红花玉兰和白玉兰冷应激代谢组学分析
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摘要
为探究红花玉兰和白玉兰低温胁迫下的代谢表型特性,以红花玉兰和白玉兰茎尖为材料,采用GC-MS检测技术,分别检测了两个物种在常温20℃和低温4℃下的代谢物质,共检测出代谢物323种,对所有样本进行了多元统计分析。将处理间代谢物进行了四次比对分析,并对差异代谢物进行了统计和归类,筛选出了相关性最高的代谢通路,总结分析了两个物种在低温胁迫下的代谢表型。结果显示,两个物种具有十分明显的代谢表型特性,常温下白玉兰较红花玉兰积累了更丰富的基础代谢物质;在低温胁迫下,红花玉兰氨基酸类和糖类相关物质代谢显著,白玉兰仅氨基酸类物质代谢显著。本研究为红花玉兰抗寒良种的选育和分子改良提供了代谢依据。
Taking shoot tips of Magnolia wufengensis and Magnolia denudata as materials, GC-MS detection technique was used to investigate the metabolic phenotypes of Magnolia wufengensis and Magnolia denudata under cold stress. The metabolites of the two species were detected at 20℃ and 4℃, a total of 323 kinds of plant metabolites had been detected and multivariate statistical analysis was performed on all the samples. Four times of comparison analysis were respectively carried out. And then the metabolites were statistically classified and the most relevant metabolic pathways were screened out. In this research, the metabolic phenotypes of the two species were summarized and analyzed under cold stress. The results showed that the two species had very obvious metabolic phenotypic characteristics: Magnolia denudata accumulated more abundant basal metabolites at room temperature.The amino acids and carbohydrate-related substances of Magnolia wufengensis were significantly metabolized under low temperature stress, while in Magnolia denudata only amino acids that were metabolized distinctly. This study might provide a metabolic basis for breeding and molecular improvement of cold-resistant Magnolia wufengensis varieties.
Taking shoot tips of Magnolia wufengensis and Magnolia denudata as materials, GC-MS detection technique was used to investigate the metabolic phenotypes of Magnolia wufengensis and Magnolia denudata under cold stress. The metabolites of the two species were detected at 20℃ and 4℃, a total of 323 kinds of plant metabolites had been detected and multivariate statistical analysis was performed on all the samples. Four times of comparison analysis were respectively carried out. And then the metabolites were statistically classified and the most relevant metabolic pathways were screened out. In this research, the metabolic phenotypes of the two species were summarized and analyzed under cold stress. The results showed that the two species had very obvious metabolic phenotypic characteristics: Magnolia denudata accumulated more abundant basal metabolites at room temperature.The amino acids and carbohydrate-related substances of Magnolia wufengensis were significantly metabolized under low temperature stress, while in Magnolia denudata only amino acids that were metabolized distinctly. This study might provide a metabolic basis for breeding and molecular improvement of cold-resistant Magnolia wufengensis varieties.
