铜胁迫对不同抗性种群海州香薷酸性转化酶基因启动子甲基化的影响
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摘要
采用亚硫酸氢盐测序技术,检测了Cu胁迫对海州香薷(Elsholtzia haichowensis Sun)Cu抗性和非抗性种群酸性转化酶基因启动子甲基化的影响。结果表明,海州香薷液泡转化酶基因(v INV)和细胞壁转化酶基因(cw INV)的启动子在CG位点分别表现出超甲基化和低甲基化的现象。酸性转化酶基因启动子CHG和CHH位点的甲基化状态受Cu胁迫影响较大。Cu胁迫下,v INV和cw INV启动子分别有1个CHG和6个CHH位点的甲基化状态在Cu抗性种群和非抗性种群之间表现出较大差异。抗Cu种群中这些甲基化差异位点对Cu胁迫不敏感,但在非抗性种群中这些位点的甲基化水平在Cu胁迫后出现大幅上升或下降。有些甲基化差异位点位于或者临近预测的启动子顺式作用元件区域,可能参与Cu胁迫下酸性转化酶基因的表达调控。
In the present study, bisulfite sequencing(BS-seq) was performed to detect alterations in DNA methylation of acid invertase gene promoters in Cu-tolerant(MP) and nontolerant populations(NMP) of metallophyte Elsholtzia haichowensis under Cu treatment. We found that the vacuolar invertase gene(v INV) promoter of E. haichowensis was generally methylated in the CG site,whereas the cell wall invertase gene(cw INV) promoter displayed hypomethylation at the CG site. Methylation changes at a special CHG site of the v INV promoter and six CHH sites of the cw INV promoter induced by Cu stress were observed between the two populations. In addition,CG methylation was relatively conserved in the v INV and cw INV promoters under Cu stress,whereas methylation at the CHG and CHH sites was more prone to perturbation. Under Cu stress,methylation of these individual sites remained stable in the MP,but declined or increased drastically in the NMP. This implied that MP might have evolved modulating mechanisms due to long-term Cu stress,including changes in methylation patterns in specific DNA sequences and cytosine sites.
In the present study, bisulfite sequencing(BS-seq) was performed to detect alterations in DNA methylation of acid invertase gene promoters in Cu-tolerant(MP) and nontolerant populations(NMP) of metallophyte Elsholtzia haichowensis under Cu treatment. We found that the vacuolar invertase gene(v INV) promoter of E. haichowensis was generally methylated in the CG site,whereas the cell wall invertase gene(cw INV) promoter displayed hypomethylation at the CG site. Methylation changes at a special CHG site of the v INV promoter and six CHH sites of the cw INV promoter induced by Cu stress were observed between the two populations. In addition,CG methylation was relatively conserved in the v INV and cw INV promoters under Cu stress,whereas methylation at the CHG and CHH sites was more prone to perturbation. Under Cu stress,methylation of these individual sites remained stable in the MP,but declined or increased drastically in the NMP. This implied that MP might have evolved modulating mechanisms due to long-term Cu stress,including changes in methylation patterns in specific DNA sequences and cytosine sites.
引文
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[27]Jimenez-Arias D,Borges AA,Luis JC,Valdes F,Sandalio LM,Perez JA.Priming effect of menadione sodium bisulphite against salinity stress in Arabidopsis involves epigenetic changes in genes controlling proline metabolism[J].Environ Exp Bot,2015,120:23-30.
[28]Boyko A,Kovalchuk I.Epigenetic control of plant stress response[J].Environ Mol Mutagen,2008,49(1):61-72.
[29]Uthup TK,Ravindran M,Bini K,Thakurdas S.Divergent DNA methylation patterns associated with abiotic stress in Hevea brasiliensis[J].Mol Plant,2011,4(6):996-1013.
[30]Chatterjee R,Vinson C.Cp G methylation recruits sequence specific transcription factors essential for tissue specific gene expression[J].BBA-Gene Regul Mech,2012,1819(7):763-770.
[31]Lee GH,Sohn SH,Park EY,Park YD.Aberrant promoter methylation occurred from multicopy transgene and SU(VAR)3-9 homolog 9(SUVH9)gene in transgenic Nicotiana benthamiana[J].Funct Plant Biol,2012,39(9):764-773.
