铝胁迫下金沙柚3个WRKY基因的生物信息学与表达分析
|
摘要
WRKY是植物中特有的具有锌指结构的转录因子家族之一,在植物非生物逆境胁迫应答中起着重要的调控作用。为研究金沙柚WRKY在铝胁迫条件下的作用,研究在前期构建的金沙柚铝胁迫转录组数据基础上,利用生物信息学方法从金沙柚根中得到了能显著响应铝胁迫的3个WRKY基因(Cg4g001470、Cg7g003890和Cg6g010690),开放阅读框长度分别为966、933和993 bp,各编码321、310和330个氨基酸,分别命名为CgWRKY40、CgWRKY70-1和CgWRKY70-2,氨基酸序列和结构分析显示,这3个基因编码的蛋白分别属于WRKY蛋白的IIa、III a和III a类型,PSORT亚细胞定位预测均定位于细胞核。实时荧光定量PCR分析表明,3个WRKY基因在金沙柚的嫩根、老根、茎和叶的表达量存在较大差异,在老根的表达量均最高;在铝胁迫处理后,均能出现不同程度的上调表达,相对表达量分别为处理前的4.58、1.52和12.06倍。在铝胁迫条件下,水杨酸能较大幅度的调控3个WRKY基因的上调表达,分别为对照的133.80、33.39和114.23倍;过氧化氢调控3个WRKY基因出现显著上调表达,分别为对照的86.40倍、7.93倍和102.32倍;水杨酸与过氧化氢共同作用下能调控3个WRKY基因出现不同程度的上调表达,分别为对照的51.81倍、1.51倍、37.72倍。上述研究表明,3个WRKY基因均能参与铝胁迫响应,且均可以被水杨酸、过氧化氢、水杨酸与过氧化氢调控上调表达。
WRKY is a large family of plant?specific transcription factors with zinc finger motif,which plays an important role in regulating plantresponse to abiotic stress.In this study,3 WRKY genes(Cg4 g001470,Cg7 g003890 and Cg6 g010690 were identified,named CgWRKY40,CgWRKY70-1 and CgWRKY70-2),which could significantly respond to aluminum stress,using bioinformatics method from transcriptome database of Jinsha pomelo.The open reading frame lengths of the three WRKY genes were 966,933 and 993 bp,and en?coded 321,310 and 330 amino acids,which belonged to WRKY protein type IIa,III,and III a,respectively.PSORT predicted that these three proteins were all localized in the nucleus.Real-time fluorescent quantitative PCR analysis showed that the expression levels of the three WRKY genes varied in the tender roots,old roots,stems and leaves of Jinsha pomelo tissues,and the highest expression level was in the old roots.The expression pattern of the three genes were various after aluminum stress treatment,and up-regulated to 4.58,1.52 and4.58 times that of the control.The three genes could be up-regulated by salicylic acid under aluminum stress,and the relative expression increased up to 133.80,33.39 and 114.23 times that of the control,respectively.All the three genes could be up-regulated by hydrogen peroxide under aluminum stress,and their relative expres?sion increased up to 86.40,7.93,and 102.32 times that of the control.The three genes could be up-regulated by salicylic acid and hydrogen peroxide under aluminum stress,and their relative expression increased up to51.81,1.51,37.72 times that of the control.The research results showed that the three WRKY genes were in?volved in aluminum stress response,and could be up-regulated by salicylic acid,hydrogen peroxide,salicylic acid and hydrogen peroxide.
