水稻茉莉酸甲酯和白叶枯病菌诱导型启动子的克隆及功能鉴定
|
摘要
诱导型启动子能灵活、精确的调控外源基因的表达,推动了植物基因工程技术的发展。在前期研究中,我们发现了一个萜类合成酶基因LOC_Os02g36140,该基因的表达受茉莉酸甲酯(MeJA)诱导。在本研究中,我们通过PCR技术分离了该基因5'端的2 272 bp启动子序列,命名为PRJA140,构建了PRJA140启动子驱动gusA基因表达的融合载体,转化水稻后对该基因的启动子活性进行分析。MeJA和白叶枯病菌处理转基因水稻后发现,和对照相比,转基因水稻根和叶片中GUS基因的表达明显上调,说明PRJA140启动子能被MeJA和白叶枯病菌诱导,是一个诱导型启动子。
Inducible promoter can flexibly and accurately regulate the expression of exogenous genes and promote the development of plant genetic engineering technology. In the previous study, we found a terpene synthase gene LOC_Os02g36140, the expression of which was induced by methyl jasmonate(MeJA). In this study,we isolated the 2272 bp promoter sequence at the 5' terminal of LOC_Os02g36140 by PCR technology, which was named as PRJA140. In addition, a fusion vector driven by PRJA140 promoter to drive gus A gene expression was constructed, and the promoter activity of this gene was analyzed after rice transformation. After the transgenic rice was treated with MeJA and bacterial blight, we found that the expression of GUS in leaves and roots of the transgenic rice was obviously increased when compared with the control, indicating the PRJA140 promoter could be induced by MeJA and bacterial blight and it might be an inducible promoter.
Inducible promoter can flexibly and accurately regulate the expression of exogenous genes and promote the development of plant genetic engineering technology. In the previous study, we found a terpene synthase gene LOC_Os02g36140, the expression of which was induced by methyl jasmonate(MeJA). In this study,we isolated the 2272 bp promoter sequence at the 5' terminal of LOC_Os02g36140 by PCR technology, which was named as PRJA140. In addition, a fusion vector driven by PRJA140 promoter to drive gus A gene expression was constructed, and the promoter activity of this gene was analyzed after rice transformation. After the transgenic rice was treated with MeJA and bacterial blight, we found that the expression of GUS in leaves and roots of the transgenic rice was obviously increased when compared with the control, indicating the PRJA140 promoter could be induced by MeJA and bacterial blight and it might be an inducible promoter.
引文
Bailey-Serres J.,and Ma W.B.,2017,Plant biology:an immunity boost combats crop disease,Nature,545:420-421
Boter M.,Ruiz-Rivero O.,Abdeen A.,and Prat S.,2004,Conserved MYC transcription factors play a key role in jasmonate signaling both in tomato and Arabidopsis,Genes&Development,18:1577-1591
Brown R.L.,Kazan K.,Mc Grath K.C.,Maclean D.J.,and Manners J.M.,2003,A role for the GCC-box in jasmonate-mediated activation of the PDF1.2 gene of Arabidopsis,Plant Physiol.,132(2):1020-1032
Bruce T.J.,Matthes M.C.,Chamberlain K.,Woodcock C.M.,Mohib A.,Webster B.,Smart L.E.,Birkett M.A.,Pickett J A.,and Napier J.A.,2008,cis-Jasmone induces Arabidopsis genes that affect the chemical ecology of multitrophic interactions with aphids and their parasitoids,Proc.Natil.Acad.Sci.USA,105(12):4553-4558
DenancéN.,Sánchez-Vallet A.,Goffner D.,and Molina A.,2013,Disease resistance or growth:the role of plant hormones in balancing immune responses and fitness costs,Frontiers in Plant Science,4:155
