玉米苯丙氨酸解氨酶家族基因的鉴定与纹枯病的抗病分析
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摘要
玉米纹枯病是中国西南玉米产区的主要病害之一,苯丙氨酸解氨酶(PAL)活性变化在不同纹枯病抗性的玉米材料中存在差异,目前对PAL家族基因还缺乏系统深入的研究,尤其是参与玉米-纹枯病菌互作的研究甚少。本研究利用RT-qPCR方法,结合芯片数据库和转录组测序结果,鉴定了玉米PAL家族基因和组织表达特征,并分析了其在纹枯病菌诱导前后抗病和感病自交系中的表达情况。结果显示,玉米基因组中共有13个PAL基因,Zm PAL2、ZmPAL3、ZmPAL5、ZmPAL8、ZmPAL9和ZmPAL10均为组成性高表达,ZmPAL1、ZmPAL4、ZmPAL6和ZmPAL7均为组织特异性低表达,而同源度极高的ZmPAL11、ZmPAL12和ZmPAL13在所有检测组织中均不表达。在强致病力纹枯病菌诱导前,10个PAL基因的本底表达量在抗病和感病自交系的叶鞘中均无明显差异,但在诱导后都增强高表达,以应对纹枯病菌的侵染。这些结果说明不同抗性的玉米材料中可能存在类似但响应程度不同的PAL家族基因参与玉米-纹枯病菌互作的机制。
Maize sheath blight is one of the most devastating diseases for stable maize production of southwest region of China. The activity of phenylalanine ammonia-lyase(PAL) was different in maize materials with different resistance to sheath blight. Currently, systematic and in-depth studies of the PAL genes family and especially its roles in interactions among maize and sheath-blight fungus are scarce. Here, we used RT-qPCR method, combined microarray data and RNA-seq results, the PAL gene members and tissue expression characteristics in maize were identified and the PAL gene expression in resistant and susceptible inbred lines of sheath blight before and after induction was analyzed. Thirteen members of PAL genes in maize genome were identified and six(ZmPAL2,ZmPAL3, ZmPAL5, ZmPAL8, ZmPAL9 and ZmPAL10) of thirteen PAL paralogs were constitutively high expressed, while four(ZmPAL1, ZmPAL4, ZmPAL6 and ZmPAL7) showed organ-specific low expression and others (ZmPAL11, ZmPAL12 and ZmPAL13) with extremely high homology were not expressed in all tested tissues. The background expressions of 10 PAL genes in the leaf sheath of resistant and susceptible inbred lines were similar difference before R. solani AG1 IA inoculation, but all of them were significantly up-regulated after infection to deal with the infection of R. solani. These results suggested that the mechanisms of the PAL family genes might involved in interactions among maize and sheath-blight fungus might be possibly common, and the levels of the response to R. solani infection are similar but infection are different among resistant and susceptible genotypes.
Maize sheath blight is one of the most devastating diseases for stable maize production of southwest region of China. The activity of phenylalanine ammonia-lyase(PAL) was different in maize materials with different resistance to sheath blight. Currently, systematic and in-depth studies of the PAL genes family and especially its roles in interactions among maize and sheath-blight fungus are scarce. Here, we used RT-qPCR method, combined microarray data and RNA-seq results, the PAL gene members and tissue expression characteristics in maize were identified and the PAL gene expression in resistant and susceptible inbred lines of sheath blight before and after induction was analyzed. Thirteen members of PAL genes in maize genome were identified and six(ZmPAL2,ZmPAL3, ZmPAL5, ZmPAL8, ZmPAL9 and ZmPAL10) of thirteen PAL paralogs were constitutively high expressed, while four(ZmPAL1, ZmPAL4, ZmPAL6 and ZmPAL7) showed organ-specific low expression and others (ZmPAL11, ZmPAL12 and ZmPAL13) with extremely high homology were not expressed in all tested tissues. The background expressions of 10 PAL genes in the leaf sheath of resistant and susceptible inbred lines were similar difference before R. solani AG1 IA inoculation, but all of them were significantly up-regulated after infection to deal with the infection of R. solani. These results suggested that the mechanisms of the PAL family genes might involved in interactions among maize and sheath-blight fungus might be possibly common, and the levels of the response to R. solani infection are similar but infection are different among resistant and susceptible genotypes.