WRKY is a large family of plant?specific transcription factors with zinc finger motif,which plays an important role in regulating plantresponse to abiotic stress.In this study,3 WRKY genes(Cg4 g001470,Cg7 g003890 and Cg6 g010690 were identified,named CgWRKY40,CgWRKY70-1 and CgWRKY70-2),which could significantly respond to aluminum stress,using bioinformatics method from transcriptome database of Jinsha pomelo.The open reading frame lengths of the three WRKY genes were 966,933 and 993 bp,and en?coded 321,310 and 330 amino acids,which belonged to WRKY protein type IIa,III,and III a,respectively.PSORT predicted that these three proteins were all localized in the nucleus.Real-time fluorescent quantitative PCR analysis showed that the expression levels of the three WRKY genes varied in the tender roots,old roots,stems and leaves of Jinsha pomelo tissues,and the highest expression level was in the old roots.The expression pattern of the three genes were various after aluminum stress treatment,and up-regulated to 4.58,1.52 and4.58 times that of the control.The three genes could be up-regulated by salicylic acid under aluminum stress,and the relative expression increased up to 133.80,33.39 and 114.23 times that of the control,respectively.All the three genes could be up-regulated by hydrogen peroxide under aluminum stress,and their relative expres?sion increased up to 86.40,7.93,and 102.32 times that of the control.The three genes could be up-regulated by salicylic acid and hydrogen peroxide under aluminum stress,and their relative expression increased up to51.81,1.51,37.72 times that of the control.The research results showed that the three WRKY genes were in?volved in aluminum stress response,and could be up-regulated by salicylic acid,hydrogen peroxide,salicylic acid and hydrogen peroxide.
引文
[1]Eulgem T,Rushton P J,Robatzek S,et al.The WRKY superfamily of plant transcription factors[J].Trends in Plant Science,2000,5(5):199-206.
[2]郝中娜,陶荣祥.WRKY转录因子超家族的研究[J].生命科学,2006,18(2):175-179.Hao Z N,Tao R X.Study of WRKY transcription factor superfamily[J].Chinese Bulletin of Life Sciences,2006,18(2):175-179.
[3]Rushton P J,Somssich I E,Ringler P,et al.WRKY transcription factors[J].Plant Signaling&Behavior,2010,15(5):247-258.
[4]Ulker B,Somssich I E.WRKY transcription factors:from DNA binding towards biological function[J].Current Opinion inPlant Biology,2004,7(5):491-498.
[5]Zhang Y,Wang L.The WRKY transcription factor superfamily:its origin in eukaryotes and expansion in plants[J].Bmc Evo?lutionary Biology,2005,5(1):1-12.
[6]Chen L,Song Y,Li S,et al.The role of WRKY transcription factors in plant abiotic stresses[J].Biochim Biophys Acta,2012,1819(2):120-128.
[7]Chen H,Lai Z,Shi J,et al.Roles of Arabidopsis WRKY18,WRKY40 and WRKY60 transcription factors in plant responsesto abscisic acid and abiotic stress[J].Bmc Plant Biology,2010,10(1):281.
[8]Vives-Peris V,Marmaneu D,Gómez-Cadenas A,et al.Characterization of citrus,WRKY transcription factors and their re?sponses to phytohormones and abiotic stresses[J].Biologia Plantarum,2017:1-12.
[9]Jiang Y,Deyholos M K.Functional characterization of Arabidopsis NaCl-inducible WRKY25 and WRKY33 transcriptionfactors in abiotic stresses[J].Plant Molecular Biology,2009,69(1-2):91-105.
[10]Gong X,Zhang J,Hu J,et al.FcWRKY70,a WRKY protein of Fortunella crassifolia,functions in drought tolerance and mod?ulates putrescine synthesis by regulating arginine decarboxylase gene[J].Plant Cell&Environment,2015,38(11):2248-2262.
[11]Dang F F,Wang Y N,Yu L,et al.CaWRKY40,a WRKY protein of pepper,plays an important role in the regulation of toler?ance to heat stress and resistance to Ralstonia solanacearum infection[J].Plant Cell&Environment,2013,36(4):757-774.
[12]Mehtap?ahin-?evik,Gloria A Moore.Identification of a drought-and cold-stress inducible WRKY gene in the cold-hardyCitrus relative Poncirus trifoliata[J].New Zealand Journal of Crop&Horticultural Science,2013,41(2):57-68.
[13]Zhou L,Wang N N,Gong S Y,et al.Overexpression of a cotton(Gossypium hirsutum)WRKY gene,GhWRKY34,in Arabi?dopsis,enhances salt-tolerance of the transgenic plants[J].Plant Physiology&Biochemistry,2015,96:311-320.
