基于GC-MS技术对不同产地稻米的代谢组学分析
|
摘要
基于气相色谱-质谱(gas chromatography-mass spectrometry,GC-MS)联用技术对建三江和响水地区的60份稻米样本进行代谢组学研究。利用主成分分析和正交偏最小二乘-判别分析等多元统计分析方法,对建三江和响水的稻米样本进行检测,在样本中共检测出143个峰,鉴定出包括氨基酸、脂肪酸、核苷酸、有机酸、多元醇、糖类等在内的39个代谢产物。实验考察产地对水稻代谢组的影响,结果表明建三江地区和响水地区稻米代谢产物在数量和含量上均存在显著差异。对显著变化(P<0.05,VIP>1)的差异代谢物进行鉴定,共筛选出16种差异代谢产物。实验表明,利用GC-MS技术,结合多元统计分析的方法,可用于不同地区稻米代谢产物差异的研究,为稻米产地溯源提供理论依据。
Based on gas chromatography-mass spectrometry(GC-MS), metabolomics studies were conducted on 60 rice varieties growing in Jiansanjiang and Xiangshui, Heilongjiang. By applying the multivariate statistical analysis methods principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA), we examined rice samples collected from the two growing areas. A total of 143 peaks were detected in the samples, and a total of 39 metabolites including amino acids, fatty acids, nucleotides, organic acids, polyols, sugars were identified. The effect of the geographical origin on rice metabolome was evaluated. The results showed significant differences in both the kinds and quantities of metabolites in rice samples from different growing areas. A total of 16 differential metabolites with significant changes(P < 0.05, variable importance in the projection(VIP) > 1) were identified. The results showed that GC-MS combined with multivariate statistical analysis could be used to study the differences in the metabolite profiles of rice samples collected from different growing regions. This study provides the basis for studying the geographical traceability of rice.
Based on gas chromatography-mass spectrometry(GC-MS), metabolomics studies were conducted on 60 rice varieties growing in Jiansanjiang and Xiangshui, Heilongjiang. By applying the multivariate statistical analysis methods principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA), we examined rice samples collected from the two growing areas. A total of 143 peaks were detected in the samples, and a total of 39 metabolites including amino acids, fatty acids, nucleotides, organic acids, polyols, sugars were identified. The effect of the geographical origin on rice metabolome was evaluated. The results showed significant differences in both the kinds and quantities of metabolites in rice samples from different growing areas. A total of 16 differential metabolites with significant changes(P < 0.05, variable importance in the projection(VIP) > 1) were identified. The results showed that GC-MS combined with multivariate statistical analysis could be used to study the differences in the metabolite profiles of rice samples collected from different growing regions. This study provides the basis for studying the geographical traceability of rice.
引文
[1]NICHOLSON J K,CONNELLY J,LINDON J C,et al.Metabonomics:a platform for studying drug toxicity and gene function[J].Nature Review Drug Discovery,2002,1(2):153-161.DOI:10.1038/nrd728.
[2]REZZI S,RAMADAN Z,MARTIN F P J,et al.Human metabolic phenotypes link directly to specific dietary preferences in healthy individuals[J].Journal of Proteome Research,2007,6(11):4469-4477.DOI:10.1021/pr070431h.
[3]杨军,宋硕林,PEREZ J C,等.代谢组学及其应用[J].生物工程学报,2005,21(1):1-5.DOI:10.13345/j.cjb.2005.01.001.
[4]徐旻,林东海,刘昌孝.代谢组学研究现状与展望[J].医学学报,2005,40(9):769-774.DOI:10.16438/j.0513-4870.2005.09.001.
[5]唐惠儒,王玉兰.代谢组学研究[J].生命科学,2007,19(3):272-280.DOI:10.3969/j.issn.1004-0374.2007.03.005.
[6]黄强,尹沛源,路鑫,等.色谱-质谱联用技术在代谢组学中的应用[J].色谱,2009,27(5):566-572.DOI:10.3321/j.issn:1000-8713.2009.05.007.
