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不同花色品种蝴蝶兰花色素苷含量分析及相关基因表达研究

关键词: 基因表达  

Abstract: In order to clarify the roles of anthocyanin biosynthetic genes in the anthocyanin biosynthesis and pigmentation of Phalaenopsis flower, the expression of 7 structural genes PAL, 4CL, CHS, DFR, F3H, F35H, ANS and 3 regulating genes were quantified at different developmental stages in the red, yellow and white flowered Phalaenopsis cultivars. The results showed that expression profiles of the 10 genes are cultivar dependent and varied at different developmental stages. All 10 gene transcripts are lower in leaves compared to flower. In Huangjinjia, the expressions of four genes PAL、4CL、CHS,F3H and F35H are high and reach the peak at the stage Ⅲ. In Kongganghongyin, DFR, CHS, F3H, F35H expressed high at the stage Ⅲ. While in Fulexiyang and Konggangfengye, CHS, DFR, F3H, F35H and ANS expressed low in three stages compared to another 4 cultivars. F3H transcript level was the lowest among the 10 genes studied in the 6 cultivars. CHS transcript was detected in all stage of flower in 6 cultivars. The low expression of F3H gene in the 6 varieties indicates that there is no direct correlation between anthocyanin synthesis and F3H. The expression of bHLH was high in the 6 cultivars of different developmental stages, while the accumulation of MYB transcript was very low in 6 cultivar. These results indicated that the expression of CHS, DFR, F35H and ANS genes are necessary in Phalaenopsis anthocyanin biosynthesis. The transcription factor bHLH expression may have greater impact on Phalaenopsis flower color formation.

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[J].Current Opinion in Biotechnology, 2008, 19(2):190-197 [31]Katsumoto Y, Fukuchi MizutaniM, Fukui Y, Brugliera F, Holton TA, KaranM, Nakamura N, Yonekura Sakakibara K, Togami J, Pigeaire A, Tao G Q, Nehra N S, Lu C Y, Dyson B K, Tsuda S, Ashikari T, Kusumi T, Mason J G, Tanaka Y.Engineering of the rose flavonoid biosynthetic pathway successfully generated blue hued flowers accumulating delphinidin[J]. [J].Plant Cell Physiol, 2007, 48(11):1589-1600 [32]Kumar V1, Yadav SK.Overexpression of CsANR increased flavan-3-ols and decreased anthocyanins in transgenic tobacco[J].[J].Mol Biotechnol, 2013, 54(2):426-435

