菊花基因组DNA甲基化水平对根系硝态氮吸收的影响
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摘要
以菊花的扦插生根苗为试材,在缺氮和适氮水培条件下,分别使用甲基化抑制剂(5-aza C)和甲基供体(SAM)处理菊花根系,测定处理后基因组DNA甲基化水平变化、根系构型相关参数、根系硝态氮含量以及NO3-转运蛋白基因Cm NRT1.1、Cm NRT2.1、Cm NAR2.1的相对表达量。结果表明,在适氮条件下,5-aza C处理的菊花根系DNA甲基化水平降低,根系总直径、总表面积及总体积增大;根系NO3-含量与对照相比显著增加,根系NO3-转运蛋白基因Cm NRT1.1、Cm NRT2.1、Cm NAR2.1相对表达量也有所升高。据此推测,降低菊花根系基因组DNA甲基化水平可以提高根系NO3-转运相关基因的表达,进而促进根系对NO3-的吸收。
Using the rooted cuttings of chrysanthemum(Chrysanthemum morifolium) as test materials, the chrysanthemum roots system was treated with methylation inhibitor(5-aza C) and methyl donor(SAM) under the condition of nitorgen deficiency and suitable nitrogen hydroponics. The changes of DNA methylation level, root system configuration parameters, the contents of NO3-and the relative expression levels of NO3-transporter gene Cm NRT1.1, Cm NRT2.1 and Cm NAR2.1 were measured. The results showed that the DNA methylation level with 5-aza C treatment was reduced; the total root diameter, total surface area and total volume of root system increased. In addition, the contents of NO3-was significantly increased and the relative expression levels of Cm NRT1.1, Cm NR2.1 and Cm NAR2.1 in root system were also increased. It is conjectured that the reduction of DNA methylation level in the genome of chrysanthemum could increase the expression of NO3-transport related genes, and then promote the absorption of NO3-in the root system.
Using the rooted cuttings of chrysanthemum(Chrysanthemum morifolium) as test materials, the chrysanthemum roots system was treated with methylation inhibitor(5-aza C) and methyl donor(SAM) under the condition of nitorgen deficiency and suitable nitrogen hydroponics. The changes of DNA methylation level, root system configuration parameters, the contents of NO3-and the relative expression levels of NO3-transporter gene Cm NRT1.1, Cm NRT2.1 and Cm NAR2.1 were measured. The results showed that the DNA methylation level with 5-aza C treatment was reduced; the total root diameter, total surface area and total volume of root system increased. In addition, the contents of NO3-was significantly increased and the relative expression levels of Cm NRT1.1, Cm NR2.1 and Cm NAR2.1 in root system were also increased. It is conjectured that the reduction of DNA methylation level in the genome of chrysanthemum could increase the expression of NO3-transport related genes, and then promote the absorption of NO3-in the root system.
引文
Bucher E,Reinders J,Mirouze M(2012).Epigenetic control of transposon transcription and mobility in Arabidopsis.Curr Opin Plant Biol,15(5):503-510
Chen F(2009).5-aza C on the development and DNA methylation of wheat(Triticum aestivum L.)(dissertation).Kaifeng:Henan University(in Chinese with English abstract)[陈芳(2009).5-氮杂胞苷对小麦生长发育及DNA甲基化的影响(学位论文).开封:河南大学]
Cheng JF,Dai TB,Jing Q,et al(2007).Root morphological and physiological characteristics in relation to nitrogen absorption efficiency in different rice genotypes.Acta Pedol Sin,(2):266-272(in Chinese with English abstract)[程建峰,戴廷波,荆奇等(2007).不同水稻基因型的根系形态生理特性与高效氮素吸收.土壤学报,(2):266-272]
Fieldes MA,Schaeffer SM,Krech MJ,et al(2005).DNA hypomethylation in 5-azacytidine induced early flowering lines of flax.Theor Appl Genet,111(1):136-149