[7]CALINGACION M N,BOUALAPHANH C,DAYGON V D,et al.A genomics and multi-platform metabolomics appro/ach to identify new traits of rice quality in traditional and improved varieties[J].Metabolomics,2012,8(7):771-783.DOI:10.1007/s11306-011-0374-4.
[8]MATSUDA F,OIKAWA A,KUSANO M,et al.Dissection of genotype-phenotype associations in rice grains using metabolome quantitative trait loci analysis[J].Plant Journal,2012,70(4):624-636.DOI:10.1111/j.1365-313X.2012.04903.x.
[9]CHEN W,GUO Z,WANG W,et al.A novel integrated method for large-scale detection,identification,and quantification of widelytargeted metabolites:application in the study of rice metabolomics[J].Molecular Plant,2013,6:1769-1780.DOI:10.1093/mp/sst080.
[10]JUNG E S,LEE S,LIM S H,et al.Metabolite profiling of the shortterm responses of rice leaves(Oryza sativa cv.Ilmi)cultivated under different LED lights and its correlations with antioxidant activities[J].Plant Science,2013,210:61-69.DOI:10.1016/j.plantsci.2013.05.004.
[11]YANG Z,NAKABAYASHI R,OKAZAKI Y,et al.Toward better annotation in plant metabolomics:isolation and structure elucidation of 36 specialized metabolites from Oryza sativa(rice)by using MS/MS and NMR analyses[J].Metabolomics,2014,10:543-555.DOI:10.1007/s11306-013-0619-5.
[12]王娜娜,冯昕鞾,孙玉萍,等.气相色谱结合化学计量学区分大米贮藏时间与产地[J].分析测试学报,2013,32(10):1227-1231.DOI:10.3969/j.issn.1004-4957.2013.10.014.
[13]王鹏飞,王倩倩,李先恩,等.GC-MS技术在延胡索块茎代谢产物研究中的应用[J].植物学报,2012(2):149-154.DOI:10.3724/SP.J.1259.2012.00149.
[14]CHAN E C,KOH P K,MAL M,et al.Metabolic profiling of human colorectal cancer using high-resolution magic angle spinning nuclear magnetic resonance(HR-MAS NMR)spectroscopy and gas chromatography mass spectrometry(GC/MS)[J].Journal of Proteome Research,2009,8(1):352-361.DOI:10.1021/pr8006232.
[15]STYCZYNSKI M P,MOXLEY J F,TONG L V,et al.Systematic identification of conserved metabolites in GC/MS data for metabolomics and biomarker discovery[J].Analytical Chemistry,2007,79(3):966-973.DOI:10.1016/0165-2370(82)80005-3.
[16]ZHANG Q,WANG G,DU Y,et al.GC/MS analysis of the rat urine for metabonomic research[J].Journal of Chromatography B Analytical Technologies in the Biomedical&Life Sciences,2007,854(1/2):20-25.DOI:10.1016/j.jchromb.2007.03.048.
[17]LUEDEMANN A,STRASSBURG K,ERBAN A,et al.TagFinder for the quantitative analysis of gas chromatography-mass spectrometry(GC-MS)-based metabolite profiling experiments[J].Bioinformatics,2008,24(5):732-737.DOI:10.1093/bioinformatics/btn023.
[18]PONGSUWAN W,BAMBA T,YONETANI T,et al.Quality prediction of Japanese green tea using pyrolyzer coupled GC/MSbased metabolic fingerprinting[J].Jounral of Agricultural and Food Chemistry,2008,56(3):744-750.
[19]SHU X L,FRANK T,SHU Q Y,et al.Metabolite profiling of germinating rice seeds[J].Jounral of Agricultural and Food Chemistry,2008,56(24):11612-11620.DOI:10.1021/jf802671p.