[1]王有国, 李枝林, 孙慧晶, 郭红, 张广辉.兰科植物花色形成与变异分子机理[J]. [J].分子植物育种网络版, 2011,, 9(80):1584-1590 [2]Wang Youguo, Li Zhilin, Sun Huijing, Guo Hong, Zhang Guanghui.Molecular Mechanism of Flower Color Formation and Variation in Orichidaceae[J].[J].Fenzi Zhiwu Yuzhong (Online), http://mpb. chinese.sophiapublisher.com, 2011, 9(80):1584-1590 [3]. [4]Weiss D.Regulation of flower pigmentation and growth: Multiple signaling pathways control anthocyanin synthesis in expanding petals[J].Physiologia Plantarum, 2000, 110(2):152-157 [5]李崇晖,任 羽,黄素荣,等.蝴蝶石斛兰花色表型及类黄酮成分分析[J].园艺学报, 2013, 40(1):107-116 [6]LI Chong-hui,REN Yu,HUANG Su-rong,HUANG Shao-hua,and YANG Guang-sui.Floral Colors of Phalaenopsis Type Dendrobium and Their Flavonoid Composition[J].Acta Horticulturae Sinica, 2013, 40(1):107-116 [7]NAKATSUKA T,NISHIHARA M,MISHIBA K,YAMAMURA S..Temporal expression of flavonoid biosynthesis—related genes regulates flower pigmentation in gentian plants[J].[J].Plant Sci, 2005, 168(5):1309-1318 [8]GONG ZH ZH,YAMAZAKI M, SUGIYAMA M, el a1..Cloning and molecular analysis of structural genes involved in anthocyanin biosynthesis and expressed in a forma—specific manner in Perilla frutescens[J].[J].Plant Mol Biol., 1997, 35(6):915-927 [9]HONDA C,KOTODA N,WADA M,KONDO S,et a1..Anthocyanin biosynthetic genes are coordinately expressed during red coloration in apple skin[J].[J].Plant Physiology and Biochemistry, 2002, 40:955-962 [10]KIM S G.BINZEL M L,PARK S H,YO0 K S,PIKE L M..Inactivation of DFR(Dihydroflavonol 4-reductase)gene transcription results in blockage of anthocyanin production in yellow onions(Allium cepa)[J].[J].Molecular Breeding, 2004, 14(3):253-263 [11]FEYISSA D N,LOVDAL T,OLSEN K M, SLIMESTAD R,LILI O C..The endogenous GL3,but not EGL3,gene is necessary for anthocyanin accumulation as induced by nitrogen depletion in Arabidopsis rosette stage leaves[J].[J].Planta, 2009, 230:747-754 [12]陈素梅,朱喜荣,陈发棣,罗火林,吕国胜,房伟民,章镇.不同花色菊花品种花色素结构基因的表达特性[J].西北植物学报, 2010, 30(3):453-458 [13]CH EN Sum ei, ZHU Xirong, CH EN Fadi, LUO H uolin, Lv Guosheng, FANG Weiming,ZHANG Zhen.Expression Profiles of Anthocyanin Biosynthetic Genes in Chrysanthemum Cultivars with Different Flower Colors[J]. [J].Aeta Bot.Borea1. Occident.Sin, 2010, 30(3):453-458 [14]Han YY1, Ming F, Wang W, Wang JW, Ye MM, Shen DL.Molecular evolution and functional specialization of chalcone synthase superfamily from Phalaenopsis orchid[J]. [J].Genetica, 2006, 128(1-3):429-438 [15]Wang J, Ming F., Han Y., and Shen D., . Flavonoid-3,5’-hydroxylase from Phalaenopsis: A novel member of cytochrome P450s, its cDNA cloning, endogenous expression and molecular modeling[J][J].Biotechnology Letters, 2006, 28(5):327-334 [16]Chen WH, Hsu CY, Cheng HY, Chang H, Chen HH, Ger MJ.Downregulation of putative UDP-glucose: flavonoid 3-O-glucosyltransferase gene alters flower coloring in Phalaenopsis[J]. [J].Plant Cell Rep, 2011, 30(6):1007-1017 [17]Wang L1, Albert NW, Zhang H, Arathoon S, Boase MR, Ngo H, Schwinn KE, Davies KM, Lewis DH.Temporal and spatial regulation of anthocyanin biosynthesis provide diverse flower colour intensities and patterning in Cymbidium orchid[J].[J].Planta., 2014, 240(5):983-1002 [18]韩科厅,赵 莉,唐杏姣,胡 可,戴思兰.菊花花青素苷合成关键基因表达与花色表型的关系[J].园艺学报, 2012, 39(3):516-524 [19]HAN Ke-ting, ZHAO Li, TANG Xing-jiao, HU Ke, DAI Si-lan.The Relationship Between the Expression of Key Genes in Anthocyanin Biosynthesis and the Color of Chrysanthemum[J].[J].Acta Horticulturae Sinica, 2012, 39(3):516-524 [20]Mudalige-Jayawickrama RG., Champagne M.M., Hieber A.D., and Kuehnle A.R., .Cloning and characterization of two anthocyanin biosynthetic genes from Dendrobium orchid[J].[J].J. Am. Soc. Hort. Sci, 2005, 130:611-618 [21]Pitakdantham W.Sutabutra T,Chiemsombat P.,and Pitaksutheepong C.,Isolation and characterization of chalcone synthase gene isolated from Dendrobium Sonia Earsakul[J].Pakistan Journal of Biological Sciences, 2010, 13(20):1000-1005 [22]Tanaka Y.Sasaki N,and Ohmiya A.,Biosynthesis of plant pigments: Anthocyanins,betalains and carotenoids[J].The Plant Journal, 2008, 54(4):733-749 [23]Gonzalez A, Zhao M, Leavitt JM, Lloyd AM.Regulation of the anthocyanin biosynthetic pathway by the TTG1bHLHMyb transcriptional complex in Arabidopsis seedlings[J].Plant J, 2008, 53(5):814-827 [24]Vimolmangkang S, Han Y, Wei G, Korban SS.An apple MYB transcription factor, MdMYB3, is involved in regulation of anthocyanin biosynthesis and flower development[J]. [J].BMC Plant Biol.., 2013, 7(13):176-189 [25]Griesbach R J., and Klein T.M., .In situ geneticcomplementation of a flower color mutant in Doritis pulcherrima (Orchidaceael) [J]. [J].Lindleyana, 1993, 8:223-226 [26]Ma H.and Pooler M,Ratio of Myc and Myb transcription factors regulates anthocyanin production in orchid flowers[J].J. Amer. Soc. Hort. Sci., 2008, 133(1):133-138 [27]Bailey PC, Martin C, Toledo-Ortiz G, Quail PH, Huq E, Heim MA, Jakoby M, Werber M, Weisshaar B.Update on the basic helix-loop-helix transcription factor gene family in Arabidopsis thaliana[J].Plant Cell, 2003, 15(11):2497-2501 [28]Li X, Duan X, Jiang H, Sun Y, Tang Y, Yuan Z, Guo J, Liang W, Chen L, Yin J et al Genome-wide analysis of basic helix-loophelix transcription factor family in rice and Arabidopsis[J].Genome-wide analysis of basic helix-loophelix transcription factor family in rice and Arabidopsis[J].[J].Plant Physiol,, 2006, 141(4):1167-1184 [29]Albert N.W.,Arathoon S,Collette V.E,Schwinn K.E.,Jameson P.E.,Lewis D.H.,Zhang H.B.,and Davies K.M.,Activation of anthocyanin synthesis in Cymbidium orchids: variability between known regulators[J].Plant Cell Tissue and Organ Culture, 2010, 100(3):355-360 [30]Tanaka Y, Ohmiya A.Seeing is believing: Engineering anthocyanin and carotenoid biosynthetic pathways[J]. [J].Current Opinion in Biotechnology, 2008, 19(2):190-197 [31]Katsumoto Y, Fukuchi MizutaniM, Fukui Y, Brugliera F, Holton TA, KaranM, Nakamura N, Yonekura Sakakibara K, Togami J, Pigeaire A, Tao G Q, Nehra N S, Lu C Y, Dyson B K, Tsuda S, Ashikari T, Kusumi T, Mason J G, Tanaka Y.Engineering of the rose flavonoid biosynthetic pathway successfully generated blue hued flowers accumulating delphinidin[J]. [J].Plant Cell Physiol, 2007, 48(11):1589-1600 [32]Kumar V1, Yadav SK.Overexpression of CsANR increased flavan-3-ols and decreased anthocyanins in transgenic tobacco[J].[J].Mol Biotechnol, 2013, 54(2):426-435

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