‘纽荷尔’脐橙与‘尤力克’柠檬种间体细胞杂种的代谢特征分析
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摘要
利用GC–MS(气相色谱—质谱)测定柑橘种间体细胞杂种——‘纽荷尔’脐橙(Citrus sinensis Osbeck)+‘尤力克’柠檬[Citrus limon(L.)Burm. F.]和其两个融合亲本果实的初生代谢物含量,包括可溶性糖、有机酸、氨基酸和脂肪酸;利用LC–MS(液相色谱—质谱)测定果实类黄酮含量,综合评估体细胞杂种果实的代谢特征。结果表明,体细胞杂种初生代谢物组成总体偏向‘尤力克’柠檬,苹果酸、柠檬酸和脂肪酸含量均高于两亲本,大多数氨基酸含量介于两亲本之间。体细胞杂种中共检测到48种类黄酮,整体偏向柠檬亲本;其中有33种物质含量介于两亲本之间,15种高于或低于两亲本。
The levels of primary metabolite in the fruit pulp of somatic hybrid [Citrus sinensis Osbeck‘Newhall'+ Citrus limon(L.)Burm. F.‘Eureka'] and both parental species were quantitatively evaluated by GC–MS(gas chromatography-mass spectrometry)and flavonoid contents were investigated by LC–MS(Liquid chromatography-mass spectrometry). The results indicated the allotetraploid was closer to the lemon parent in the accumulation of primary metabolites. The levels of malic acid,citric acid and four fatty acids were higher than those of the diploid parents,while the contents of most amino acids in the allotetraploid showed an additive manner. Additionally,a total of 48 flavonoids were detected in the allotetraploid. The contents of 33 flavonoids were intermediate between both parents. Meanwhile,the levels of 15 flavonoids were transgressive. The allotetraploid was closer to the lemon parent in flavonoid composition.
The levels of primary metabolite in the fruit pulp of somatic hybrid [Citrus sinensis Osbeck‘Newhall'+ Citrus limon(L.)Burm. F.‘Eureka'] and both parental species were quantitatively evaluated by GC–MS(gas chromatography-mass spectrometry)and flavonoid contents were investigated by LC–MS(Liquid chromatography-mass spectrometry). The results indicated the allotetraploid was closer to the lemon parent in the accumulation of primary metabolites. The levels of malic acid,citric acid and four fatty acids were higher than those of the diploid parents,while the contents of most amino acids in the allotetraploid showed an additive manner. Additionally,a total of 48 flavonoids were detected in the allotetraploid. The contents of 33 flavonoids were intermediate between both parents. Meanwhile,the levels of 15 flavonoids were transgressive. The allotetraploid was closer to the lemon parent in flavonoid composition.
引文
Aleza P,Juárez J,Cuenca J,Ollitrault P,Navarro L.2012.Extensive citrus triploid hybrid production by 2x×4x sexual hybridizations and parent-effect on the length of the juvenile phase.Plant Cell Reports,31(9):1723-1735.
Alonzo G,Fatta Del Bosco S,Palazzolo E,Saiano F,Tusa N.2000.Citrus somatic hybrid leaf essential oil.Flavour and Fragrance Journal,15(4):258-262.
Bassene J B,Berti L,Carcouet E,Dhuique-Mayer C,Fanciullino A L,Bouffin J,Froelicher Y.2008.Influence of mitochondria origin on fruit quality in a citrus cybrid.Journal of Agricultural and Food Chemistry,56(18):8635-8640.
Bassene J B,Berti L,Costantino G,Carcouet E,Kamiri M,Tomi F,Froelicher Y.2009a.Inheritance of characters involved in fruit quality in a citrus interspecific allotetraploid somatic hybrid.Journal of Agricultural and Food Chemistry,57:5065-5070.
Bassene J B,Frorlicher Y,Dhuique-Mayer C,Mouhaya W,Ferrer R M,Ancill O G,Morillon R,Navarro L,Ollitrault P.2009b.Non-additive phenotype and transcriptomic inheritance in a citrus allotetraploid somatic hybrid between C.reticulata and C.limon:the case of pulp carotenoid biosynthesis pathway.Plant Cell Reports,28(11):1689-1697.
Birchler J A.2012.Insights from paleogenomic and population studies into the consequences of dosage sensitive gene expression in plants.Current Opinion in Plant Biology,15(5):544-548.
