巴西橡胶树APX基因家族成员的鉴定及表达分析
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摘要
活性氧是一类含氧的化学活性小分子,会对细胞的结构和功能产生危害。抗坏血酸过氧化物酶(APX)是活性氧清除系统的关键成员之一,对于维持植物体内活性氧代谢平衡有着重要作用。通过生物信息学方法,本研究从橡胶树全基因组序列中鉴定出8个HbAPXs。进化树分析显示这些成员可归为2个亚类;基因结构结果显示每个成员至少含有4个内含子;转录组数据分析表明HbAPXs在橡胶树不同组织、叶片不同发育时期及乙烯利处理后不同阶段胶乳中的表达存在差异,其中HbAPX3和HbAPX8可能在叶片发育和胶乳活性氧代谢调控中起关键作用。橡胶树HbAPXs的鉴定为深入探索橡胶树生长发育及响应外界胁迫中的分子机制提供一定基础。
Reactive oxygen species(ROS) are a kind of chemically avtive small molecules containing oxygen, and harm to cell structure as well as function. Ascorbate peroxidase(APX) is a key member of ROS scavenging system, playing an important role in maintaining the balance of ROS metabolism in plant. In the present research, eight Hb APXs were identified from the whole genome sequence of rubber tree by using bioinformatics method. Phylogenetic tree analysis showed that the these members could be classified into two subgroups. The results of gene structure showed that each member contained at least four introns. Transcriptome data analysis showed that the expression pattern of Hb APXs was different in various tissues of Hevea brasiliensis, different development stages of leaves, different time-point of latex after ethephon treatment. Further analysis indicated Hb APX3 and Hb APX8 might be key members for leaf development and regulation of latex ROS metabolism. The identifying of Hb APXs would provide a basis for further exploring the the molecular mechanism of growth and development of Hevea brasiliensis and its response to stresses.
Reactive oxygen species(ROS) are a kind of chemically avtive small molecules containing oxygen, and harm to cell structure as well as function. Ascorbate peroxidase(APX) is a key member of ROS scavenging system, playing an important role in maintaining the balance of ROS metabolism in plant. In the present research, eight Hb APXs were identified from the whole genome sequence of rubber tree by using bioinformatics method. Phylogenetic tree analysis showed that the these members could be classified into two subgroups. The results of gene structure showed that each member contained at least four introns. Transcriptome data analysis showed that the expression pattern of Hb APXs was different in various tissues of Hevea brasiliensis, different development stages of leaves, different time-point of latex after ethephon treatment. Further analysis indicated Hb APX3 and Hb APX8 might be key members for leaf development and regulation of latex ROS metabolism. The identifying of Hb APXs would provide a basis for further exploring the the molecular mechanism of growth and development of Hevea brasiliensis and its response to stresses.
引文
Apel K.,and Hirt H.,2004,Reactive oxygen species:metabolism,oxidative stress,and signal transduction,Annu.Rev.Plant Biol.,55:373-399
Bailey-Serres J.,and Mittler R.,2006,The roles of reactive oxygen species in plant cells,Plant Physiol.,141(2):311
Caverzan A.,Bonifacio A.,Carvalho F.E.,Andrade C.M.,Passaia G.,Schünemann M.,Maraschin F.S.,Martins M.O.,Teixeira F.K.,Rauber R.,Margis R.,Silveira J.A.,and Margis-Pinheiro M.,2014,The knockdown of chloroplastic ascorbate peroxidases reveals its regulatory role in the photosynthesis and protection under photo-oxidative stress in rice,Plant Sci.,214:74-87
Chao J.Q.,Chen Y.Y.,Yang S.G.,and Tian W.M.,2016,Cloning and expressional analysis of HbCCoAOMT in rubber tree(Hevea brasiliensis Muell.Arg.),Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),37(12):1-7(晁金泉,陈月异,杨署光,田维敏,2016,巴西橡胶树HbCCoAOMT基因克隆及其表达分析,热带作物学报,37(12):1-7)
Chao J.,Yang S.,Chen Y.,and Tian W.M.,2016,Evaluation of reference genes for quantitative real-Time PCR analysis of the gene expression in laticifers on the basis of latex flow in rubber tree(Hevea brasiliensis Muell.Arg.),Front.Plant Sci.,7:1149