引文
Fraser C.M.,and Chapple C.,2011,The phenylpropanoid pathway in Arabidopsis,Arabidopsis Book,9:e0152
Gai J.T.,Shen J.K.,and Wang P.,2016,Identification and sequence analysis of PAL gene family in major crops,Jiangsu Nongye Kexue(Agricultural Sciences of Jiangsu),44(6):45-49(盖江涛,沈建凯,王鹏,2016,主要作物中PAL基因家族的鉴定和序列分析,江苏农业科学,44(6):45-49)
Gao W.C.,Li Y.,Zhang Y.C.,Zhang Z.M.,Pan G.T.,and Lin H.J.,2016,Cloning of maize WRKY76 and its expression patterns involved in resistance to maize banded sheath blight,Guangxi Zhiwu(Guihaia),36(3):273-279(高文成,李玥,张银超,张志明,潘光堂,林海建,2016,玉米转录因子WRKY76克隆及抗纹枯病表达模式分析,广西植物,36(3):273-279)
Huang J.,Gu M.,Lai Z.,Fan B.,Shi K.,Zhou Y.H.,Yu J.Q.,and Chen Z.,2010,Functional analysis of the Arabidopsis PALgene family in plant growth,development,and response to environmental stress,Plant Physiol.,153(4):1526-1538
Jin Q.C.,Ye H.Z.,and Zhang M.,2003,Relationship between the activity of PAL and resistance of corn to maize sheath blight,Sichuan Nongye Daxue Xuebao(Journal of Sichuan Agricultural University),21(2):116-118(金庆超,叶华智,张敏,2003,苯丙氨酸解氨酶活性与玉米对纹枯病抗性的关系,四川农业大学学报,21(2):116-118)
Kawamata S.,Shimobarai K.,Imura Y.,Ozaki M.,Ichinose Y.,Shiraishi T.,Kunoh H.,and Yamada T.,1997,Temporal and spatial pattern of expression of the pea phenylalanine ammonia-lyase genel promoter in transgenic tobacco,Plant Cell Physiol.,38(7):792-803
La Camera S.,Gouzerh G.,Dhondt S.,Hoffmann L.,Fritig B.,Legrand M.,and Heitz T.,2004,Metabolic reprogramming in plant innate immunity:the contributions of phenylpropanoid and oxylipin pathways,Immunol.Rev.,198(1):267-284
Liu L.,Ma Y.Y.,Zhang Z.M.,Leng P.F.,Pan G.T.,and Zhao M.J.,2009,Analysis function of different defense enzymes on maize,Yumi Kexue(Journal of Maize Sciences),17(3):99-102,106(刘丽,马永毅,张志明,冷鹏飞,潘光堂,赵茂俊,2009,玉米不同防卫酶系对纹枯病作用的研究,玉米科学,17(3):99-102,106)
Ma Y.N.,Han C.,Chen J.Y.,Li H.Y.,He K.,Liu A.X.,and Li D.H.,2015,Fungal cellulase is an elicitor but its enzymatic activity is not required for its elicitor activity,Mol.Plant Pathol.,16(1):14-26
MacDonald M.J.,and D'Cunha G.B.,2007,A modern view of phenylalanine ammonia lyase,Biochem.and Cell Biol.,85(3):273-282
Santiago L.J.M.,Louro R.P.,and Oliveira D.E.D.,2000,Compartmentation of phenolic compounds and phenylalanine ammonia-lyase in leaves of Phyllanthus tenellus Roxb.and their induction by copper sulphate,Ann.Bot.,86(5):1023-1032
Sekhon R.S.,Lin H.,Childs K.L.,Hansey C.N.,Buell C.R.,De Leon N.,and and Kaeppler S.M.,2011,Genome-wide atlas of transcription during maize development,Plant J.,66(4):553-563
Sun H.Y.,Quan X.L.,Fu S.,Wu S.Q.,2016,Research progress of phenylalanine ammonia-lyase(PAL)gene in Arabidopsis thaliana,Yanbian Daxue Nongxue Xuebao(Agricultural Science Journal of Yanbian University),38(1):88-92(孙海燕,全雪丽,付爽,吴松权,2016,拟南芥苯丙氨酸解氨酶(PAL)基因的研究进展,延边大学农学学报,38(1):88-92)
Wang S.M.,Zhao X.L.,Ma Y.N.,and Liu A.X.,2016,Histological changes and expression of defense related genes of maize during the interaction with Rhizoctoniasolani AG1-IAYumi Kexue(Journal of Maize Sciences),24(5):37-42(王绍敏,赵新兰,马亚男,刘爱新,2016,Rhizoctoniasolan AG1-IA侵染玉米过程中组织病理学变化及防卫反应基因的表达,玉米科学,24(5):37-42)