Chen Jia-jing,Peng Zhao-xin,Shi Mei-yan,Xu Juan.2016.Advances in on flavonoid composition and metabolism in citrus.Acta Horticulturae Sinica,43(2):384-400.(in Chinese)陈嘉景,彭昭欣,石梅艳,徐娟.2016.柑橘中类黄酮的组成与代谢研究进展.园艺学报,43(2):384-400.
Cohen H,Fait A,Tel-Zur N.2013.Morphological,cytological and metabolic consequences of autopolyploidization in Hylocereus(Cactaceae)species.BMC Plant Biology,13(1):173-186.
Gancel A L,Olle’D,Ollitrault P,Luro F,Brillouet J M.2002.Leaf and peel volatile compounds of an interspecific citrus somatic hybrid[Citrus aurantifolia(Christm.)Swing+Citrus paradisi Macfayden].Flavor and Fragrance Journal,17(6):416-424.
Gancel A L,Ollitrault P,Froelicher Y,Tomi F,Jacquemond C,Luro F,Brillouet J M.2003.Leaf volatile compounds of seven citrus somatic tetraploid hybrids sharing willow leaf mandarin(Citrus deliciosa Ten)as their common parent.Journal of Agricultural and Food Chemistry,51(20):6006-6013.
Gancel A L,Ollitrault P,Froelicher Y,Tomi F,Jacquemond C,Luro F,Brillouet J M.2005.Leaf volatile compounds of six citrus somatic allotetraploid hybrids originating from various combinations of lime,lemon,citron,sweet orange,and grapefruit.Journal of Agricultural and Food Chemistry,53(6):2224-2230.
Grosser J W,Gmitter F G Jr.2011.Protoplast fusion for production of tetraploids and triploids:applications for scion and rootstock breeding in citrus.Plant Cell,Tissue and Organ Culture,104(3):343-357.
Grosser J W,Gmitter F G Jr,Tusa N,Chandler J L.1990.Somatic hybrid plants from sexually incompatible woody species:Citrus reticulata and Citropsis gilletiana.Plant Cell Reports,8(11):656-659.
Guo Wen-wu,Deng Xiu-xin,Hu Chun-gen.1999.Somatic hybrid plant regeneration between navel orange and lemon via protoplast electro-fusion.Journal of Wuhan Botanical Research,17(2):127-129.(in Chinese)郭文武,邓秀新,胡春根.1999.脐橙与柠檬种间细胞电融合再生杂种植株.武汉植物学研究,17(2):127-129.
Guo W W,Xiao S X,Deng X X.2013.Somatic cybrid production via protoplast fusion for citrus improvement.Scientia Horticulturae,163:20-26.
Lavania U C,Srivastava S,Lavania S,Basu S,Misra N K,Mukai Y.2012.Autopolyploidy differentially influences body size in plants,but facilitates enhanced accumulation of secondary metabolites,causing increased cytosine methylation.The Plant Journal,71(4):539-549.
Li A,Liu D,Wu J,Zhao X,Hao M,Geng S,Yan J,Jiang X,Zhang L,Wu J,Yin L,Zhang R,Wu L,Zheng Y,Mao L.2014.mRNA and small RNA transcriptomes reveal insights into dynamic homoeolog regulation of allopolyploid heterosis in nascent hexaploid wheat.Plant Cell,26(5):1878-1900.
Recupero G R,Russo G,Recupero S.2005.New promising Citrus triploid hybrids selected from crosses between monoembryonic diploid female and tetraploid male parents.HortScience,40(3):516-520.
Tan F Q,Tu H,Liang W J,Long J M,Wu X M,Zhang H Y and Guo W W.2015.Comparative metabolic and transcriptional analysis of a doubled diploid and its diploid citrus rootstock(C.junos cv.Ziyang Xiangcheng)suggests its potential value for stress resistance improvement.BMCPlant Biology,15(1):89-103.
Tan F Q,Tu H,Wang R,Wu X M,Xie K D,Chen J J,Zhang H Y,Xu J,Guo W W.2017.Metabolic adaptation following genome doubling in citrus doubled diploids revealed by non-targeted metabolomics.Metabolomics,13(11):143-155.
Tusa N,Abbate L,Renda A,Ruberto G.2007.Polyphenols distribution in juices from Citrus allotetraploid somatic hybrids and their sexual hybrids.Journal of Agricultural and Food Chemistry,55(22):9089-9094.
Wang J,Tian L,Lee H S,Wei N E,Jiang H,Watson B,Madlung A,Osborn T C,Doerge R W,Comai L.2006.Genomewide nonadditive gene regulation in Arabidopsis allotetraploids.Genetics,172(1):507-517.
