ABB  Vol.6 No.12 , December 2015
Transforming the Snapdragon Aurone Biosynthetic Genes into Petunia Alters Coloration Patterns in Transgenic Flowers
Abstract: Aurones belong to a small class of flavonoids that provide yellow color in some floricultural plants including snapdragon. To explore novel flower coloration, two full-length cDNAs encoding chalcone 4'-O-glucosyltransferase (designated as SRY4'CGT) and aureusidin synthase (designated as SRYAS1) in the aurone biosynthetic pathway were cloned from yellow flowers of snapdragon (Antirrhinum majus cv. Ribbon Yellow). Binary vectors were constructed and transformed separately into Petunia hybrida harboring blue flowers. Only a few flowers in 4 out of 9 transgenic SRY4'CGT plants showed variegated blue-white sectors; as time passed, amounts of variegated flowers and proportion of white sectors in the background blue color of the new-born flowers gradually increased, until finally, the petal color was completely white in all late-born flowers. In contrast, a few flowers in 3 out of 13 transgenic SRYAS1 plants showed variegated blue-white sectors; but, the amounts of variegated flowers did not increase over the whole flowering stage, and no complete white flowers were observed. RNA samples isolated from blue and white sectors of T1 transgenic SRY4'CGT plants were analyzed by reverse transcription-PCR, transgenic SRY4'CGT transcripts were detected in both sectors; however, transcripts of an upstream gene, chalcone synthase (CHS), were abundantly detected in the blue sectors but largely reduced in the white sectors, suggesting that the expression of CHS gene was suppressed in white sectors of transgenic plants. Furthermore, HPLC coupled with mass spectrometry demonstrated cyandin, malvidin and their derivatives were absent in white sectors, causing the white phenotype. Our findings may be attractive to molecular breeders.
Cite this paper: Wang, C. , Chin, Y. , Lin, C. , Chen, P. and To, K. (2015) Transforming the Snapdragon Aurone Biosynthetic Genes into Petunia Alters Coloration Patterns in Transgenic Flowers. Advances in Bioscience and Biotechnology, 6, 702-722. doi: 10.4236/abb.2015.612073.

[1]   Bartley, G.E. and Scolnik, P.A. (1995) Plant Carotenoids: Pigments for Photoprotection, Visual Attraction, and Human Health. Plant Cell, 7, 1027-1038.

[2]   To, K.Y. and Wang, C.K. (2006) Molecular Breeding of Flower Color. In: Teixeira da Silva, J.A., Ed., Floriculture, Ornamental and Plant Biotechnology, Volume 1, Global Science Books, London, 300-310.

[3]   Nishihara, M. and Nakatsuka, T. (2011) Genetic Engineering of Flavonoid Pigments to Modify Flower Color in Floricultural Plants. Biotechnology Letters, 33, 433-441.

[4]   Schwinn, K.E. and Davies, K.M. (2004) Flavonoids. In: Davies, K.M., Ed., Plant Pigments and Their Manipulations, Blackwell Publishing Ltd., Oxford, 92-149.

[5]   Tanaka, Y., Katsumoto, Y., Brugliera, F. and Mason, J. (2005) Genetic Engineering in Floriculture. Plant Cell, Tissue and Organ Culture, 80, 1-24.

[6]   Tanaka, Y., Brugliera, F., Kalc, G., Senior, M., Dyson, B., Nakamura, N., Katsumoto, Y. and Chandler, S. (2010) Flower Color Modification by Engineering of the Flavonoid Biosynthetic Pathway: Practical Perspectives. Bioscience, Biotechnology and Biochemistry, 74, 1760-1769.

[7]   Vogt, T. (2010) Phenylpropanoid Biosynthesis. Molecular Plant, 3, 2-20.

[8]   Chandler, S.F. and Brugliera, F. (2011) Genetic Modification in Floriculture. Biotechnology Letter, 33, 207-214.