[14]Wang X,Zeng J,Li Y,et al.Expression of TaWRKY44,a wheat WRKY gene,in transgenic tobacco confers multiple abioticstress tolerances[J].Front Plant Sci,2015,6:615.
[15]周鹏飞,贾瑞瑞,陈善春,等.柑橘4个WRKY转录因子基因的克隆及其响应柑橘溃疡病菌侵染的表达分析[J].园艺学报,2017,44(3):452-462.Zhou P F,Jia R R,Chen S C,et al.Cloning and expression analysis of four citrus WRKY Genes responding to Xanthomonasaxonopodis pv. citri[J].Acta Horticulturae Sinica,2017,44(3):452-462.
[16]Sun L M,Pei Y X,Liu Z Q.Relationship between H_2S signal and WRKY in ABA regulating root growth and stomatal move?ment[J].Bulletin of Botanical Research,2016,36(1):97-104.
[17]Gong X Q,Hu J B,Liu J H.Cloning and characterization of FcWRKY40,A WRKY transcription factor from Fortunella crassi?folia,linked to oxidative stress tolerance[J].Plant Cell Tissue&Organ Culture,2014,119(1):1-14.
[18]Li S J,Yin X R,Wang W L,et al.Citrus CitNAC62 cooperates with CitWRKY1 to participate in citric acid degradation viaup-regulation of CitAco3[J].Journal of Experimental Botany,2017,68(13):3419.
[19]Chen L S.Physiological responses and tolerance of citrus to aluminum toxicity[J].Journal of Plant Physiology and MolecularBiology,2006,32(2):143-55.
[20]Lin Z,Myhre D L.Differential response of citrus rootstocks to aluminum levels in nutrient solutions:I.Plant growth[J].Jour?nal of Plant Nutrition,1991a,14:1223-1238.
[21]Lin Z,Myhre D L.Differential response of citrus rootstocks to aluminum levels in nutrient solutions:II.Plant mineral concen?trations[J].Journal of Soil Science and Plant Nutrition,1991b,14:1239-1254.
[22]Yang L T,Jiang H X,Tang N,et al.Mechanisms of aluminum-tolerance in two species of citrus:secretion of organic acid an?ions and immobilization of aluminum by phosphorus in roots[J].Plant Science,2011,180:521-530.
[23]Wang X,Xu Y,Zhang S,et al.Genomic analyses of primitive,wild and cultivated citrus provide insights into asexual repro?duction[J].Nature Genetics,2017,49(5):765-772.
[24]Yang L T,Qi Y P,Jiang H X,et al.Roles of organic acid anion secretion in aluminium tolerance of higher plants[J].BiomedResearch International,2012,2013(1):173682.
[25]Guo P,Qi Y P,Yang L T,et al.Root adaptive responses to aluminum-treatment revealed by RNA-Seq in two citrus specieswith different aluminum-tolerance[J].Frontiers in Plant Science,2017,8(210).
[26]李学鑫,白志川,岳欢,等.甜橙WRKY转录因子序列的生物信息学分析[J].北方园艺,2013,(1):119-123.Li X X,Bai Z C,Yue H,et al.Bioinformatics analysis of orange WRKY transcription factor sequence[J].Northern Horticul?ture,2013,(1):119-123.
[27]Silva S.Aluminium toxicity targets in plants[J].Journal of Botany,2012,2012:1-8.
[28]Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Del?ta C(T))Method[J].Methods,2001,25(4):402-408.
[29]Poschenrieder C,Gunse B,Corrales I,et al.A glance into aluminum toxicity and resistance in plants[J].Science of the TotalEnvironment,2008,400(1-3):356-368.
[30]Arunakumara K K I U,Walpola B C,Yoon M H.How do citrus crops cope with aluminum toxicity?[J].Korean Journal ofSoil Science and Fertilizer,2012,45(6):928-935.