引文
Aa J.Y.,2010,Analysis of metabolomics data:principal component analysis,Zhongguo Linchuang Yaolixue Yu Zhiliaoxue(Chinese Journal of Clinical Pharmacology and Therapeutics),15(5):481-489(阿基业,2010,代谢组学数据处理方法-主成分分析,中国临床药理学与治疗学,15(5):481-489)
Benina M.,Obata T.,Mehterov N.,Ivanov I.,Petrov V.,Toneva V.,Fernie A.R.,and Gechev T.S.,2013,Comparative metabolic profiling of Haberlea rhodopensis,Thellungiella halophyla,and Arabidopsis thaliana exposed to low temperature,Front.Plant Sci.,4:499
Bino R.J.,Hall R.D.,Fiehn O.,Kopka J.,Saito K.,Draper J.,Nikolau B.J.,Mendes P.,Roessner-Tunali U.,Beale M.H.,Trethewey R.N.,Lange B.M.,Wurtele E.S.,and Sumner L.W.,2004,Potential of metabolomics as a functional genomics tool,Trends Plant Sci.,9(9):418-425
Cook D.,Fowler S.,Fiehn O.,and Thomashow M.F.,2004,Aprominent role for the CBF cold response pathway in configuring the low-temperature metabolome of Arabidopsis,Proc.Natl.Acad.Sci.USA,101(42):15243-15248
Duan L.X.,and Qi X.Q.,2015,GC-MS-based plant metabolomics researches,Shengming Kexue(Chinese Bulletin of Life Sciences),27(8):971-977(段礼新,漆小泉,2015,基于GC-MS的植物代谢组学研究,生命科学,27(8):971-977)
Fiehn O.,2002,Metabolomics-the link between genotypes and phenotypes,Plant Molecular Biology,48(1-2):155-171
Gavaghan C.L.,Holmes E.,Lenz E.,Wilson I.D.,and Nicholson J.K.,2000,An NMR-based metabonomic approach to investigate the biochemical consequences of genetic strain differences:application to the C57BL10J and Alpk:ApfCD mouse,FEBS Lett.,484(3):169-174
Gilmour S.J.,Sebolt A.M.,Salazar M.P.,Everard J.D.,and Thomashow M.F.,2000,Overexpression of the Arabidopsis CBF3 transcriptional activator mimics multiple biochemical changes associated with cold acclimation,Plant Physiol.,124(4):1854-1865
Gong Q.,Li P.,Ma S.,Indu Rupassara S.,and Bohnert H.J.,2005,Salinity stress adaptation competence in the extremophile Thellungiella halophila in comparison with its relative Arabidopsis thaliana,Plant J.,44(5):826-839
Guy C.L.,Huber J.L.,and Huber S.C.,1992,Sucrose phosphate synthase and sucrose accumulation at low temperature,Plant Physiol.,100(1):502-508
Jaglo-Ottosen K.R.,Gilmour S.J.,Zarka D.G.,Schabenberger O.,and Thomashow M.F.,1998,Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance,Science,280(5360):104-106
Jonsson P.,Johansson A.I.,Gullberg J.,Trygg J.,A J.,Grung B.,Marklund S.,Sj觟str觟m M.,Antti H.,and Moritz T.,2005,High-throughput data analysis for detecting and identifying differences between samples in GC/MS-based metabolomic analyses,Anal.Chem.,77(17):5635-5642
Kishitani S.,Watanabe K.,Yasuda S.,Arakawa K.,and Takabe T.,1994,Accumulation of glycinebetaine during cold acclimation and freezing tolerance in leaves of winter and spring barley plants,Plant,Cell&Environment,17(1):89-95
Koster K.L.,and Lynch D.V.,1992,Solute accumulation and compartmentation during the cold acclimation of Puma rye,Plant Physiol.,98(1):108-113
