Gene cloning and expression analysis of limonene synthase in Syringa oblata and S.oblata var.alba
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摘要
Syringa species are important ornamentals with strong floral scent,of which monoterpenes are the main component.In this study,a new monoterpene synthase gene,named SoLIM,was collected from the flowers of Syringa oblata and S.oblata var.alba using a homologous cloning method.The full-length cDNA of SoLIM was1746 bp and encoded 581 amino acids.Sequence analysis showed that SoLIM contained the DDxxD and RRx8 W motifs,which are two typical conserved monoterpene synthase motifs,and was thus classified as belonging to the Tpsb subfamily.Using quantitative reverse-transcription PCR,SoLIM was significantly expressed in the petals and pistils of S.oblata and S.oblata var.alba,respectively.SoLIM expression peaked earlier than the D-limonene emissions in the diurnal experiments,but occurred later when D-limonene had peaked during the flowering phase,indicating that differences in SoLIM gene expression and D-limonene emissions existed.The synthesis of floral scent is thus associated with diverse regulatory mechanisms that require further investigation.
Syringa species are important ornamentals with strong floral scent,of which monoterpenes are the main component.In this study,a new monoterpene synthase gene,named SoLIM,was collected from the flowers of Syringa oblata and S.oblata var.alba using a homologous cloning method.The full-length cDNA of SoLIM was1746 bp and encoded 581 amino acids.Sequence analysis showed that SoLIM contained the DDxxD and RRx8 W motifs,which are two typical conserved monoterpene synthase motifs,and was thus classified as belonging to the Tpsb subfamily.Using quantitative reverse-transcription PCR,SoLIM was significantly expressed in the petals and pistils of S.oblata and S.oblata var.alba,respectively.SoLIM expression peaked earlier than the D-limonene emissions in the diurnal experiments,but occurred later when D-limonene had peaked during the flowering phase,indicating that differences in SoLIM gene expression and D-limonene emissions existed.The synthesis of floral scent is thus associated with diverse regulatory mechanisms that require further investigation.
Syringa species are important ornamentals with strong floral scent,of which monoterpenes are the main component.In this study,a new monoterpene synthase gene,named SoLIM,was collected from the flowers of Syringa oblata and S.oblata var.alba using a homologous cloning method.The full-length cDNA of SoLIM was1746 bp and encoded 581 amino acids.Sequence analysis showed that SoLIM contained the DDxxD and RRx8 W motifs,which are two typical conserved monoterpene synthase motifs,and was thus classified as belonging to the Tpsb subfamily.Using quantitative reverse-transcription PCR,SoLIM was significantly expressed in the petals and pistils of S.oblata and S.oblata var.alba,respectively.SoLIM expression peaked earlier than the D-limonene emissions in the diurnal experiments,but occurred later when D-limonene had peaked during the flowering phase,indicating that differences in SoLIM gene expression and D-limonene emissions existed.The synthesis of floral scent is thus associated with diverse regulatory mechanisms that require further investigation.
