两个玉米自交系对纹枯病的抗病反应机制初探
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摘要
玉米纹枯病是由立枯丝核菌(Rhizoctonia solani Kühn)引起的一种重要病害。具有优良性状的自交系是开展玉米抗病品种选育的重要种质资源。本试验以西南地区两个优良玉米自交系R08与18-599R为材料,采用人工接种R.solani菌丝融合群AG1-IA,对成株期玉米进行了抗性鉴定,并对病原菌诱导后苗期玉米叶片上的活性氧(ROS)、抗病相关基因(PR1、ZmDREB2A、ZmERF3和WIP1)表达及细胞坏死情况进行了动态检测。结果显示,R08对纹枯病表现为中抗,而18-599R为高感材料;受R.solani侵染后,玉米叶片ROS的积累在R08中较18-599R多;两个自交系中基因表达量及表达时间存在显著差异;R08叶片出现显著的细胞坏死反应,而18-599R相对较轻。本研究表明两个自交系对R.solani的抗性差异主要体现在相关抗病反应的时间和强度上,这为进一步研究玉米纹枯病抗病机理提供了依据。
Maize banded leaf and sheath blight(BLSB)is an important disease caused by Rhizoctonia solani Kühn,and screening and breeding of resistant maize germplasms is one of major strategies for disease control.In this study,after inoculation with R.solani AG1-IA,the resistance of two inbred maize cultivars R08 and18-599 R was evaluated,and defense responses,including production of reactive oxygen species(ROS),expression of defenserelated genes(PR1,ZmDREB2 A,ZmERF3 and WIP1)and cell death,were examined on the detached leaves of maize seedlings at different indicated time points.The results showed that R08 was susceptible to AG1-IA,whereas 18-599 R was highly susceptible.Reactive oxygen species(ROS)was accumulated in a higher level at indicated time points on the detached leaves of R08 than that of 18-599 R.Expression of four defense-related genes was induced differently in timing and aptitude in the two inbred lines.Eventually,cell death appeared significantly in the leaves of R08 as compared to 18-599 R.These data indicate that the difference of two maize inbred lines in resistance might be not of qualitative,but of quantitative nature,which provides significant basis for uncovering the resistance mechanism of maize to R.solani.
Maize banded leaf and sheath blight(BLSB)is an important disease caused by Rhizoctonia solani Kühn,and screening and breeding of resistant maize germplasms is one of major strategies for disease control.In this study,after inoculation with R.solani AG1-IA,the resistance of two inbred maize cultivars R08 and18-599 R was evaluated,and defense responses,including production of reactive oxygen species(ROS),expression of defenserelated genes(PR1,ZmDREB2 A,ZmERF3 and WIP1)and cell death,were examined on the detached leaves of maize seedlings at different indicated time points.The results showed that R08 was susceptible to AG1-IA,whereas 18-599 R was highly susceptible.Reactive oxygen species(ROS)was accumulated in a higher level at indicated time points on the detached leaves of R08 than that of 18-599 R.Expression of four defense-related genes was induced differently in timing and aptitude in the two inbred lines.Eventually,cell death appeared significantly in the leaves of R08 as compared to 18-599 R.These data indicate that the difference of two maize inbred lines in resistance might be not of qualitative,but of quantitative nature,which provides significant basis for uncovering the resistance mechanism of maize to R.solani.
引文
[1] 邹军顺,李新果,马文峰.中国玉米种植产业发展现状分析及政策建议[J].粮食科技与经济,2014,39(1):13-15.
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[3] 肖炎农,李建生,郑用链,等.湖北省玉米纹枯病病原丝核菌的种类和致病性[J].菌物系统,2002,21(3):419-424.
[4] 夏海波,伍恩宇,于金凤.黄淮海地区夏玉米纹枯病菌的融合群鉴定[J].菌物学报,2008,27(3):360-367.
[5] 牛福芳,董明明,赵新兰,等.我国部分地区玉米丝核菌组成及其致病类型分析[J].植物保护学报,2009,36(4):289-294.
[6] 李菊,夏海波,于金凤.中国东北地区玉米纹枯病菌的融合群鉴定[J].菌物学报,2011,30(3):392-399.