Finnegan EJ,Peacock WJ,Dennis ES(1996).Reduced DNAmethylation in Arabidopsis thaliana results in abnormal plant development.Proc Natl Acad Sci USA,93(16):8449-8454
Gonzalez M,Li F(2012).DNA replication,RNAi and epigenetic inheritance.Epigenetics,7(1):14-19
Guo YH,Yu YY,Wen LZ,et al(2017).Up-regulation of Cm NRTs and Cm ANR1 genes expression contribute to root configuration changes for efficient capturing NO3-in the roots of chrysanthemum.Sci Hortic,225:438-444
Gutzat R,Scheid OM(2012).Epigenetic responses to stress:triple defense.Curr Opin Plant Biol,15(5):568-573
Huang ZF,Zheng CS,Zhang CH,et al(2009).Effects of controlled release fertilizer on available nutrient utilization rate and growth and ornamental quality of chrysanthemum.Shandong Agric Sci,10:70-73(in Chinese with English abstract)[黄在范,郑成淑,张翠华等(2009).控释肥对土壤有效养分利用率及菊花生长和观赏品质的影响.山东农业科学,10:70-73]
King GJ(1995).Morphological development in Brassica oleracea is modulated by in vivo treatment with 5-azacytidine.J Hortic Sci,70(2):333-342
Law JA,Jacobsen SE(2010).Establishing,maintaining and modifying DNA methylation patterns in plants and animals.Nat Rev Genet,11(3):204
Law RD,Suttle JC(2005).Chromatin remodeling in plant cell culture:patterns of DNA methylation and histone H3 and H4 acetylation vary during growth of asynchronous potato cell suspensions.Plant Physiol Bioch,43(6):527-534
Liang D,Zhang ZJ,Wu Hl,et al(2014).Single-base-resolution methylomes of Populus trichocarpa reveal the association between DNA methylation and drought stress.BMC Genet,15(1):S9
Li ML,Zeng GW,Zhu ZJ(2003).Analysis of effects of5-azacytidine on promoting flowering in non-heading chinese cabbage.J Zhejiang Univ,29(3):287-290(in Chinese with English abstract)[李梅兰,曾广文,朱祝军(2003).5-氮杂胞苷促进白菜开花的效应分析.浙江大学学报,29(3):287-290]
Li XL,Lin ZX,Nie YC,et al(2009).MSAP analysis of epigenetic changes in cotton(Gossypium hirsutum L.)under salt stress.Acta Agron Sin,(4):588-596(in Chinese with English abstract)[李雪林,林忠旭,聂以春等(2009).盐胁迫下棉花基因组DNA表观遗传变化的MSAP分析.作物学报,(4):588-596]
Li XY,Wang XF,He K,et al(2008).High-resolution mapping of epigenetic modifications of the rice genome uncovers interplay between DNA methylation,histone methylation,and gene expression.Plant Cell,20(2):259-276
Li ZA,Du YQ,Li J,et al(2016).Effects of dimethyl sulfoxide on Arabidopsis thaliana DNA methylation.Plant Physiol J,52(01):117-124(in Chinese with English abstract)[李忠爱,杜亚琼,李杰等(2016).二甲基亚砜对拟南芥基因组DNA甲基化的影响.植物生理学报,52(01):117-124]
Li ZL,Wang HT,Chen RJ,et al(2014).Studying genomic methylation of Arabidopsis thaliana seedlings under cadmium stress using MSAP.J Agro-Environ Sci,33(01):28-36(in Chinese with English abstract)[李照令,王鹤潼,陈瑞娟等(2014).运用MSAP研究镉胁迫对拟南芥幼苗基因甲基化的影响.农业环境科学学报,33(01):28-36]
Liu B(2008).Epigenetic variation and crop genetic improvement.Jilin Agric Univ,30(4):386-393(in Chinese with English abstract)[刘宝(2008).表观遗传变异与作物遗传改良.吉林农业大学学报,30(4):386-393]
Luo JP,Wang M,La HG(2017).Mapping and functional analysis of a candidate gene involved in DNA demethylation process in Arabidopsis thaliana.Plant Physiol J,53(04):721-728(in Chinese with English abstract)[罗俊鹏,王敏,腊红桂(2017).一个参与拟南芥DNA去甲基化过程候选基因的定位与功能分析.植物生理学报,53(04):721-728]