[20]LYTOVCHENKO A,BELEGGIA R,SCHAUER N,et al.Application of GC-MS for the detection of lipophilic compounds in diverse plant tissues[J].Plant Methods,2009,5(1):4.DOI:10.1186/1746-4811-5-4.
[21]何翠薇,陈玉萍,覃洁萍,等.木薯茎杆及叶化学成分初步研究[J].时珍国医国药,2011,22(4):908-909.
[22]HONG Z Y,LIN Z B,LIU Y,et al.Innovative microwave-assisted oximation and silylation procedures for metabolomic analysis of plasma samples using gas chromatography-mass spectrometry[J].Journalof Chromatography A,2012,1254:14-22.DOI:10.1016/J.CHROMA.2012.07.033.
[23]田福林,李红,刘成雁,等.GC-MS法对不同产地大米的快速鉴定[J].分析测试学报,2011(9):1059-1062.DOI:10.3969/j.issn.1004-4957.2011.09.020.
[24]王玲.转基因水稻的代谢组学研究[D].北京:北京化工大学,2013.
[25]何秀全,谭德冠,孙雪飘,等.应用GC-MS技术分离鉴定木薯叶片代谢物的极性组分[J].热带作物学报,2012,33(3):422-426.DOI:10.3969/j.issn.1000-2561.2012.03.006.
[26]王俊,许多宽,肖勇,等.基于化学指标的烟叶产区正交偏最小二乘判别分析[C]//中国烟草学会2017年学术年会.2017:9.DOI:10.13496/j.issn.1007-5119.2017.01.016.
[27]MORITA S,WADA H,MATSUE Y.Countermeasures for heat damage in rice grain quality under climate change[J].Plant Production Science,2016,19(1):1-11.DOI:10.1080/1343943x.2015.1128114.
[28]程建华,桑志红,李海静,等.基于GC-TOF/MS技术的转Bt基因大米代谢组学研究[J].分析测试学报,2016,35(10):1217-1224.DOI:10.3969/j.issn.1004-4957.2016.10.001.
[29]KAPLAN F,KOPKA J,SUNG D Y,et al.Transcript and metabolite profiling during cold acclimation of Arabidopsis reveals an intricate relationship of cold-regulated gene expression with modifications in metabolite content[J].Plant Journal,2007,50(6):967-981.DOI:10.1111/j.1365-313X.2007.03100.x.
[30]周晋军,刘蓬,尹静静,等.脯氨酸代谢途径在调控水稻phyB突变体干旱胁迫耐性中的作用[J].山东农业科学,2014,46(3):1-4.DOI:10.14083/j.issn.1001-4942.2014.03.020.
[31]LUGAN R,NIOGRET M F,LEPORT L,et al.Metabolome and water homeostasis analysis of Thellungiella salsuginea suggests that dehydration tolerance is a key response to osmotic stress in this halophyte[J].Plant Journal,2010,64(2):215-229.DOI:10.1111/j.1365-313X.2010.04323.x.
[32]谢兆辉.氨酰-tRNA合成酶维持翻译忠实性的机制[J].中国生物化学与分子生物学报,2011,27(2):110-115.DOI:10.13865/j.cnki.cjbmb.2011.02.002.
[33]张玉玉.植物中氨酰tRNA合成酶及相应tRNA的统计分析[D].北京:中国科学院北京基因组研究所,2013.
[34]钱瑞,张壮志,孙娟,等.温度和光照条件对裙带菜配子体生长的影响[J].河北渔业,2016(5):9-11;16.
[2]REZZI S,RAMADAN Z,MARTIN F P J,et al.Human metabolic phenotypes link directly to specific dietary preferences in healthy individuals[J].Journal of Proteome Research,2007,6(11):4469-4477.DOI:10.1021/pr070431h.
[3]杨军,宋硕林,PEREZ J C,等.代谢组学及其应用[J].生物工程学报,2005,21(1):1-5.DOI:10.13345/j.cjb.2005.01.001.
