彩叶草的查尔酮异构酶（CHI）基因的遗传转化及其功能验证
|
摘要
类黄酮是植物重要的次生代谢产物,亦称维生素P,常与维生素C伴存,普遍存在于蔬菜、水果、花和谷物中的天然色素,因多呈黄色而称生物类黄酮。参与植物花色的形成、UV防护、抵抗病原体、花粉发育、植物激素的运输以及豆科植物根瘤菌的相互作用,具有十分重要的生理功能。至今已分离鉴定出的有4000多种。许多研究证实,类黄酮在人体中具有多种生物活性,包括抗氧化、自由基清除、抗病毒、抗发炎、血管舒张活性等,在某些情况下也有金属螯合特性。查尔酮异构酶(ChalconeIsomerase,CHI,EC 5.5.1.6)是类黄酮合成途径早期阶段的关键酶之一。是植物花色素苷生物合成和豆科异黄酮植物抗毒索合成所必需的酶,以单体的形式普遍存在于大多数植物中,其蛋白的三维折叠结构在植物中具有唯一性。已被看成一种植物特有的基因标记。因此,CHI基因调控机制的研究对于了解豆科植物异黄酮生物合成特性、增加药用植物类黄酮的生物合成和花色形成的分子机制有重要的意义。
1、CHI基因的正向和反向植物表达载体的构建
用X-ba I和Sma I双酶切p~(UCm-CHI)和中间载体p~(TriplEx2),回收、连接目的片段与载体。构建中间载体pT-CHIs和pT-CHIa重组质粒。用X-ba I和Sma I双酶切pT-CHIs和pT-CHIa和表达载体质粒pC2301-35S-Nos,回收、连接目的片段与载体,分别构建成正反向植物表达载体pC2301-CHIs和pC2301-CHIa。采用冻融法把p~(C2301-CHIs)和p~(C2301-CHIa)分别转化农杆菌LBA4404。
2、根癌农杆菌介导的CHI基因对烟草的遗传转化
将-80℃下保存的农杆菌菌种LBA4404接种到YEB+Km 50mg/L+Sm125mg/L的培养基平板上,27-28℃倒置培养2-3d。菌落长出后,挑取单菌落接种丁含Km 50mg/L,Sm 125mg/L的液体YEB培养基中,27-28℃,180rpm振荡培养至OD600为0.6-0.8。取OD_(600)为0.6-0.8的农杆菌菌液1-2μl,加入到20-25ml的YEB中,摇菌至OD_(600)约为0.8,备用。
取烟草的无菌苗叶片,切成5mmx5mm大小,浸入备好的农杆菌菌液(OD_(600)=0.2)中感染5-10min。其间不断振荡使菌液与叶片充分接触。滤去菌液。取出叶片,用无菌滤纸吸干叶片表面多余的菌液,转入铺有滤纸的共培养培养基MS+2.0mg/L 6-BA+0.5mg/L NAA中。叶片背面朝向培养基。暗培养2-3d。共培养结束后,转入选择分化培养基MS+2.0mg/L 6-BA+0.5mg/L IAA+200mg/L Km+500mg/LCef中进行选择培养,20d继代一次,直到分化出芽(需要45-60d)。当分生不定芽达1cm以上时,切下不定芽,转入生根筛选培养基1/2MS+2.0mg/L 6-BA+0.5mg/L IAA+100mg/L Km+200mg/L Cef进行生根筛选。用农杆菌(含有正向表达载体质粒P~(C2301-CHIs))转化烟草(W38),所得转化株系命名为W~+。用农杆菌(含有反向表达载体质粒P~(C2301-CHIa))转化烟草(W38),,所得转化株系命名为w~-。未转化的烟草对照株系命名为W~0。统计结果,W~+获得23株存活的抗性苗:W~-获得20株存活的抗性苗:W~0获得10株。
3、转基因植株的鉴定和转基因植株类黄酮、花色素苷含量测定
将转化植株的叶片浸于GUS染液中染色,再对已染上色的植株进行PCR检测。得到检测结果为阳性的转基因植株W~+为12株,W~-为13株。
对PCR检测结果为阳性的转化植株取移栽后2个月的植株叶片进行类黄酮含量和花色素苷含量的测定。结果表明,W~+类黄酮含量比w~0略高一些,最高的可以比对照的烟草高50%左右,而又有个别的转正向基因的2—3个植株的类黄酮含量低于对照烟草;W~-类黄酮含量则低于对照烟草。W~+花色素苷含量比W~0明显高一些,最高的是对照的烟草花色素苷含量的2倍:W~-花色素苷含量则略低于对照烟草。
Flavonoids are pliant-specific secondary metabolites derived from the phenylpropanoid pathway,they always been called Vitamin P and they are accompany with Vitamin C.They are the natural pigment in vegetable,fruit,flower and corn,because they always assume yellow,so they been called bioflavonoids. They are involved in various biological processes during plant growth and development,including flower pigmentation,protection against UV irradiation,defense,pollen development,male fertility,cell cycle regulation,and auxin transport.They also play key roles in symbiotic interactions between plants and microbes.There are about 4000 kinds of flavonoid have been aparted until now.A lot of researches have indicated that flavonoids have multi-biologically active in our body,including anti-oxidation characteristic、clearing free radicals、resisting virus、resisting inflammation、vein expanding and so on、in some special time they can chelated metal hydronium.Chalcone isomerase(CHI,EC 5.5.1.6)is an important enzyme in the phenylpropanoid pathway.CHI is involved in a very early step of the flavonoid and isoflavonoid pathway,it can be found in most plants by monomer and the three-dimensional collapse structure of albumen is exclusive and it has been as proper gene badge in plants.So it has an important signification for learning isoflavonoid biosynthesis characteristic of leguminous plants,increasing the flavonoid biosynthesis of officinal plants and the molecular mechanism of flower colour.
1、Construction of plant expressional vector of sense and antisense CHI gene
p~(UCm-CHI)and p~(TriplEx2)digested with X-ba I and Sma I was constructed for mediated vector p(T-CHI)in which has correct direction,p~(T-CHI)and p~(C2301-35S-Nos)digested with X-ba I and Sma I was Constructed for plant expressional vector p~(C2301-CHIs),and p~(C2301-CHIa)of sense and antisense CHI gene.p~(C2301-CHIs)and p~(C2301-CHIa)were transferred into LBA4404,respectively.
2、Introduction of CHI gene into Petunia mediated by Agrobacterium tumejaekns
Put Agrobaterium that preserved in -80℃refrigeratory in substrate YEB+Km 50mg/L+Sm125mg/L, and convert-cultivate at 27-28℃for 2-3 days.Choose single tribe into liquid substrate of YEB for surging at 180rpm until OD_(600)adjusted to 0.6-0.8,and then take 1-2μl Agrobaterium liquid into 20-25ml YEB, convert-cultivate until OD_(600)adjusted to 0.8 for use.
Young leaves of were excised from sterile seedlings of tobacco,cut into 5×5 mm~2,dipped into Agrobaterium(OD adjusted to 0.2)for 15min.Patted dry on the filter paper,transferred to MS+2.0mg/L 6-BA+0.5mg/L NAA medium and co-cultured for 2-3 days in darkness.After co-cultivation,transferred to MS+2.0mg/L 6-BA+0.5mg/L IAA+200mg/L Km+500mg/L Cef for selection culture.The resistant shoots were rooted on 1/2MS+2.0mg/L 6-BA+0.5mg/L IAA+100mg/L Km+200mg/L Cef and then transferred the plantlets with normal roots to field for further analysis..The tobacco transformed by p~(C2301-CHIs)was named W+ and the tobacco transformed by p~(C2301-CHIa)was named W~-.Untransformed tobacco is control W~0. Statistic results showed that the resistant shoots W+ has23 lines;W~- has 18 lines and W~0 has 10 lines.