Lu XK,Wang DL,Yin ZJ,et al(2014).Genomic DNA methylation polymorphism analysis of cotton under Na Cl and Na2CO3 stress.Sci Agric Sin Chin,47(16):3132-3142(in Chinese with English abstract)[陆许可,王德龙,阴祖军等(2014).Na Cl和Na2CO3对不同棉花基因组的DNA甲基化影响.中国农业科学,47(16):3132-3142]
M?ller AL,Pedas P,Andersen B,et al(2011).Responses of barley root and shoot proteomes to long-term nitrogen deficiency,short-term nitrogen starvation and ammonium.Plant Cell Environ,34(12):2024-2037
Naydenov M,Baev V,Apostolova E,et al(2015).High-temperature effect on genes engaged in DNA methylation and affected by DNA methylation in Arabidopsis.Plant Physiol Bioch,87:102-108
Nie LJ(2008).DNA methylation alteration of chrysanthemum(Dendranthema grandiflorum)during in vitro growth by artificial inducement(dissertation).Kaifeng:Henan University(in Chinese with English abstract)[聂丽娟(2008).离体条件下人工诱导菊花DNA甲基化变化(学位论文).开封:河南大学]
Orsel M,Chopin F,Leleu O,et al(2006).Characterization of a two-component high-affinity nitrate uptake system in Arabidopsis.Physiology and protein-protein interaction.Plant Physiol,142(3):1304-1317
Plett D,Toubia J,Garnett T,et al(2010).Dichotomy in the NRT gene families of dicots and grass species.PLo SONE,5(12):152-158
Secco D,Whelan J,Rouached H,et al(2017).Nutrient stress-induced chromatin changes in plants.Curr Opin Plant Biol,39:1-7
Tian SB,Lin GY,Zheng CB,et al(2011).Cloning and expression of Cm CO and Cm FT of floral development genes in chrysanthemum.Acta Hortic Sin,38(6):1129-1138(in Chinese with English abstract)[田素波,林桂玉,郑成淑等(2011).菊花花发育基因Cm CO和Cm FT的克隆与表达分析.园艺学报,38(6):1129-1138]
Vanyushin BF(2006).DNA methylation in plants.Curr Top Microbiol,301(1):67
Wang XC,Chen L,Yang YJ,et al(2006).Advances in plant nitrogen nutrient genotypic difference.J Eco-Agric,14(03):12-15(in Chinese with English abstract)[王新超,陈亮,杨亚军等(2006).植物氮素营养基因型差异研究进展.中国生态农业学报,14(03):12-15]
Wang YM,Chen FJ(2004).Response of root morphology to nitrate supply and its contribution to nitrogen uptake in maize.J Plant Nutr,27(12):2189-2202
Wang ZC,Nie LJ,He YX(2009).The effect of 5-aza C cytidine to the DNA methylation and morphogenesis character of chrysanthemum during in vitro growth.Acta Hortic Sin,36(12):1783-1790(in Chinese with English abstract)[王子成,聂丽娟,何艳霞(2009).离体条件下5-氮杂胞嘧啶核苷对菊花DNA甲基化和表型性状的影响.园艺学报,36(12):1783-1790]
Wu YJ(2012).Epigenetics mechanism studies of maize seedling root under treatment of nitrogen deficiency and re-adding nitrogen(dissertation).Yangling:Northest A&F University(in Chinese with English abstract)[武永军(2012).缺氮复氮处理下玉米幼苗根系的表观遗传机制研究(学位论文).杨凌:西北农林科技大学]
Xiong LZ,Xu CG,Saghai Maroof MA,et al(1999).Patterns of cytosine methylation in an elite rice hybrid and its parental lines,detected by a methylation-sensitive amplification polymorphism technique.Mol Gen Genet,261(3):439-446
Xu BH(2005).Studies of the effect of DNA methylation on the growth and development of the primary root in the Triticum aestivum(dissertation).Beijing:Capital Normal University(in Chinese with English abstract)[徐宝华(2005).DNA甲基化对小麦根系早期生长发育影响的研究(学位论文).北京:首都师范大学]
Chen F(2009).5-aza C on the development and DNA methylation of wheat(Triticum aestivum L.)(dissertation).Kaifeng:Henan University(in Chinese with English abstract)[陈芳(2009).5-氮杂胞苷对小麦生长发育及DNA甲基化的影响(学位论文).开封:河南大学]
