Abstract
Anthocyanin is the primary pigment contributing to red, violet, and blue flower color formation. The solubility of anthocyanins is enhanced by UDP glucose: flavonoid 3-O-glucosyltransferase (UFGT) through transfer of the glucosyl moiety from UDP-glucose to 3-hydroxyl group to produce the first stable pigments. To assess the possibility that UFGT is involved in the flower color formation in Phalaenopsis, the transcriptional activities of PeUFGT3, and other flower color-related genes in developing red or white flower buds were examined using RT-PCR analysis. In contrast with chalcone synthase, chalcone isomerase, and anthocyanidin synthase genes, PeUFGT3 transcriptional activity was higher expressed in the red color of Phalaenopsis cultivars. In the red labellum of Phalaenopsis ‘Luchia Lady’, PeUFGT3 also showed higher expression levels than that in the white perianth. PeUFGT3 was predominantly expressed in the red region of flower among various Phalaenopsis cultivars. To investigate the role of PeUFGT3 in red flower color formation, PeUFGT3 was specifically knocked down using RNA interference technology via virus inducing gene silencing in Phalaenopsis. The PeUFGT3-suppressed Phalaenopsis exhibited various levels of flower color fading that was well correlated with the extent of reduced level of PeUFGT3 transcriptional activity. Furthermore, there was a significant decrease in anthocyanin content in the PeUFGT3-suppressed Phalaenopsis flowers. The decrease of anthocyanin content due to PeUFGT3 gene silencing possibly caused the faded flower color in PeUFGT3-suppressed Phalaenopsis. Consequently, these results suggested that the glycosylation-related gene PeUFGT3 plays a critical role in red color formation in Phalaenopsis.
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Abbreviations
- ANL1 :
-
Anthocyaninless1
- ANS:
-
Anthocyanidin synthase
- CHI:
-
Chalcone isomerase
- CHS:
-
Chalcone synthase
- CymMV:
-
Cymbidium mosaic virus
- DFR:
-
Dihydroflavonol 4-reductase
- EST:
-
Expressed sequence tags
- F3′H:
-
Flavonoid 3′-hydroxylase
- F3′,5′H:
-
Flavonoid 3′,5′-hydroxylase
- GT:
-
Glycosyltransferase
- PCR:
-
Polymerase chain reaction
- RT-PCR:
-
Reverse transcription polymerase chain reaction
- RNAi:
-
RNA interference
- SiRNA:
-
Short interfering RNA
- UFGT:
-
UDP glucose: flavonoid 3-O-glucosyltransferase
- VIGS:
-
Virus-induced gene silencing
References
Aida R, Yoshida K, Kondo T, Kishimoto S, Shibata M (2000) Copigmentation gives bluer flowers on transgenic torenia plants with the antisense dihydroflavonol-4-reductase gene. Plant Sci 160:49–56
Forkmann G (1991) Flavonoids as flower pigments: the formation of the natural spectrum and its extension by genetic engineering. Plant Breed 106:1–26
Forkmann G, Martens S (2001) Metabolic engineering and applications of flavonoids. Curr Opin Biotechnol 12:155–160
Fukuchi-Mizutani M, Okuhara H, Fukui Y, Nakao M, Katsumoto Y, Yonekura-Sakakibara K, Kusumi T, Hase T, Tanaka Y (2003) Biochemical and molecular characterization of a novel UDP-glucose: anthocyanin 3′-O-glucosyltransferase, a key enzyme for blue anthocyanin biosynthesis, from Gentian. Plant Physiol 132:1652–1663
Fukusaki E, Kawasaki K, Kajiyama S, An C-I, Suzuki K, Tanaka Y, Kobayashi A (2004) Flower color modulations of Torenia hybrida by downregulation of chalcone synthase genes with RNA interference. J Biotechnol 111:229–240
Grotewold E (2006) The genetics and biochemistry of floral pigments. Annu Rev Plant Biol 57:761–780