引文
Alonso WR,Rajaonarivony JI,Gershenzon J,Croteau R(1992)Purification of 4S-limonene synthase,a monoterpene cyclase from the glandular trichomes of peppermint(Mentha 9 piperita)and Spearmint(Mentha spicata).Biol Chem 267:7582-7587
Byun-McKay A,Godard KA,Toudefallah M,Martin DM,Alfaro R,King J,Bohlmann J,Plant AL(2006)Wound-induced terpene synthase gene expression in Sitka Spruce that exhibit resistance or susceptibility to attack by the white Pine weevil.Plant Physiol140:1009-1021
Chappell J(1995)The biochemistry and molecular biology of isoprenoid metabolism.Plant Physiol 107:1-6
Dudareva N,Cseke L,Blanc VM,Pichersky E(1996)Evolution of floral scent in Clarkia:novel patterns of S-linalool synthase gene expression in the C.breweri flower.Plant Cell 8:1137-1148
Dudareva N,Murfitt LM,Mann CJ,Gorenstein N,Kolosova N,Kish CM,Bonham C,Wood K(2000)Developmental regulation of methyl benzoate biosynthesis and emission in Snapdragon flowers.Plant Cell 12:949-961
Dudareva N,Martin D,Kish CM,Kolosova N,Gorenstein N,Faldt J,Miller B,Bohlmann J(2003)(E)-b-Ocimene and myrcene synthase genes of floral scent biosynthesis in Snapdragon:function and expression of three terpene synthase genes of a new terpene synthase subfamily.Plant Cell 15:1227-1241
Fu JX,Hou D,Zhang C,Bao ZY,Zhao HB,Hu SQ(2017)The emission of the floral scent of four Osmanthus fragrans cultivars in response to different temperatures.Molecules 22:430
Hu ZH,Zhang HX,Leng PS,Zhao J,Wang WH,Wang SD(2013)The emission of floral scent from Lilium‘siberia’in response to light intensity and temperature.Acta Physiol Plant35:1691-1700
Hu ZH,Li TJ,Zheng J,Leng PS,Yang K,Zhang KZ(2016)A new monoterpene synthase gene involved in the monoterpene production from Lilium‘siberia’.J Anim Plant Sci26:1389-1398
Huang KF,Lee YR,Tseng YH,Wang SY,Chu FH(2015)Cloning and functional characterization of a monoterpene synthase gene from Eleutherococcus trifoliatus.Holzforschung 69:163-171
Iijima Y,Davidovich-Rikanati D,Fridman E,Gang DR,Bar E,Lewinsohn E,Pichersky E(2004)The biochemical and molecular basis for the divergent patterns in the biosynthesis of terpenes and phenylpropenes in the peltate glands of three cultivars of Basil.Plant Physiol 136:3724-3736
Kim SY,An HR,Park PM,Baek YS,Kwon OK,Park SY,Park PH(2016)Analysis of floral scent patterns in flowering stages and floral organs of Maxillaria using an electronic nose.Flower Res J24:171-180
Knudsen JT,Tollsten L,Bergstrom LG(1993)Floral scents-a checklist of volatile compounds isolated by head-space techniques.Phytochemistry 33:253-280
Knudsen JT,Eriksson R,Gershenzon J,Stahl B(2006)Diversity and distribution of floral scent.Bot Rev 72:1-120
Kolosova N,Gorenstein N,Kish CM,Dudareva N(2001)Regulation of circadian methyl benzoate emission in diurnally and nocturnally emitting plants.Plant Cell 13:2333-2347
Li TJ,Hu ZH,Zheng J,Leng PS,Yang K(2016)Diurnal changes of monoterpenes emission and Li-mTPS expression in Lilium‘siberia’.Acta Agric Boreali Occident Sin 25:763-769
McGarvey DJ,Croteau R(1995)Terpenoid metabolism.Plant Cell7:1015-1026
Mendoza-Poudereux I,Munoz-Bertomeu J,Navarro A,Arrillaga I,Segura J(2014)Enhanced levels of S-linalool by metabolic engineering of the terpenoid pathway in spike lavender leaves.Metab Eng 23:136-144
ParePW,Tumlinson JH(1997)De novo biosynthesis of volatiles induced by insect herbivory in cotton plants.Plant Physiol114:1161-1167