Chao J.,Zhang S.,Chen Y.,and Tian W.M.,2015,Cloning,heterologous expression and characterization of ascorbate peroxidase(APX)gene in laticifer cells of rubber tree(Hevea brasiliensis Muell.Arg.),Plant Physiol.Biochem.,97:331-338
Chew O.,Whelan J.,and Millar A.H.,2003,Molecular definition of the ascorbateglutathione cycle in Arabidopsis mitochondria reveals dual targeting of antioxidant defenses in plants,J.Biol.Chem.,278(47):46869-46877
Corpas F.J.,Barroso J.B.,and del Río L.A.,2001,Peroxisomes as a source of reactive oxygen species and nitric oxide signal molecules in plant cells,Trends Plant Sci.,6(4):145-150
Dauphinee A.N.,Fletcher J.I.,Denbigh G.L.,Lacroix C.R.,and Gunawardena A.H.L.A.N.,2017,Remodelling of lace plant leaves:antioxidants and ROS are key regulators of programmed cell death,Planta,246(1):133-147
Dou J.H.,Yu S.X.,Fan S.L.,Pang C.Y.,and Song M.Z.,2010,SOD and Plant Stress Resistance,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),8(2):359-364(窦俊辉,喻树迅,范术丽,庞朝友,宋美珍,2010,SOD与植物胁迫抗性,分子植物育种,8(2):359-364)
Fang Y.,Mei H.,Zhou B.,Xiao X.,Yang M.,Huang Y.,Long X.,Hu S.,and Tang C.,2016,De novo transcriptome analysis reveals distinct defense mechanisms by young and mature leaves of Hevea brasiliensis(Para Rubber Tree),Sci.Rep.,6:33151
Ishikawa T.,and Shigeoka S.,2008,Recent advances in ascorbate biosynthesis and the physiological significance of ascorbate peroxidase in photosynthesizing organisms,Biosci.Biotechnol.Biochem.,72(5):1143-1154
Khanna-Chopra R.,Jajoo A.,and Semwal V.K.,2011,Chloroplasts and mitochondria have multiple heat tolerant isozymes of SOD and APX in leaf and inflorescence in Chenopodium album,Biochem.and Biophys.Res.Commun.,412(4):522-525
Kovacs F.A.,Sarath G.,Woodworth K.,Twigg P.,and Tobias C.M.,2013,Abolishing activity against ascorbate in a cytosolic ascorbate peroxidase from switchgrass,Phytochemistry,94:45-52
Najami N.,Janda T.,Barriah W.,Kayam G.,Tal M.,Guy M.,and Volokita M.,2008,Ascorbate peroxidase gene family in tomato:its identification and characterization,Mol.Genet.Genomics,279(2):171-182
Narendra S.,Venkataramani S.,Shen G.,Wang J.,Pasapula V.,Lin Y.,Kornyeyev D.,Holaday A.S.,and Zhang H.,2006,The Arabidopsis ascorbate peroxidase 3 is a peroxisomal membrane-bound antioxidant enzyme and is dispensable for Arabidopsis growth and development,J.Exp.Bot.,57(12):3033-3042
Pandey S.,Fartyal D.,Agarwal A.,Shukla T.,James D.,Kaul T.,Negi Y.K.,Arora S.,and Reddy M.K.,2017,Abiotic stress tolerance in plants:myriad roles of ascorbate peroxidase,Front.Plant Sci.,8:581
Tang C.,Huang D.,Yang J.,Liu S.,Sakr S.,Li H.,Zhou Y.,and Qin Y.,2010,The sucrose transporter HbSUT3 plays an active role in sucrose loading to laticifer and rubber productivity in exploited trees of Hevea brasiliensis(para rubber tree),Plant Cell Environ.,33(10):1708-1720
Teixeira F.K.,Menezes-Benavente L.,Galv觔o V.C.,Margis R,and Margis-Pinheiro M.,2006,Rice ascorbate peroxidase gene family encodes functionally diverse isoforms localized in different subcellular compartments,Planta,224(2):300-314
Trapnell C.,Roberts A.,Goff L.,Pertea G.,Kim D.,Kelley D.R.,Pimentel H.,Salzberg S.L.,Rinn J.L.,and Pachter L.,2012,Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks,Nat.Protoc.,7(3):562-578
Wang Y.X.,Du J.Z.,Hao Y.S.,Zhang H.H.,Wang X.Q.,and Sun Y.,2017,Over-expression of GhAPX gene in the improvement of salt tolerance in Populus sp.,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(7):2579-2583(王亦学,杜建中,郝曜山,张欢欢,王晓清,孙毅,2017,过量表达GhAPX基因提高中林美荷杨耐盐性,分子植物育种,15(7):2579-2583)