3、The identification of transformed plants and the mensuration of flavonoids and anthocynnin in transgenic tobacco
Put the leaf of transformed tobaccos into GUS,and then identify transformed masculing tobaccos by common PCR.The PCR results suggested that transgenic W~+ has 12 lines;transgenic W~- has 13 lines.
The flavonoids and anthocyanin were exracted after transplant for 2 month ago of tramsforned plants which were PCR positive results.W~0 are controls.The results showed that the flavonoids of W~+ was more than W~0,the highest is nearly up to 1.5 folds,but 2-3 transformed plant is lower than the controls.The flavonoids of W~- was lower than W~0.However,the content of anthocynnin in transgenic tobacco leaves increase remarkably,the highest is up to 2 folds;the content of anthocynnin of W~-is lower than the controls.
1、CHI基因的正向和反向植物表达载体的构建
用X-ba I和Sma I双酶切p~(UCm-CHI)和中间载体p~(TriplEx2),回收、连接目的片段与载体。构建中间载体pT-CHIs和pT-CHIa重组质粒。用X-ba I和Sma I双酶切pT-CHIs和pT-CHIa和表达载体质粒pC2301-35S-Nos,回收、连接目的片段与载体,分别构建成正反向植物表达载体pC2301-CHIs和pC2301-CHIa。采用冻融法把p~(C2301-CHIs)和p~(C2301-CHIa)分别转化农杆菌LBA4404。
2、根癌农杆菌介导的CHI基因对烟草的遗传转化
将-80℃下保存的农杆菌菌种LBA4404接种到YEB+Km 50mg/L+Sm125mg/L的培养基平板上,27-28℃倒置培养2-3d。菌落长出后,挑取单菌落接种丁含Km 50mg/L,Sm 125mg/L的液体YEB培养基中,27-28℃,180rpm振荡培养至OD600为0.6-0.8。取OD_(600)为0.6-0.8的农杆菌菌液1-2μl,加入到20-25ml的YEB中,摇菌至OD_(600)约为0.8,备用。
取烟草的无菌苗叶片,切成5mmx5mm大小,浸入备好的农杆菌菌液(OD_(600)=0.2)中感染5-10min。其间不断振荡使菌液与叶片充分接触。滤去菌液。取出叶片,用无菌滤纸吸干叶片表面多余的菌液,转入铺有滤纸的共培养培养基MS+2.0mg/L 6-BA+0.5mg/L NAA中。叶片背面朝向培养基。暗培养2-3d。共培养结束后,转入选择分化培养基MS+2.0mg/L 6-BA+0.5mg/L IAA+200mg/L Km+500mg/LCef中进行选择培养,20d继代一次,直到分化出芽(需要45-60d)。当分生不定芽达1cm以上时,切下不定芽,转入生根筛选培养基1/2MS+2.0mg/L 6-BA+0.5mg/L IAA+100mg/L Km+200mg/L Cef进行生根筛选。用农杆菌(含有正向表达载体质粒P~(C2301-CHIs))转化烟草(W38),所得转化株系命名为W~+。用农杆菌(含有反向表达载体质粒P~(C2301-CHIa))转化烟草(W38),,所得转化株系命名为w~-。未转化的烟草对照株系命名为W~0。统计结果,W~+获得23株存活的抗性苗:W~-获得20株存活的抗性苗:W~0获得10株。
3、转基因植株的鉴定和转基因植株类黄酮、花色素苷含量测定
将转化植株的叶片浸于GUS染液中染色,再对已染上色的植株进行PCR检测。得到检测结果为阳性的转基因植株W~+为12株,W~-为13株。
对PCR检测结果为阳性的转化植株取移栽后2个月的植株叶片进行类黄酮含量和花色素苷含量的测定。结果表明,W~+类黄酮含量比w~0略高一些,最高的可以比对照的烟草高50%左右,而又有个别的转正向基因的2—3个植株的类黄酮含量低于对照烟草;W~-类黄酮含量则低于对照烟草。W~+花色素苷含量比W~0明显高一些,最高的是对照的烟草花色素苷含量的2倍:W~-花色素苷含量则略低于对照烟草。
Flavonoids are pliant-specific secondary metabolites derived from the phenylpropanoid pathway,they always been called Vitamin P and they are accompany with Vitamin C.They are the natural pigment in vegetable,fruit,flower and corn,because they always assume yellow,so they been called bioflavonoids. They are involved in various biological processes during plant growth and development,including flower pigmentation,protection against UV irradiation,defense,pollen development,male fertility,cell cycle regulation,and auxin transport.They also play key roles in symbiotic interactions between plants and microbes.There are about 4000 kinds of flavonoid have been aparted until now.A lot of researches have indicated that flavonoids have multi-biologically active in our body,including anti-oxidation characteristic、clearing free radicals、resisting virus、resisting inflammation、vein expanding and so on、in some special time they can chelated metal hydronium.Chalcone isomerase(CHI,EC 5.5.1.6)is an important enzyme in the phenylpropanoid pathway.CHI is involved in a very early step of the flavonoid and isoflavonoid pathway,it can be found in most plants by monomer and the three-dimensional collapse structure of albumen is exclusive and it has been as proper gene badge in plants.So it has an important signification for learning isoflavonoid biosynthesis characteristic of leguminous plants,increasing the flavonoid biosynthesis of officinal plants and the molecular mechanism of flower colour.
1、Construction of plant expressional vector of sense and antisense CHI gene
p~(UCm-CHI)and p~(TriplEx2)digested with X-ba I and Sma I was constructed for mediated vector p(T-CHI)in which has correct direction,p~(T-CHI)and p~(C2301-35S-Nos)digested with X-ba I and Sma I was Constructed for plant expressional vector p~(C2301-CHIs),and p~(C2301-CHIa)of sense and antisense CHI gene.p~(C2301-CHIs)and p~(C2301-CHIa)were transferred into LBA4404,respectively.
2、Introduction of CHI gene into Petunia mediated by Agrobacterium tumejaekns
Put Agrobaterium that preserved in -80℃refrigeratory in substrate YEB+Km 50mg/L+Sm125mg/L, and convert-cultivate at 27-28℃for 2-3 days.Choose single tribe into liquid substrate of YEB for surging at 180rpm until OD_(600)adjusted to 0.6-0.8,and then take 1-2μl Agrobaterium liquid into 20-25ml YEB, convert-cultivate until OD_(600)adjusted to 0.8 for use.
Young leaves of were excised from sterile seedlings of tobacco,cut into 5×5 mm~2,dipped into Agrobaterium(OD adjusted to 0.2)for 15min.Patted dry on the filter paper,transferred to MS+2.0mg/L 6-BA+0.5mg/L NAA medium and co-cultured for 2-3 days in darkness.After co-cultivation,transferred to MS+2.0mg/L 6-BA+0.5mg/L IAA+200mg/L Km+500mg/L Cef for selection culture.The resistant shoots were rooted on 1/2MS+2.0mg/L 6-BA+0.5mg/L IAA+100mg/L Km+200mg/L Cef and then transferred the plantlets with normal roots to field for further analysis..The tobacco transformed by p~(C2301-CHIs)was named W+ and the tobacco transformed by p~(C2301-CHIa)was named W~-.Untransformed tobacco is control W~0. Statistic results showed that the resistant shoots W+ has23 lines;W~- has 18 lines and W~0 has 10 lines.