Fernandez-Calvo P.,Chini A.,Fernandez-Barbero G.,Chico J.M.,Gimenez-Ibanez S.,Geerinck J.,Eeckhout D.,Schweizer F.,Godoy M.,Franco-Zorrilla J.M.,Pauwels L.,Witters E.,Puga M.I.,Paz-Ares J.,Goossens A.,Reymond P.,De Jaeger G.,and Solano R.,2011,The Arabidopsis b HLHtranscription factors MYC3 and MYC4 are targets of JAZrepressors and act additively with MYC2 in the activation of jasmonate responses,The Plant Cell,23(2):701-715
Gao L.,Cao Y.,Xia Z.,Jiang G.,Liu G.,Zhang W.,and Zhai W.,2013,Do transgenesis and marker-assisted backcross breeding produce substantially equivalent plants?-A comparative study of transgenic and backcross rice carrying bacterial blight resistant gene Xa21,BMC Genomics,14:738
Hiei Y.,Ohta S.,Komari T.,and Kumashiro T.,1994,Efficient transformation of rice(Oryza sativa L.)mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA,Plant J.,6(2):271-282
Kauffman S.A.,1973,Control circuits for determination and transdetermination,Science,181(4097):310-318
Lescot M.,Dehais P.,Thijs G.,Marchal K.,Moreau Y.,Van de Peer Y.,Rouze P.,and Rombauts S.,2002,PlantC ARE,a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences,Nucleic Acids Res.,30(1):325-327
Li T.,Chen X.,Zhong X.,Zhao Y.,Liu X.,Zhou S.,Cheng S.,and Zhou D.X.,2013,Jumonji C domain protein JMJ705-mediated removal of histone H3 lysine 27 trimethylation is involved in defense-related gene activation in rice,The Plant Cell,25(11):4725-4736
Memelink J.,2009,Regulation of gene expression by jasmonate hormones,Phytochemistry,70(13-14):1560-1570
Miyamoto K.,Shimizu T.,Lin F.,Sainsbury F.,Thuenemann E.,Lomonossoff G.,Nojiri H.,Yamane H.,and Okada K.,2012,Identification of an E-box motif responsible for the expression of jasmonic acid-induced chitinase gene Os Chia4a in rice,J.Plant Physiol.,169(6):621-627
Nakashima K.,Tran L.S.,Van Nguyen D.,Fujita M.,Maruyama K.,Todaka D.,Ito Y.,Hayashi N.,Shinozaki K.,and Yamaguchi-Shinozaki K.,2007,Functional analysis of a NAC-type transcription factor OsN AC6 involved in abiotic and biotic stress-responsive gene expression in rice,Plant J.,51(4):617-630
Oh S.J.,Song S.I.,Kim Y.S.,Jang H.J.,Kim S.Y.,Kim M.,Kim Y.K.,Nahm B.H.,and Kim J.K.,2005,Arabidopsis CBF3/DREB1A and ABF3 in transgenic rice increased tolerance to abiotic stress without stunting growth,Plant Physiol.,138(1):341-351
Qi T.,Song S.,Ren Q.,Wu D.,Huang H.,Chen Y.,Fan M.,Peng W.,Ren C.,and Xie D.,2011,The Jasmonate-ZIM-domain proteins interact with the WD-Repeat/b HLH/MYB complexes to regulate Jasmonate-mediated anthocyanin accumulation and trichome initiation in Arabidopsis thaliana,The Plant Cell,23(5):1795-1814
Thines B.,Katsir L.,Melotto M.,Niu Y.,Mandaokar A.,Liu G.,Nomura K.,He S.Y.,Howe G.A.,and Browse J.,2007,JAZrepressor proteins are targets of the SCFCOI1complex during jasmonate signalling,Nature,448(7154):661-665
Tian D.,Traw M.B.,Chen J.Q.,Kreitman M.,and Bergelson J.,2003,Fitness costs of R-gene-mediated resistance in Arabidopsis thaliana,Nature,423(6935):74-77
Uji Y.,Taniguchi S.,Tamaoki D.,Shishido H.,Akimitsu K.,and Gomi K.,2016,Overexpression of Os MYC2 results in the up-regulation of early JA-rresponsive genes and bacterial blight resistance in rice,Plant&Cell Physiology,57(9):1814-1827