Han YY, Ming F, Wang W, Wang JW, Ye MM, Shen DL (2006) Molecular evolution and functional specialization of chalcone synthase superfamily from Phalaenopsis Orchid. Genetica 128:429–438
Harborne JB, Williams CA (1998) Anthocyanins and other flavonoids. Nat Prod Rep 15:631–652
Harborne JB, Williams CA (2000) Advances in flavonoid research since 1992. Phytochemistry 55:481–504
Harborne JB, Williams CA (2001) Anthocyanins and other flavonoids. Nat Prod Rep 18:310–333
Hieber AD, Mudalige-Jayawickrama RG, Kuehnle AR (2006) Color genes in the orchid Oncidium Gower Ramsey: identification, expression, and potential genetic instability in an interspecific cross. Planta 223:521–531
Holsters M, de Waele D, Depicker A, Messens E, van Montagu M, Schell J (1978) Transfection and transformation of Agrobacterium tumefaciens. Mol Gen Genet 163:181–187
Holton TA, Cornish EC (1995) Genetics and biochemistry of anthocyanin biosynthesis. Plant Cell 7:1071–1083
Hung KT, Cheng DG, Hsu YT, Kao CH (2008) Abscisic acid-induced hydrogen peroxide is required for anthocyanin accumulation in leaves of rice seedlings. J Plant Physiol 165:1280–1287
Jackson D, Roberts K, Martin C (1992) Temporal and spatial control of expression of anthocyanin biosynthetic genes in developing flowers of Antirrhinum majus. Plant J 2:425–434
Kahn RA, Bak S, Svendsen I, Halkier BA, Moller BL (1997) Isolation and reconstitution of cytochrome P450ox and in vitro reconstitution of the entire biosynthetic pathway of the cyanogenic glucoside dhurrin from sorghum. Plant Physiol 115:1661–1670
Kobayashi S, Ishimaru M, Ding CK, Yakushiji H, Goto N (2001) Comparison of UDP-glucose: flavonoid 3-O-glucosyltransferase (UFGT) gene sequences between white grapes (Vitis vinifera) and their sports with red skin. Plant Sci 160:543–550
Kobayashi S, Ishimaru M, Hiraoka K, Honda C (2002) Myb-related genes of the Kyoho grape (Vitis labruscana) regulate anthocyanin biosynthesis. Planta 215:924–933
Koes R, Verweij W, Quattrocchio F (2005) Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. Trends Plant Sci 10:236–242
Kreuz K, Tommasini R, Martinoia E (1996) Old enzymes for a new job (herbicide detoxification in plants). Plant Physiol 111:349–353
Kubo A, Aono M, Nakajima N, Saji H, Tanaka K, Kondo N (1999) Differential responses in activity of antioxidant enzymes to different environmental stresses in Arabidopsis thaliana. J Plant Res 112:279–290
Kubo H, Nawa N, Lupsea SA (2007) Anthocyaninless1 gene of Arabidopsis thaliana encodes a UDP-glucose: flavonoid-3-O-glucosyltransferase. J Plant Res 120:445–449
Kuehnle AR, Lewis DH, Markham KR, Mitchell KA, Davies KM, Jordan BR (1997) Floral flavonoids and pH in Dendrobium orchid species and hybrids. Euphytica 95:187–194
Liew CF, Goh CJ, Loh CS, Lim SH (1998a) Cloning and characterization of full-length cDNA clones encoding chalcone synthase from the orchid Bromheadia finlaysoniana. Plant Physiol Biochem 36:647–656
Liew CF, Loh CS, Goh CJ, Lim SH (1998b) The isolation, molecular characterization and expression of dihydroflavonol 4-reductase cDNA in the orchid, Bromheadia finlaysoniana. Plant Sci 135:161–169
Lo Piero AR, Consoli A, Puglisi I, Orestano G, Recupero GR, Petrone G (2005) Anthocyaninless cultivars of sweet orange lack to express the UDP-glucose flavonoid 3-O-glucosyl transferase. J Plant Biochem Biotechnol 14:9–14
Lu HC, Chen HH, Tsai WC, Chen WH, Su HJ, Chang DCN, Yeh HH (2007) Strategies for functional validation of genes involved in reproductive stages of orchids. Plant Physiol 143:558–569
Martin C, Gerats T (1992) The control of flower coloration. In: Jordan B (ed) Molecular biology of flowers. CIB International, Wallingford, pp 219–255