[32]Ay N,Janack B,Fischer A,Reuter G,Humbeck K.Alterations of histone modifications at the senescenceassociated gene Hv S40 in barley during senescence[J].Plant Mol Biol,2015,89(1-2):127-141.
[33]Richards EJ.Inherited epigenetic variation-revisiting soft inheritance[J].Nat Rev Genet,2006,7(5):395-401.
[34]Akimoto K,Katakami H,Kim HJ,Ogawa E,Sano CM,et al.Epigenetic inheritance in rice plants[J].Ann BotLondon,2007,100(2):205-217.
[2]Tang SR,Wilke BM,Huang CY.The uptake of copper by plants dominantly growing on copper mining spoils along the Yangtze River,the People's Republic of China[J].Plant Soil,1999,209(2):225-232.
[3]Liu C,Xu ZR,Cai SW,Zhang L,Xiong ZT.c DNA cloning,heterologous expression and characterization of a cell wall invertase from copper tolerant population of Elsholtzia haichowensis[J].Biologia,2015,70(8):1063-1069.
[4]Cai SW,Xiong ZT,Li L,Li MJ,Zhang L,Liu C,Xu ZR.Differential responses of root growth,acid invertase activity and transcript level to copper stress in two contrasting populations of Elsholtzia haichowensis[J].Ecotoxicology,2014,23(1):76-91.
[5]Huang WX,Cao Y,Huang LJ,Ren C,Xiong ZT.Differential expression of acid invertase genes in roots of metallicolous and non-metallicolous populations of Rumex japonicus under copper stress[J].Chemosphere,2011,84(10):1432-1439.
[6]Xiong ZT,Wang T,Liu K,Zhang ZZ,Gan JH,et al.Differential invertase activity and root growth between Cu-tolerant and non-tolerant populations in Kummerowia stipulacea under Cu stress and nutrient deficiency[J].Environ Exp Bot,2008,62(1):17-27.
[7]Xu ZR,Liu C,Cai SW,Zhang L,Xiong ZT.Heterologous expression and comparative characterization of vacuolar invertases from Cu-tolerant and non-tolerant populations of Elsholtzia haichowensis[J].Plant Cell Rep,2015,34(10):1781-1790.
[8]Chen M,LüSL,Meng YJ.Epigenetic performers in plants[J].Dev Growth Differ,2010,52(6):555-566.
[9]崔国鹏,王圣洁,覃瑞,刘虹,龚汉雨,等.极性生长的细胞中胞嘧啶甲基化的动态变化及其对GABA信号的响应[J].植物科学学报,2015,33(4):521-527.Cui GP,Wang SJ,Qin R,Liu H,Gong HY,et al.Dynamic changes of methylcytosine in polarized cell growth and their response to GABA signals[J].Plant Science Journal,2015,33(4):521-527.
[10]Song QX,Lu X,Li QT,Chen H,Hu XY,et al.Genomewide analysis of DNA methylation in soybean[J].Mol Plant,2013,6(6):1961-1974.
[11]Eprintsev AT,Fedorin DN,Karabutova LA,Igamberdiev AU.Expression of genes encoding subunits A and B of succinate dehydrogenase in germinating maize seeds is regulated by methylation of their promoters[J].J Plant Physiol,2016,205:33-40.
[12]邓莹,高乐旋,朱珠,杨继.不同水陆环境中喜旱莲子草甲基化调控因子表达水平和差异表达基因启动子区甲基化状态分析[J].植物科学学报,2014,32(5):475-486.Deng Y,Gao LX,Zhu Z,Yang J.Differential expression of DNA methylation regulating factors and dynamic methylation patterns of Alternanthera philoxeroides under different water treatments[J].Plant Science Journal,2014,32(5):475-486.
[13]Kumar M,Bijo AJ,Baghel RS,Reddy CRK,Jha B.Selenium and spermine alleviate cadmium induced toxicity in the red seaweed Gracilaria dura by regulating antioxidants and DNA methylation[J].Plant Physiol Bioch,2012,51:129-138.
[14]Aina R,Sgorbati S,Santagostino A,Labra M,Ghiani A,Citterio S.Specific hypomethylation of DNA is induced by heavy metals in white clover and industrial hemp[J].Physiol Plant,2004,121(3):472-480.
[15]Greco M,Chiappetta A,Bruno L,Bitonti MB.In Posidonia oceanica cadmium induces changes in DNA methylation and chromatin patterning[J].J Exp Bot,2012,63(2):695-709.