Xiao X.Z.,Luo S.Q.,Xu W.X.,Wu X.D.,and Liu S.Z.,1999,Effect of microcut system on yield and physiological status of hevea clone PR107,Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),20(1):8-12(校现周,罗世巧,许闻献,魏小弟,刘实忠,1999,微割对橡胶无性系PR107产量及生理状况的影响,热带作物学报,20(1):8-12)
Yang C.,Li W.,Cao J.,Meng F.,Yu Y.,Huang J.,Jiang L.,Liu M.,Zhang Z.,Chen X.,Miyamoto K.,Yamane H.,Zhang J.,Chen S.,and Liu J.,2017,Activation of ethylene signaling pathways enhances disease resistance by regulating ROSand phytoalexin production in rice,Plant J.,89(2):338-353
Yao Y.,He R.J.,Xie Q.L.,Zhao X.H.,Deng X.M.,He J.B.,Song L.,He J.,Marchant A.,Chen X.Y.,and Wu A.M.,2017,ETHYLENE RESPONSE FACTOR 74(ERF74)plays an essential role in controlling a respiratory burst oxidase homolog D(RbohD)-dependent mechanism in response to different stresses in Arabidopsis,New Phytol.,213(4):1667-1681
Zhang H.,Li A.,Zhang Z.,Huang Z.,Lu P.,Zhang D.,Liu X.,Zhang Z.F.,and Huang R.,2016,Ethylene response factor TERF1,regulated by ETHYLENE-INSENSITIVE3-like factors,functions in reactive oxygen species(ROS)scavenging in Tobacco(Nicotiana tabacum L.),Sci.Rep.,6:29948
Zhu J.,and Zhang Z.,2009,Ethylene stimulation of latex production in Hevea brasiliensis,Plant Signal.Behav.,4(11):1072-1074
Zou Z.,Yang L.,Gong J.,Mo Y.,Wang J.,Cao J.,An F.,and Xie G.,2016,Genome-wide identification of Jatropha curcas aquaporin genes and the comparative analysis provides insights into the gene family expansion and evolution in Hevea brasiliensis,Front Plant Sci.,7:395
Bailey-Serres J.,and Mittler R.,2006,The roles of reactive oxygen species in plant cells,Plant Physiol.,141(2):311
Caverzan A.,Bonifacio A.,Carvalho F.E.,Andrade C.M.,Passaia G.,Schünemann M.,Maraschin F.S.,Martins M.O.,Teixeira F.K.,Rauber R.,Margis R.,Silveira J.A.,and Margis-Pinheiro M.,2014,The knockdown of chloroplastic ascorbate peroxidases reveals its regulatory role in the photosynthesis and protection under photo-oxidative stress in rice,Plant Sci.,214:74-87
Chao J.Q.,Chen Y.Y.,Yang S.G.,and Tian W.M.,2016,Cloning and expressional analysis of HbCCoAOMT in rubber tree(Hevea brasiliensis Muell.Arg.),Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),37(12):1-7(晁金泉,陈月异,杨署光,田维敏,2016,巴西橡胶树HbCCoAOMT基因克隆及其表达分析,热带作物学报,37(12):1-7)
Chao J.,Yang S.,Chen Y.,and Tian W.M.,2016,Evaluation of reference genes for quantitative real-Time PCR analysis of the gene expression in laticifers on the basis of latex flow in rubber tree(Hevea brasiliensis Muell.Arg.),Front.Plant Sci.,7:1149
Chao J.,Zhang S.,Chen Y.,and Tian W.M.,2015,Cloning,heterologous expression and characterization of ascorbate peroxidase(APX)gene in laticifer cells of rubber tree(Hevea brasiliensis Muell.Arg.),Plant Physiol.Biochem.,97:331-338
Chew O.,Whelan J.,and Millar A.H.,2003,Molecular definition of the ascorbateglutathione cycle in Arabidopsis mitochondria reveals dual targeting of antioxidant defenses in plants,J.Biol.Chem.,278(47):46869-46877
Corpas F.J.,Barroso J.B.,and del Río L.A.,2001,Peroxisomes as a source of reactive oxygen species and nitric oxide signal molecules in plant cells,Trends Plant Sci.,6(4):145-150
Dauphinee A.N.,Fletcher J.I.,Denbigh G.L.,Lacroix C.R.,and Gunawardena A.H.L.A.N.,2017,Remodelling of lace plant leaves:antioxidants and ROS are key regulators of programmed cell death,Planta,246(1):133-147
Dou J.H.,Yu S.X.,Fan S.L.,Pang C.Y.,and Song M.Z.,2010,SOD and Plant Stress Resistance,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),8(2):359-364(窦俊辉,喻树迅,范术丽,庞朝友,宋美珍,2010,SOD与植物胁迫抗性,分子植物育种,8(2):359-364)
Fang Y.,Mei H.,Zhou B.,Xiao X.,Yang M.,Huang Y.,Long X.,Hu S.,and Tang C.,2016,De novo transcriptome analysis reveals distinct defense mechanisms by young and mature leaves of Hevea brasiliensis(Para Rubber Tree),Sci.Rep.,6:33151