3、The identification of transformed plants and the mensuration of flavonoids and anthocynnin in transgenic tobacco
Put the leaf of transformed tobaccos into GUS,and then identify transformed masculing tobaccos by common PCR.The PCR results suggested that transgenic W~+ has 12 lines;transgenic W~- has 13 lines.
The flavonoids and anthocyanin were exracted after transplant for 2 month ago of tramsforned plants which were PCR positive results.W~0 are controls.The results showed that the flavonoids of W~+ was more than W~0,the highest is nearly up to 1.5 folds,but 2-3 transformed plant is lower than the controls.The flavonoids of W~- was lower than W~0.However,the content of anthocynnin in transgenic tobacco leaves increase remarkably,the highest is up to 2 folds;the content of anthocynnin of W~-is lower than the controls.
引文
[1]程金水主编.园林植物遗传育种.北京:中国林业出版社,2000
[2][日]安田齐著,傅玉兰译,花色的生理生物化学.北京:中国林业出版社,1989
[3][日]安田齐著,张承志,佟丽译.花色之谜.北京:中国林业出版社,1989
[4]于晓南 张启翔.观赏植物的花色素苷与花色.林业科学,2002,vol.38,No.3:147-153
[5]Jayaram C,Peter A P.Anthocyanion pigmentation and transposable elements in Maize Aleurome.Plant Breeding Reviews,1990,8:91 - 137
[6]Wheldale M.On the formation of anthocyanion.J Genet,1911,1:133-157
[7]Madhuri G Arhula R R.Plant biotechnology of flavonoida.Plant Biotech.1999,16(3):179-199
[8]Martin C.Gerats T The control of flower coloration,In:Jordan B R(ed)The molecular biology of flowering Redwood Books,U.K.,1993,219-225
[9]Kondo T K,Yoshida A,Nakagawa T et al.Structural basis of blue-color development in flower petals from Commeliaa communis.Nature,1992.358:515-518
[10]Mazza G,Miniati E.Anthocyanions in fruits,vegetables and grains.USA:CRC Press,1993,1-2829
[11]Strack D,Wray V.Anthocyanins,In:Harborne J B(ed).Method in plant biochemistry.Vol 1.Academic Press USA,1989,325-35
[12]Pietta PG.Flavonoids as antioxidants.J Nat Prod.2000.63:1035-1042.
[13]张德权,台建祥,付勤.生物类黄酮的研究及应用概况[J].食品与发酵工业,1999,25(6):52-57
[14]Wayner DDM,Burton GW,Ingold KU,Barclay LRC,Locke SJ.The relative contributions of vitamin E,urate,ascorbate and proteins to the total peroxyl radical-trapping antioxidant activity of human blood plasma.Biochim et Biophys Acta,1987,924(3):408-419
[15]Halliweli B.How to characterize a biological antioxidant.Free Radic Res Commult.1990,9:1-32
[16]Jovanovic SV,Steeden S,Tosic M,Marjanovic B,Simicg MG.Flavonoids as antioxidants.J Am Chem SOC,1994,116:4846-4851
[17]Terao J,Piskula M,Yao Q.Protective effect of epicatechin,epicatechin gallate,and quercetin on lipid peroxidation in phospholipid bilayers.Archives Biochem & Biophys.1994,308(I):278-284
[18]Mangiapane H,Thomson J,Salter A,Brown S,Bell GD,White DA.The inhibition of the oxidation of low density lipoprotein by(+)-catechin,a naturally occurring flavonoid.Biochem Pharmacol,1992,43(3):445-450
[19]Buettner GR.The pecking order of free radicals and antioxidants:lipid peroxidation,[alpha]一tocopherol,and ascorbate.Archives Biochem&Biophys,1993,300(2):535-543
[20]Jovanovic SV.Jankovic I.Josimovic L.Electron-transfer reactions of alkyl peroxy radicals.J Am Chem SOC,1992,114:9018-9021
[21]Rice-Evans CA.Miller NJ,Paganga G.Structure-antioxidant activitv relationships of flavonoids and phenolic acids.Free Radic Biof & Med.1996,20(7):933-956
[22]Cotelle N,Bernie JL,Hcnichart JP,Catteau JP,Gaydou E,Wallet JC.Scavenger and antioxidant properties of ten synthetic flavones.Free Radic Biol &Med,1992,13(3):211-219
[23]Zhou YC,Zheng RL.Phenolic compounds and an analog as superoxide anion scavengers and anti oxidants.Biochem Pharmacol,1991,42(6):1177-1179
[24]Avila M,Velasco J,Cansado J,Notario V.Quercetin mediates the down-regulation of mutant p53 in the human breast cancer cell line MDA-M13468.Cancer Res,1994,54(9):2424-2428
[25]da Silva EL,Piskula MK,Yamamoto N,Moon J-H,Terao J.Quercetin metabolites inhibit copper ion -induced lipid peroxidation in rat plasma.FEBS Lett,1998,430(3):405-408
[26]Hertog MGL,Feskens EJM,Kromhout D,Hollman PCH,Katan MB.Dietary antioxidant flavonoids and risk of coronary heart disease:the Zutphen Elderly Study.The Lancet,1993,342(8878):1007-1011
[27]Levites Y,Youdim MBH,Maor G,Mandel S.Attenuation of 6-hydroxydopamine(6-OHDA)-induced nuclear factor-kappaB(NF-kappaB)activation and cell death by tea extracts in neuronal cultures.Biochem Pharmacol,2002,63:21-29
[28]Kootstra A.Protection from UV-B-induced DNA damage by flavonoids.Plant Mol Biol,1994,26:771-774
[29]Stapleton AE,Walbot V.Flavonoids can protect maize DNA from the induction of ultraviolet radiation damage.Plant Physiol,1994,105(3):881-889
[30]姚新生.天然药物化学[M].北京:人民出版社,1996:194
[31]吴东方,周本宏,罗顺德.银杏叶黄酮对肝脏MDA生成的影响[J].中国中药杂志,1997,28(6):348-350
[32]晏文洁,李家璞.类黄酮抗氧化力与其结构之关系[J].台湾农业化学与食品科学,2000,38(1):80-88
[33]Siess M H.The influence of food flavonoids on the activity of some hepstic microsomal monooxygenases in rats[J].Food Chem Toxic,1982,20:883-889
[34]天津轻工业学院与无锡轻工业学院.食品生物化学[M].北京:中国轻工业出版社,2005
[35]Satu S Pekkarinen,I Marina Heinonen,Anu lhopla.Flavonoids quercetin,myrice-tin,kaemferol and(+)-catechin as antioxidants in methyl linoleate[J].Journal of the Science of Food and Agriculture,1999,79:499-506
[36]Martens S,Axel M.Flavones and flavone synthases[J].Phytochemistry,2005,66:2399-2407
[37]Dixon RA,Ferreira D.Genistein[J].Phytochemistry,2002,60:205-211
[38]张华峰,王瑛,黄宏文.类黄酮生物合成途径的进化及其在淫羊藿中的研究展望[J].中草药
[39]Yu O,Shi J,Hession AO,Maxwell CA.McGonigh B.Metabolic engineering to increase isoflavone biosynthesis in soybean seed[J].Phytochemistry,2003,63(7):753-763
[40]Yu O,Jung W,Shi J,Croes RA,Fader GM,McGonigle B,Odell GT.Production of the lsoflavones Genistein and Daidzein in Non-Legume Dicot and Monocot Tissues[J].Plant Physiology,2000,124:781-793
[41]Luczkiewicz M,Kokotkiewiez A.Co-cultures of shoots and hairy roots of Genista tinctoria L.for synthesis and biotransformation of large amounts of phytoestrogens[J].Plant Science,2005,169:86,2-871
[42]Federici E,Touche A,Choquart S,Avanti O,Fay L,Oford E,Courtois D.High isoflavone content and estrogenic activity of 25 year-old Glycine max tissue cultures[J].Phytochemistry,2003,64,717-724