Cheng JF,Dai TB,Jing Q,et al(2007).Root morphological and physiological characteristics in relation to nitrogen absorption efficiency in different rice genotypes.Acta Pedol Sin,(2):266-272(in Chinese with English abstract)[程建峰,戴廷波,荆奇等(2007).不同水稻基因型的根系形态生理特性与高效氮素吸收.土壤学报,(2):266-272]
Fieldes MA,Schaeffer SM,Krech MJ,et al(2005).DNA hypomethylation in 5-azacytidine induced early flowering lines of flax.Theor Appl Genet,111(1):136-149
Finnegan EJ,Peacock WJ,Dennis ES(1996).Reduced DNAmethylation in Arabidopsis thaliana results in abnormal plant development.Proc Natl Acad Sci USA,93(16):8449-8454
Gonzalez M,Li F(2012).DNA replication,RNAi and epigenetic inheritance.Epigenetics,7(1):14-19
Guo YH,Yu YY,Wen LZ,et al(2017).Up-regulation of Cm NRTs and Cm ANR1 genes expression contribute to root configuration changes for efficient capturing NO3-in the roots of chrysanthemum.Sci Hortic,225:438-444
Gutzat R,Scheid OM(2012).Epigenetic responses to stress:triple defense.Curr Opin Plant Biol,15(5):568-573
Huang ZF,Zheng CS,Zhang CH,et al(2009).Effects of controlled release fertilizer on available nutrient utilization rate and growth and ornamental quality of chrysanthemum.Shandong Agric Sci,10:70-73(in Chinese with English abstract)[黄在范,郑成淑,张翠华等(2009).控释肥对土壤有效养分利用率及菊花生长和观赏品质的影响.山东农业科学,10:70-73]
King GJ(1995).Morphological development in Brassica oleracea is modulated by in vivo treatment with 5-azacytidine.J Hortic Sci,70(2):333-342
Law JA,Jacobsen SE(2010).Establishing,maintaining and modifying DNA methylation patterns in plants and animals.Nat Rev Genet,11(3):204
Law RD,Suttle JC(2005).Chromatin remodeling in plant cell culture:patterns of DNA methylation and histone H3 and H4 acetylation vary during growth of asynchronous potato cell suspensions.Plant Physiol Bioch,43(6):527-534
Liang D,Zhang ZJ,Wu Hl,et al(2014).Single-base-resolution methylomes of Populus trichocarpa reveal the association between DNA methylation and drought stress.BMC Genet,15(1):S9
Li ML,Zeng GW,Zhu ZJ(2003).Analysis of effects of5-azacytidine on promoting flowering in non-heading chinese cabbage.J Zhejiang Univ,29(3):287-290(in Chinese with English abstract)[李梅兰,曾广文,朱祝军(2003).5-氮杂胞苷促进白菜开花的效应分析.浙江大学学报,29(3):287-290]
Li XL,Lin ZX,Nie YC,et al(2009).MSAP analysis of epigenetic changes in cotton(Gossypium hirsutum L.)under salt stress.Acta Agron Sin,(4):588-596(in Chinese with English abstract)[李雪林,林忠旭,聂以春等(2009).盐胁迫下棉花基因组DNA表观遗传变化的MSAP分析.作物学报,(4):588-596]
Li XY,Wang XF,He K,et al(2008).High-resolution mapping of epigenetic modifications of the rice genome uncovers interplay between DNA methylation,histone methylation,and gene expression.Plant Cell,20(2):259-276
Li ZA,Du YQ,Li J,et al(2016).Effects of dimethyl sulfoxide on Arabidopsis thaliana DNA methylation.Plant Physiol J,52(01):117-124(in Chinese with English abstract)[李忠爱,杜亚琼,李杰等(2016).二甲基亚砜对拟南芥基因组DNA甲基化的影响.植物生理学报,52(01):117-124]
Li ZL,Wang HT,Chen RJ,et al(2014).Studying genomic methylation of Arabidopsis thaliana seedlings under cadmium stress using MSAP.J Agro-Environ Sci,33(01):28-36(in Chinese with English abstract)[李照令,王鹤潼,陈瑞娟等(2014).运用MSAP研究镉胁迫对拟南芥幼苗基因甲基化的影响.农业环境科学学报,33(01):28-36]
Liu B(2008).Epigenetic variation and crop genetic improvement.Jilin Agric Univ,30(4):386-393(in Chinese with English abstract)[刘宝(2008).表观遗传变异与作物遗传改良.吉林农业大学学报,30(4):386-393]