Qin Y,Yang XX,Leng PS,Hu ZH(2015)Analysis of floral volatile components from 6 Syringa plants collected by dynamic headspace sampling by ATD-GC/MS.Acta Bot Boreal Occident Sin 35:2078-2088
Shimada T,Endo T,Fujii H,Omura M(2005)Isolation and characterization of a new D-limonene synthase gene with a different expression pattern in Citrus unshiu Marc.Sci Hortic105:507-512
Su GZ,Cao Y,Li C,Yu XL,Gao XL,Tu PF,Chai XY(2015)Phytochemical and pharmacological progress on the genus Syringa.Chem Cent J 9:2
Sun HN,Zhang T,Fan QQ,Qi XY,Zhang F,Fang WM,Jiang JF,Chen FD,Chen SM(2015)Identification of floral scent in Chrysanthemum cultivars and wild relatives by gas chromatography-mass spectrometry.Molecules 20:5346-5359
Wang JH,Dudareva N,Bhakta S,Raguso RA,Pichersky E(1997)Floral scent production in Clarkia breweri(Onagraceae):II.Localization and developmental modulation of the enzyme S-adenosyl-L-methionine:(iso)eugenol O-methyltransferase and phenylpropanoid emission.Plant Physiol 114:213-221
Wang HT,Yu X,Liu Y,Liang CY,Li WL(2013)Analysis of genetic variability and relationships among Mentha L.using the limonene synthase gene,LS.Gene 524:246-252
Yang XX,Zhao J,Zheng J,Leng PS,Li XL,Hu ZH,Liu JB,Meng X(2016)Analysis of floral scent emitted from Syringa plants.J For Res 27:273-281
Yue YC,Yu RC,Fan YP(2014)Characterization of two monoterpene synthases involved in floral scent formation in Hedychium coronarium.Planta 240:745-762
Zhang JH,Sun HL,Chen SY,Zeng L,Wang TT(2017)Anti-fungal activity,mechanism studies on a-phellandrene and nonanal against Penicillium cyclopium.Bot Stud 58:13
Zhao J,Hu ZH,Leng PS,Zhang HX,Cheng FY(2012)Fragrance composition in six tree peony cultivars.Kor J Hort Sci Technol30:617-625
Byun-McKay A,Godard KA,Toudefallah M,Martin DM,Alfaro R,King J,Bohlmann J,Plant AL(2006)Wound-induced terpene synthase gene expression in Sitka Spruce that exhibit resistance or susceptibility to attack by the white Pine weevil.Plant Physiol140:1009-1021
Chappell J(1995)The biochemistry and molecular biology of isoprenoid metabolism.Plant Physiol 107:1-6
Dudareva N,Cseke L,Blanc VM,Pichersky E(1996)Evolution of floral scent in Clarkia:novel patterns of S-linalool synthase gene expression in the C.breweri flower.Plant Cell 8:1137-1148
Dudareva N,Murfitt LM,Mann CJ,Gorenstein N,Kolosova N,Kish CM,Bonham C,Wood K(2000)Developmental regulation of methyl benzoate biosynthesis and emission in Snapdragon flowers.Plant Cell 12:949-961
Dudareva N,Martin D,Kish CM,Kolosova N,Gorenstein N,Faldt J,Miller B,Bohlmann J(2003)(E)-b-Ocimene and myrcene synthase genes of floral scent biosynthesis in Snapdragon:function and expression of three terpene synthase genes of a new terpene synthase subfamily.Plant Cell 15:1227-1241
Fu JX,Hou D,Zhang C,Bao ZY,Zhao HB,Hu SQ(2017)The emission of the floral scent of four Osmanthus fragrans cultivars in response to different temperatures.Molecules 22:430
Hu ZH,Zhang HX,Leng PS,Zhao J,Wang WH,Wang SD(2013)The emission of floral scent from Lilium‘siberia’in response to light intensity and temperature.Acta Physiol Plant35:1691-1700
Hu ZH,Li TJ,Zheng J,Leng PS,Yang K,Zhang KZ(2016)A new monoterpene synthase gene involved in the monoterpene production from Lilium‘siberia’.J Anim Plant Sci26:1389-1398