[31]Chen R F,Zhang F L,Zhang Q M,et al.Aluminium–phosphorus interactions in plants growing on acid soils:does phospho?rus always alleviate aluminium toxicity[J].Journal of the Science of Food and Agriculture,2012,92(5):995-1000.
[32]张觅,罗小英,白文钦,等.香橙苹果酸脱氢酶CjMDH基因表达特性及转基因烟草的耐铝性分析[J].园艺学报,2008,12:1751-1758.Zhang M,Luo X Y,Bai W Q,et al.Characterization of malate dehydrogenase gene from citrus junos and its transgenic tobac?co’s tolerance to aluminium toxicity[J].Acta Horticulturae Sinica,2008,12:1751-1758.
[33]向小华,吴新儒,晁江涛,等.普通烟草WRKY基因家族的鉴定及表达分析[J].遗传,2016,38(9):840-856.Xiang X H,Wu X R,Chao J T,et al.Genome wide identification and expression analysis of the WRKY gene family in com?mon tobacco(Nicotiana tabacum L.)[J].Hereditas,2016,38(9):840-856.
[34]Hong Y,Wang M,Liu S,et al.Transcriptome-wide identification and expression profiles of the WRKY transcription factorfamily in Broomcorn millet(Panicum miliaceum L.)[J].Bmc Genomics,2016,17(1):343.
[35]Fan X,Guo Q,Xu P,et al.Transcriptome-Wide identification of salt-responsive members of the WRKY gene family in Gossyp?ium aridum[J].Plos One,2015,10(5):e0126148.
[36]Ali M A,Azeem F,Nawaz M A,et al.Transcription factors WRKY11 and WRKY17 are involved in abiotic stress responses inArabidopsis[J].Journal of Plant Physiology,2018,226:12.
[37]Guo R,Qiao H,Zhao J,et al.The grape VlWRKY3 gene promotes abiotic and biotic stress tolerance in transgenic Arabidopsisthaliana[J].Frontiers in Plant Science,2018,9.
[38]Okay S,Derelli E,Unver T.Transcriptome-wide identification of bread wheat WRKY transcription factors in response todrought stress[J].Molecular Genetics&Genomics Mgg,2014,289(5):765-781.
[39]Zhou Q Y,Tian A G,Zou H F,et al.Soybean WRKY-type transcription factor genes,GmWRKY13,GmWRKY21,and Gm?WRKY54,confer differential tolerance to abiotic stresses in transgenic Arabidopsis plants[J].Plant Biotechnology Journal,2008,6(5):486-503.
[40]Yang G Y,Zhang W H,Sun Y D,et al.Two novel WRKY genes from Juglans regia,JrWRKY6,and JrWRKY53,are involvedin abscisic acid-dependent stress responses[J].Biologia Plantarum,2017,61(4):611-621.
[41]Jiang Y,Deyholos M K.Comprehensive transcriptional profiling of NaCl-stressed Arabidopsis roots reveals novel classes ofresponsive genes[J].Bmc Plant Biology,2006,6(1):25.
[42]Wei Y,Shi H,Xia Z,et al.Genome-Wide identification and expression analysis of the WRKY gene gamily in Cassava[J].Front Plant Sci,2016,7(2):25.
[43]Dou L,Zhang X,Pang C,et al.Genome-wide analysis of the WRKY gene family in cotton[J].Molecular Genetics&Genom?ics,2014,289(6):1103-1121.
[44]Yanbing G,Zhirui J,Fumei C,et al.Genome-wide identification and expression analysis of the WRKY gene family in peach[J].Yi Chuan,2016,38(3):254-270.
[45]Jiang J,Ma S,Ye N,et al.WRKY transcription factors in plant responses to stresses[J].Journal of Integrative Plant Biology,2016,59(2):86.
[46]Cheng Y,Yao Z,Meiying R,et al.Function of WRKY transcription factors in plant responses to biotic/abiotic stresses[J].Molecular Plant Breeding,2016,14(11):3214-3223.