Wang S.M.,Zhao X.L.,Ma Y.N.,and Liu A.X.,2017,Effects of different Rhizoctonia spp.on lognin metabolism,total phenols content and expression of defense related genes in maize,Gansu Nongye Daxuebao(Journal of Gansu Agricultural University),52(5):55-60(王绍敏,赵新兰,马亚男,刘爱新,2017,不同致病力Rhizoctonia菌株对玉米木质素代谢、总酚含量及相关基因表达的影响,甘肃农大学报,52(5):55-60)
Xu W.L.,Pei X.L.,Jing Z.G.,and Xiong Z.L.,2015,Genome-wide identification and expression analysis of chalcone synthase gene in pepper,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(8):1747-1752(徐婉莉,裴徐梨,荆赞革,熊自立,2015,辣椒查尔酮合成酶基因家族全基因组鉴定及表达特征分析,基因组学与应用生物学,34(8):1747-1752)
Yang J.P.,Tang H.T.,Yang J.X.,Li X.,Chen D.Q.,Zhang B.,Shi Y.G.,and Huang Y.X.,2005,Identification and inheritance of resistance of maize germplasm to sheath blight(Rhizoctonia solani),Zhiwu Bingli Xuebao(Acta Phytopa Thologica Sinica),35(2):174-178(杨俊品,唐海涛,杨家秀,李晓,陈德全,张彪,石永刚,黄宜祥,2005,抗玉米纹枯病材料的鉴定及抗性遗传研究,植物病理学报,35(2):174-178)
Zhang S.S.,2015,Preliminary study on the MYB111 and MYB148 transcription factors regulating the phenylpropa noid metabolism in maize,Thesis for M.S.,Research Institute Sichuan Agricultural University,Supervisor:Huang Y.B.,pp.13-33(张爽爽,2015,玉米MYB111和MYB148转录因子调控苯丙烷代谢途径的初步研究,硕士学位论文,四川农业大学研究生院,导师:黄玉碧,pp.13-33)
Zhang X.,and Liu C.J.,2015,Multifaceted regulations of gateway enzyme phenylalanine ammonia-lyase in the biosynthesis of phenylpropanoids.,Mol.Plant,8(1):17-27
Zhang Y.,Butelli E.,and Martin C.,2014,Engineering anthocyanin biosynthesis in plants,Curr.Opin.Plant Biol.,19:81-90
Gai J.T.,Shen J.K.,and Wang P.,2016,Identification and sequence analysis of PAL gene family in major crops,Jiangsu Nongye Kexue(Agricultural Sciences of Jiangsu),44(6):45-49(盖江涛,沈建凯,王鹏,2016,主要作物中PAL基因家族的鉴定和序列分析,江苏农业科学,44(6):45-49)
Gao W.C.,Li Y.,Zhang Y.C.,Zhang Z.M.,Pan G.T.,and Lin H.J.,2016,Cloning of maize WRKY76 and its expression patterns involved in resistance to maize banded sheath blight,Guangxi Zhiwu(Guihaia),36(3):273-279(高文成,李玥,张银超,张志明,潘光堂,林海建,2016,玉米转录因子WRKY76克隆及抗纹枯病表达模式分析,广西植物,36(3):273-279)
Huang J.,Gu M.,Lai Z.,Fan B.,Shi K.,Zhou Y.H.,Yu J.Q.,and Chen Z.,2010,Functional analysis of the Arabidopsis PALgene family in plant growth,development,and response to environmental stress,Plant Physiol.,153(4):1526-1538
Jin Q.C.,Ye H.Z.,and Zhang M.,2003,Relationship between the activity of PAL and resistance of corn to maize sheath blight,Sichuan Nongye Daxue Xuebao(Journal of Sichuan Agricultural University),21(2):116-118(金庆超,叶华智,张敏,2003,苯丙氨酸解氨酶活性与玉米对纹枯病抗性的关系,四川农业大学学报,21(2):116-118)
Kawamata S.,Shimobarai K.,Imura Y.,Ozaki M.,Ichinose Y.,Shiraishi T.,Kunoh H.,and Yamada T.,1997,Temporal and spatial pattern of expression of the pea phenylalanine ammonia-lyase genel promoter in transgenic tobacco,Plant Cell Physiol.,38(7):792-803
La Camera S.,Gouzerh G.,Dhondt S.,Hoffmann L.,Fritig B.,Legrand M.,and Heitz T.,2004,Metabolic reprogramming in plant innate immunity:the contributions of phenylpropanoid and oxylipin pathways,Immunol.Rev.,198(1):267-284