[7] 荣廷昭,李晚枕,杨克诚,等.西南生态区玉米育种[M].北京:中国农业出版社,2003:187-191.
[8] 黄天述,叶华智,王晓鸣,等.玉米种质资源对纹枯病的抗性鉴定与评价[J].植物遗传资源学报,2005,6(3):291-295.
[9] Nürnberger T.Signal perception in plant pathogen defense[J].Cellular and Molecular Life Sciences,1999,55(2):167-182.
[10]Thordal-Christensen H.Fresh insights into processes of nonhost resistance[J].Current Opinion in Plant Biology,2003,6(4):351-357.
[11]苏顺宗,吴玲,张晓霞,等.以玉米自交系18-599为受体的转基因再生体系优化[J].四川农业大学学报,2013,31(2):122-124.
[12]熊尧宇,李芦江,文水清,等.玉米自交系08-641不同选择方向回交改良后代主要性状的配合力[J].玉米科学,2012,20(1):10-14.
[13]龙书生,曹远林,李亚玲,等.小麦抗条锈病过敏性坏死反应中的活性氧代谢[J].西北农林科技大学学报,2009,37(11):334-336.
[14]张小莉,王鹏程,宋纯鹏.植物细胞过氧化氢的测定方法[J].植物学报,2009,44(1):103-106.
[15]Chintamanani S,Hulbert S H,Johal G S,et al.Identification of a maize locus that modulates the hypersensitive defense response,using mutant-assisted gene identification and characterization[J].Genetics,2010,184:813-825.
[16]Nguyen H T,Leipner J,Stamp P,et al.Low temperature stress in maize(Zea mays L.)induces genes involved in photosynthesis and signal transduction as studied by suppression subtractive hybridization[J].Plant Physiology and Biochemistry,2009,47:116-122.
[17]杨爱国,潘光堂,叶华智,等.玉米自交系纹枯病抗性鉴定及抗病资源筛选[J].植物保护,2003,29(1):25-28.
[18]Wisser R J,Balint-Kurti P J,Nelson R J.The genetic architecture of disease resistance in maize:a synthesis of published studies[J].Phytopathology,2006,96:120-129.
[19]赵茂俊.抗玉米纹枯病种质资源筛选及QTL定位[D].成都:四川农业大学,2004.
[20]Levine A,Tenhaken R,Dixon R,et al.H2O2from the oxidative burst orchestrates the plant hypersensitive disease resistance response[J].Cell,1994,79:583.
[21]Rohrmeier T,Lehle L.WIP1,a wound-inducible gene from maize with homology to Bowman–Birk proteinase inhibitors[J].Plant Molecular Biology,1993,22:783-792.
[22]练云,刘允军,王国英.玉米受伤诱导基因wip1的启动子克隆机表达分析[J].中国农业科学,2014,47(14):2889-2896.
[23]Gutterson N,Reuber T L.Regulation of disease resistance pathways by AP2/ERF transcription factors[J].Current in Opinion Plant Biology,2004,7(4):465-471.
[24]Pre M,Atallah M,Champion A,et al.The AP2/ERF domain transcription factor ORA59integrates jasmonic acid and ethylene signals in plant defense[J].Plant Physiology,2008,147(3):1347-1357.
[25]Anderson J P,Lichtenzveig J,Gleason C,et al.The B-3ethylene response factor MtERF1-1mediates resistance to a subset of root pathogens in Medicago truncatula without adversely affecting symbiosis with Rhizobia[J].Plant Physiology,2010,154(2):861-873.
[26]Zhang Gaiyuan,Chen Ming,Li Liancheng,et al.Overexpression of the soybean GmERF3gene,an AP2/ERF type transcription factor for increased tolerances to salt,drought,and diseases in transgenic tobacco[J].Journal of Experimental Botany,2009,60(13):3781-3796.
[27]Liang Chen,Zhang Zhengyan,Liang Hongxia,et al.Over expression of TiERF1enhances resistance to sharp eyespot in transgenic wheat[J].Journal of Experimental Botany,2008,59(15):4195-4204.
