UV-B辐射对神农香菊萜类物质合成及其相关基因表达的影响
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摘要
该研究以神农香菊为材料,用强度为400μw·cm~(-2)的紫外光UV-B对其进行辐射处理,辐射时间分别为0、0.5、1、2、4 h,探讨了UV-B辐射对神农香菊萜类物质合成及其相关基因表达的影响。结果表明:(1)较短时间的UV-B辐射对神农香菊萜类物质合成相关基因表达量有明显的促进作用。与对照相比,0.5、1、2、4 h处理对相关基因表达量均有不同程度的提高;在2 h处理下HMGR、DXR、TPS、GPS基因的相对表达量达到最大值,在4 h处理下FPS和DXS的相对表达量达到最大值,其中FPS基因表达量变化最显著,为对照的69倍。(2) MVA途径中,去氢白菖烯、杜松萜烯的含量与FPS基因表达量4 h内持续上升的变化趋势保持一致,1-石竹烯与HMGR的变化趋势保持一致,表现为先升高后降低。(3) MEP合成途径中,α-侧柏酮、崖柏酮、β-侧柏酮的含量呈现与DXR、GPS、TPS基因表达量相同的变化趋势,桉树脑在UV-B辐射4 h内持续上升,与DXS基因的变化一致。由此可以推断,UV-B辐射通过影响各自途径中一些关键基因的表达量,进而影响了神农香菊萜类物质的合成量。
In this study,Chrysanthemum indicum var. aromaticum was used as material. We examined the changes of biosynthesis of terpenoid and expression of related genes in C. indicum var. aromaticum following the treatment of different durations( 0,0.5,1,2,4 h) of 400 μw·cm~(-2) UV-B radiation. The results were as follows:( 1) The short time of UVB radiation could significantly promote the expression of related genes in C. indicum var. aromaticum. Compared with the control,0.5,1,2,4 h treatment of UV-B radiation increased the expression of related genes. Meanwhile,relative expressions of HMGR,DXR,TPS,GPS reached the peak after two hours' treatment of UV-B radiation. As well as the relative expressions of FPS and DXS reached the maximum at four hours' treatment. A significant increase in FPS gene expression was found when plant exposed to four hours of UV-B duration( 69 times compared with control).( 2) In mevalonate pathway( MVA),the contents of calamenene,cadinene increased following the change of FPS gene expression during four hours of UV-B radiation. The change of the content of 1-caryophyllene was consistent with the change of HMGR gene expression,which increased first and then decreased.( 3) In the pathway of 2-methyl-D-erythritol-4-phosphate( MEP),the contents of α-thujone,thujone and β-thujone showed the same trend as the expressions of DXR,TPS and GPS. The content of cajuputole generally increased during four hours' treatment responding to the change of DXS gene expression. In summary,the data indicate that UV-B radiation can change the regulation of terpenoid biosynthesis in C. indicum var. aromaticum by affecting some key genes' expressions.
In this study,Chrysanthemum indicum var. aromaticum was used as material. We examined the changes of biosynthesis of terpenoid and expression of related genes in C. indicum var. aromaticum following the treatment of different durations( 0,0.5,1,2,4 h) of 400 μw·cm~(-2) UV-B radiation. The results were as follows:( 1) The short time of UVB radiation could significantly promote the expression of related genes in C. indicum var. aromaticum. Compared with the control,0.5,1,2,4 h treatment of UV-B radiation increased the expression of related genes. Meanwhile,relative expressions of HMGR,DXR,TPS,GPS reached the peak after two hours' treatment of UV-B radiation. As well as the relative expressions of FPS and DXS reached the maximum at four hours' treatment. A significant increase in FPS gene expression was found when plant exposed to four hours of UV-B duration( 69 times compared with control).( 2) In mevalonate pathway( MVA),the contents of calamenene,cadinene increased following the change of FPS gene expression during four hours of UV-B radiation. The change of the content of 1-caryophyllene was consistent with the change of HMGR gene expression,which increased first and then decreased.( 3) In the pathway of 2-methyl-D-erythritol-4-phosphate( MEP),the contents of α-thujone,thujone and β-thujone showed the same trend as the expressions of DXR,TPS and GPS. The content of cajuputole generally increased during four hours' treatment responding to the change of DXS gene expression. In summary,the data indicate that UV-B radiation can change the regulation of terpenoid biosynthesis in C. indicum var. aromaticum by affecting some key genes' expressions.
