水稻OsWRKY78转录因子响应盐胁迫的表达与功能研究
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摘要
以OsWRKY78基因及其相应的RNAi转基因水稻为研究对象,分析OsWRKY78转录因子响应盐胁迫的表达和功能,研究WRKY转录因子参与水稻耐盐的机制。结果表明:水稻OsWRKY78基因启动子中存在30多个与非生物胁迫相关的顺式调控元件。基因表达和GUS组织化学染色分析表明OsWRKY78的表达受盐诱导。抑制OsWRKY78基因表达可显著增强水稻在种子萌发和小苗生长阶段的耐盐性,一定程度上是通过调节OsLEA3、OsRAB21等与逆境相关基因的表达来实现的。
To study the mechanism of WRKY transcription factor involved in salt tolerance of rice, OsWRKY78 gene and its RNAi transgenic rice were used for analyzing the expression and functions of OsWRKY78 in response to salt stress. Sequence analysis indicated that more than thirty cis-elements related to abiotic stresses existed in OsWRKY78 promoter. Expression and GUS histochemical assays demonstrated that the transcription of OsWRKY78 was induced by salt. Moreover, repression of OsWRKY78 expression could enhance rice salt tolerance remarkably during seed germination and seedling stages, which might be partly because of the expression regulation of a number of stress-related genes, including OsLEA3 and OsRAB21.
To study the mechanism of WRKY transcription factor involved in salt tolerance of rice, OsWRKY78 gene and its RNAi transgenic rice were used for analyzing the expression and functions of OsWRKY78 in response to salt stress. Sequence analysis indicated that more than thirty cis-elements related to abiotic stresses existed in OsWRKY78 promoter. Expression and GUS histochemical assays demonstrated that the transcription of OsWRKY78 was induced by salt. Moreover, repression of OsWRKY78 expression could enhance rice salt tolerance remarkably during seed germination and seedling stages, which might be partly because of the expression regulation of a number of stress-related genes, including OsLEA3 and OsRAB21.
引文
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[2] PEETHAMBARAN P K,GLENZ R,HONINGER S,et al.Salt-inducible expression of OsJAZ8 improves resilience against salt-stress [J].BMC Plant Biol,2018,18(1):311.
[3] JAMIL A,RIAZ S,ASHRAF M,et al.Gene expression profiling of plants under salt stress [J].Crit Rev Plant Sci,2011,30(5):435-458.
[4] KRISHNAMURTHY S L,SHARMA P C,BATRA V,et al.Effect of salinity and use of stress indices of morphological and physiological traits at the seedling stage in rice [J].Indian J Exp Biol,2016,54(12):843-850.
[5] LI J,BESSEAU S,TORONEN P,et al.Defense-related transcription factors WRKY70 and WRKY54 modulate osmotic stress tolerance by regulating stomatal aperture in Arabidopsis [J].New Phytol,2013,200(2):457-472.
[6] TRIPATHI P,RABARA R C,RUSHTON P J.A systems biology perspective on the role of WRKY transcription factors in drought responses in plants [J].Planta,2014,239(2):255-266.
[7] VIANA V E,BUSANELLO C,DA MAIA L C,et al.Activation of rice WRKY transcription factors:an army of stress fighting soldiers[J].Curr Opin Plant Biol,2018,45:268-275.
[8] ZHOU Q Y,TIAN A G,ZOU H F,et al.Soybean WRKY-type transcription factor genes,GmWRKY13,GmWRKY21,and GmWRKY54,confer differential tolerance to abiotic stresses in transgenic Arabidopsis plants.GmWRKY54 conferred salt and drought tolerance [J].Plant Biotechnol J,2008,6(5):486-503.
[9] SONG H,WANG P,HOU L,et al.Global analysis of WRKY genes and their response to dehydration and salt stress in soybean [J].Front Plant Sci,2016,7:9.
[10] SHI W Y,DU Y T,MA J,et al.The WRKY transcription factor GmWRKY12 confers drought and salt tolerance in soybean [J].Int J Mol Sci,2018,19(12):E4087.
[11] XU Z,RAZA Q,XU L,et al.GmWRKY49,a salt-responsive nuclear protein,improved root length and governed better salinity tolerance in transgenic Arabidopsis [J].Front Plant Sci,2018,9:809.