[47]Banerjee A,Roychoudhury A.WRKY proteins:signaling and regulation of expression during abiotic stress responses[J].TheScientific World Journal,2015(807560):807560.
[48]Deng-Wei J,Sang X L,Shu B,et al.Expression analysis of maize WRKY transcription factor genes under abiotic stress[J].Guangdong Agricultural Sciences,2017,44(1):15-22.
[49]Xin P F,Gao C S,Cheng C H,et al.Identification and characterization of hemp WRKY transcription factors in response toabiotic stresses[J].Biologia Plantarum,2016,60(3):489-495.
[50]Tang Y,Kuang J F,Wang F Y,et al.Molecular characterization of PR,and WRKY,genes during SA-and MeJA-induced re?sistance against Colletotrichum musae,in banana fruit[J].Postharvest Biology&Technology,2013,79(1):62-68.
[51]Zhang J Y,Tong Z G,Gao Z H,et al.Expression of MhWRKY1 gene induced by the elicitors SA,MeJA,ACC and the applering spot pathogen[J].Scientia Agricultura Sinica,2011,44(5):990-999.
[52]Costa A,Drago I,Behera S,et al.H2O2in plant peroxisomes:an in vivo analysis uncovers a Ca2+-dependent scavenging sys?tem[J].Plant Journal for Cell&Molecular Biology,2010,62(5):760-772.
[53]Scarpeci T E,Zanor M I,Carrillo N,et al.Generation of superoxide anion in chloroplasts of Arabidopsis thaliana during ac?tive photosynthesis:a focus on rapidly induced genes[J].Plant Molecular Biology,2008,66(4):361-378.
[54]Scarpeci T E,Zanor M I,Muellerroeber B,et al.Overexpression of AtWRKY30 enhances abiotic stress tolerance during earlygrowth stages in Arabidopsis thaliana[J].Plant Molecular Biology,2013,83(3):265-277.
[55]Babitha K C,Ramu S V,Pruthvi V,et al.Co-expression of AtbHLH17 and AtWRKY28 confers resistance to abiotic stress inArabidopsis[J].Transgenic Research,2013,22(2):327-341.
[56]Lei Z,Liu G,Meng X,et al.A WRKY,gene from Tamarix hispida,ThWRKY4,mediates abiotic stress responses by modulat?ing reactive oxygen species and expression of stress-responsive genes[J].Plant Molecular Biology,2013,82(4-5):303-320.
[57]Liu B,Hong Y B,Zhang Y F,et al.Tomato WRKY transcriptional factor SlDRW1 is required for disease resistance againstBotrytis cinerea and tolerance to oxidative stress[J].Plant Science an International Journal of Experimental Plant Biology,2014,227(5):145-156.
[2]郝中娜,陶荣祥.WRKY转录因子超家族的研究[J].生命科学,2006,18(2):175-179.Hao Z N,Tao R X.Study of WRKY transcription factor superfamily[J].Chinese Bulletin of Life Sciences,2006,18(2):175-179.
[3]Rushton P J,Somssich I E,Ringler P,et al.WRKY transcription factors[J].Plant Signaling&Behavior,2010,15(5):247-258.
[4]Ulker B,Somssich I E.WRKY transcription factors:from DNA binding towards biological function[J].Current Opinion inPlant Biology,2004,7(5):491-498.
[5]Zhang Y,Wang L.The WRKY transcription factor superfamily:its origin in eukaryotes and expansion in plants[J].Bmc Evo?lutionary Biology,2005,5(1):1-12.
[6]Chen L,Song Y,Li S,et al.The role of WRKY transcription factors in plant abiotic stresses[J].Biochim Biophys Acta,2012,1819(2):120-128.
[7]Chen H,Lai Z,Shi J,et al.Roles of Arabidopsis WRKY18,WRKY40 and WRKY60 transcription factors in plant responsesto abscisic acid and abiotic stress[J].Bmc Plant Biology,2010,10(1):281.
[8]Vives-Peris V,Marmaneu D,Gómez-Cadenas A,et al.Characterization of citrus,WRKY transcription factors and their re?sponses to phytohormones and abiotic stresses[J].Biologia Plantarum,2017:1-12.