[4]徐旻,林东海,刘昌孝.代谢组学研究现状与展望[J].医学学报,2005,40(9):769-774.DOI:10.16438/j.0513-4870.2005.09.001.
[5]唐惠儒,王玉兰.代谢组学研究[J].生命科学,2007,19(3):272-280.DOI:10.3969/j.issn.1004-0374.2007.03.005.
[6]黄强,尹沛源,路鑫,等.色谱-质谱联用技术在代谢组学中的应用[J].色谱,2009,27(5):566-572.DOI:10.3321/j.issn:1000-8713.2009.05.007.
[7]CALINGACION M N,BOUALAPHANH C,DAYGON V D,et al.A genomics and multi-platform metabolomics appro/ach to identify new traits of rice quality in traditional and improved varieties[J].Metabolomics,2012,8(7):771-783.DOI:10.1007/s11306-011-0374-4.
[8]MATSUDA F,OIKAWA A,KUSANO M,et al.Dissection of genotype-phenotype associations in rice grains using metabolome quantitative trait loci analysis[J].Plant Journal,2012,70(4):624-636.DOI:10.1111/j.1365-313X.2012.04903.x.
[9]CHEN W,GUO Z,WANG W,et al.A novel integrated method for large-scale detection,identification,and quantification of widelytargeted metabolites:application in the study of rice metabolomics[J].Molecular Plant,2013,6:1769-1780.DOI:10.1093/mp/sst080.
[10]JUNG E S,LEE S,LIM S H,et al.Metabolite profiling of the shortterm responses of rice leaves(Oryza sativa cv.Ilmi)cultivated under different LED lights and its correlations with antioxidant activities[J].Plant Science,2013,210:61-69.DOI:10.1016/j.plantsci.2013.05.004.
[11]YANG Z,NAKABAYASHI R,OKAZAKI Y,et al.Toward better annotation in plant metabolomics:isolation and structure elucidation of 36 specialized metabolites from Oryza sativa(rice)by using MS/MS and NMR analyses[J].Metabolomics,2014,10:543-555.DOI:10.1007/s11306-013-0619-5.
[12]王娜娜,冯昕鞾,孙玉萍,等.气相色谱结合化学计量学区分大米贮藏时间与产地[J].分析测试学报,2013,32(10):1227-1231.DOI:10.3969/j.issn.1004-4957.2013.10.014.
[13]王鹏飞,王倩倩,李先恩,等.GC-MS技术在延胡索块茎代谢产物研究中的应用[J].植物学报,2012(2):149-154.DOI:10.3724/SP.J.1259.2012.00149.
[14]CHAN E C,KOH P K,MAL M,et al.Metabolic profiling of human colorectal cancer using high-resolution magic angle spinning nuclear magnetic resonance(HR-MAS NMR)spectroscopy and gas chromatography mass spectrometry(GC/MS)[J].Journal of Proteome Research,2009,8(1):352-361.DOI:10.1021/pr8006232.
[15]STYCZYNSKI M P,MOXLEY J F,TONG L V,et al.Systematic identification of conserved metabolites in GC/MS data for metabolomics and biomarker discovery[J].Analytical Chemistry,2007,79(3):966-973.DOI:10.1016/0165-2370(82)80005-3.
[16]ZHANG Q,WANG G,DU Y,et al.GC/MS analysis of the rat urine for metabonomic research[J].Journal of Chromatography B Analytical Technologies in the Biomedical&Life Sciences,2007,854(1/2):20-25.DOI:10.1016/j.jchromb.2007.03.048.
[17]LUEDEMANN A,STRASSBURG K,ERBAN A,et al.TagFinder for the quantitative analysis of gas chromatography-mass spectrometry(GC-MS)-based metabolite profiling experiments[J].Bioinformatics,2008,24(5):732-737.DOI:10.1093/bioinformatics/btn023.