Xie Kai-dong,Wang Hui-qin,Wang Xiao-pei,Liang Wu-jun,Xie Zong-zhou,Yi Hua-lin,Deng Xiu-xin,Grosser J W,Guo Wen-wu.2013.Extensive citrus triploid breeding by crossing monoembryonic diploid females with allotetraploid male parents.Scientia Agricultura Sinica,46(21):4550-4557.(in Chinese)解凯东,王惠芹,王晓培,梁武军,谢宗周,伊华林,邓秀新,Grosser J W,郭文武.2013.单胚性二倍体为母本与异源四倍体杂交大规模创制柑橘三倍体.中国农业科学,46(21):4550-4557.
Xie Kai-dong,Wang Xiao-pei,Wang Hui-qin,Liang Wu-jun,Xie Zong-zhou,Guo Da-yong,Yi Hua-lin,Deng Xiu-xin,Grosser J W,Guo Wen-wu.2014.High efficient and extensive production of triploid citrus plants by crossing polyembryonic diploids with tetraploids.Acta Horticulturae Sinica,41(4):613-620.(in Chinese)解凯东,王晓培,王惠芹,梁武军,谢宗周,郭大勇,伊华林,邓秀新,Grosser J W,郭文武.2014.以柑橘多胚性二倍体母本倍性杂交培育三倍体.园艺学报,41(4):613-620.
Xu C G,Tang T X,Chen R,Liang C H,Liu X Y,Wu C L,Yang Y S,Yang D P,Wu H.2014.A comparative study of bioactive secondary metabolite production in diploid and tetraploid Echinacea purpurea(L.).Moench.Plant Cell Tissue&Organ Culture,116(3):323-332
Yang J,Liu D,Wang X,Ji C,Cheng F,Liu B,Hu Z,Ju Y,Yao P,Li X.2016.The genome sequence of allopolyploid Brassica juncea and analysis of differential homoeolog gene expression influencing selection.Nature Genetics,48(10):1225-1232.
Yoo M J,Szadkowski E,Wendel J F.2013.Homoeolog expression bias and expression level dominance in allopolyploid cotton.Heredity,110(2):171-180.
Yu X,Wang X,Hyldgaard B,Zhu Z,Zhou R,Kj?r K H,Ouzounis T,Lou Q,Li J,Cai Q,Rosenqvist E,Ottosen C,Chen J.2018.Allopolyploidization in Cucumis contributes to delayed leaf maturation with repression of redundant homoeologous genes.The Plant Journal,94(2):393-404.
Yun Z,Gao H,Liu P,Liu S,Luo T,Jin S,Xu Q,Xu J,Cheng Y J,Deng X X.2013.Comparative proteomic and metabolomic profiling of citrus fruit with enhancement of disease resistance by postharvest heat treatment.BMC Plant Biology,13(1):44-60.
Zhang J,Wang X,Yu O,Tang J J,Gu X G,Wan X C,Fang C B.2010.Metabolic profiling of strawberry(Fragaria×ananassa Duch.)during fruit development and maturation.Journal of Experimental Botany,62(3):1103-1118.
Zheng Bei-bei,Xie Zong-zhou,Guo Wen-wu.2013.Parental dominant inheritance of fruit carotenoids,sugars and organic acids in a citrus interspecific allotetraploid somatic hybrid between Bonnaza navel orange and rough lemon.Acta Horticulturae Sinica,40(7):1262-1268.(in Chinese)郑蓓蓓,谢宗周,郭文武.2013.脐橙与粗柠檬体细胞杂种果实类胡萝卜素、糖酸遗传的亲本偏向性.园艺学报,40(7):1262-1268.
Alonzo G,Fatta Del Bosco S,Palazzolo E,Saiano F,Tusa N.2000.Citrus somatic hybrid leaf essential oil.Flavour and Fragrance Journal,15(4):258-262.
Bassene J B,Berti L,Carcouet E,Dhuique-Mayer C,Fanciullino A L,Bouffin J,Froelicher Y.2008.Influence of mitochondria origin on fruit quality in a citrus cybrid.Journal of Agricultural and Food Chemistry,56(18):8635-8640.
Bassene J B,Berti L,Costantino G,Carcouet E,Kamiri M,Tomi F,Froelicher Y.2009a.Inheritance of characters involved in fruit quality in a citrus interspecific allotetraploid somatic hybrid.Journal of Agricultural and Food Chemistry,57:5065-5070.