[43]Wang XL,Shin KH,Hur HG,Kim SI.Enhanced biosynthesis of dihydrodaidzein and dihydrogenistein by a newly isolated bovine rumen anaerobic bacterium[J].Journal of Biotechnology,2005,115:261-269
[44]Wiilits MG,Giovanni M,Prata RTN,Kramer CM,Luca VD,Steffens JC.Graser G.Biowfermentation of modified flavonoids:an example of in vivo diversification of secondary metabolites [J].Phytochemistry,2004,65:31-41
[45]Yan Y,Chemler J,Huang L,Martens S,Koffas MA.Metabolic engineering of anthocyanin biosynthesis in Eseheriehia coli[J].Appl Environ Mierobiol,2005,71(7):3617-3623
[46]Shindo K,Kagiyama Y,Nakamura R,Hara A,Ikenaga H,Furukawa K,Misawa N.Enzymatic synthesis of novel antioxidant flavonoids by Esehefiehia coil cells expressing modified metabolic genes involved in biphenyl catabolism[J].Journal of Molecular Catalysis B:Enzymatic,2003.23:9-16
[47]Jiang H,Wood KV,Morgan JA.Metabolic engineering of the phenylpropanoid pathway in Saccharomyces cerevisiae[J],Appl Environ Microbiol,2005,71(6):2962-2969
[48]Jack Sullivan.Anthocyanin.CPNewsletter,1998,27(3):26-28
[49]刘玲,李霞,金同铭.高压液相色谱法在花色素苷分析中的应用.北京农业科学,1998,16(2):30-34
[50]Michael N Clifford.Review anthocyanins-nature,occurrence and dietary burden.Journal of the Sicence of Food and Agriculture,2000(80):1063-1072
[51]Iversen C.Degradation of C-vitamin and anthicyanins in black-currant juice,in Proceedings of the Symposium on Polyphenols and Anthocyanins as Food Colorants and Antioxidants.Ed by Pfannhauser W and Strigl A.EU,Brussels,Belgium,1996:34-34
[52]张昭其,庞学群.真菌花色素苷酶研究进展.微生物学通报,2001,28(6):86-89
[53]庞学群等.pH值和温度对荔枝果皮花色素苷稳定性的影响.园艺学报,2001,28(1):25-30
[54]王华等.葡萄酒中花色素苷研究现状.中外葡萄与葡萄酒,2002(2):25-28
[55]A.Lukton,C.O.Chichester,G.Mackinney.The Breakdown of strawberry anthocyanin pigment.Food technology,September,1956(10):427-431
[56]庞学群等.氧化还原物质对荔枝果皮花色素苷稳定性的影响.华南农业大学学报,2001,22(2):15-17
[57]Huang H T,J.Agr,food chem.1995,3(2):141-146
[58]Mol J,Jenkins G.J.,Schafer E.&Weiss D.Signal perception,transduction,and gene expression involved in anthocyanin biosynthesis.Crit Rev Plant Sci,1996,15:525-557
[59]Brandt K,Giannini A,Lercari B.Photomorphogenic responses to UV radiation Ⅲ:a comparative study of UV2B effects on anthocyanin and flavonoid accumulation in wild-type and aurea mutant of tomato(Lycopersicon esculent um Mill.).Photochem Photobiol,1995,62:1081-1087
[60]Krizek DT,Britz SJ,Mirecki RM.Inhibitory effects of ambient levels of solar UV-A and UV-B radiation on growth of cv.New Red Fire lettuce.Physiol Plant,1998,103:1-7
[61]Graham TL.Flavonoid and flavonol glycoside metabolism in A rabi dopsis.Plant Physiol Biochem,1998,36:135-144
[62]Christie PJ,Alfenito MR,Walbot V.Impact of low-temperaturestress on general phenylpropanoid and anthocyanin pathways:enhancement of transcript abundance and anthocyanin pigmentation in maize seedlings.Planta,1994,194:541-549
[63]Shichijo C,Hamada T,Hiraoka M et al.Enhancement of red-light-induced anthocyanin synthesis in sorghum first internodes by moderate low temperature given in the preirradiation culture period.Planta,1993,191:238-245
[64]Tholakalabavi A,Zwiazek JJ,Thorpe TA.Osmotically-stressed poplar cell cultures:anthocyanin accumulation,deaminase activity,and solute composition.J Plant Physiol,1997,151:489-496
[65]Kumar A,Sood A,Palni L.M.S.and Guptal A.K.In vitro propagation of Gladiolus hybridus Hort.:Synergistic effect of heat shock and sucrose on morphogenesis Micropropagation of gladiolus.Plant Cell Tussue Organ Cult.,1999,57:105-112
[66]Nagira Y and Ozeki Y.A system in which anthocyanin synthesis is induced in regenerated torenia shoots.J Plant Res,2004,117:377-383
[67]Solfanelli C,Poggi A,Loreti E,Alpi A and Perata P.Sucrose-Specific Induction of the Anthocyanin Biosynthetic Pathway in Arabidopsis.Plant Physiol.,2006,140(2):637-646
[68]孙明霞,王宝增,范海,赵可大。叶片中的花色素苷及其对植物适应环境的意义。植物生理学通讯,2003,39(6):688-694
[69]Dixon RA,Xie DY,Sharma SB.Proanthocyanidins-a final frontier in flavonoid research? New Phytologist,2005,165(1):9-28
[70]张石宝,胡虹,李树云.花卉基因工程研究进展Ⅰ:花色.云南植物研究,2001,23(4):479-487
[71]王关林,方宏筠.植物基因工程原理与技术.北京:科学出版社,1998
[72]Kreuzaler F.Flavanone synthase from petroselium hortense,molecular weight,subunit coorposition,size of messenger RNA and absence of pantethenyi residuce.Eru J Biochem.,1979,99-114
[73]Koes RE,Spelt CE,Van der Elzen P J M.Cloning and molecular characterization of the chalcone synthase multigene family of petunia hybrid.Gene,1989,81:245-257
[74]Koes RE,Spelt CE,Mol TNM.The chalcone synthase multigene family of petunia hybrid,(V30):Diferential light-regulated expression during flower development and UV light induction.Plant Molecular Biology,1 989,12:2 13-225
[75]林泉。色素基因的表达和调控.见:许智宏,刘春明主编.植物发育的分子机理,北京:科学出版社1998,107-110
[76]Birch,R.G.Plant transformation:problem and strategies for practical application.Annu Rev Plant Physiol Plant Mol Biol.,1997,48:297-326
[77]Holton T.A.,Brugilera,F.Cloning and expression of cytochrome P450 genes controlling flower colour.Nature,1993,Nov18,366(6452):276-279
[78]Brugliera F,Barri-Rewell G,Holton TA,et al.Isolation and characterization of a flavonoid3'-hydroxylasec DNA clone corresponding to the Htl locus of Petunia hybrida.Plant J,1999,19:441-51
[79]Kitada C,Gong Z,Tanaka Y,et al.Differential expression of two cytochrome P450s involved in the biosynthesis of flavones and anthocyanins in chemo-varietal forms of Perilla frutescens.Plant Cell Physiology,2001,42:1 338-1344
[80]UeyamaY,Suzuki K,Fukuchi-Mizutani M,et al.Molecular and biochemical characterization of torenia flavonoid3 '-hydroxylase and flavone synthase Ⅱ and modification of flower color by modulating the expression of these genes.Plant Science,2002,163:2 53-263
[81]Toda K,Yang DJ,Yamanaka N,et al.A single-base deletion in soybean flavonoid 3'-hydroxylase gene is associated with gray pubescence color.Plant Mol Biol,2002,50:187-196
[82]van Tunen A J,Hartman SA,Mur LA,Mol JN(1989).Regulation of chalcone isomerase(CHl)gene expression in petunia hydria:the use of alternative promoters in corolla,anthers and pollen.Plant Mo l Biol,12:539-551