[9]Jiang Y,Deyholos M K.Functional characterization of Arabidopsis NaCl-inducible WRKY25 and WRKY33 transcriptionfactors in abiotic stresses[J].Plant Molecular Biology,2009,69(1-2):91-105.
[10]Gong X,Zhang J,Hu J,et al.FcWRKY70,a WRKY protein of Fortunella crassifolia,functions in drought tolerance and mod?ulates putrescine synthesis by regulating arginine decarboxylase gene[J].Plant Cell&Environment,2015,38(11):2248-2262.
[11]Dang F F,Wang Y N,Yu L,et al.CaWRKY40,a WRKY protein of pepper,plays an important role in the regulation of toler?ance to heat stress and resistance to Ralstonia solanacearum infection[J].Plant Cell&Environment,2013,36(4):757-774.
[12]Mehtap?ahin-?evik,Gloria A Moore.Identification of a drought-and cold-stress inducible WRKY gene in the cold-hardyCitrus relative Poncirus trifoliata[J].New Zealand Journal of Crop&Horticultural Science,2013,41(2):57-68.
[13]Zhou L,Wang N N,Gong S Y,et al.Overexpression of a cotton(Gossypium hirsutum)WRKY gene,GhWRKY34,in Arabi?dopsis,enhances salt-tolerance of the transgenic plants[J].Plant Physiology&Biochemistry,2015,96:311-320.
[14]Wang X,Zeng J,Li Y,et al.Expression of TaWRKY44,a wheat WRKY gene,in transgenic tobacco confers multiple abioticstress tolerances[J].Front Plant Sci,2015,6:615.
[15]周鹏飞,贾瑞瑞,陈善春,等.柑橘4个WRKY转录因子基因的克隆及其响应柑橘溃疡病菌侵染的表达分析[J].园艺学报,2017,44(3):452-462.Zhou P F,Jia R R,Chen S C,et al.Cloning and expression analysis of four citrus WRKY Genes responding to Xanthomonasaxonopodis pv. citri[J].Acta Horticulturae Sinica,2017,44(3):452-462.
[16]Sun L M,Pei Y X,Liu Z Q.Relationship between H_2S signal and WRKY in ABA regulating root growth and stomatal move?ment[J].Bulletin of Botanical Research,2016,36(1):97-104.
[17]Gong X Q,Hu J B,Liu J H.Cloning and characterization of FcWRKY40,A WRKY transcription factor from Fortunella crassi?folia,linked to oxidative stress tolerance[J].Plant Cell Tissue&Organ Culture,2014,119(1):1-14.
[18]Li S J,Yin X R,Wang W L,et al.Citrus CitNAC62 cooperates with CitWRKY1 to participate in citric acid degradation viaup-regulation of CitAco3[J].Journal of Experimental Botany,2017,68(13):3419.
[19]Chen L S.Physiological responses and tolerance of citrus to aluminum toxicity[J].Journal of Plant Physiology and MolecularBiology,2006,32(2):143-55.
[20]Lin Z,Myhre D L.Differential response of citrus rootstocks to aluminum levels in nutrient solutions:I.Plant growth[J].Jour?nal of Plant Nutrition,1991a,14:1223-1238.
[21]Lin Z,Myhre D L.Differential response of citrus rootstocks to aluminum levels in nutrient solutions:II.Plant mineral concen?trations[J].Journal of Soil Science and Plant Nutrition,1991b,14:1239-1254.
[22]Yang L T,Jiang H X,Tang N,et al.Mechanisms of aluminum-tolerance in two species of citrus:secretion of organic acid an?ions and immobilization of aluminum by phosphorus in roots[J].Plant Science,2011,180:521-530.
[23]Wang X,Xu Y,Zhang S,et al.Genomic analyses of primitive,wild and cultivated citrus provide insights into asexual repro?duction[J].Nature Genetics,2017,49(5):765-772.
