温度和紫外照射对紫苏FAD7基因表达的影响
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摘要
植物ω-3脂肪酸脱氢酶(ω-3FAD)基因是催化亚油酸转化为α-亚麻酸(ALA)的关键酶基因,通过调节该基因的表达,可以提高植物ALA的含量。为了研究温度和紫外照射对紫苏PfFAD7的影响,通过RTPCR方法分析了紫苏地上组织的特异性表达和温度、紫外胁迫下紫苏叶片和茎中PfFAD7基因的积累情况。分析表明,PfFAD7基因在紫苏全植株中均有表达,但在叶片中表达量最高,温度和紫外照射均影响PfFAD7基因的表达,低温可诱导PfFAD7基因的表达,而高温则抑制PfFAD7基因的表达;UV-B照射下,PfFAD7基因在叶片和茎中表达量均表现为先升高再降低。本试验对于紫苏ω-3脂肪酸脱氢酶的研究有利于高水平ALA的积累,更有利于紫苏资源的开发和利用,同时对于进一步了解不饱和脂肪酸的积累和代谢过程以及关键基因PfFAD7在此过程中的功能提供了依据。
Plant ω-3 fatty acid dehydrogenase(ω-3 FAD) gene is a key enzyme gene that catalyzes the conversion of linoleic acid to α-linolenic acid(ALA). Plants α-linolenic acid(ALA) content could be increased by djusting the expression of this gene. In order to study the effect of temperature and ultraviolet irradiation on the PfFAD7 of Perilla frutescens, this study analyzed the specific expression of aboveground tissue in Perilla frutescens and the accumulation condition of gene PfFAD7 in stem and leaves of Perilla frutescens under temperature and ultraviolet irradiation stress by RT-PCR. The result showed that PfFAD7 was expressed in the whole plant of Perilla frutescens, but expressed highest in the leaves. And both the temperature and ultraviolet irradiation could affect the expression of PfFAD7 gene. Low temperature might induce the expression of PfFAD7, while the high temperature might inhibit the expression of PfFAD7. Under UV-B irradiation, the expression of PfFAD7 in leaves and stems increased first and then decreased. The study of ω-3 fatty acid dehydrogenase in Perilla frutescens might be beneficial to the accumulation of high level ALA, which might be more favorable to the development and utilization of perilla resources. And this study might provide a basis for further understanding the accumulation and metabolism of unsaturated fatty acids and the function of the key gene PfFAD7 in this process.
Plant ω-3 fatty acid dehydrogenase(ω-3 FAD) gene is a key enzyme gene that catalyzes the conversion of linoleic acid to α-linolenic acid(ALA). Plants α-linolenic acid(ALA) content could be increased by djusting the expression of this gene. In order to study the effect of temperature and ultraviolet irradiation on the PfFAD7 of Perilla frutescens, this study analyzed the specific expression of aboveground tissue in Perilla frutescens and the accumulation condition of gene PfFAD7 in stem and leaves of Perilla frutescens under temperature and ultraviolet irradiation stress by RT-PCR. The result showed that PfFAD7 was expressed in the whole plant of Perilla frutescens, but expressed highest in the leaves. And both the temperature and ultraviolet irradiation could affect the expression of PfFAD7 gene. Low temperature might induce the expression of PfFAD7, while the high temperature might inhibit the expression of PfFAD7. Under UV-B irradiation, the expression of PfFAD7 in leaves and stems increased first and then decreased. The study of ω-3 fatty acid dehydrogenase in Perilla frutescens might be beneficial to the accumulation of high level ALA, which might be more favorable to the development and utilization of perilla resources. And this study might provide a basis for further understanding the accumulation and metabolism of unsaturated fatty acids and the function of the key gene PfFAD7 in this process.