[12] NIU C F,WEI W,ZHOU Q Y,et al.Wheat WRKY genes TaWRKY2 and TaWRKY19 regulate abiotic stress tolerance in transgenic Arabidopsis plants [J].Plant Cell Environ,2012,35(6):1156-1170.
[13] WANG C,DENG P,CHEN L,et al.A wheat WRKY transcription factor TaWRKY10 confers tolerance to multiple abiotic stresses in transgenic tobacco [J].PLos ONE,2013,8(6):e65120.
[14] WANG X,ZENG J,LI Y,et al.Expression of TaWRKY44,a wheat WRKY gene,in transgenic tobacco confers multiple abiotic stress tolerances [J].Front Plant Sci,2015,6:615.
[15] YOUSFI F E,MAKHLOUFI E,MARANDE W,et al.Comparative analysis of WRKY genes potentially involved in salt stress responses in Triticum turgidum L.ssp.durum [J].Front Plant Sci,2017,7:2034.
[16] SHI W,HAO L,LI J,et al.The Gossypium hirsutum WRKY gene GhWRKY39-1 promotes pathogen infection defense responses and mediates salt stress tolerance in transgenic Nicotiana benthamiana[J].Plant Cell Rep,2014,33(3):483-498.
[17] YAN H,JIA H,CHEN X,et al.The cotton WRKY transcription factor GhWRKY17 functions in drought and salt stress in transgenic Nicotiana benthamiana through ABA signaling and the modulation of reactive oxygen species production [J].Plant Cell Physiol,2014,55(12):2060-2076.
[18] ZHOU L,WANG N N,GONG S Y,et al.Overexpression of a cotton (Gossypium hirsutum) WRKY gene,GhWRKY34,in Arabidopsis enhances salt-tolerance of the transgenic plants [J].Plant Physiol Biochem,2015,96:311-320.
[19] CHU X,WANG C,CHEN X,et al.The cotton WRKY gene GhWRKY41 positively regulates saltand drought stress tolerance in transgenic Nicotiana benthamiana [J].PLoS ONE,2015,10(11):e0143022.
[20] CHINNUSAMY V,JAGENDORF A,ZHU J K.Understanding and improving salt tolerance in plants [J].Crop Sci,2005,45:437-448.
[21] 周根友,翟彩娇,邓先亮,等.盐逆境对水稻产量、光合特性及品质的影响 [J].中国水稻科学,2018,32(2):146-154.
[22] QIU Y,YU D.Over-expression of the stress-induced OsWRKY45 enhances disease resistance and drought tolerance in Arabidopsis [J].Environ Exp Bot,2009,65(1):35-47.
[23] PILLAI S E,KUMAR C,PATEL H K,et al.Overexpression of a cell wall damage induced transcription factor,OsWRKY42,leads to enhanced callose deposition and tolerance to salt stress but does not enhance tolerance to bacterial infection [J].BMC Plant Biol,2018,18(1):177.
[24] GANGULY M,DATTA K,ROYCHOUDHURY A,et al.Overexpression of Rab16A gene in indica rice variety for generating enhanced salt tolerance [J].Plant Signal Behav,2012,7(4):502-509.
[25] XIAO B,HUANG Y,TANG N,et al.Over-expression of a LEA gene in rice improves drought resistance under the field conditions [J].Theor Appl Genet,2007,115(1):35-46.
[26] YAO L,CHENG X,GU Z,et al.The AWPM-19 family protein OsPM1 mediates abscisic acid influx and drought response in rice [J].Plant Cell,2018,30(6):1258-1276.
[27] FENG X J,LI J R,QI S L,et al.Light affects salt stress-induced transcriptional memory of P5CS1 in Arabidopsis [J].Proc Natl Acad Sci USA,2016,113(51):E8335-E8343.
[28] ZHANG C Q,XU Y,LU Y,et al.The WRKY transcription factor OsWRKY78 regulates stem elongation and seed development in rice [J].Planta,2011,234(3):541-554.
[29] JIANG J,MA S,YE N,et al.WRKY transcription factors in plant responses to stresses [J].J Integr Plant Biol,2017,59(2):86-101.
[30] VIANA V E,BUSANELLD C,DA MAIA L C,et al.Activation of rice WRKY transcription factors:an army of stress fighting soldiers [J].Curr Opin Plant Biol,2018,45:268-275.
[31] SHEN H,LIU C,ZHANG Y,et al.OsWRKY30 is activated by MAP kinases to confer drought tolerance in rice [J].Plant Mol Bio,2012,80(3):241-253.