Huang KF,Lee YR,Tseng YH,Wang SY,Chu FH(2015)Cloning and functional characterization of a monoterpene synthase gene from Eleutherococcus trifoliatus.Holzforschung 69:163-171
Iijima Y,Davidovich-Rikanati D,Fridman E,Gang DR,Bar E,Lewinsohn E,Pichersky E(2004)The biochemical and molecular basis for the divergent patterns in the biosynthesis of terpenes and phenylpropenes in the peltate glands of three cultivars of Basil.Plant Physiol 136:3724-3736
Kim SY,An HR,Park PM,Baek YS,Kwon OK,Park SY,Park PH(2016)Analysis of floral scent patterns in flowering stages and floral organs of Maxillaria using an electronic nose.Flower Res J24:171-180
Knudsen JT,Tollsten L,Bergstrom LG(1993)Floral scents-a checklist of volatile compounds isolated by head-space techniques.Phytochemistry 33:253-280
Knudsen JT,Eriksson R,Gershenzon J,Stahl B(2006)Diversity and distribution of floral scent.Bot Rev 72:1-120
Kolosova N,Gorenstein N,Kish CM,Dudareva N(2001)Regulation of circadian methyl benzoate emission in diurnally and nocturnally emitting plants.Plant Cell 13:2333-2347
Li TJ,Hu ZH,Zheng J,Leng PS,Yang K(2016)Diurnal changes of monoterpenes emission and Li-mTPS expression in Lilium‘siberia’.Acta Agric Boreali Occident Sin 25:763-769
McGarvey DJ,Croteau R(1995)Terpenoid metabolism.Plant Cell7:1015-1026
Mendoza-Poudereux I,Munoz-Bertomeu J,Navarro A,Arrillaga I,Segura J(2014)Enhanced levels of S-linalool by metabolic engineering of the terpenoid pathway in spike lavender leaves.Metab Eng 23:136-144
ParePW,Tumlinson JH(1997)De novo biosynthesis of volatiles induced by insect herbivory in cotton plants.Plant Physiol114:1161-1167
Qin Y,Yang XX,Leng PS,Hu ZH(2015)Analysis of floral volatile components from 6 Syringa plants collected by dynamic headspace sampling by ATD-GC/MS.Acta Bot Boreal Occident Sin 35:2078-2088
Shimada T,Endo T,Fujii H,Omura M(2005)Isolation and characterization of a new D-limonene synthase gene with a different expression pattern in Citrus unshiu Marc.Sci Hortic105:507-512
Su GZ,Cao Y,Li C,Yu XL,Gao XL,Tu PF,Chai XY(2015)Phytochemical and pharmacological progress on the genus Syringa.Chem Cent J 9:2
Sun HN,Zhang T,Fan QQ,Qi XY,Zhang F,Fang WM,Jiang JF,Chen FD,Chen SM(2015)Identification of floral scent in Chrysanthemum cultivars and wild relatives by gas chromatography-mass spectrometry.Molecules 20:5346-5359
Wang JH,Dudareva N,Bhakta S,Raguso RA,Pichersky E(1997)Floral scent production in Clarkia breweri(Onagraceae):II.Localization and developmental modulation of the enzyme S-adenosyl-L-methionine:(iso)eugenol O-methyltransferase and phenylpropanoid emission.Plant Physiol 114:213-221
Wang HT,Yu X,Liu Y,Liang CY,Li WL(2013)Analysis of genetic variability and relationships among Mentha L.using the limonene synthase gene,LS.Gene 524:246-252
Yang XX,Zhao J,Zheng J,Leng PS,Li XL,Hu ZH,Liu JB,Meng X(2016)Analysis of floral scent emitted from Syringa plants.J For Res 27:273-281
Yue YC,Yu RC,Fan YP(2014)Characterization of two monoterpene synthases involved in floral scent formation in Hedychium coronarium.Planta 240:745-762
Zhang JH,Sun HL,Chen SY,Zeng L,Wang TT(2017)Anti-fungal activity,mechanism studies on a-phellandrene and nonanal against Penicillium cyclopium.Bot Stud 58:13
Zhao J,Hu ZH,Leng PS,Zhang HX,Cheng FY(2012)Fragrance composition in six tree peony cultivars.Kor J Hort Sci Technol30:617-625