引文
Berberich T.,Harada M.,Sugawara K.,Kodama H.,Iba K.,and Ksano T.,1998,Two maize genes encodingω-3 fatty acid desaturase and their differential expression to temperature,Plant Mol.Biol.,36(2):297-306
Brandle J.R.,Campbell W.F.,Sisson W.B.,and Caldwell M.M.,1977,Net photosynthesis,electron transport capacity,and ultrastructure of Pisum sativum L.exposed to ultraviolet-Bradiation,Plant Physiol.,60(1):165-169
Christine J.,2000,Letter regarding dietary supplement health claim forω-3 fatty acids and coronary heart disease,Food and Drug Administration,12:2005
Dias A.S.,Barreiro M.G.,Campos P.S.,Ramalho J.C.,and Lidon F.C.,2010,Wheat cellular membrane thermotolerance under heat stress,J.Agron.Crop Sci.,196(2):100-108
Domínguez T.,Hernandez M.L.,Pennycooke J.C.,Jiménez P.,Martínez-Rivas J.M.,Sanz C.,Stockinger E.J.,S觃nchez-Ser rano J.J.,and Sanmartín M.,2010,Increasingω-3 desaturase expression in tomato results in altered aroma profile and enhanced resistance to cold stress,Plant Physiol.,153(2):655-665
Falcone D.L.,Ogas J.P.,and Somerville C.R.,2004,Regulation of membrane fatty acid composition by temperature in mutants of Arabidopsis with alterations in membrane lipid composition,BMC Plant Biol.,4(1):17
Kris-Etherton P.M.,Hecker K.D.,and Binkoski A.E.,2004,Polyunsaturated fatty acids and cardiovascular health,Nutr.Rev.,62(11):414-426
Kodama H.,Horiguchi G.,Nishiuchi T.,Nishimura M.,and Iba K.,1995,Fatty acid desaturation during chilling acclimation is one of the factors involved in conferring low-temperature tolerance to young tobacco leaves,Plant Physiol.,107(4):1177-1185
Lunn J.,and Theobald H.E.,2006,The health effects of dietary unsaturated fatty acids,Nutr.Bull.,31(3):178-224
Lai Z.C.,Lai G.T.,Zhang Q.L.,and Lin Y.L.,2013,Molecular cloning and functional analysis of fatty acid desaturase in the wild banana(Musa spp.,AB group),Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),34(10):1947-1954(赖志宸,赖恭梯,张群林,林玉玲,2013,野生蕉(Musa spp.,AB group)抗寒基因FAD7的分子克隆与功能分析,热带作物学报,34(10):1947-1954)
Livak K.J.,and Schmittgen T.D.,2001,Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CTmethod,Methods,25(4):402-408
Mikami K.,and Murata N.,2003,Membrane fluidity and the perception of environmental signals in cyanobacteria and plants,Prog.Lipid Res.,42(6):527-543
Murakami Y.,Tsuyama M.,Kobayashi Y.,Kodama H.,and Iba K.,2000,Trienoic fatty acids and plant tolerance of high temperature,Science,287(5452):476-479
Nishiuchi T.,Nakamura T.,Abe T.,Kodama H.,Nishimura M.,and Iba K.,1995,Tissue-specific and light responsive regulation of the promoter region of the Arabidopsis thaiiana chloroplastω-3 fatty acid desaturase gene(FAD7),Plant Mol.Biol.,29(3):599-609
Pearcy R.W.,1978,Effect of growth temperature on the fatty acid composition of the leaf lipids in Atriplex lentiformis(Torr.)Wats.,Plant Physiol.,61(4):484-486
Tang S.Y.,Guan R.Z.,Zhang H.S.,and Huang J.,2007,Cloning and expression analysis of three cDNAs encodingω-3 fatty acid desaturases from Descurainia sophia,Biotechnol.Lett.,29(9):1417-1424
Tang X.D.,An L.Z.,and Wang X.L.,1998,Effects of enhanced UV-B radiation on properties of microsomal membrane of broad bean leaves,Zhiwu Shengli Xuebao(Acta Phytophysiologica Sinica),24(2):171-176(唐旭东,安黎哲,王勋陵,1998,增强UV-B辐射对蚕豆叶片微粒体膜一些性质的影响,植物生理学报,24(2):171-176)