[83]van Tunen AJ,Koes RE,Spelt CE,van der Krol AR,Stuitje AR,Mol JN(1988).Cloning of the two ehalcone flavanone isomerase genes from Petunia hybrid:coordinate,light-regulated and differential expression of flavonoid genes.EMBO J,7:1257-1263
[84]杨运英,廖伟平,梁建遴.彩叶草及其在园林中的应用.广东农业科学,2005,6:45-47
[85]蒋武生,芦翠乔,杨延明,吴丁.彩叶草的试管快速繁殖.河南农业科学,1991,4:30-31
[86]段黄金,高疆生.彩叶草的组培快繁技术研究.中国农学通报,2001.17(1):25-27
[87]黄海帆.李萍,贺爱利.彩叶草组织培养.植物生理学通讯,2003,39(4):339
[88]关日强,黄卓烈,朱志凯,张璞.彩叶草红色素的理化性质.植物资源与环境学报,2001,10(2):56-58
[89]刘士哲,林东教,唐淑军,罗健.利用漂浮植物修复系统栽培风四草、彩叶草和茉莉净化富营养化污水的研究.应用生态学报,2004,15(7):1261-1265
[90]Gensheimer M,Mushegian A.Chalcone isomerase family and fold:no longer unique to plants.Protein Sci,2004,13(2):540-544
[91]Mehdy M C,Lamb C J.Chalcone isomerase cDNA cloning and mRNA induction by fungal elicitor,wonding and infection.EMBO J,1988,6:1527-1533
[92]Herles C,Braune A,Blaut M.First bacterial chalcone isomerase isolated from Eubacterium ramulus.Arch Microbiol,2004,181:428-434
[93]Shimada N,Aoki T,Sato S,Nakamura Y,Tabata S,Ayabe S.A cluster of genes encodes the two types of chalcone isomerase involved in the biosynthesis of general flavonoids and legume-specific 5-deoxy(iso)flavonoids in Lotus japonicus.Plant Physiol,2003,131:941-951
[94]Druka A,Kudma D,Rostoks N,Brueggeman R,Von Wettstein D,Kleinhofs A.Chalcone isomerase gene from rice(Oryza sativa)and barley(Hordeum vulgare):physical,genetic and mutation mapping.Gene,2003,302(1-2):171-178
[95]许本波.甘蓝型油菜类黄酮途径CHI、F3H和F3'H基因家族的克隆及在黄、黑籽之间的差异表达.西南大学博士学位论文,2006
[96]Jez J M,Bowman M E,Dixon R A,Noel J P.Structure and mechanism of the evolutionarily unique plant enzyme chalcone isomerase.Nat Struct Biol,2000,7:786-791
[97]Jez J M,Noel J P.Reaction mechnism of chalcone isomerase pH dependence,diffusion control,and product binding differences.J Bio Chem,2002,277(2):1361-1369
[98]Bednar RA,Hadcock JR.Purification and characterization of chalcone isomerase from soybeans,J Biol Chem,1988,263:9582-9588
[99]McKhann HI,Hirsch AM.Isolation of chalcone synthase and chalcone isomerase cDNAs from alfalfa (Medicago sativa L.):highest transcript levels occur in young roots and root tips.Plant Mol Biol,1994,24:767-777
[100]Goormachtig S,Lievens S,Herman S,Van Montagu M,Holsters M.Chalcone reductasehomologous transcripts accumulate during development of stem-borne nodules on the tropical legume Sesbania rostrata.Planta,1999,209:45-52
[101]Kim HK,Jang YH,Baek IS,Lee JH.,Park M J.Polymorphism and expression of isoflavone synthase genes from soybean cultivars.Mol Cells,2005,19:67-73
[102]Yang WC,Canter Cremers HC J,Hogendijk P,Katinakis P,Wijffelman CA.In-situ localization of chalcone synthase mRNA in pea root nodule development.Plant J,1992,2:143-151
[103]Koornneef M.Mutations affecting the testa colour in Arabidopsis.Arabidopsis Inf Serv,1990,27:1-4
[104]van Tunen A J,Mur LA,Recourt K,Gerats AG,Mol JN.Regulation and manipulation of flavonoid gene expression in anthers of petunia:the molecular basis of the Po mutation.Plant Cell,1991,3:39-48
[105]Forkmann G,Dangelmayr B.Genetic control of chalcone isomerase activity in flowers of Dianthus caryophyllus.Biochem Genet,1980,18:519-527
[106]Kuhn B,Forkmann G,Seyfert W.Genetic control of chalcone-flavanone isomerase activity in Callistephus chinensis.Planta,1978,38:199-203
[107]Miyajima I.,Maehara T,Kage T,Fujieda K.Identification of the main agent causing yellow color of yellow - flowered cyclamen mutant.J Jpn Soc Hort Sci,1991,60:409-414
[108]Itoh Y,Higcta D,Suzuki A,Yoshida H,Ozeki Y.Excision of transposable elements from the chalcone isomerase and dihydroflavonol 4-reductase genes may contribute to the variegation of the yellow-flowered carnation(Dianthus caryophyllus).Plant Cell Physiol,1992,43:578-585
[109]Reuber S,Jende-Strid B,Wray V,Weissenbock G.Accumulation of the chalcone isosalipurposide in primary leaves of barley flavonoid mutants indicates a defective chalcone isomerase.Physiol Plant,1997,101:827-832
[110]Kim S,Jones R,Yoo KS,Deroles SC.Gold color in onions(Allium cepa):a natural mutation of the chalcone isomerase gene resulting in a premature stop codon.Mol Genet Genomics,2004,272:411-419
[111]Moustafa E,Wong E.Purification and properties of chalcone-flavanone isomerase from soya bean seed.Phytochemistry,1967,6:625-632
[112]祝钦泷.转查尔酮异构酶(CHI)基因矮牵牛花色改变及其花器官变异的研究.西南农业大学硕士学位论文,2004
[113]Nishihara M,Nakatsuka T,Yamamura S.Flavonoid components and flower color change in transgenic tobacco plants by suppression of chalcone isomerase gene.FEBS Lett,2005,579(27):6074-6078
[114]Li FX,Jin ZP,Qu WQ,Zhao DX,Ma FS.Cloning of a cDNA encoding the Saussurea medusa chalcone isomerase and its expression in transgenic tobacco.Plant Physiol Biochem,2006,44:455-461
[115]Li FX,Jin ZP,Zhao DX,Cheng LQ,Fu CX,Ma FS.Overexpression of the Saussurea medusa chalcone isomerase gene in Saussurea involucrata hairy root cultures enhances their biosynthesis of apigenin.Phytochem,2006,67:553-560
[2][日]安田齐著,傅玉兰译,花色的生理生物化学.北京:中国林业出版社,1989
[3][日]安田齐著,张承志,佟丽译.花色之谜.北京:中国林业出版社,1989
[4]于晓南 张启翔.观赏植物的花色素苷与花色.林业科学,2002,vol.38,No.3:147-153
[5]Jayaram C,Peter A P.Anthocyanion pigmentation and transposable elements in Maize Aleurome.Plant Breeding Reviews,1990,8:91 - 137
[6]Wheldale M.On the formation of anthocyanion.J Genet,1911,1:133-157
[7]Madhuri G Arhula R R.Plant biotechnology of flavonoida.Plant Biotech.1999,16(3):179-199
[8]Martin C.Gerats T The control of flower coloration,In:Jordan B R(ed)The molecular biology of flowering Redwood Books,U.K.,1993,219-225
[9]Kondo T K,Yoshida A,Nakagawa T et al.Structural basis of blue-color development in flower petals from Commeliaa communis.Nature,1992.358:515-518
[10]Mazza G,Miniati E.Anthocyanions in fruits,vegetables and grains.USA:CRC Press,1993,1-2829
[11]Strack D,Wray V.Anthocyanins,In:Harborne J B(ed).Method in plant biochemistry.Vol 1.Academic Press USA,1989,325-35
[12]Pietta PG.Flavonoids as antioxidants.J Nat Prod.2000.63:1035-1042.