Bassene J B,Frorlicher Y,Dhuique-Mayer C,Mouhaya W,Ferrer R M,Ancill O G,Morillon R,Navarro L,Ollitrault P.2009b.Non-additive phenotype and transcriptomic inheritance in a citrus allotetraploid somatic hybrid between C.reticulata and C.limon:the case of pulp carotenoid biosynthesis pathway.Plant Cell Reports,28(11):1689-1697.
Birchler J A.2012.Insights from paleogenomic and population studies into the consequences of dosage sensitive gene expression in plants.Current Opinion in Plant Biology,15(5):544-548.
Chen Jia-jing,Peng Zhao-xin,Shi Mei-yan,Xu Juan.2016.Advances in on flavonoid composition and metabolism in citrus.Acta Horticulturae Sinica,43(2):384-400.(in Chinese)陈嘉景,彭昭欣,石梅艳,徐娟.2016.柑橘中类黄酮的组成与代谢研究进展.园艺学报,43(2):384-400.
Cohen H,Fait A,Tel-Zur N.2013.Morphological,cytological and metabolic consequences of autopolyploidization in Hylocereus(Cactaceae)species.BMC Plant Biology,13(1):173-186.
Gancel A L,Olle’D,Ollitrault P,Luro F,Brillouet J M.2002.Leaf and peel volatile compounds of an interspecific citrus somatic hybrid[Citrus aurantifolia(Christm.)Swing+Citrus paradisi Macfayden].Flavor and Fragrance Journal,17(6):416-424.
Gancel A L,Ollitrault P,Froelicher Y,Tomi F,Jacquemond C,Luro F,Brillouet J M.2003.Leaf volatile compounds of seven citrus somatic tetraploid hybrids sharing willow leaf mandarin(Citrus deliciosa Ten)as their common parent.Journal of Agricultural and Food Chemistry,51(20):6006-6013.
Gancel A L,Ollitrault P,Froelicher Y,Tomi F,Jacquemond C,Luro F,Brillouet J M.2005.Leaf volatile compounds of six citrus somatic allotetraploid hybrids originating from various combinations of lime,lemon,citron,sweet orange,and grapefruit.Journal of Agricultural and Food Chemistry,53(6):2224-2230.
Grosser J W,Gmitter F G Jr.2011.Protoplast fusion for production of tetraploids and triploids:applications for scion and rootstock breeding in citrus.Plant Cell,Tissue and Organ Culture,104(3):343-357.
Grosser J W,Gmitter F G Jr,Tusa N,Chandler J L.1990.Somatic hybrid plants from sexually incompatible woody species:Citrus reticulata and Citropsis gilletiana.Plant Cell Reports,8(11):656-659.
Guo Wen-wu,Deng Xiu-xin,Hu Chun-gen.1999.Somatic hybrid plant regeneration between navel orange and lemon via protoplast electro-fusion.Journal of Wuhan Botanical Research,17(2):127-129.(in Chinese)郭文武,邓秀新,胡春根.1999.脐橙与柠檬种间细胞电融合再生杂种植株.武汉植物学研究,17(2):127-129.
Guo W W,Xiao S X,Deng X X.2013.Somatic cybrid production via protoplast fusion for citrus improvement.Scientia Horticulturae,163:20-26.
Lavania U C,Srivastava S,Lavania S,Basu S,Misra N K,Mukai Y.2012.Autopolyploidy differentially influences body size in plants,but facilitates enhanced accumulation of secondary metabolites,causing increased cytosine methylation.The Plant Journal,71(4):539-549.
Li A,Liu D,Wu J,Zhao X,Hao M,Geng S,Yan J,Jiang X,Zhang L,Wu J,Yin L,Zhang R,Wu L,Zheng Y,Mao L.2014.mRNA and small RNA transcriptomes reveal insights into dynamic homoeolog regulation of allopolyploid heterosis in nascent hexaploid wheat.Plant Cell,26(5):1878-1900.
Recupero G R,Russo G,Recupero S.2005.New promising Citrus triploid hybrids selected from crosses between monoembryonic diploid female and tetraploid male parents.HortScience,40(3):516-520.
Tan F Q,Tu H,Liang W J,Long J M,Wu X M,Zhang H Y and Guo W W.2015.Comparative metabolic and transcriptional analysis of a doubled diploid and its diploid citrus rootstock(C.junos cv.Ziyang Xiangcheng)suggests its potential value for stress resistance improvement.BMCPlant Biology,15(1):89-103.