引文
BROUN P,2004.Transcription factors as tools for metabolic engineering in plants[J].Curr Opin Plant Biol,7(2):202-209.
CHAPPELL J,1995.The biochemistry and molecular biology of isoprenoid metabolism[J].Plant Physiol,107(1):1-6.
DOLZHENKO Y,BERTEA CM,OCCHIPINTI A,et al.,2010.UV-B modulates the interplay between terpenoids and flavonoids in peppermint(Mentha×piperita L.)[J].J Photochem Photobiol B-Biol,100(2):67-75.
DUDAREVA N,ANDERSSON S,ORLOVA I,et al.,2005.The nonmevalonate pathway supports both monoterpene and sesquiterpene formation in snapdragon flowers[J].Proc Natl Acad Sci USA,102(3):933-938.
ENFISSI EMA,FRASER PD,LOIS LM,et al.,2005.Metabolic engineering of the mevalonate and non-mevalonate isopentenyl diphosphate-forming path ways for the production of health-promoting isoprenoids in tomato[J].Plant Biotechnol J,3(1):17-27.
HARTMANN U,VALENTINE WJ,CHRISTIE JM,et al.,1998.Identification of UV/blue light-response elements in the Arabidopsis thaliana chalcone synthase promoter using a homologous protoplast transient expression system[J].Plant Mol Biol,36(5):741-754.
HE M,LIU YZ,ZHANG R,et al.,2015.Dendranthema indicum var.aromaticum epidermal hairs and glandular hair secretions[J].J NE For Univ,43(9):117-120.[何淼,刘颖竹,张蕊,等,2015.神农香菊表皮毛特征及腺毛分泌物成分[J].东北林业大学学报,43(9):117-120.]
HU B,HONG L,LIU X,et al.,2012.Comparative study of the tissue-specific distribution of ABA from Arachis hypogaea L.and expression of the 9-CIS epoxycarotenoid dioxygenase1(Ah NCED1)during plant development[J].Biotechnol Biotechnol Equip,26(5):3201-3205.
JI BZ,LIU SW,TIAN L,et al.,2007.Advances in studies on juvenile hormone biosynthesis[J].Chin Bull Life Sci,19(1):90-96.[嵇保中,刘曙雯,田铃,等,2007.保幼激素生物合成研究进展[J].生命科学,19(1):90-96.]
LIANG ZS,FANG YM,YANG DF,2017.Biosynthesis segulation and metabolic engineering of terpenoids in plants[J].J Zhejiang Sci-Technol Univ(Nat Sci Ed),37(2):255-264.[梁宗锁,方誉民,杨东风,2017.植物萜类化合物生物合成与调控及其代谢工程研究进展[J].浙江理工大学学报(自然科学版),37(2):255-264.]
LIVAK KJ,SCHMITTGEN TD,2001.Analysis of relative gene expression data using real-time quantitative PCR and the2-ΔΔCTmethod[J].Methods,25(4):402-408.
LIU QH,CHEN YL,LI GY,1986.The research of introduction and propagation of Dendranthema indicum var.aromaticum[J].J Wuhan Bot Res,4(1):105-106.[刘启红,陈英兰,李广彦,1986.神农香菊引种和繁殖研究[J].武汉植物学研究,4(1):105-106.]
LIU QH,ZHANG SP,1983.A new variety of Dendranthema Gaertn from Shennongjia of Hubei[J].J Wuhan Bot Res,1(2):237-238.[刘启宏,张树藩,1983.神农架菊属一新变种[J].武汉植物学研究,1(2):237-238.]
MALHOTRA N,KUMAR V,SOOD H,et al.,2014.Multiple genes of mevalonate and non-mevalonate pathways contribute to high aconites content in an endangered medicinal herb,Aconitum heterophyllum Wall[J].Phytochemistry,108(12):26-34.