Martin C, Gerats T (1993) Control of pigment biosynthesis genes during petal development. Plant Cell 5:1253–1264
Mudalige RG, Kuehnle AR (2004) Orchid biotechnology in production and improvement. Hort Sci 39:11–17
Nakajima JI, Yoshikazu T, Yamazak M, Saito K (2001) Reaction mechanism from leucoanthocyanidin to anthocyanidin 3-glucoside, a key reaction for coloring in anthocyanin biosynthesis. J Biol Chem 276:25797–25803
Nishihara M, Nakatsuka T, Yamamura S (2005) Flavonoid components and flower color change in transgenic tobacco plants by suppression of chalcone isomerase gene. FEBS Lett 579:6074–6078
Nishimura G (1981) Comparative morphology of Cattleya and Phalaenopsis (Orchidaceae) seedlings. Bot Gaz 142:360–365
O’Neill SD, Nadeau JA, Zhang XS, Bui AQ, Halevy AH (1993) Interorgan regulation of ethylene biosynthetic genes by pollination. Plant Cell 5:419–432
Saito K, Yamazaki M (2002) Biochemistry and molecular biology of the late-stage of biosynthesis of anthocyanin: lessons from Perilla frutescens as a model plant. New Phytol 155:9–23
Springob K, Nakajima J, Yamazaki M, Saito K (2003) Recent advances in the biosynthesis and accumulation of anthocyanins. Nat Prod Rep 20:288–303
Suzuki K-I, Xue H-M, Tanaka Y, Fukui Y, Fukuchi-Mizutani M, Murakami Y, Katsumoto Y, Tsuda S, Kusumi T (2000) Flower color modifications of Torenia hybrida by cosuppression of anthocyanin biosynthesis genes. Mol Breed 6:239–246
Taguchi G, Fujikawa S, Yazawa T, Kodaira R, Hayashida N, Shimosaka M, Okazaki M (2000) Scopoletin uptake from culture medium and accumulation in the vacuoles after conversion to scopolin in 2, 4-d-treated tobacco cells. Plant Sci 151:153–161
Tsai WC, Hsiao YY, Lee SH, Tung CW, Wang DP, Wang HC, Chen WH, Chen HH (2006) Expression analysis of the ESTs derived from the flower buds of Phalaenopsis equestris. Plant Sci 170:426–432
Tsuda S, Fukui Y, Nakamura N, Katsumoto Y, Yonekura-Sakakibara K, Fukuchi-Mizutani M, Ohira K, Ueyama Y, Ohkawa H, Holton TA, Kusumi T, Tanaka Y (2004) Flower color modification of Petunia hybrida commercial varieties by metabolic engineering. Plant Biotechnol 21:377–386
Vogt T, Jones P (2000) Glycosyltransferases in plant natural product synthesis: characterization of a supergene family. Trends Plant Sci 5:380–386
Wang J, Ming F, Han Y, Shen D (2006) Flavonoid-3′, 5′-hydroxylase from Phalaenopsis: a novel member of cytochrome P450s, its cDNA cloning, endogenous expression and molecular modeling. Biotechnol Lett 28:327–334
Werner C, Matile P (1985) Accumulation of coumarylglucosides in vacuoles of barley mesophyll protoplasts. J Plant Physiol 118:237–249
Williams CA, Grayer RJ (2004) Anthocyanins and other flavonoids. Nat Prod Rep 21:539–573
Yeh HH, Tian T, Rubio L, Crawford B, Falk BW (2000) Asynchronous accumulation of lettuce infectious yellows virus RNAs 1 and 2 and identification of an RNA 1 trans enhancer of RNA 2 accumulation. J Virol 74:5762–5768
Acknowledgments
The authors like to thank Dr. H. H. Yeh, Department of Plant Pathology and Microbiology, National Taiwan University for providing the pCymMV-pro60 vector. This work was supported by grants to M.-J. Ger from the National Science Council of Taiwan, Republic of China.
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Chen, WH., Hsu, CY., Cheng, HY. et al. Downregulation of putative UDP-glucose: flavonoid 3-O-glucosyltransferase gene alters flower coloring in Phalaenopsis . Plant Cell Rep 30, 1007–1017 (2011). https://doi.org/10.1007/s00299-011-1006-1
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DOI: https://doi.org/10.1007/s00299-011-1006-1