[16]Erturk FA,Agar G,Arslan E,Nardemir G.Analysis of genetic and epigenetic effects of maize seeds in response to heavy metal(Zn)stress[J].Environ Sci Pollut R,2015,22(13):10291-10297.
[17]丁国华,郭丹蒂,关旸,刘保东,池春玉.重金属铅镉对濒危植物中华水韭(Isoetes sinensis)DNA甲基化的影响[J].农业环境科学学报,2017,36(2):246-249.Ding GH,Guo DD,Guan Y,Liu BD,Chi CY.Effect of Pb and Cd on DNA methylation of Isoetes sinensis,a rare plant[J].Journal of Agro-Environment Science,2017,36(2):246-249.
[18]Feng SJ,Liu XS,Tao H,Tan SK,Chu SS,et al.Variation of DNA methylation patterns associated with gene expression in rice(Oryza sativa)exposed to cadmium[J].Plant Cell Environ,2016,39(12):2629-2649.
[19]Ou XF,Zhang YH,Xu CM,Lin XY,Zang Q,et al.Transgenerational inheritance of modified DNA methylation patterns and enhanced tolerance induced by heavy metal stress in rice(Oryza sativa L.)[J].PLo S One,2012,7(9):1-11.
[20]Uddin MS,Sun WL,He XH,da Silva JAT,Cheng Q.An improved method to extract DNA from mango Mangifera indica[J].Biologia,2014,69(2):133-138.
[21]Clark SJ,Statham A,Stirzaker C,Molloy PL,Frommer M.DNA methylation:Bisulphite modification and analysis[J].Nat Protoc,2006,1(5):2353-2364.
[22]Gruntman E,Qi YJ,Slotkin RK,Roeder T,Martienssen RA,Sachidanandam R.Kismeth:Analyzer of plant methylation states through bisulfite sequencing[J].BMC Bioinformatics,2008,9:371.
[23]Meyer P.DNA methylation systems and targets in plants[J].Febs Letts,2011,585(13):2008-2015.
[24]Bird AP.Cp G-rich islands and the function of DNA methylation[J].Nature,1986,321(6067):209-213.
[25]Cokus SJ,Feng SH,Zhang XY,Chen ZG,Merriman B,et al.Shotgun bisulphite sequencing of the Arabidopsis genome reveals DNA methylation patterning[J].Nature,2008,452(7184):215-219.
[26]Rombauts S,Florquin K,Lescot M,Marchal K,Rouze P,Van de Peer Y.Computational approaches to identify promoters and cis-regulatory elements in plant genomes[J].Plant Physiol,2003,132(3):1162-1176.
[27]Jimenez-Arias D,Borges AA,Luis JC,Valdes F,Sandalio LM,Perez JA.Priming effect of menadione sodium bisulphite against salinity stress in Arabidopsis involves epigenetic changes in genes controlling proline metabolism[J].Environ Exp Bot,2015,120:23-30.
[28]Boyko A,Kovalchuk I.Epigenetic control of plant stress response[J].Environ Mol Mutagen,2008,49(1):61-72.
[29]Uthup TK,Ravindran M,Bini K,Thakurdas S.Divergent DNA methylation patterns associated with abiotic stress in Hevea brasiliensis[J].Mol Plant,2011,4(6):996-1013.
[30]Chatterjee R,Vinson C.Cp G methylation recruits sequence specific transcription factors essential for tissue specific gene expression[J].BBA-Gene Regul Mech,2012,1819(7):763-770.
[31]Lee GH,Sohn SH,Park EY,Park YD.Aberrant promoter methylation occurred from multicopy transgene and SU(VAR)3-9 homolog 9(SUVH9)gene in transgenic Nicotiana benthamiana[J].Funct Plant Biol,2012,39(9):764-773.
[32]Ay N,Janack B,Fischer A,Reuter G,Humbeck K.Alterations of histone modifications at the senescenceassociated gene Hv S40 in barley during senescence[J].Plant Mol Biol,2015,89(1-2):127-141.
[33]Richards EJ.Inherited epigenetic variation-revisiting soft inheritance[J].Nat Rev Genet,2006,7(5):395-401.
[34]Akimoto K,Katakami H,Kim HJ,Ogawa E,Sano CM,et al.Epigenetic inheritance in rice plants[J].Ann BotLondon,2007,100(2):205-217.
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