MARTIN DM,CHIANG A,LUND ST,et al.,2012.Biosynthesis of wine aroma:Transcript profiles of hydroxymethylbutenyl diphosphate reductase,geranyl diphosphate synthase,and linalool/nerolidol synthase parallel monoterpenol glycoside accumulation in Gewürztraminer grapes[J].Planta,236(3):919-929.
MORRIS WL,DUCREUX LJM,HEDDEN P,et al.,2006.Overexpression of a bacterial 1-deoxy-D-xylulose-5-phosphate synthase gene in potato tubers perturbs the isoprenoid metabolic network:Implication for the control of the tuber life cycle[J].J Exp Bot,57(12):3007-3018.
PU XH,YUE XL,AN LZ,2017.The response of plant to UV-B radiation[J].Sci Sin(Vitae),47(8):818-828.[蒲晓宏,岳修乐,安黎哲,2017.植物对UV-B辐射的响应与调控机制[J].中国科学:生命科学,47(8):818-828.]
QI Y,XING YY,ZHENG H,et al.,2014.UV-A and UV-Binvolved in induction and regulation of anthocyanin biosynthesis in cabbage[J].J Chin Agric Univ,19(2):86-94.[齐艳,邢燕霞,郑禾,等,2014.UV-A和UV-B提高甘蓝幼苗花青素含量以及调控基因表达分析[J].中国农业大学学报,19(2):86-94.]
SAFRANY J,HAASZ V,MATE Z,et al.,2008.Identification of a novel cis-regulatory element for UV-B-induced transcription in Arabidopsis[J].Plant J,54(3):402-414.
SEARLES PS,FLINT SD,CALDWELL MM,2001.A meta-analysis of plant field studies simulating stratospheric ozone depletion[J].Oecologia,127(1):1-10.
TANG L,MA X,ZHOU ZQ,2014.An evolutionary study of plant terpene synthases[J].J SW Univ(Nat Sci Ed),36(4):89-96.[唐亮,马香,周志钦,2014.植物萜类合成酶的进化研究[J].西南大学学报(自然科学版),36(4):89-96.]
ZHANG L,LIN SQ,HU YL,et al.,2013.Cloning of DXRgene and its expression analysis during fruit ripening in loquat[J].J Fruit Sci,30(4):563-566.[张玲,林顺权,胡又厘,等,2013.枇杷DXR基因的克隆及其在果实成熟过程中的表达分析[J].果树学报,30(4):563-566.]
CHAPPELL J,1995.The biochemistry and molecular biology of isoprenoid metabolism[J].Plant Physiol,107(1):1-6.
DOLZHENKO Y,BERTEA CM,OCCHIPINTI A,et al.,2010.UV-B modulates the interplay between terpenoids and flavonoids in peppermint(Mentha×piperita L.)[J].J Photochem Photobiol B-Biol,100(2):67-75.
DUDAREVA N,ANDERSSON S,ORLOVA I,et al.,2005.The nonmevalonate pathway supports both monoterpene and sesquiterpene formation in snapdragon flowers[J].Proc Natl Acad Sci USA,102(3):933-938.
ENFISSI EMA,FRASER PD,LOIS LM,et al.,2005.Metabolic engineering of the mevalonate and non-mevalonate isopentenyl diphosphate-forming path ways for the production of health-promoting isoprenoids in tomato[J].Plant Biotechnol J,3(1):17-27.
HARTMANN U,VALENTINE WJ,CHRISTIE JM,et al.,1998.Identification of UV/blue light-response elements in the Arabidopsis thaliana chalcone synthase promoter using a homologous protoplast transient expression system[J].Plant Mol Biol,36(5):741-754.
HE M,LIU YZ,ZHANG R,et al.,2015.Dendranthema indicum var.aromaticum epidermal hairs and glandular hair secretions[J].J NE For Univ,43(9):117-120.[何淼,刘颖竹,张蕊,等,2015.神农香菊表皮毛特征及腺毛分泌物成分[J].东北林业大学学报,43(9):117-120.]
HU B,HONG L,LIU X,et al.,2012.Comparative study of the tissue-specific distribution of ABA from Arachis hypogaea L.and expression of the 9-CIS epoxycarotenoid dioxygenase1(Ah NCED1)during plant development[J].Biotechnol Biotechnol Equip,26(5):3201-3205.