Liu L.,Ma Y.Y.,Zhang Z.M.,Leng P.F.,Pan G.T.,and Zhao M.J.,2009,Analysis function of different defense enzymes on maize,Yumi Kexue(Journal of Maize Sciences),17(3):99-102,106(刘丽,马永毅,张志明,冷鹏飞,潘光堂,赵茂俊,2009,玉米不同防卫酶系对纹枯病作用的研究,玉米科学,17(3):99-102,106)
Ma Y.N.,Han C.,Chen J.Y.,Li H.Y.,He K.,Liu A.X.,and Li D.H.,2015,Fungal cellulase is an elicitor but its enzymatic activity is not required for its elicitor activity,Mol.Plant Pathol.,16(1):14-26
MacDonald M.J.,and D'Cunha G.B.,2007,A modern view of phenylalanine ammonia lyase,Biochem.and Cell Biol.,85(3):273-282
Santiago L.J.M.,Louro R.P.,and Oliveira D.E.D.,2000,Compartmentation of phenolic compounds and phenylalanine ammonia-lyase in leaves of Phyllanthus tenellus Roxb.and their induction by copper sulphate,Ann.Bot.,86(5):1023-1032
Sekhon R.S.,Lin H.,Childs K.L.,Hansey C.N.,Buell C.R.,De Leon N.,and and Kaeppler S.M.,2011,Genome-wide atlas of transcription during maize development,Plant J.,66(4):553-563
Sun H.Y.,Quan X.L.,Fu S.,Wu S.Q.,2016,Research progress of phenylalanine ammonia-lyase(PAL)gene in Arabidopsis thaliana,Yanbian Daxue Nongxue Xuebao(Agricultural Science Journal of Yanbian University),38(1):88-92(孙海燕,全雪丽,付爽,吴松权,2016,拟南芥苯丙氨酸解氨酶(PAL)基因的研究进展,延边大学农学学报,38(1):88-92)
Wang S.M.,Zhao X.L.,Ma Y.N.,and Liu A.X.,2016,Histological changes and expression of defense related genes of maize during the interaction with Rhizoctoniasolani AG1-IAYumi Kexue(Journal of Maize Sciences),24(5):37-42(王绍敏,赵新兰,马亚男,刘爱新,2016,Rhizoctoniasolan AG1-IA侵染玉米过程中组织病理学变化及防卫反应基因的表达,玉米科学,24(5):37-42)
Wang S.M.,Zhao X.L.,Ma Y.N.,and Liu A.X.,2017,Effects of different Rhizoctonia spp.on lognin metabolism,total phenols content and expression of defense related genes in maize,Gansu Nongye Daxuebao(Journal of Gansu Agricultural University),52(5):55-60(王绍敏,赵新兰,马亚男,刘爱新,2017,不同致病力Rhizoctonia菌株对玉米木质素代谢、总酚含量及相关基因表达的影响,甘肃农大学报,52(5):55-60)
Xu W.L.,Pei X.L.,Jing Z.G.,and Xiong Z.L.,2015,Genome-wide identification and expression analysis of chalcone synthase gene in pepper,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(8):1747-1752(徐婉莉,裴徐梨,荆赞革,熊自立,2015,辣椒查尔酮合成酶基因家族全基因组鉴定及表达特征分析,基因组学与应用生物学,34(8):1747-1752)
Yang J.P.,Tang H.T.,Yang J.X.,Li X.,Chen D.Q.,Zhang B.,Shi Y.G.,and Huang Y.X.,2005,Identification and inheritance of resistance of maize germplasm to sheath blight(Rhizoctonia solani),Zhiwu Bingli Xuebao(Acta Phytopa Thologica Sinica),35(2):174-178(杨俊品,唐海涛,杨家秀,李晓,陈德全,张彪,石永刚,黄宜祥,2005,抗玉米纹枯病材料的鉴定及抗性遗传研究,植物病理学报,35(2):174-178)
Zhang S.S.,2015,Preliminary study on the MYB111 and MYB148 transcription factors regulating the phenylpropa noid metabolism in maize,Thesis for M.S.,Research Institute Sichuan Agricultural University,Supervisor:Huang Y.B.,pp.13-33(张爽爽,2015,玉米MYB111和MYB148转录因子调控苯丙烷代谢途径的初步研究,硕士学位论文,四川农业大学研究生院,导师:黄玉碧,pp.13-33)
Zhang X.,and Liu C.J.,2015,Multifaceted regulations of gateway enzyme phenylalanine ammonia-lyase in the biosynthesis of phenylpropanoids.,Mol.Plant,8(1):17-27
Zhang Y.,Butelli E.,and Martin C.,2014,Engineering anthocyanin biosynthesis in plants,Curr.Opin.Plant Biol.,19:81-90