[32] TAO Z,KOU Y,LIU H,et al.OsWRKY45 alleles playdifferent roles in abscisic acid signalling and salt stress tolerance but similar roles in drought and cold tolerance in rice [J].J Exp Bot,2011,62(14):4863-4874.
[33] PENG X,WANG H,JANG J C,et al.OsWRKY80-OsWRKY4 module as a positive regulatory circuitin rice resistance against Rhizoctonia solani [J].Rice,2016,9(1):63.
[34] HUANG F J,LI J,LI R,et al.The transcription factor OsWRKY45 negatively modulates the resistance of rice to the brown plant hopper Nilapar vatalugens [J].Int J Mol Sci,2016,17(6):697.
[35] BAO W,WANG X,CHEN M,et al.A WRKY transcription factor,PcWRKY33,from Polygonum cuspidatum reduces salt tolerance in transgenic Arabidopsis thaliana [J].Plant Cell Rep,2018,37(7):1033-1048.
[2] PEETHAMBARAN P K,GLENZ R,HONINGER S,et al.Salt-inducible expression of OsJAZ8 improves resilience against salt-stress [J].BMC Plant Biol,2018,18(1):311.
[3] JAMIL A,RIAZ S,ASHRAF M,et al.Gene expression profiling of plants under salt stress [J].Crit Rev Plant Sci,2011,30(5):435-458.
[4] KRISHNAMURTHY S L,SHARMA P C,BATRA V,et al.Effect of salinity and use of stress indices of morphological and physiological traits at the seedling stage in rice [J].Indian J Exp Biol,2016,54(12):843-850.
[5] LI J,BESSEAU S,TORONEN P,et al.Defense-related transcription factors WRKY70 and WRKY54 modulate osmotic stress tolerance by regulating stomatal aperture in Arabidopsis [J].New Phytol,2013,200(2):457-472.
[6] TRIPATHI P,RABARA R C,RUSHTON P J.A systems biology perspective on the role of WRKY transcription factors in drought responses in plants [J].Planta,2014,239(2):255-266.
[7] VIANA V E,BUSANELLO C,DA MAIA L C,et al.Activation of rice WRKY transcription factors:an army of stress fighting soldiers[J].Curr Opin Plant Biol,2018,45:268-275.
[8] ZHOU Q Y,TIAN A G,ZOU H F,et al.Soybean WRKY-type transcription factor genes,GmWRKY13,GmWRKY21,and GmWRKY54,confer differential tolerance to abiotic stresses in transgenic Arabidopsis plants.GmWRKY54 conferred salt and drought tolerance [J].Plant Biotechnol J,2008,6(5):486-503.
[9] SONG H,WANG P,HOU L,et al.Global analysis of WRKY genes and their response to dehydration and salt stress in soybean [J].Front Plant Sci,2016,7:9.
[10] SHI W Y,DU Y T,MA J,et al.The WRKY transcription factor GmWRKY12 confers drought and salt tolerance in soybean [J].Int J Mol Sci,2018,19(12):E4087.
[11] XU Z,RAZA Q,XU L,et al.GmWRKY49,a salt-responsive nuclear protein,improved root length and governed better salinity tolerance in transgenic Arabidopsis [J].Front Plant Sci,2018,9:809.
[12] NIU C F,WEI W,ZHOU Q Y,et al.Wheat WRKY genes TaWRKY2 and TaWRKY19 regulate abiotic stress tolerance in transgenic Arabidopsis plants [J].Plant Cell Environ,2012,35(6):1156-1170.
[13] WANG C,DENG P,CHEN L,et al.A wheat WRKY transcription factor TaWRKY10 confers tolerance to multiple abiotic stresses in transgenic tobacco [J].PLos ONE,2013,8(6):e65120.
[14] WANG X,ZENG J,LI Y,et al.Expression of TaWRKY44,a wheat WRKY gene,in transgenic tobacco confers multiple abiotic stress tolerances [J].Front Plant Sci,2015,6:615.
[15] YOUSFI F E,MAKHLOUFI E,MARANDE W,et al.Comparative analysis of WRKY genes potentially involved in salt stress responses in Triticum turgidum L.ssp.durum [J].Front Plant Sci,2017,7:2034.
[16] SHI W,HAO L,LI J,et al.The Gossypium hirsutum WRKY gene GhWRKY39-1 promotes pathogen infection defense responses and mediates salt stress tolerance in transgenic Nicotiana benthamiana[J].Plant Cell Rep,2014,33(3):483-498.