[13]张德权,台建祥,付勤.生物类黄酮的研究及应用概况[J].食品与发酵工业,1999,25(6):52-57
[14]Wayner DDM,Burton GW,Ingold KU,Barclay LRC,Locke SJ.The relative contributions of vitamin E,urate,ascorbate and proteins to the total peroxyl radical-trapping antioxidant activity of human blood plasma.Biochim et Biophys Acta,1987,924(3):408-419
[15]Halliweli B.How to characterize a biological antioxidant.Free Radic Res Commult.1990,9:1-32
[16]Jovanovic SV,Steeden S,Tosic M,Marjanovic B,Simicg MG.Flavonoids as antioxidants.J Am Chem SOC,1994,116:4846-4851
[17]Terao J,Piskula M,Yao Q.Protective effect of epicatechin,epicatechin gallate,and quercetin on lipid peroxidation in phospholipid bilayers.Archives Biochem & Biophys.1994,308(I):278-284
[18]Mangiapane H,Thomson J,Salter A,Brown S,Bell GD,White DA.The inhibition of the oxidation of low density lipoprotein by(+)-catechin,a naturally occurring flavonoid.Biochem Pharmacol,1992,43(3):445-450
[19]Buettner GR.The pecking order of free radicals and antioxidants:lipid peroxidation,[alpha]一tocopherol,and ascorbate.Archives Biochem&Biophys,1993,300(2):535-543
[20]Jovanovic SV.Jankovic I.Josimovic L.Electron-transfer reactions of alkyl peroxy radicals.J Am Chem SOC,1992,114:9018-9021
[21]Rice-Evans CA.Miller NJ,Paganga G.Structure-antioxidant activitv relationships of flavonoids and phenolic acids.Free Radic Biof & Med.1996,20(7):933-956
[22]Cotelle N,Bernie JL,Hcnichart JP,Catteau JP,Gaydou E,Wallet JC.Scavenger and antioxidant properties of ten synthetic flavones.Free Radic Biol &Med,1992,13(3):211-219
[23]Zhou YC,Zheng RL.Phenolic compounds and an analog as superoxide anion scavengers and anti oxidants.Biochem Pharmacol,1991,42(6):1177-1179
[24]Avila M,Velasco J,Cansado J,Notario V.Quercetin mediates the down-regulation of mutant p53 in the human breast cancer cell line MDA-M13468.Cancer Res,1994,54(9):2424-2428
[25]da Silva EL,Piskula MK,Yamamoto N,Moon J-H,Terao J.Quercetin metabolites inhibit copper ion -induced lipid peroxidation in rat plasma.FEBS Lett,1998,430(3):405-408
[26]Hertog MGL,Feskens EJM,Kromhout D,Hollman PCH,Katan MB.Dietary antioxidant flavonoids and risk of coronary heart disease:the Zutphen Elderly Study.The Lancet,1993,342(8878):1007-1011
[27]Levites Y,Youdim MBH,Maor G,Mandel S.Attenuation of 6-hydroxydopamine(6-OHDA)-induced nuclear factor-kappaB(NF-kappaB)activation and cell death by tea extracts in neuronal cultures.Biochem Pharmacol,2002,63:21-29
[28]Kootstra A.Protection from UV-B-induced DNA damage by flavonoids.Plant Mol Biol,1994,26:771-774
[29]Stapleton AE,Walbot V.Flavonoids can protect maize DNA from the induction of ultraviolet radiation damage.Plant Physiol,1994,105(3):881-889
[30]姚新生.天然药物化学[M].北京:人民出版社,1996:194
[31]吴东方,周本宏,罗顺德.银杏叶黄酮对肝脏MDA生成的影响[J].中国中药杂志,1997,28(6):348-350
[32]晏文洁,李家璞.类黄酮抗氧化力与其结构之关系[J].台湾农业化学与食品科学,2000,38(1):80-88
[33]Siess M H.The influence of food flavonoids on the activity of some hepstic microsomal monooxygenases in rats[J].Food Chem Toxic,1982,20:883-889
[34]天津轻工业学院与无锡轻工业学院.食品生物化学[M].北京:中国轻工业出版社,2005
[35]Satu S Pekkarinen,I Marina Heinonen,Anu lhopla.Flavonoids quercetin,myrice-tin,kaemferol and(+)-catechin as antioxidants in methyl linoleate[J].Journal of the Science of Food and Agriculture,1999,79:499-506
[36]Martens S,Axel M.Flavones and flavone synthases[J].Phytochemistry,2005,66:2399-2407
[37]Dixon RA,Ferreira D.Genistein[J].Phytochemistry,2002,60:205-211
[38]张华峰,王瑛,黄宏文.类黄酮生物合成途径的进化及其在淫羊藿中的研究展望[J].中草药
[39]Yu O,Shi J,Hession AO,Maxwell CA.McGonigh B.Metabolic engineering to increase isoflavone biosynthesis in soybean seed[J].Phytochemistry,2003,63(7):753-763
[40]Yu O,Jung W,Shi J,Croes RA,Fader GM,McGonigle B,Odell GT.Production of the lsoflavones Genistein and Daidzein in Non-Legume Dicot and Monocot Tissues[J].Plant Physiology,2000,124:781-793
[41]Luczkiewicz M,Kokotkiewiez A.Co-cultures of shoots and hairy roots of Genista tinctoria L.for synthesis and biotransformation of large amounts of phytoestrogens[J].Plant Science,2005,169:86,2-871
[42]Federici E,Touche A,Choquart S,Avanti O,Fay L,Oford E,Courtois D.High isoflavone content and estrogenic activity of 25 year-old Glycine max tissue cultures[J].Phytochemistry,2003,64,717-724
[43]Wang XL,Shin KH,Hur HG,Kim SI.Enhanced biosynthesis of dihydrodaidzein and dihydrogenistein by a newly isolated bovine rumen anaerobic bacterium[J].Journal of Biotechnology,2005,115:261-269
[44]Wiilits MG,Giovanni M,Prata RTN,Kramer CM,Luca VD,Steffens JC.Graser G.Biowfermentation of modified flavonoids:an example of in vivo diversification of secondary metabolites [J].Phytochemistry,2004,65:31-41
[45]Yan Y,Chemler J,Huang L,Martens S,Koffas MA.Metabolic engineering of anthocyanin biosynthesis in Eseheriehia coli[J].Appl Environ Mierobiol,2005,71(7):3617-3623
[46]Shindo K,Kagiyama Y,Nakamura R,Hara A,Ikenaga H,Furukawa K,Misawa N.Enzymatic synthesis of novel antioxidant flavonoids by Esehefiehia coil cells expressing modified metabolic genes involved in biphenyl catabolism[J].Journal of Molecular Catalysis B:Enzymatic,2003.23:9-16
