.2004.Systematic acquired resistance[J].Annual Reviews of Phytopathol,42(4):185-209.
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 in Arabidopsis[J].Plant and Cell Physiology,53(5):901-911.
He Y Q,Zhang,H H,Sun Z T,Li J M,Hong G J,Zhu Q S,Zhu X B,Mac Farlane S,Yan F B,Chen J P.2017.Jasmonic acid-mediated defense suppresses brassinosteroid-mediated susceptibility to Rice black streaked dwarf virus infection in rice[J].New Phytologist,214(1):388-399.
Lee Y H,Hong J K.2014.Differential defence responses of susceptible and resistant kimchi cabbage cultivars to anthracnose,black spot and black rot diseases[J].Plant Pathology,64(2):406-415.
LemariéS,Robert-Seilaniantz A,Lariagon C,Lemoine J,Marnet N,Jubault M,Manzanares-Dauleux M J,Gravot A.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 Physiol,56(11):2158-2168.
Mei C,Qi M,Sheng G,Yang Y.2006.Inducible overexpression of a rice allene oxide synthase gene increases the endogenous jasmonic acid level,PR gene expression,and host resistance to fungal infection[J].Molecular Plant Microbe Interactions,19(10):1127-1137.
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].PLo S One,11(12):e0167295.
Oka K,Kobayashi M,Mitsuhara I,Seo S.2013.Jasmonic acid negatively regulates resistance to Tobacco mosaic virus in tobacco[J].Plant&Cell Physiology,54(12):1999-2010.
Seo S,Seto H,Yamakawa H,Ohashi Y.2001.Transient accumulation of jasmonic acid during the synchronized hypersensitive cell death in tobacco mosaic virus-infected tobacco leaves[J].Molecular Plant Microbe Interactions,14(2):261-264.
Thaler J S.1999.Jasmonate-inducible plant defences cause increased parasitism of herbivores[J].Nature,399:686-688.
Walters D,Cowley T,Mitchell A.2002.Methyl jasmonate alters polyamine metabolism and induces systemic protection against powdery mildew infection in barley seedlings[J].Journal of Experimental Botany,53(369):747-756.
Zhang S,Li X,Sun Z,Shao S,Hu L,Ye M,Zhou Y,Xia X,Yu J,Shi K.2015.Antagonism between phytohormone signalling underliesthe variation in disease susceptibility of tomato plants under elevated CO[2J].Journal of Experimental Botany,66(7):1951-1963.
Zhu F,Xi D H,Yuan S,Xu F,Zhang D W,Lin H H.2014.Salicylic acid and jasmonic acid are essential for systemic resistance against Tobacco mosaic virus in nicotiana benthamiana[J].Molecular Plant Microbe Interactions,27(6):567-577.