Wunn J.,Kloti A.,Burkhardt P.K.,Biswas G.C.,Launis K.,Iglesias V.A.,and Potrykus I.,1996,Transgenic indica rice breeding line IR58 expressing a synthetic cry IA(b)gene from Bacillus thuringiensis provides effective insect pest control,Biotechnology(NY),14(2):171-176
Xu G.,Yuan M.,Ai C.,Liu L.,Zhuang E.,Karapetyan S.,Wang S.,and Dong X.,2017,u ORF-mediated translation allows engineered plant disease resistance without fitness costs,Nature,545:491-494
Yamada S.,Kano A.,Tamaoki D.,Miyamoto A.,Shishido H.,Miyoshi S.,Taniguchi S.,Akimitsu K.,and Gomi K.,2012,Involvement of OsJ AZ8 in jasmonate-induced resistance to bacterial blight in rice,Plant&Cell Physiology,53(12):2060-2072
Zhang X.,Li M.J.,Zhang B.,Yin X.M.,Wang M.L.,and Xia X.J.,2016,Possibility study on improving salt tolerance of rice by overexpressing PHD-finger transcription factor gene Os Msr16,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(7):1820-1827(张新,李明娟,张斌,殷绪明,王曼玲,夏新界,2016,水稻PHD-finger转录因子基因Os Msr16增强耐盐性的可能性研究,基因组学与应用生物学,35(7):1820-1827)
Zheng H.,Lin S.,Zhang Q.,Lei Y.,Hou L.,and Zhang Z.,2010,Functional identification and regulation of the Pt Drl02 gene promoter from triploid white poplar,Plant Cell Reports,29(5):449-460
Boter M.,Ruiz-Rivero O.,Abdeen A.,and Prat S.,2004,Conserved MYC transcription factors play a key role in jasmonate signaling both in tomato and Arabidopsis,Genes&Development,18:1577-1591
Brown R.L.,Kazan K.,Mc Grath K.C.,Maclean D.J.,and Manners J.M.,2003,A role for the GCC-box in jasmonate-mediated activation of the PDF1.2 gene of Arabidopsis,Plant Physiol.,132(2):1020-1032
Bruce T.J.,Matthes M.C.,Chamberlain K.,Woodcock C.M.,Mohib A.,Webster B.,Smart L.E.,Birkett M.A.,Pickett J A.,and Napier J.A.,2008,cis-Jasmone induces Arabidopsis genes that affect the chemical ecology of multitrophic interactions with aphids and their parasitoids,Proc.Natil.Acad.Sci.USA,105(12):4553-4558
DenancéN.,Sánchez-Vallet A.,Goffner D.,and Molina A.,2013,Disease resistance or growth:the role of plant hormones in balancing immune responses and fitness costs,Frontiers in Plant Science,4:155
Fernandez-Calvo P.,Chini A.,Fernandez-Barbero G.,Chico J.M.,Gimenez-Ibanez S.,Geerinck J.,Eeckhout D.,Schweizer F.,Godoy M.,Franco-Zorrilla J.M.,Pauwels L.,Witters E.,Puga M.I.,Paz-Ares J.,Goossens A.,Reymond P.,De Jaeger G.,and Solano R.,2011,The Arabidopsis b HLHtranscription factors MYC3 and MYC4 are targets of JAZrepressors and act additively with MYC2 in the activation of jasmonate responses,The Plant Cell,23(2):701-715
Gao L.,Cao Y.,Xia Z.,Jiang G.,Liu G.,Zhang W.,and Zhai W.,2013,Do transgenesis and marker-assisted backcross breeding produce substantially equivalent plants?-A comparative study of transgenic and backcross rice carrying bacterial blight resistant gene Xa21,BMC Genomics,14:738
Hiei Y.,Ohta S.,Komari T.,and Kumashiro T.,1994,Efficient transformation of rice(Oryza sativa L.)mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA,Plant J.,6(2):271-282
Kauffman S.A.,1973,Control circuits for determination and transdetermination,Science,181(4097):310-318
Lescot M.,Dehais P.,Thijs G.,Marchal K.,Moreau Y.,Van de Peer Y.,Rouze P.,and Rombauts S.,2002,PlantC ARE,a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences,Nucleic Acids Res.,30(1):325-327