Wang H.S.,Yu C.,Tang X.F.,Zhu Z.J.,Ma N.N.,and Meng Q.W.,2014,A tomato endoplasmic reticulum(ER)-typeω-3fatty acid desaturase(LeFAD3)functions in early seedling tolerance to salinity stress,Plant Cell Rep.,33(1):131-142
Wang J.W.,2006,Cloning and characterization of a rice(Oryza sativa L.)gene encoding a temperature-dependent chloroplastω-3 fatty acid desaturase,Dissertation for Ph.D.,Fudan University,Supervisor:Shen D.L.,pp.1-81(王敬文,2006,水稻ω-3脂肪酸脱氢酶的基因克隆和功能研究,博士学位论文,复旦大学,导师:沈大棱,pp.1-81)
Yu C.,Wang H.S.,Yang S.,Tang X.F.,Duan M.,and Meng Q.W.,2009,Overexperssion of endoplasmic reticulcumω-3 fatty acid desaturase gene improves chilling tolerance in tomato,Plant Physiol.Biochem.,47(11):1102-1112
Yoon S.H.,and Noh S.,2011,Positional distribution of fatty acids in perilla(Perilla frutescens L.)oil,J.Am.Oil Chem.Soc.,88(1):157-158
Yang J.H.,Chen T.,and Wang X.L.,2000,The influence of enhanced ultraviolet-B radiation on chloroplast membrane composition and membrane fluidity in wheat leaves,Zhiwu Shengtai Xuebao(Chinese Journal of Plant Ecology),24(1):102-105(杨景宏,陈拓,王勋陵,2000,增强紫外线B辐射对小麦叶绿体膜组分和膜流动性的影响,植物生态学报,24(1):102-105)
Zhou Z.,Wang M.J.,Hu J.J.,Lu M.Z.,and Wang J.H.,2010,Improve freezing tolerance in transgenic poplar by overexpressing aω-3 fatty acid desaturase gene,Mol.Breed.,25(4):571-579
Zhang M.,Barg R.,Yin M.,Gueta-Dahan Y.,Leikin-Frenkel A.,Salts Y.,Shabtai S.,and Ben-Hayyim G.,2005,Modulated fatty acid desaturation via overexpression of two distinctω-3 desaturases differentially alters tolerance to various abiotic stresses in transgenic tobacco cells and plants,Plant J.,44(3):361-371
Zheng H.Q.,Rowland O.,and Kunst L.,2005,Disruptions of the Arabidopsis enoyl co A reductase gene reveal an essential role for very long chain fatty acid synthesis in cell expansion during plant morphogenesis,Plant Cell,5(17):1467-1481
Brandle J.R.,Campbell W.F.,Sisson W.B.,and Caldwell M.M.,1977,Net photosynthesis,electron transport capacity,and ultrastructure of Pisum sativum L.exposed to ultraviolet-Bradiation,Plant Physiol.,60(1):165-169
Christine J.,2000,Letter regarding dietary supplement health claim forω-3 fatty acids and coronary heart disease,Food and Drug Administration,12:2005
Dias A.S.,Barreiro M.G.,Campos P.S.,Ramalho J.C.,and Lidon F.C.,2010,Wheat cellular membrane thermotolerance under heat stress,J.Agron.Crop Sci.,196(2):100-108
Domínguez T.,Hernandez M.L.,Pennycooke J.C.,Jiménez P.,Martínez-Rivas J.M.,Sanz C.,Stockinger E.J.,S觃nchez-Ser rano J.J.,and Sanmartín M.,2010,Increasingω-3 desaturase expression in tomato results in altered aroma profile and enhanced resistance to cold stress,Plant Physiol.,153(2):655-665
Falcone D.L.,Ogas J.P.,and Somerville C.R.,2004,Regulation of membrane fatty acid composition by temperature in mutants of Arabidopsis with alterations in membrane lipid composition,BMC Plant Biol.,4(1):17
Kris-Etherton P.M.,Hecker K.D.,and Binkoski A.E.,2004,Polyunsaturated fatty acids and cardiovascular health,Nutr.Rev.,62(11):414-426
Kodama H.,Horiguchi G.,Nishiuchi T.,Nishimura M.,and Iba K.,1995,Fatty acid desaturation during chilling acclimation is one of the factors involved in conferring low-temperature tolerance to young tobacco leaves,Plant Physiol.,107(4):1177-1185
Lunn J.,and Theobald H.E.,2006,The health effects of dietary unsaturated fatty acids,Nutr.Bull.,31(3):178-224