Tan F Q,Tu H,Wang R,Wu X M,Xie K D,Chen J J,Zhang H Y,Xu J,Guo W W.2017.Metabolic adaptation following genome doubling in citrus doubled diploids revealed by non-targeted metabolomics.Metabolomics,13(11):143-155.
Tusa N,Abbate L,Renda A,Ruberto G.2007.Polyphenols distribution in juices from Citrus allotetraploid somatic hybrids and their sexual hybrids.Journal of Agricultural and Food Chemistry,55(22):9089-9094.
Wang J,Tian L,Lee H S,Wei N E,Jiang H,Watson B,Madlung A,Osborn T C,Doerge R W,Comai L.2006.Genomewide nonadditive gene regulation in Arabidopsis allotetraploids.Genetics,172(1):507-517.
Xie Kai-dong,Wang Hui-qin,Wang Xiao-pei,Liang Wu-jun,Xie Zong-zhou,Yi Hua-lin,Deng Xiu-xin,Grosser J W,Guo Wen-wu.2013.Extensive citrus triploid breeding by crossing monoembryonic diploid females with allotetraploid male parents.Scientia Agricultura Sinica,46(21):4550-4557.(in Chinese)解凯东,王惠芹,王晓培,梁武军,谢宗周,伊华林,邓秀新,Grosser J W,郭文武.2013.单胚性二倍体为母本与异源四倍体杂交大规模创制柑橘三倍体.中国农业科学,46(21):4550-4557.
Xie Kai-dong,Wang Xiao-pei,Wang Hui-qin,Liang Wu-jun,Xie Zong-zhou,Guo Da-yong,Yi Hua-lin,Deng Xiu-xin,Grosser J W,Guo Wen-wu.2014.High efficient and extensive production of triploid citrus plants by crossing polyembryonic diploids with tetraploids.Acta Horticulturae Sinica,41(4):613-620.(in Chinese)解凯东,王晓培,王惠芹,梁武军,谢宗周,郭大勇,伊华林,邓秀新,Grosser J W,郭文武.2014.以柑橘多胚性二倍体母本倍性杂交培育三倍体.园艺学报,41(4):613-620.
Xu C G,Tang T X,Chen R,Liang C H,Liu X Y,Wu C L,Yang Y S,Yang D P,Wu H.2014.A comparative study of bioactive secondary metabolite production in diploid and tetraploid Echinacea purpurea(L.).Moench.Plant Cell Tissue&Organ Culture,116(3):323-332
Yang J,Liu D,Wang X,Ji C,Cheng F,Liu B,Hu Z,Ju Y,Yao P,Li X.2016.The genome sequence of allopolyploid Brassica juncea and analysis of differential homoeolog gene expression influencing selection.Nature Genetics,48(10):1225-1232.
Yoo M J,Szadkowski E,Wendel J F.2013.Homoeolog expression bias and expression level dominance in allopolyploid cotton.Heredity,110(2):171-180.
Yu X,Wang X,Hyldgaard B,Zhu Z,Zhou R,Kj?r K H,Ouzounis T,Lou Q,Li J,Cai Q,Rosenqvist E,Ottosen C,Chen J.2018.Allopolyploidization in Cucumis contributes to delayed leaf maturation with repression of redundant homoeologous genes.The Plant Journal,94(2):393-404.
Yun Z,Gao H,Liu P,Liu S,Luo T,Jin S,Xu Q,Xu J,Cheng Y J,Deng X X.2013.Comparative proteomic and metabolomic profiling of citrus fruit with enhancement of disease resistance by postharvest heat treatment.BMC Plant Biology,13(1):44-60.
Zhang J,Wang X,Yu O,Tang J J,Gu X G,Wan X C,Fang C B.2010.Metabolic profiling of strawberry(Fragaria×ananassa Duch.)during fruit development and maturation.Journal of Experimental Botany,62(3):1103-1118.
Zheng Bei-bei,Xie Zong-zhou,Guo Wen-wu.2013.Parental dominant inheritance of fruit carotenoids,sugars and organic acids in a citrus interspecific allotetraploid somatic hybrid between Bonnaza navel orange and rough lemon.Acta Horticulturae Sinica,40(7):1262-1268.(in Chinese)郑蓓蓓,谢宗周,郭文武.2013.脐橙与粗柠檬体细胞杂种果实类胡萝卜素、糖酸遗传的亲本偏向性.园艺学报,40(7):1262-1268.