[17] YAN H,JIA H,CHEN X,et al.The cotton WRKY transcription factor GhWRKY17 functions in drought and salt stress in transgenic Nicotiana benthamiana through ABA signaling and the modulation of reactive oxygen species production [J].Plant Cell Physiol,2014,55(12):2060-2076.
[18] ZHOU L,WANG N N,GONG S Y,et al.Overexpression of a cotton (Gossypium hirsutum) WRKY gene,GhWRKY34,in Arabidopsis enhances salt-tolerance of the transgenic plants [J].Plant Physiol Biochem,2015,96:311-320.
[19] CHU X,WANG C,CHEN X,et al.The cotton WRKY gene GhWRKY41 positively regulates saltand drought stress tolerance in transgenic Nicotiana benthamiana [J].PLoS ONE,2015,10(11):e0143022.
[20] CHINNUSAMY V,JAGENDORF A,ZHU J K.Understanding and improving salt tolerance in plants [J].Crop Sci,2005,45:437-448.
[21] 周根友,翟彩娇,邓先亮,等.盐逆境对水稻产量、光合特性及品质的影响 [J].中国水稻科学,2018,32(2):146-154.
[22] QIU Y,YU D.Over-expression of the stress-induced OsWRKY45 enhances disease resistance and drought tolerance in Arabidopsis [J].Environ Exp Bot,2009,65(1):35-47.
[23] PILLAI S E,KUMAR C,PATEL H K,et al.Overexpression of a cell wall damage induced transcription factor,OsWRKY42,leads to enhanced callose deposition and tolerance to salt stress but does not enhance tolerance to bacterial infection [J].BMC Plant Biol,2018,18(1):177.
[24] GANGULY M,DATTA K,ROYCHOUDHURY A,et al.Overexpression of Rab16A gene in indica rice variety for generating enhanced salt tolerance [J].Plant Signal Behav,2012,7(4):502-509.
[25] XIAO B,HUANG Y,TANG N,et al.Over-expression of a LEA gene in rice improves drought resistance under the field conditions [J].Theor Appl Genet,2007,115(1):35-46.
[26] YAO L,CHENG X,GU Z,et al.The AWPM-19 family protein OsPM1 mediates abscisic acid influx and drought response in rice [J].Plant Cell,2018,30(6):1258-1276.
[27] FENG X J,LI J R,QI S L,et al.Light affects salt stress-induced transcriptional memory of P5CS1 in Arabidopsis [J].Proc Natl Acad Sci USA,2016,113(51):E8335-E8343.
[28] ZHANG C Q,XU Y,LU Y,et al.The WRKY transcription factor OsWRKY78 regulates stem elongation and seed development in rice [J].Planta,2011,234(3):541-554.
[29] JIANG J,MA S,YE N,et al.WRKY transcription factors in plant responses to stresses [J].J Integr Plant Biol,2017,59(2):86-101.
[30] VIANA V E,BUSANELLD C,DA MAIA L C,et al.Activation of rice WRKY transcription factors:an army of stress fighting soldiers [J].Curr Opin Plant Biol,2018,45:268-275.
[31] SHEN H,LIU C,ZHANG Y,et al.OsWRKY30 is activated by MAP kinases to confer drought tolerance in rice [J].Plant Mol Bio,2012,80(3):241-253.
[32] TAO Z,KOU Y,LIU H,et al.OsWRKY45 alleles playdifferent roles in abscisic acid signalling and salt stress tolerance but similar roles in drought and cold tolerance in rice [J].J Exp Bot,2011,62(14):4863-4874.
[33] PENG X,WANG H,JANG J C,et al.OsWRKY80-OsWRKY4 module as a positive regulatory circuitin rice resistance against Rhizoctonia solani [J].Rice,2016,9(1):63.
[34] HUANG F J,LI J,LI R,et al.The transcription factor OsWRKY45 negatively modulates the resistance of rice to the brown plant hopper Nilapar vatalugens [J].Int J Mol Sci,2016,17(6):697.
[35] BAO W,WANG X,CHEN M,et al.A WRKY transcription factor,PcWRKY33,from Polygonum cuspidatum reduces salt tolerance in transgenic Arabidopsis thaliana [J].Plant Cell Rep,2018,37(7):1033-1048.