Lee Y.P.,Babakov A.,de Boer B.,Zuther E.,and Hincha D.K.,2012,Comparison of freezing tolerance,compatible solutes and polyamines in geographically diverse collections of Thellungiella sp.and Arabidopsis thaliana accessions,BMCPlant Biol.,12:131
Li M.Y.,2017,Plasma proteomic s and metabolomics in patients with chronic atrial fibrillation in rheumatic heart disease,Dissertation for Ph.D.,Peking Union Medical College,Supervisor:Liu X.C.,pp.12-15(李明阳,2017,风湿性心脏病慢性房颤患者的血浆蛋白质组学和代谢组学研究,博士学位论文,北京协和医学院,导师:刘晓程,pp.12-15)
Li T.,Liang J.,Xie L.,Xie X.D.,Wei S.S.,and Nong Z.X.,2013,Effects of low temperature stress on physiological and biochemical properties of different families of Acacia crassicarpa,Dongbei Linye Daxue Xuebao(Journal of Northeast Forestry University),41(2):6-9(李婷,梁机,谢乐,谢秀丹,韦素桑,农忠献,2013,低温胁迫对厚荚相思不同家系抗寒生理生化特征的影响,东北林业大学学报,41(2):6-9)
Liang D.W.,Ma L.Y.,Jia Z.K.,Hao Y.,Huai H.M.,Zhu Z.L.,and Wang X.L.,2010,Study on the influences of naturel cooling on the cold tolerance physiological indicators of Magnolia wufengensis,Linye Keji Kaifa(China Forestry Science and Technology),24(2):23-26(梁大伟,马履一,贾忠奎,郝跃,怀慧明,朱仲龙,王晓玲,2010,自然降温对红花玉兰抗寒生理指标的影响,林业科技开发,24(2):23-26)
Luedemann A.,Strassburg K.,Erban A.,and Kopka J.,2008,TagFinder for the quantitative analysis of gas chromatography-mass spectrometry(GC-MS)-based metabolite profiling experiments,Bioinformatics,24(5):732-737
Ma L.Y.,Wang L.R.,He S.C.,Liu X.,and Wang X.Q.,2006a,Anew species of Magnolia(Magnoliaceae)from Hubei China,Zhiwu Yanjiu(Bulletin of Botanical Research),26(1):4-8(马履一,王罗荣,贺随超,刘鑫,王希群,2006a,中国木兰科木兰属---新种,植物研究,26(1):4-8)
Ma L.Y.,Wang L.R.,He S.C.,Liu X.,and Wang X.Q.,2006b,Anew variety of Magnolia(Magnoliaceae)from Hubei China,Zhiwu Yanjiu(Bulletin of Botanical Research),26(5):516-519(马履一,王罗荣,贺随超,刘鑫,王希群,2006b,中国木兰科木兰属一新变种,植物研究,26(5):516-519)
Medina J.,CataláR.,and Salinas J.,2011,The CBFs:three Arabidopsis transcription factors to cold acclimate,Plant Sci.,180(1):3-11
Murelli C.,Rizza F.,Albini F.M.,Dulio A.,Terzi V.,and Cattivelli L.,1995,Metabolic changes associated with cold-acclimation in contrasting cultivars of barley,Physiol.Plant.,94(1):87-93
Nielsen J.,and Oliver S.,2005,The next wave in metabolome analysis,Trends Biotechnol.,23(11):544-546
Obata T.,and Fernie A.R.,2012,The use of metabolomics to dissect plant responses to abiotic stresses,Cell.Mol.Life Sci.,69(19):3225-3243
Sanchez D.H.,Pieckenstain F.L.,Escaray F.,Erban A.,Kraemer U.,Udvardi M.K.,and Kopka J.,2011,Comparative ionomics and metabolomics in extremophile and glycophytic lotus species under salt stress challenge the metabolic preadaptation hypothesis,Plant Cell Environ.,34(4):605-617
Taji T.,Ohsumi C.,Iuchi S.,Seki M.,Kasuga M.,Kobayashi M.,Yamaguchi-Shinozaki K.,and Shinozaki K.,2002,Important roles of drought-and cold-inducible genes for galactinol synthase in stress tolerance in Arabidopsis thaliana,Plant J.,29(4):417-426
Thomashow M.F.,2010,Molecular basis of plant cold acclimation:insights gained from studying the CBF cold response pathway,Plant Physiol.,154(2):571-577