[24]Yang L T,Qi Y P,Jiang H X,et al.Roles of organic acid anion secretion in aluminium tolerance of higher plants[J].BiomedResearch International,2012,2013(1):173682.
[25]Guo P,Qi Y P,Yang L T,et al.Root adaptive responses to aluminum-treatment revealed by RNA-Seq in two citrus specieswith different aluminum-tolerance[J].Frontiers in Plant Science,2017,8(210).
[26]李学鑫,白志川,岳欢,等.甜橙WRKY转录因子序列的生物信息学分析[J].北方园艺,2013,(1):119-123.Li X X,Bai Z C,Yue H,et al.Bioinformatics analysis of orange WRKY transcription factor sequence[J].Northern Horticul?ture,2013,(1):119-123.
[27]Silva S.Aluminium toxicity targets in plants[J].Journal of Botany,2012,2012:1-8.
[28]Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Del?ta C(T))Method[J].Methods,2001,25(4):402-408.
[29]Poschenrieder C,Gunse B,Corrales I,et al.A glance into aluminum toxicity and resistance in plants[J].Science of the TotalEnvironment,2008,400(1-3):356-368.
[30]Arunakumara K K I U,Walpola B C,Yoon M H.How do citrus crops cope with aluminum toxicity?[J].Korean Journal ofSoil Science and Fertilizer,2012,45(6):928-935.
[31]Chen R F,Zhang F L,Zhang Q M,et al.Aluminium–phosphorus interactions in plants growing on acid soils:does phospho?rus always alleviate aluminium toxicity[J].Journal of the Science of Food and Agriculture,2012,92(5):995-1000.
[32]张觅,罗小英,白文钦,等.香橙苹果酸脱氢酶CjMDH基因表达特性及转基因烟草的耐铝性分析[J].园艺学报,2008,12:1751-1758.Zhang M,Luo X Y,Bai W Q,et al.Characterization of malate dehydrogenase gene from citrus junos and its transgenic tobac?co’s tolerance to aluminium toxicity[J].Acta Horticulturae Sinica,2008,12:1751-1758.
[33]向小华,吴新儒,晁江涛,等.普通烟草WRKY基因家族的鉴定及表达分析[J].遗传,2016,38(9):840-856.Xiang X H,Wu X R,Chao J T,et al.Genome wide identification and expression analysis of the WRKY gene family in com?mon tobacco(Nicotiana tabacum L.)[J].Hereditas,2016,38(9):840-856.
[34]Hong Y,Wang M,Liu S,et al.Transcriptome-wide identification and expression profiles of the WRKY transcription factorfamily in Broomcorn millet(Panicum miliaceum L.)[J].Bmc Genomics,2016,17(1):343.
[35]Fan X,Guo Q,Xu P,et al.Transcriptome-Wide identification of salt-responsive members of the WRKY gene family in Gossyp?ium aridum[J].Plos One,2015,10(5):e0126148.
[36]Ali M A,Azeem F,Nawaz M A,et al.Transcription factors WRKY11 and WRKY17 are involved in abiotic stress responses inArabidopsis[J].Journal of Plant Physiology,2018,226:12.
[37]Guo R,Qiao H,Zhao J,et al.The grape VlWRKY3 gene promotes abiotic and biotic stress tolerance in transgenic Arabidopsisthaliana[J].Frontiers in Plant Science,2018,9.
[38]Okay S,Derelli E,Unver T.Transcriptome-wide identification of bread wheat WRKY transcription factors in response todrought stress[J].Molecular Genetics&Genomics Mgg,2014,289(5):765-781.
[39]Zhou Q Y,Tian A G,Zou H F,et al.Soybean WRKY-type transcription factor genes,GmWRKY13,GmWRKY21,and Gm?WRKY54,confer differential tolerance to abiotic stresses in transgenic Arabidopsis plants[J].Plant Biotechnology Journal,2008,6(5):486-503.
[40]Yang G Y,Zhang W H,Sun Y D,et al.Two novel WRKY genes from Juglans regia,JrWRKY6,and JrWRKY53,are involvedin abscisic acid-dependent stress responses[J].Biologia Plantarum,2017,61(4):611-621.