Lai Z.C.,Lai G.T.,Zhang Q.L.,and Lin Y.L.,2013,Molecular cloning and functional analysis of fatty acid desaturase in the wild banana(Musa spp.,AB group),Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),34(10):1947-1954(赖志宸,赖恭梯,张群林,林玉玲,2013,野生蕉(Musa spp.,AB group)抗寒基因FAD7的分子克隆与功能分析,热带作物学报,34(10):1947-1954)
Livak K.J.,and Schmittgen T.D.,2001,Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CTmethod,Methods,25(4):402-408
Mikami K.,and Murata N.,2003,Membrane fluidity and the perception of environmental signals in cyanobacteria and plants,Prog.Lipid Res.,42(6):527-543
Murakami Y.,Tsuyama M.,Kobayashi Y.,Kodama H.,and Iba K.,2000,Trienoic fatty acids and plant tolerance of high temperature,Science,287(5452):476-479
Nishiuchi T.,Nakamura T.,Abe T.,Kodama H.,Nishimura M.,and Iba K.,1995,Tissue-specific and light responsive regulation of the promoter region of the Arabidopsis thaiiana chloroplastω-3 fatty acid desaturase gene(FAD7),Plant Mol.Biol.,29(3):599-609
Pearcy R.W.,1978,Effect of growth temperature on the fatty acid composition of the leaf lipids in Atriplex lentiformis(Torr.)Wats.,Plant Physiol.,61(4):484-486
Tang S.Y.,Guan R.Z.,Zhang H.S.,and Huang J.,2007,Cloning and expression analysis of three cDNAs encodingω-3 fatty acid desaturases from Descurainia sophia,Biotechnol.Lett.,29(9):1417-1424
Tang X.D.,An L.Z.,and Wang X.L.,1998,Effects of enhanced UV-B radiation on properties of microsomal membrane of broad bean leaves,Zhiwu Shengli Xuebao(Acta Phytophysiologica Sinica),24(2):171-176(唐旭东,安黎哲,王勋陵,1998,增强UV-B辐射对蚕豆叶片微粒体膜一些性质的影响,植物生理学报,24(2):171-176)
Wang H.S.,Yu C.,Tang X.F.,Zhu Z.J.,Ma N.N.,and Meng Q.W.,2014,A tomato endoplasmic reticulum(ER)-typeω-3fatty acid desaturase(LeFAD3)functions in early seedling tolerance to salinity stress,Plant Cell Rep.,33(1):131-142
Wang J.W.,2006,Cloning and characterization of a rice(Oryza sativa L.)gene encoding a temperature-dependent chloroplastω-3 fatty acid desaturase,Dissertation for Ph.D.,Fudan University,Supervisor:Shen D.L.,pp.1-81(王敬文,2006,水稻ω-3脂肪酸脱氢酶的基因克隆和功能研究,博士学位论文,复旦大学,导师:沈大棱,pp.1-81)
Yu C.,Wang H.S.,Yang S.,Tang X.F.,Duan M.,and Meng Q.W.,2009,Overexperssion of endoplasmic reticulcumω-3 fatty acid desaturase gene improves chilling tolerance in tomato,Plant Physiol.Biochem.,47(11):1102-1112
Yoon S.H.,and Noh S.,2011,Positional distribution of fatty acids in perilla(Perilla frutescens L.)oil,J.Am.Oil Chem.Soc.,88(1):157-158
Yang J.H.,Chen T.,and Wang X.L.,2000,The influence of enhanced ultraviolet-B radiation on chloroplast membrane composition and membrane fluidity in wheat leaves,Zhiwu Shengtai Xuebao(Chinese Journal of Plant Ecology),24(1):102-105(杨景宏,陈拓,王勋陵,2000,增强紫外线B辐射对小麦叶绿体膜组分和膜流动性的影响,植物生态学报,24(1):102-105)
Zhou Z.,Wang M.J.,Hu J.J.,Lu M.Z.,and Wang J.H.,2010,Improve freezing tolerance in transgenic poplar by overexpressing aω-3 fatty acid desaturase gene,Mol.Breed.,25(4):571-579
Zhang M.,Barg R.,Yin M.,Gueta-Dahan Y.,Leikin-Frenkel A.,Salts Y.,Shabtai S.,and Ben-Hayyim G.,2005,Modulated fatty acid desaturation via overexpression of two distinctω-3 desaturases differentially alters tolerance to various abiotic stresses in transgenic tobacco cells and plants,Plant J.,44(3):361-371
Zheng H.Q.,Rowland O.,and Kunst L.,2005,Disruptions of the Arabidopsis enoyl co A reductase gene reveal an essential role for very long chain fatty acid synthesis in cell expansion during plant morphogenesis,Plant Cell,5(17):1467-1481
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