[28]Sels J,Mathys J,De Coninck B M,et al.Plant pathogenesisrelated(PR)proteins:A focus on PR peptides[J].Plant Physiology and Biochemistry,2008,46(11):941-950.
[29]van Loon L C.Induced resistance in plants and the role of pathogenesis-related proteins[J].European Journal of Plant Pathology,1997,103(9):753-765.
[30]Chen Zhixiang,Malamy J,Henning J,et al.Induction,modification,and transduction of the salicylic acid signal in plant defense responses[J].Proceedings of the National Academy of Sciences of the United States of America,1995,92(10):4134-4137.
[31]乐美旺,陈实,潘庆华,等.水稻和稻瘟病菌互作中的信号传导及防御反应基因诱导表达的研究[J].植物病理学报,2007,37(1):42-49.
[32]王晨芳.小麦与条锈病互作过程中活性氧迸发的组织学和细胞化学研究[D].杨凌:西北农林科技大学,2008.
[33]van loon L C,van Kammen A.Polyacrylamide disc electrophoresis of the soluble leaf proteins from Nicotiana tabacum var.‘Samsun’and‘Samsun NN’Ⅱ.Change in protein constitution after infection with tobacco mosaic virus[J].Virology,1970,40:190-211.
[34]Morris S W,Vernooij B,Titatarn S,et al.Induced resistance responses in maize[J].Molecular Plant Microbe Interaction,1998,11:643-658.
[35]Mittler R,Vanderauwera S.ROS signaling:the new wave?[J].Trends in Plant Science,2011,16(6):300-309.
[36]Zheng Aiping,Lin Runmao,Zhang Danhua,et al.The evolution and pathogenic mechanisms of the rice sheath blight pathogen[J].Nature Communications,2013,4:1424.
[2] 唐海涛,荣延昭,杨俊品.玉米纹枯病研究进展[J].玉米科学,2004,12(1):93-96.
[3] 肖炎农,李建生,郑用链,等.湖北省玉米纹枯病病原丝核菌的种类和致病性[J].菌物系统,2002,21(3):419-424.
[4] 夏海波,伍恩宇,于金凤.黄淮海地区夏玉米纹枯病菌的融合群鉴定[J].菌物学报,2008,27(3):360-367.
[5] 牛福芳,董明明,赵新兰,等.我国部分地区玉米丝核菌组成及其致病类型分析[J].植物保护学报,2009,36(4):289-294.
[6] 李菊,夏海波,于金凤.中国东北地区玉米纹枯病菌的融合群鉴定[J].菌物学报,2011,30(3):392-399.
[7] 荣廷昭,李晚枕,杨克诚,等.西南生态区玉米育种[M].北京:中国农业出版社,2003:187-191.
[8] 黄天述,叶华智,王晓鸣,等.玉米种质资源对纹枯病的抗性鉴定与评价[J].植物遗传资源学报,2005,6(3):291-295.
[9] Nürnberger T.Signal perception in plant pathogen defense[J].Cellular and Molecular Life Sciences,1999,55(2):167-182.
[10]Thordal-Christensen H.Fresh insights into processes of nonhost resistance[J].Current Opinion in Plant Biology,2003,6(4):351-357.
[11]苏顺宗,吴玲,张晓霞,等.以玉米自交系18-599为受体的转基因再生体系优化[J].四川农业大学学报,2013,31(2):122-124.
[12]熊尧宇,李芦江,文水清,等.玉米自交系08-641不同选择方向回交改良后代主要性状的配合力[J].玉米科学,2012,20(1):10-14.
[13]龙书生,曹远林,李亚玲,等.小麦抗条锈病过敏性坏死反应中的活性氧代谢[J].西北农林科技大学学报,2009,37(11):334-336.
[14]张小莉,王鹏程,宋纯鹏.植物细胞过氧化氢的测定方法[J].植物学报,2009,44(1):103-106.
[15]Chintamanani S,Hulbert S H,Johal G S,et al.Identification of a maize locus that modulates the hypersensitive defense response,using mutant-assisted gene identification and characterization[J].Genetics,2010,184:813-825.
[16]Nguyen H T,Leipner J,Stamp P,et al.Low temperature stress in maize(Zea mays L.)induces genes involved in photosynthesis and signal transduction as studied by suppression subtractive hybridization[J].Plant Physiology and Biochemistry,2009,47:116-122.