[47]Jiang H,Wood KV,Morgan JA.Metabolic engineering of the phenylpropanoid pathway in Saccharomyces cerevisiae[J],Appl Environ Microbiol,2005,71(6):2962-2969
[48]Jack Sullivan.Anthocyanin.CPNewsletter,1998,27(3):26-28
[49]刘玲,李霞,金同铭.高压液相色谱法在花色素苷分析中的应用.北京农业科学,1998,16(2):30-34
[50]Michael N Clifford.Review anthocyanins-nature,occurrence and dietary burden.Journal of the Sicence of Food and Agriculture,2000(80):1063-1072
[51]Iversen C.Degradation of C-vitamin and anthicyanins in black-currant juice,in Proceedings of the Symposium on Polyphenols and Anthocyanins as Food Colorants and Antioxidants.Ed by Pfannhauser W and Strigl A.EU,Brussels,Belgium,1996:34-34
[52]张昭其,庞学群.真菌花色素苷酶研究进展.微生物学通报,2001,28(6):86-89
[53]庞学群等.pH值和温度对荔枝果皮花色素苷稳定性的影响.园艺学报,2001,28(1):25-30
[54]王华等.葡萄酒中花色素苷研究现状.中外葡萄与葡萄酒,2002(2):25-28
[55]A.Lukton,C.O.Chichester,G.Mackinney.The Breakdown of strawberry anthocyanin pigment.Food technology,September,1956(10):427-431
[56]庞学群等.氧化还原物质对荔枝果皮花色素苷稳定性的影响.华南农业大学学报,2001,22(2):15-17
[57]Huang H T,J.Agr,food chem.1995,3(2):141-146
[58]Mol J,Jenkins G.J.,Schafer E.&Weiss D.Signal perception,transduction,and gene expression involved in anthocyanin biosynthesis.Crit Rev Plant Sci,1996,15:525-557
[59]Brandt K,Giannini A,Lercari B.Photomorphogenic responses to UV radiation Ⅲ:a comparative study of UV2B effects on anthocyanin and flavonoid accumulation in wild-type and aurea mutant of tomato(Lycopersicon esculent um Mill.).Photochem Photobiol,1995,62:1081-1087
[60]Krizek DT,Britz SJ,Mirecki RM.Inhibitory effects of ambient levels of solar UV-A and UV-B radiation on growth of cv.New Red Fire lettuce.Physiol Plant,1998,103:1-7
[61]Graham TL.Flavonoid and flavonol glycoside metabolism in A rabi dopsis.Plant Physiol Biochem,1998,36:135-144
[62]Christie PJ,Alfenito MR,Walbot V.Impact of low-temperaturestress on general phenylpropanoid and anthocyanin pathways:enhancement of transcript abundance and anthocyanin pigmentation in maize seedlings.Planta,1994,194:541-549
[63]Shichijo C,Hamada T,Hiraoka M et al.Enhancement of red-light-induced anthocyanin synthesis in sorghum first internodes by moderate low temperature given in the preirradiation culture period.Planta,1993,191:238-245
[64]Tholakalabavi A,Zwiazek JJ,Thorpe TA.Osmotically-stressed poplar cell cultures:anthocyanin accumulation,deaminase activity,and solute composition.J Plant Physiol,1997,151:489-496
[65]Kumar A,Sood A,Palni L.M.S.and Guptal A.K.In vitro propagation of Gladiolus hybridus Hort.:Synergistic effect of heat shock and sucrose on morphogenesis Micropropagation of gladiolus.Plant Cell Tussue Organ Cult.,1999,57:105-112
[66]Nagira Y and Ozeki Y.A system in which anthocyanin synthesis is induced in regenerated torenia shoots.J Plant Res,2004,117:377-383
[67]Solfanelli C,Poggi A,Loreti E,Alpi A and Perata P.Sucrose-Specific Induction of the Anthocyanin Biosynthetic Pathway in Arabidopsis.Plant Physiol.,2006,140(2):637-646
[68]孙明霞,王宝增,范海,赵可大。叶片中的花色素苷及其对植物适应环境的意义。植物生理学通讯,2003,39(6):688-694
[69]Dixon RA,Xie DY,Sharma SB.Proanthocyanidins-a final frontier in flavonoid research? New Phytologist,2005,165(1):9-28
[70]张石宝,胡虹,李树云.花卉基因工程研究进展Ⅰ:花色.云南植物研究,2001,23(4):479-487
[71]王关林,方宏筠.植物基因工程原理与技术.北京:科学出版社,1998
[72]Kreuzaler F.Flavanone synthase from petroselium hortense,molecular weight,subunit coorposition,size of messenger RNA and absence of pantethenyi residuce.Eru J Biochem.,1979,99-114
[73]Koes RE,Spelt CE,Van der Elzen P J M.Cloning and molecular characterization of the chalcone synthase multigene family of petunia hybrid.Gene,1989,81:245-257
[74]Koes RE,Spelt CE,Mol TNM.The chalcone synthase multigene family of petunia hybrid,(V30):Diferential light-regulated expression during flower development and UV light induction.Plant Molecular Biology,1 989,12:2 13-225
[75]林泉。色素基因的表达和调控.见:许智宏,刘春明主编.植物发育的分子机理,北京:科学出版社1998,107-110
[76]Birch,R.G.Plant transformation:problem and strategies for practical application.Annu Rev Plant Physiol Plant Mol Biol.,1997,48:297-326
[77]Holton T.A.,Brugilera,F.Cloning and expression of cytochrome P450 genes controlling flower colour.Nature,1993,Nov18,366(6452):276-279
[78]Brugliera F,Barri-Rewell G,Holton TA,et al.Isolation and characterization of a flavonoid3'-hydroxylasec DNA clone corresponding to the Htl locus of Petunia hybrida.Plant J,1999,19:441-51
[79]Kitada C,Gong Z,Tanaka Y,et al.Differential expression of two cytochrome P450s involved in the biosynthesis of flavones and anthocyanins in chemo-varietal forms of Perilla frutescens.Plant Cell Physiology,2001,42:1 338-1344
[80]UeyamaY,Suzuki K,Fukuchi-Mizutani M,et al.Molecular and biochemical characterization of torenia flavonoid3 '-hydroxylase and flavone synthase Ⅱ and modification of flower color by modulating the expression of these genes.Plant Science,2002,163:2 53-263
[81]Toda K,Yang DJ,Yamanaka N,et al.A single-base deletion in soybean flavonoid 3'-hydroxylase gene is associated with gray pubescence color.Plant Mol Biol,2002,50:187-196