Ishikawa T.,and Shigeoka S.,2008,Recent advances in ascorbate biosynthesis and the physiological significance of ascorbate peroxidase in photosynthesizing organisms,Biosci.Biotechnol.Biochem.,72(5):1143-1154
Khanna-Chopra R.,Jajoo A.,and Semwal V.K.,2011,Chloroplasts and mitochondria have multiple heat tolerant isozymes of SOD and APX in leaf and inflorescence in Chenopodium album,Biochem.and Biophys.Res.Commun.,412(4):522-525
Kovacs F.A.,Sarath G.,Woodworth K.,Twigg P.,and Tobias C.M.,2013,Abolishing activity against ascorbate in a cytosolic ascorbate peroxidase from switchgrass,Phytochemistry,94:45-52
Najami N.,Janda T.,Barriah W.,Kayam G.,Tal M.,Guy M.,and Volokita M.,2008,Ascorbate peroxidase gene family in tomato:its identification and characterization,Mol.Genet.Genomics,279(2):171-182
Narendra S.,Venkataramani S.,Shen G.,Wang J.,Pasapula V.,Lin Y.,Kornyeyev D.,Holaday A.S.,and Zhang H.,2006,The Arabidopsis ascorbate peroxidase 3 is a peroxisomal membrane-bound antioxidant enzyme and is dispensable for Arabidopsis growth and development,J.Exp.Bot.,57(12):3033-3042
Pandey S.,Fartyal D.,Agarwal A.,Shukla T.,James D.,Kaul T.,Negi Y.K.,Arora S.,and Reddy M.K.,2017,Abiotic stress tolerance in plants:myriad roles of ascorbate peroxidase,Front.Plant Sci.,8:581
Tang C.,Huang D.,Yang J.,Liu S.,Sakr S.,Li H.,Zhou Y.,and Qin Y.,2010,The sucrose transporter HbSUT3 plays an active role in sucrose loading to laticifer and rubber productivity in exploited trees of Hevea brasiliensis(para rubber tree),Plant Cell Environ.,33(10):1708-1720
Teixeira F.K.,Menezes-Benavente L.,Galv觔o V.C.,Margis R,and Margis-Pinheiro M.,2006,Rice ascorbate peroxidase gene family encodes functionally diverse isoforms localized in different subcellular compartments,Planta,224(2):300-314
Trapnell C.,Roberts A.,Goff L.,Pertea G.,Kim D.,Kelley D.R.,Pimentel H.,Salzberg S.L.,Rinn J.L.,and Pachter L.,2012,Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks,Nat.Protoc.,7(3):562-578
Wang Y.X.,Du J.Z.,Hao Y.S.,Zhang H.H.,Wang X.Q.,and Sun Y.,2017,Over-expression of GhAPX gene in the improvement of salt tolerance in Populus sp.,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),15(7):2579-2583(王亦学,杜建中,郝曜山,张欢欢,王晓清,孙毅,2017,过量表达GhAPX基因提高中林美荷杨耐盐性,分子植物育种,15(7):2579-2583)
Xiao X.Z.,Luo S.Q.,Xu W.X.,Wu X.D.,and Liu S.Z.,1999,Effect of microcut system on yield and physiological status of hevea clone PR107,Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),20(1):8-12(校现周,罗世巧,许闻献,魏小弟,刘实忠,1999,微割对橡胶无性系PR107产量及生理状况的影响,热带作物学报,20(1):8-12)
Yang C.,Li W.,Cao J.,Meng F.,Yu Y.,Huang J.,Jiang L.,Liu M.,Zhang Z.,Chen X.,Miyamoto K.,Yamane H.,Zhang J.,Chen S.,and Liu J.,2017,Activation of ethylene signaling pathways enhances disease resistance by regulating ROSand phytoalexin production in rice,Plant J.,89(2):338-353
Yao Y.,He R.J.,Xie Q.L.,Zhao X.H.,Deng X.M.,He J.B.,Song L.,He J.,Marchant A.,Chen X.Y.,and Wu A.M.,2017,ETHYLENE RESPONSE FACTOR 74(ERF74)plays an essential role in controlling a respiratory burst oxidase homolog D(RbohD)-dependent mechanism in response to different stresses in Arabidopsis,New Phytol.,213(4):1667-1681
Zhang H.,Li A.,Zhang Z.,Huang Z.,Lu P.,Zhang D.,Liu X.,Zhang Z.F.,and Huang R.,2016,Ethylene response factor TERF1,regulated by ETHYLENE-INSENSITIVE3-like factors,functions in reactive oxygen species(ROS)scavenging in Tobacco(Nicotiana tabacum L.),Sci.Rep.,6:29948
Zhu J.,and Zhang Z.,2009,Ethylene stimulation of latex production in Hevea brasiliensis,Plant Signal.Behav.,4(11):1072-1074
Zou Z.,Yang L.,Gong J.,Mo Y.,Wang J.,Cao J.,An F.,and Xie G.,2016,Genome-wide identification of Jatropha curcas aquaporin genes and the comparative analysis provides insights into the gene family expansion and evolution in Hevea brasiliensis,Front Plant Sci.,7:395