Li T.,Chen X.,Zhong X.,Zhao Y.,Liu X.,Zhou S.,Cheng S.,and Zhou D.X.,2013,Jumonji C domain protein JMJ705-mediated removal of histone H3 lysine 27 trimethylation is involved in defense-related gene activation in rice,The Plant Cell,25(11):4725-4736
Memelink J.,2009,Regulation of gene expression by jasmonate hormones,Phytochemistry,70(13-14):1560-1570
Miyamoto K.,Shimizu T.,Lin F.,Sainsbury F.,Thuenemann E.,Lomonossoff G.,Nojiri H.,Yamane H.,and Okada K.,2012,Identification of an E-box motif responsible for the expression of jasmonic acid-induced chitinase gene Os Chia4a in rice,J.Plant Physiol.,169(6):621-627
Nakashima K.,Tran L.S.,Van Nguyen D.,Fujita M.,Maruyama K.,Todaka D.,Ito Y.,Hayashi N.,Shinozaki K.,and Yamaguchi-Shinozaki K.,2007,Functional analysis of a NAC-type transcription factor OsN AC6 involved in abiotic and biotic stress-responsive gene expression in rice,Plant J.,51(4):617-630
Oh S.J.,Song S.I.,Kim Y.S.,Jang H.J.,Kim S.Y.,Kim M.,Kim Y.K.,Nahm B.H.,and Kim J.K.,2005,Arabidopsis CBF3/DREB1A and ABF3 in transgenic rice increased tolerance to abiotic stress without stunting growth,Plant Physiol.,138(1):341-351
Qi T.,Song S.,Ren Q.,Wu D.,Huang H.,Chen Y.,Fan M.,Peng W.,Ren C.,and Xie D.,2011,The Jasmonate-ZIM-domain proteins interact with the WD-Repeat/b HLH/MYB complexes to regulate Jasmonate-mediated anthocyanin accumulation and trichome initiation in Arabidopsis thaliana,The Plant Cell,23(5):1795-1814
Thines B.,Katsir L.,Melotto M.,Niu Y.,Mandaokar A.,Liu G.,Nomura K.,He S.Y.,Howe G.A.,and Browse J.,2007,JAZrepressor proteins are targets of the SCFCOI1complex during jasmonate signalling,Nature,448(7154):661-665
Tian D.,Traw M.B.,Chen J.Q.,Kreitman M.,and Bergelson J.,2003,Fitness costs of R-gene-mediated resistance in Arabidopsis thaliana,Nature,423(6935):74-77
Uji Y.,Taniguchi S.,Tamaoki D.,Shishido H.,Akimitsu K.,and Gomi K.,2016,Overexpression of Os MYC2 results in the up-regulation of early JA-rresponsive genes and bacterial blight resistance in rice,Plant&Cell Physiology,57(9):1814-1827
Wunn J.,Kloti A.,Burkhardt P.K.,Biswas G.C.,Launis K.,Iglesias V.A.,and Potrykus I.,1996,Transgenic indica rice breeding line IR58 expressing a synthetic cry IA(b)gene from Bacillus thuringiensis provides effective insect pest control,Biotechnology(NY),14(2):171-176
Xu G.,Yuan M.,Ai C.,Liu L.,Zhuang E.,Karapetyan S.,Wang S.,and Dong X.,2017,u ORF-mediated translation allows engineered plant disease resistance without fitness costs,Nature,545:491-494
Yamada S.,Kano A.,Tamaoki D.,Miyamoto A.,Shishido H.,Miyoshi S.,Taniguchi S.,Akimitsu K.,and Gomi K.,2012,Involvement of OsJ AZ8 in jasmonate-induced resistance to bacterial blight in rice,Plant&Cell Physiology,53(12):2060-2072
Zhang X.,Li M.J.,Zhang B.,Yin X.M.,Wang M.L.,and Xia X.J.,2016,Possibility study on improving salt tolerance of rice by overexpressing PHD-finger transcription factor gene Os Msr16,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(7):1820-1827(张新,李明娟,张斌,殷绪明,王曼玲,夏新界,2016,水稻PHD-finger转录因子基因Os Msr16增强耐盐性的可能性研究,基因组学与应用生物学,35(7):1820-1827)
Zheng H.,Lin S.,Zhang Q.,Lei Y.,Hou L.,and Zhang Z.,2010,Functional identification and regulation of the Pt Drl02 gene promoter from triploid white poplar,Plant Cell Reports,29(5):449-460