[18]PONGSUWAN W,BAMBA T,YONETANI T,et al.Quality prediction of Japanese green tea using pyrolyzer coupled GC/MSbased metabolic fingerprinting[J].Jounral of Agricultural and Food Chemistry,2008,56(3):744-750.
[19]SHU X L,FRANK T,SHU Q Y,et al.Metabolite profiling of germinating rice seeds[J].Jounral of Agricultural and Food Chemistry,2008,56(24):11612-11620.DOI:10.1021/jf802671p.
[20]LYTOVCHENKO A,BELEGGIA R,SCHAUER N,et al.Application of GC-MS for the detection of lipophilic compounds in diverse plant tissues[J].Plant Methods,2009,5(1):4.DOI:10.1186/1746-4811-5-4.
[21]何翠薇,陈玉萍,覃洁萍,等.木薯茎杆及叶化学成分初步研究[J].时珍国医国药,2011,22(4):908-909.
[22]HONG Z Y,LIN Z B,LIU Y,et al.Innovative microwave-assisted oximation and silylation procedures for metabolomic analysis of plasma samples using gas chromatography-mass spectrometry[J].Journalof Chromatography A,2012,1254:14-22.DOI:10.1016/J.CHROMA.2012.07.033.
[23]田福林,李红,刘成雁,等.GC-MS法对不同产地大米的快速鉴定[J].分析测试学报,2011(9):1059-1062.DOI:10.3969/j.issn.1004-4957.2011.09.020.
[24]王玲.转基因水稻的代谢组学研究[D].北京:北京化工大学,2013.
[25]何秀全,谭德冠,孙雪飘,等.应用GC-MS技术分离鉴定木薯叶片代谢物的极性组分[J].热带作物学报,2012,33(3):422-426.DOI:10.3969/j.issn.1000-2561.2012.03.006.
[26]王俊,许多宽,肖勇,等.基于化学指标的烟叶产区正交偏最小二乘判别分析[C]//中国烟草学会2017年学术年会.2017:9.DOI:10.13496/j.issn.1007-5119.2017.01.016.
[27]MORITA S,WADA H,MATSUE Y.Countermeasures for heat damage in rice grain quality under climate change[J].Plant Production Science,2016,19(1):1-11.DOI:10.1080/1343943x.2015.1128114.
[28]程建华,桑志红,李海静,等.基于GC-TOF/MS技术的转Bt基因大米代谢组学研究[J].分析测试学报,2016,35(10):1217-1224.DOI:10.3969/j.issn.1004-4957.2016.10.001.
[29]KAPLAN F,KOPKA J,SUNG D Y,et al.Transcript and metabolite profiling during cold acclimation of Arabidopsis reveals an intricate relationship of cold-regulated gene expression with modifications in metabolite content[J].Plant Journal,2007,50(6):967-981.DOI:10.1111/j.1365-313X.2007.03100.x.
[30]周晋军,刘蓬,尹静静,等.脯氨酸代谢途径在调控水稻phyB突变体干旱胁迫耐性中的作用[J].山东农业科学,2014,46(3):1-4.DOI:10.14083/j.issn.1001-4942.2014.03.020.
[31]LUGAN R,NIOGRET M F,LEPORT L,et al.Metabolome and water homeostasis analysis of Thellungiella salsuginea suggests that dehydration tolerance is a key response to osmotic stress in this halophyte[J].Plant Journal,2010,64(2):215-229.DOI:10.1111/j.1365-313X.2010.04323.x.
[32]谢兆辉.氨酰-tRNA合成酶维持翻译忠实性的机制[J].中国生物化学与分子生物学报,2011,27(2):110-115.DOI:10.13865/j.cnki.cjbmb.2011.02.002.
[33]张玉玉.植物中氨酰tRNA合成酶及相应tRNA的统计分析[D].北京:中国科学院北京基因组研究所,2013.
[34]钱瑞,张壮志,孙娟,等.温度和光照条件对裙带菜配子体生长的影响[J].河北渔业,2016(5):9-11;16.