Wanner L.A.,and Junttila O.,1999,Cold-induced freezing tolerance in Arabidopsis,Plant Physiol.,120(2):391-400
Wu W.T.,Chen F.J.,Jing D.L.,Liu Z.X.,and Ma L.Y.,2012,Isolation and characterization of an AGAMOUS-like gene from Magnolia wufengensis(Magnoliaceae),Plant Mol.Biol.Rep.,30(3):690-698
Yang Y.,2015,Studies on the ecological adaptability and overwintering technique of Magnolia wufengensis seedlings in Beijing,Dissertation for Ph.D.,Beijing Forestry University,Supervisor:Ma L.Y.,pp.41-56(杨杨,2015,红花玉兰幼苗在北京地区生态适应性与越冬技术研究,博士学位论文,北京林业大学,导师:马履一,pp.41-56)
Yang Y.,Jia Z.,Chen F.,Sang Z.,and Ma L.,2015,Comparative analysis of natural cold acclimation and deacclimation of two Magnolia species with different winter hardiness,Acta Physiol.Plant.,37(7):129
Zhang J.B.,Jin Y.F.,Wang S.S.,Yang H.Q.,Pang T.,Li J.Y.,Gao X.,Cui M.K.,and Gong M.,2015,Effect of different temperature on sucrose metabolism in different growth stages of tobacco,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(10):2225-2244(张建波,金云峰,王莎莎,杨慧芹,逄涛,李军营,高雪,崔明昆,龚明,2015,生长温度对不同生育期烟草蔗糖代谢的影响,基因组学与应用生物学,34(10):2225-2244)
Zhou G.Y.,Hu W.X.,Xu J.H.,and Xue Q.Z.,2013,Integrating multiple"omics"analysis for plant metabolites and their functions,Zhejiang Daxue Xuebao(Journal of Zhejiang U-niversity(Agriculture and Life Sciences)),39(3):237-245(周国艳,胡望雄,徐建红,薛庆中,2013,整合多个组学(omics)分析植物代谢产物及其功能,浙江大学学报(农业与生命科学版),39(3):237-245)
Zhu Z.L.,2012,Study on the limiting factor and winter protection methods of Magnolia wufengensis after introduction to Beijing,Thesis for M.S.,Beijing Forestry University,Supervisors:Ma L.Y.,and Jia Z.K.,pp.11-18(朱仲龙,2012,北京引种红花玉兰的限制因子与越冬防寒技术研究,硕士学位论文,北京林业大学,导师:马履一,贾忠奎,pp.11-18)
Benina M.,Obata T.,Mehterov N.,Ivanov I.,Petrov V.,Toneva V.,Fernie A.R.,and Gechev T.S.,2013,Comparative metabolic profiling of Haberlea rhodopensis,Thellungiella halophyla,and Arabidopsis thaliana exposed to low temperature,Front.Plant Sci.,4:499
Bino R.J.,Hall R.D.,Fiehn O.,Kopka J.,Saito K.,Draper J.,Nikolau B.J.,Mendes P.,Roessner-Tunali U.,Beale M.H.,Trethewey R.N.,Lange B.M.,Wurtele E.S.,and Sumner L.W.,2004,Potential of metabolomics as a functional genomics tool,Trends Plant Sci.,9(9):418-425
Cook D.,Fowler S.,Fiehn O.,and Thomashow M.F.,2004,Aprominent role for the CBF cold response pathway in configuring the low-temperature metabolome of Arabidopsis,Proc.Natl.Acad.Sci.USA,101(42):15243-15248
Duan L.X.,and Qi X.Q.,2015,GC-MS-based plant metabolomics researches,Shengming Kexue(Chinese Bulletin of Life Sciences),27(8):971-977(段礼新,漆小泉,2015,基于GC-MS的植物代谢组学研究,生命科学,27(8):971-977)
Fiehn O.,2002,Metabolomics-the link between genotypes and phenotypes,Plant Molecular Biology,48(1-2):155-171
Gavaghan C.L.,Holmes E.,Lenz E.,Wilson I.D.,and Nicholson J.K.,2000,An NMR-based metabonomic approach to investigate the biochemical consequences of genetic strain differences:application to the C57BL10J and Alpk:ApfCD mouse,FEBS Lett.,484(3):169-174
Gilmour S.J.,Sebolt A.M.,Salazar M.P.,Everard J.D.,and Thomashow M.F.,2000,Overexpression of the Arabidopsis CBF3 transcriptional activator mimics multiple biochemical changes associated with cold acclimation,Plant Physiol.,124(4):1854-1865