[17]杨爱国,潘光堂,叶华智,等.玉米自交系纹枯病抗性鉴定及抗病资源筛选[J].植物保护,2003,29(1):25-28.
[18]Wisser R J,Balint-Kurti P J,Nelson R J.The genetic architecture of disease resistance in maize:a synthesis of published studies[J].Phytopathology,2006,96:120-129.
[19]赵茂俊.抗玉米纹枯病种质资源筛选及QTL定位[D].成都:四川农业大学,2004.
[20]Levine A,Tenhaken R,Dixon R,et al.H2O2from the oxidative burst orchestrates the plant hypersensitive disease resistance response[J].Cell,1994,79:583.
[21]Rohrmeier T,Lehle L.WIP1,a wound-inducible gene from maize with homology to Bowman–Birk proteinase inhibitors[J].Plant Molecular Biology,1993,22:783-792.
[22]练云,刘允军,王国英.玉米受伤诱导基因wip1的启动子克隆机表达分析[J].中国农业科学,2014,47(14):2889-2896.
[23]Gutterson N,Reuber T L.Regulation of disease resistance pathways by AP2/ERF transcription factors[J].Current in Opinion Plant Biology,2004,7(4):465-471.
[24]Pre M,Atallah M,Champion A,et al.The AP2/ERF domain transcription factor ORA59integrates jasmonic acid and ethylene signals in plant defense[J].Plant Physiology,2008,147(3):1347-1357.
[25]Anderson J P,Lichtenzveig J,Gleason C,et al.The B-3ethylene response factor MtERF1-1mediates resistance to a subset of root pathogens in Medicago truncatula without adversely affecting symbiosis with Rhizobia[J].Plant Physiology,2010,154(2):861-873.
[26]Zhang Gaiyuan,Chen Ming,Li Liancheng,et al.Overexpression of the soybean GmERF3gene,an AP2/ERF type transcription factor for increased tolerances to salt,drought,and diseases in transgenic tobacco[J].Journal of Experimental Botany,2009,60(13):3781-3796.
[27]Liang Chen,Zhang Zhengyan,Liang Hongxia,et al.Over expression of TiERF1enhances resistance to sharp eyespot in transgenic wheat[J].Journal of Experimental Botany,2008,59(15):4195-4204.
[28]Sels J,Mathys J,De Coninck B M,et al.Plant pathogenesisrelated(PR)proteins:A focus on PR peptides[J].Plant Physiology and Biochemistry,2008,46(11):941-950.
[29]van Loon L C.Induced resistance in plants and the role of pathogenesis-related proteins[J].European Journal of Plant Pathology,1997,103(9):753-765.
[30]Chen Zhixiang,Malamy J,Henning J,et al.Induction,modification,and transduction of the salicylic acid signal in plant defense responses[J].Proceedings of the National Academy of Sciences of the United States of America,1995,92(10):4134-4137.
[31]乐美旺,陈实,潘庆华,等.水稻和稻瘟病菌互作中的信号传导及防御反应基因诱导表达的研究[J].植物病理学报,2007,37(1):42-49.
[32]王晨芳.小麦与条锈病互作过程中活性氧迸发的组织学和细胞化学研究[D].杨凌:西北农林科技大学,2008.
[33]van loon L C,van Kammen A.Polyacrylamide disc electrophoresis of the soluble leaf proteins from Nicotiana tabacum var.‘Samsun’and‘Samsun NN’Ⅱ.Change in protein constitution after infection with tobacco mosaic virus[J].Virology,1970,40:190-211.
[34]Morris S W,Vernooij B,Titatarn S,et al.Induced resistance responses in maize[J].Molecular Plant Microbe Interaction,1998,11:643-658.
[35]Mittler R,Vanderauwera S.ROS signaling:the new wave?[J].Trends in Plant Science,2011,16(6):300-309.
[36]Zheng Aiping,Lin Runmao,Zhang Danhua,et al.The evolution and pathogenic mechanisms of the rice sheath blight pathogen[J].Nature Communications,2013,4:1424.