JI BZ,LIU SW,TIAN L,et al.,2007.Advances in studies on juvenile hormone biosynthesis[J].Chin Bull Life Sci,19(1):90-96.[嵇保中,刘曙雯,田铃,等,2007.保幼激素生物合成研究进展[J].生命科学,19(1):90-96.]
LIANG ZS,FANG YM,YANG DF,2017.Biosynthesis segulation and metabolic engineering of terpenoids in plants[J].J Zhejiang Sci-Technol Univ(Nat Sci Ed),37(2):255-264.[梁宗锁,方誉民,杨东风,2017.植物萜类化合物生物合成与调控及其代谢工程研究进展[J].浙江理工大学学报(自然科学版),37(2):255-264.]
LIVAK KJ,SCHMITTGEN TD,2001.Analysis of relative gene expression data using real-time quantitative PCR and the2-ΔΔCTmethod[J].Methods,25(4):402-408.
LIU QH,CHEN YL,LI GY,1986.The research of introduction and propagation of Dendranthema indicum var.aromaticum[J].J Wuhan Bot Res,4(1):105-106.[刘启红,陈英兰,李广彦,1986.神农香菊引种和繁殖研究[J].武汉植物学研究,4(1):105-106.]
LIU QH,ZHANG SP,1983.A new variety of Dendranthema Gaertn from Shennongjia of Hubei[J].J Wuhan Bot Res,1(2):237-238.[刘启宏,张树藩,1983.神农架菊属一新变种[J].武汉植物学研究,1(2):237-238.]
MALHOTRA N,KUMAR V,SOOD H,et al.,2014.Multiple genes of mevalonate and non-mevalonate pathways contribute to high aconites content in an endangered medicinal herb,Aconitum heterophyllum Wall[J].Phytochemistry,108(12):26-34.
MARTIN DM,CHIANG A,LUND ST,et al.,2012.Biosynthesis of wine aroma:Transcript profiles of hydroxymethylbutenyl diphosphate reductase,geranyl diphosphate synthase,and linalool/nerolidol synthase parallel monoterpenol glycoside accumulation in Gewürztraminer grapes[J].Planta,236(3):919-929.
MORRIS WL,DUCREUX LJM,HEDDEN P,et al.,2006.Overexpression of a bacterial 1-deoxy-D-xylulose-5-phosphate synthase gene in potato tubers perturbs the isoprenoid metabolic network:Implication for the control of the tuber life cycle[J].J Exp Bot,57(12):3007-3018.
PU XH,YUE XL,AN LZ,2017.The response of plant to UV-B radiation[J].Sci Sin(Vitae),47(8):818-828.[蒲晓宏,岳修乐,安黎哲,2017.植物对UV-B辐射的响应与调控机制[J].中国科学:生命科学,47(8):818-828.]
QI Y,XING YY,ZHENG H,et al.,2014.UV-A and UV-Binvolved in induction and regulation of anthocyanin biosynthesis in cabbage[J].J Chin Agric Univ,19(2):86-94.[齐艳,邢燕霞,郑禾,等,2014.UV-A和UV-B提高甘蓝幼苗花青素含量以及调控基因表达分析[J].中国农业大学学报,19(2):86-94.]
SAFRANY J,HAASZ V,MATE Z,et al.,2008.Identification of a novel cis-regulatory element for UV-B-induced transcription in Arabidopsis[J].Plant J,54(3):402-414.
SEARLES PS,FLINT SD,CALDWELL MM,2001.A meta-analysis of plant field studies simulating stratospheric ozone depletion[J].Oecologia,127(1):1-10.
TANG L,MA X,ZHOU ZQ,2014.An evolutionary study of plant terpene synthases[J].J SW Univ(Nat Sci Ed),36(4):89-96.[唐亮,马香,周志钦,2014.植物萜类合成酶的进化研究[J].西南大学学报(自然科学版),36(4):89-96.]
ZHANG L,LIN SQ,HU YL,et al.,2013.Cloning of DXRgene and its expression analysis during fruit ripening in loquat[J].J Fruit Sci,30(4):563-566.[张玲,林顺权,胡又厘,等,2013.枇杷DXR基因的克隆及其在果实成熟过程中的表达分析[J].果树学报,30(4):563-566.]