Gong Q.,Li P.,Ma S.,Indu Rupassara S.,and Bohnert H.J.,2005,Salinity stress adaptation competence in the extremophile Thellungiella halophila in comparison with its relative Arabidopsis thaliana,Plant J.,44(5):826-839
Guy C.L.,Huber J.L.,and Huber S.C.,1992,Sucrose phosphate synthase and sucrose accumulation at low temperature,Plant Physiol.,100(1):502-508
Jaglo-Ottosen K.R.,Gilmour S.J.,Zarka D.G.,Schabenberger O.,and Thomashow M.F.,1998,Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance,Science,280(5360):104-106
Jonsson P.,Johansson A.I.,Gullberg J.,Trygg J.,A J.,Grung B.,Marklund S.,Sj觟str觟m M.,Antti H.,and Moritz T.,2005,High-throughput data analysis for detecting and identifying differences between samples in GC/MS-based metabolomic analyses,Anal.Chem.,77(17):5635-5642
Kishitani S.,Watanabe K.,Yasuda S.,Arakawa K.,and Takabe T.,1994,Accumulation of glycinebetaine during cold acclimation and freezing tolerance in leaves of winter and spring barley plants,Plant,Cell&Environment,17(1):89-95
Koster K.L.,and Lynch D.V.,1992,Solute accumulation and compartmentation during the cold acclimation of Puma rye,Plant Physiol.,98(1):108-113
Lee Y.P.,Babakov A.,de Boer B.,Zuther E.,and Hincha D.K.,2012,Comparison of freezing tolerance,compatible solutes and polyamines in geographically diverse collections of Thellungiella sp.and Arabidopsis thaliana accessions,BMCPlant Biol.,12:131
Li M.Y.,2017,Plasma proteomic s and metabolomics in patients with chronic atrial fibrillation in rheumatic heart disease,Dissertation for Ph.D.,Peking Union Medical College,Supervisor:Liu X.C.,pp.12-15(李明阳,2017,风湿性心脏病慢性房颤患者的血浆蛋白质组学和代谢组学研究,博士学位论文,北京协和医学院,导师:刘晓程,pp.12-15)
Li T.,Liang J.,Xie L.,Xie X.D.,Wei S.S.,and Nong Z.X.,2013,Effects of low temperature stress on physiological and biochemical properties of different families of Acacia crassicarpa,Dongbei Linye Daxue Xuebao(Journal of Northeast Forestry University),41(2):6-9(李婷,梁机,谢乐,谢秀丹,韦素桑,农忠献,2013,低温胁迫对厚荚相思不同家系抗寒生理生化特征的影响,东北林业大学学报,41(2):6-9)
Liang D.W.,Ma L.Y.,Jia Z.K.,Hao Y.,Huai H.M.,Zhu Z.L.,and Wang X.L.,2010,Study on the influences of naturel cooling on the cold tolerance physiological indicators of Magnolia wufengensis,Linye Keji Kaifa(China Forestry Science and Technology),24(2):23-26(梁大伟,马履一,贾忠奎,郝跃,怀慧明,朱仲龙,王晓玲,2010,自然降温对红花玉兰抗寒生理指标的影响,林业科技开发,24(2):23-26)
Luedemann A.,Strassburg K.,Erban A.,and Kopka J.,2008,TagFinder for the quantitative analysis of gas chromatography-mass spectrometry(GC-MS)-based metabolite profiling experiments,Bioinformatics,24(5):732-737
Ma L.Y.,Wang L.R.,He S.C.,Liu X.,and Wang X.Q.,2006a,Anew species of Magnolia(Magnoliaceae)from Hubei China,Zhiwu Yanjiu(Bulletin of Botanical Research),26(1):4-8(马履一,王罗荣,贺随超,刘鑫,王希群,2006a,中国木兰科木兰属---新种,植物研究,26(1):4-8)
Ma L.Y.,Wang L.R.,He S.C.,Liu X.,and Wang X.Q.,2006b,Anew variety of Magnolia(Magnoliaceae)from Hubei China,Zhiwu Yanjiu(Bulletin of Botanical Research),26(5):516-519(马履一,王罗荣,贺随超,刘鑫,王希群,2006b,中国木兰科木兰属一新变种,植物研究,26(5):516-519)