[41]Jiang Y,Deyholos M K.Comprehensive transcriptional profiling of NaCl-stressed Arabidopsis roots reveals novel classes ofresponsive genes[J].Bmc Plant Biology,2006,6(1):25.
[42]Wei Y,Shi H,Xia Z,et al.Genome-Wide identification and expression analysis of the WRKY gene gamily in Cassava[J].Front Plant Sci,2016,7(2):25.
[43]Dou L,Zhang X,Pang C,et al.Genome-wide analysis of the WRKY gene family in cotton[J].Molecular Genetics&Genom?ics,2014,289(6):1103-1121.
[44]Yanbing G,Zhirui J,Fumei C,et al.Genome-wide identification and expression analysis of the WRKY gene family in peach[J].Yi Chuan,2016,38(3):254-270.
[45]Jiang J,Ma S,Ye N,et al.WRKY transcription factors in plant responses to stresses[J].Journal of Integrative Plant Biology,2016,59(2):86.
[46]Cheng Y,Yao Z,Meiying R,et al.Function of WRKY transcription factors in plant responses to biotic/abiotic stresses[J].Molecular Plant Breeding,2016,14(11):3214-3223.
[47]Banerjee A,Roychoudhury A.WRKY proteins:signaling and regulation of expression during abiotic stress responses[J].TheScientific World Journal,2015(807560):807560.
[48]Deng-Wei J,Sang X L,Shu B,et al.Expression analysis of maize WRKY transcription factor genes under abiotic stress[J].Guangdong Agricultural Sciences,2017,44(1):15-22.
[49]Xin P F,Gao C S,Cheng C H,et al.Identification and characterization of hemp WRKY transcription factors in response toabiotic stresses[J].Biologia Plantarum,2016,60(3):489-495.
[50]Tang Y,Kuang J F,Wang F Y,et al.Molecular characterization of PR,and WRKY,genes during SA-and MeJA-induced re?sistance against Colletotrichum musae,in banana fruit[J].Postharvest Biology&Technology,2013,79(1):62-68.
[51]Zhang J Y,Tong Z G,Gao Z H,et al.Expression of MhWRKY1 gene induced by the elicitors SA,MeJA,ACC and the applering spot pathogen[J].Scientia Agricultura Sinica,2011,44(5):990-999.
[52]Costa A,Drago I,Behera S,et al.H2O2in plant peroxisomes:an in vivo analysis uncovers a Ca2+-dependent scavenging sys?tem[J].Plant Journal for Cell&Molecular Biology,2010,62(5):760-772.
[53]Scarpeci T E,Zanor M I,Carrillo N,et al.Generation of superoxide anion in chloroplasts of Arabidopsis thaliana during ac?tive photosynthesis:a focus on rapidly induced genes[J].Plant Molecular Biology,2008,66(4):361-378.
[54]Scarpeci T E,Zanor M I,Muellerroeber B,et al.Overexpression of AtWRKY30 enhances abiotic stress tolerance during earlygrowth stages in Arabidopsis thaliana[J].Plant Molecular Biology,2013,83(3):265-277.
[55]Babitha K C,Ramu S V,Pruthvi V,et al.Co-expression of AtbHLH17 and AtWRKY28 confers resistance to abiotic stress inArabidopsis[J].Transgenic Research,2013,22(2):327-341.
[56]Lei Z,Liu G,Meng X,et al.A WRKY,gene from Tamarix hispida,ThWRKY4,mediates abiotic stress responses by modulat?ing reactive oxygen species and expression of stress-responsive genes[J].Plant Molecular Biology,2013,82(4-5):303-320.
[57]Liu B,Hong Y B,Zhang Y F,et al.Tomato WRKY transcriptional factor SlDRW1 is required for disease resistance againstBotrytis cinerea and tolerance to oxidative stress[J].Plant Science an International Journal of Experimental Plant Biology,2014,227(5):145-156.