[82]van Tunen A J,Hartman SA,Mur LA,Mol JN(1989).Regulation of chalcone isomerase(CHl)gene expression in petunia hydria:the use of alternative promoters in corolla,anthers and pollen.Plant Mo l Biol,12:539-551
[83]van Tunen AJ,Koes RE,Spelt CE,van der Krol AR,Stuitje AR,Mol JN(1988).Cloning of the two ehalcone flavanone isomerase genes from Petunia hybrid:coordinate,light-regulated and differential expression of flavonoid genes.EMBO J,7:1257-1263
[84]杨运英,廖伟平,梁建遴.彩叶草及其在园林中的应用.广东农业科学,2005,6:45-47
[85]蒋武生,芦翠乔,杨延明,吴丁.彩叶草的试管快速繁殖.河南农业科学,1991,4:30-31
[86]段黄金,高疆生.彩叶草的组培快繁技术研究.中国农学通报,2001.17(1):25-27
[87]黄海帆.李萍,贺爱利.彩叶草组织培养.植物生理学通讯,2003,39(4):339
[88]关日强,黄卓烈,朱志凯,张璞.彩叶草红色素的理化性质.植物资源与环境学报,2001,10(2):56-58
[89]刘士哲,林东教,唐淑军,罗健.利用漂浮植物修复系统栽培风四草、彩叶草和茉莉净化富营养化污水的研究.应用生态学报,2004,15(7):1261-1265
[90]Gensheimer M,Mushegian A.Chalcone isomerase family and fold:no longer unique to plants.Protein Sci,2004,13(2):540-544
[91]Mehdy M C,Lamb C J.Chalcone isomerase cDNA cloning and mRNA induction by fungal elicitor,wonding and infection.EMBO J,1988,6:1527-1533
[92]Herles C,Braune A,Blaut M.First bacterial chalcone isomerase isolated from Eubacterium ramulus.Arch Microbiol,2004,181:428-434
[93]Shimada N,Aoki T,Sato S,Nakamura Y,Tabata S,Ayabe S.A cluster of genes encodes the two types of chalcone isomerase involved in the biosynthesis of general flavonoids and legume-specific 5-deoxy(iso)flavonoids in Lotus japonicus.Plant Physiol,2003,131:941-951
[94]Druka A,Kudma D,Rostoks N,Brueggeman R,Von Wettstein D,Kleinhofs A.Chalcone isomerase gene from rice(Oryza sativa)and barley(Hordeum vulgare):physical,genetic and mutation mapping.Gene,2003,302(1-2):171-178
[95]许本波.甘蓝型油菜类黄酮途径CHI、F3H和F3'H基因家族的克隆及在黄、黑籽之间的差异表达.西南大学博士学位论文,2006
[96]Jez J M,Bowman M E,Dixon R A,Noel J P.Structure and mechanism of the evolutionarily unique plant enzyme chalcone isomerase.Nat Struct Biol,2000,7:786-791
[97]Jez J M,Noel J P.Reaction mechnism of chalcone isomerase pH dependence,diffusion control,and product binding differences.J Bio Chem,2002,277(2):1361-1369
[98]Bednar RA,Hadcock JR.Purification and characterization of chalcone isomerase from soybeans,J Biol Chem,1988,263:9582-9588
[99]McKhann HI,Hirsch AM.Isolation of chalcone synthase and chalcone isomerase cDNAs from alfalfa (Medicago sativa L.):highest transcript levels occur in young roots and root tips.Plant Mol Biol,1994,24:767-777
[100]Goormachtig S,Lievens S,Herman S,Van Montagu M,Holsters M.Chalcone reductasehomologous transcripts accumulate during development of stem-borne nodules on the tropical legume Sesbania rostrata.Planta,1999,209:45-52
[101]Kim HK,Jang YH,Baek IS,Lee JH.,Park M J.Polymorphism and expression of isoflavone synthase genes from soybean cultivars.Mol Cells,2005,19:67-73
[102]Yang WC,Canter Cremers HC J,Hogendijk P,Katinakis P,Wijffelman CA.In-situ localization of chalcone synthase mRNA in pea root nodule development.Plant J,1992,2:143-151
[103]Koornneef M.Mutations affecting the testa colour in Arabidopsis.Arabidopsis Inf Serv,1990,27:1-4
[104]van Tunen A J,Mur LA,Recourt K,Gerats AG,Mol JN.Regulation and manipulation of flavonoid gene expression in anthers of petunia:the molecular basis of the Po mutation.Plant Cell,1991,3:39-48
[105]Forkmann G,Dangelmayr B.Genetic control of chalcone isomerase activity in flowers of Dianthus caryophyllus.Biochem Genet,1980,18:519-527
[106]Kuhn B,Forkmann G,Seyfert W.Genetic control of chalcone-flavanone isomerase activity in Callistephus chinensis.Planta,1978,38:199-203
[107]Miyajima I.,Maehara T,Kage T,Fujieda K.Identification of the main agent causing yellow color of yellow - flowered cyclamen mutant.J Jpn Soc Hort Sci,1991,60:409-414
[108]Itoh Y,Higcta D,Suzuki A,Yoshida H,Ozeki Y.Excision of transposable elements from the chalcone isomerase and dihydroflavonol 4-reductase genes may contribute to the variegation of the yellow-flowered carnation(Dianthus caryophyllus).Plant Cell Physiol,1992,43:578-585
[109]Reuber S,Jende-Strid B,Wray V,Weissenbock G.Accumulation of the chalcone isosalipurposide in primary leaves of barley flavonoid mutants indicates a defective chalcone isomerase.Physiol Plant,1997,101:827-832
[110]Kim S,Jones R,Yoo KS,Deroles SC.Gold color in onions(Allium cepa):a natural mutation of the chalcone isomerase gene resulting in a premature stop codon.Mol Genet Genomics,2004,272:411-419
[111]Moustafa E,Wong E.Purification and properties of chalcone-flavanone isomerase from soya bean seed.Phytochemistry,1967,6:625-632
[112]祝钦泷.转查尔酮异构酶(CHI)基因矮牵牛花色改变及其花器官变异的研究.西南农业大学硕士学位论文,2004
[113]Nishihara M,Nakatsuka T,Yamamura S.Flavonoid components and flower color change in transgenic tobacco plants by suppression of chalcone isomerase gene.FEBS Lett,2005,579(27):6074-6078
[114]Li FX,Jin ZP,Qu WQ,Zhao DX,Ma FS.Cloning of a cDNA encoding the Saussurea medusa chalcone isomerase and its expression in transgenic tobacco.Plant Physiol Biochem,2006,44:455-461
[115]Li FX,Jin ZP,Zhao DX,Cheng LQ,Fu CX,Ma FS.Overexpression of the Saussurea medusa chalcone isomerase gene in Saussurea involucrata hairy root cultures enhances their biosynthesis of apigenin.Phytochem,2006,67:553-560