Medina J.,CataláR.,and Salinas J.,2011,The CBFs:three Arabidopsis transcription factors to cold acclimate,Plant Sci.,180(1):3-11
Murelli C.,Rizza F.,Albini F.M.,Dulio A.,Terzi V.,and Cattivelli L.,1995,Metabolic changes associated with cold-acclimation in contrasting cultivars of barley,Physiol.Plant.,94(1):87-93
Nielsen J.,and Oliver S.,2005,The next wave in metabolome analysis,Trends Biotechnol.,23(11):544-546
Obata T.,and Fernie A.R.,2012,The use of metabolomics to dissect plant responses to abiotic stresses,Cell.Mol.Life Sci.,69(19):3225-3243
Sanchez D.H.,Pieckenstain F.L.,Escaray F.,Erban A.,Kraemer U.,Udvardi M.K.,and Kopka J.,2011,Comparative ionomics and metabolomics in extremophile and glycophytic lotus species under salt stress challenge the metabolic preadaptation hypothesis,Plant Cell Environ.,34(4):605-617
Taji T.,Ohsumi C.,Iuchi S.,Seki M.,Kasuga M.,Kobayashi M.,Yamaguchi-Shinozaki K.,and Shinozaki K.,2002,Important roles of drought-and cold-inducible genes for galactinol synthase in stress tolerance in Arabidopsis thaliana,Plant J.,29(4):417-426
Thomashow M.F.,2010,Molecular basis of plant cold acclimation:insights gained from studying the CBF cold response pathway,Plant Physiol.,154(2):571-577
Wanner L.A.,and Junttila O.,1999,Cold-induced freezing tolerance in Arabidopsis,Plant Physiol.,120(2):391-400
Wu W.T.,Chen F.J.,Jing D.L.,Liu Z.X.,and Ma L.Y.,2012,Isolation and characterization of an AGAMOUS-like gene from Magnolia wufengensis(Magnoliaceae),Plant Mol.Biol.Rep.,30(3):690-698
Yang Y.,2015,Studies on the ecological adaptability and overwintering technique of Magnolia wufengensis seedlings in Beijing,Dissertation for Ph.D.,Beijing Forestry University,Supervisor:Ma L.Y.,pp.41-56(杨杨,2015,红花玉兰幼苗在北京地区生态适应性与越冬技术研究,博士学位论文,北京林业大学,导师:马履一,pp.41-56)
Yang Y.,Jia Z.,Chen F.,Sang Z.,and Ma L.,2015,Comparative analysis of natural cold acclimation and deacclimation of two Magnolia species with different winter hardiness,Acta Physiol.Plant.,37(7):129
Zhang J.B.,Jin Y.F.,Wang S.S.,Yang H.Q.,Pang T.,Li J.Y.,Gao X.,Cui M.K.,and Gong M.,2015,Effect of different temperature on sucrose metabolism in different growth stages of tobacco,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(10):2225-2244(张建波,金云峰,王莎莎,杨慧芹,逄涛,李军营,高雪,崔明昆,龚明,2015,生长温度对不同生育期烟草蔗糖代谢的影响,基因组学与应用生物学,34(10):2225-2244)
Zhou G.Y.,Hu W.X.,Xu J.H.,and Xue Q.Z.,2013,Integrating multiple"omics"analysis for plant metabolites and their functions,Zhejiang Daxue Xuebao(Journal of Zhejiang U-niversity(Agriculture and Life Sciences)),39(3):237-245(周国艳,胡望雄,徐建红,薛庆中,2013,整合多个组学(omics)分析植物代谢产物及其功能,浙江大学学报(农业与生命科学版),39(3):237-245)
Zhu Z.L.,2012,Study on the limiting factor and winter protection methods of Magnolia wufengensis after introduction to Beijing,Thesis for M.S.,Beijing Forestry University,Supervisors:Ma L.Y.,and Jia Z.K.,pp.11-18(朱仲龙,2012,北京引种红花玉兰的限制因子与越冬防寒技术研究,硕士学位论文,北京林业大学,导师:马履一,贾忠奎,pp.11-18)