Luo JP,Wang M,La HG(2017).Mapping and functional analysis of a candidate gene involved in DNA demethylation process in Arabidopsis thaliana.Plant Physiol J,53(04):721-728(in Chinese with English abstract)[罗俊鹏,王敏,腊红桂(2017).一个参与拟南芥DNA去甲基化过程候选基因的定位与功能分析.植物生理学报,53(04):721-728]
Lu XK,Wang DL,Yin ZJ,et al(2014).Genomic DNA methylation polymorphism analysis of cotton under Na Cl and Na2CO3 stress.Sci Agric Sin Chin,47(16):3132-3142(in Chinese with English abstract)[陆许可,王德龙,阴祖军等(2014).Na Cl和Na2CO3对不同棉花基因组的DNA甲基化影响.中国农业科学,47(16):3132-3142]
M?ller AL,Pedas P,Andersen B,et al(2011).Responses of barley root and shoot proteomes to long-term nitrogen deficiency,short-term nitrogen starvation and ammonium.Plant Cell Environ,34(12):2024-2037
Naydenov M,Baev V,Apostolova E,et al(2015).High-temperature effect on genes engaged in DNA methylation and affected by DNA methylation in Arabidopsis.Plant Physiol Bioch,87:102-108
Nie LJ(2008).DNA methylation alteration of chrysanthemum(Dendranthema grandiflorum)during in vitro growth by artificial inducement(dissertation).Kaifeng:Henan University(in Chinese with English abstract)[聂丽娟(2008).离体条件下人工诱导菊花DNA甲基化变化(学位论文).开封:河南大学]
Orsel M,Chopin F,Leleu O,et al(2006).Characterization of a two-component high-affinity nitrate uptake system in Arabidopsis.Physiology and protein-protein interaction.Plant Physiol,142(3):1304-1317
Plett D,Toubia J,Garnett T,et al(2010).Dichotomy in the NRT gene families of dicots and grass species.PLo SONE,5(12):152-158
Secco D,Whelan J,Rouached H,et al(2017).Nutrient stress-induced chromatin changes in plants.Curr Opin Plant Biol,39:1-7
Tian SB,Lin GY,Zheng CB,et al(2011).Cloning and expression of Cm CO and Cm FT of floral development genes in chrysanthemum.Acta Hortic Sin,38(6):1129-1138(in Chinese with English abstract)[田素波,林桂玉,郑成淑等(2011).菊花花发育基因Cm CO和Cm FT的克隆与表达分析.园艺学报,38(6):1129-1138]
Vanyushin BF(2006).DNA methylation in plants.Curr Top Microbiol,301(1):67
Wang XC,Chen L,Yang YJ,et al(2006).Advances in plant nitrogen nutrient genotypic difference.J Eco-Agric,14(03):12-15(in Chinese with English abstract)[王新超,陈亮,杨亚军等(2006).植物氮素营养基因型差异研究进展.中国生态农业学报,14(03):12-15]
Wang YM,Chen FJ(2004).Response of root morphology to nitrate supply and its contribution to nitrogen uptake in maize.J Plant Nutr,27(12):2189-2202
Wang ZC,Nie LJ,He YX(2009).The effect of 5-aza C cytidine to the DNA methylation and morphogenesis character of chrysanthemum during in vitro growth.Acta Hortic Sin,36(12):1783-1790(in Chinese with English abstract)[王子成,聂丽娟,何艳霞(2009).离体条件下5-氮杂胞嘧啶核苷对菊花DNA甲基化和表型性状的影响.园艺学报,36(12):1783-1790]
Wu YJ(2012).Epigenetics mechanism studies of maize seedling root under treatment of nitrogen deficiency and re-adding nitrogen(dissertation).Yangling:Northest A&F University(in Chinese with English abstract)[武永军(2012).缺氮复氮处理下玉米幼苗根系的表观遗传机制研究(学位论文).杨凌:西北农林科技大学]
Xiong LZ,Xu CG,Saghai Maroof MA,et al(1999).Patterns of cytosine methylation in an elite rice hybrid and its parental lines,detected by a methylation-sensitive amplification polymorphism technique.Mol Gen Genet,261(3):439-446
Xu BH(2005).Studies of the effect of DNA methylation on the growth and development of the primary root in the Triticum aestivum(dissertation).Beijing:Capital Normal University(in Chinese with English abstract)[徐宝华(2005).DNA甲基化对小麦根系早期生长发育影响的研究(学位论文).北京:首都师范大学]