[9]   Winkel-Shirley, B. (2001) Flavonoid Biosynthesis: A Colorful Model for Genetics, Biochemistry, Cell Biology, and Biotechnology. Plant Physiology, 126, 485-493.

[10]   Nakayama, T., Yonekura-Sakakibara, K., Sato, T., Kikuchi, S., Fukui, Y., Fukuchi-Mizutani, M., Ueda, T., Nakao, M., Tanaka, Y., Kusumi, T. and Nishino, T. (2000) Aureusidin Synthase: A Polyphenol Oxidase Homolog Responsible for Flower Coloration. Science, 290, 1163-1166.

[11]   Ono, E., Fukuchi-Mizutani, M., Nakamura, N., Fukui, Y., Yonekura-Sakakibara, K., Yamaguchi, M., Nakayama, T., Tanaka, T., Kusumi, T. and Tanaka, Y. (2006) Yellow Flowers Generated by Expression of the Aurone Biosynthetic Pathway. Proceedings of the National Academy of Sciences of the United States of America, 103, 11075-11080.

[12]   Wang, C.K., Chen, P.Y., Wang, H.M. and To, K.Y. (2006) Cosuppression of Tobacco Chalcone Synthase Using Petunia Chalcone Synthase Construct Results in White Flowers. Botanical Studies, 47, 71-82.

[13]   Gerats, T. and Vandenbussche, M. (2005) A Model System for Comparative Research: Petunia. Trends in Plant Science, 10, 251-256.

[14]   Lutke, W.K. (2006) Petunia (Petunia hybrid). Methods in Molecular Biology, 344, 339-349.

[15]   Murashige, T. and Skoog, F. (1962) A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Cultures. Physiologia Plantarum, 15, 473-497.

[16]   Wang, H.M., Yin, W.C., Wang, C.K. and To, K.Y. (2009) Isolation of Functional RNA from Different Tissues of Tomato Suitable for Developmental Profiling by Microarray Analysis. Botanical Studies, 50, 115-125.

[17]   Wang, C.K., Hsu, S.Y., Chen, P.Y. and To, K.Y. (2012) Transformation and Characterization of Transgenic Bidens pilosa L. Plant Cell, Tissue and Organ Culture, 109, 457-464.

[18]   Nakayama, T., Sato, T., Fukui, Y., Yonekura-Sakakibari, K., Hayashi, H., Tanaka, Y., Kusumi, T. and Nishino, T. (2001) Specificity Analysis and Mechanism of Aurone Synthesis Catalyzed by Aureusidin Synthase, a Polyphenol Oxidase Homolog Responsible for Flower Coloration. FEBS Letters, 499, 107-111.

[19]   Giusti, M.M., Rodriguez-Saona, L.E., Griffin, D. and Wrolstad, R.E. (1999) Electrospray and Tandem Mass Spectroscopy as Tools for Anthocyanin Characterization. Journal of Agricultural and Food Chemistry, 47, 4657-4664.

[20]   Schütz, K., Persike, M., Carle, R. and Schieber, A. (2006) Characterization and Quantification of Anthocyanins in Selected Artichoke (Cynara scolymus L.) Cultivars by HPLC-DAD-ESI-MSn. Analytical and Bioanalytical Chemistry, 384, 1511-1517.

[21]   Hillebrand, S., Naumann, H., Kitzinski, N., Köhler, N. and Winterhalter, P. (2009) Isolation and Characterization of Anthocyanins from Blue-Fleshed Potatoes (Solanum tuberosum L.). Food, 3, 96-101.

[22]   Zhao, H., Hu, X., Chen, X., Shi, S., Jiang, X., Chen, W. and Zhang, S. (2015) Analysis and Improved Characterization of Minor Antioxidants from Leaves of Malus doumeri Using a Combination of Major Constituents’ Knockout with High-Performance Liquid Chromatography-Diode Array Detector-Quadrupole Time-of-Flight Tandem Mass Spectrometry. Journal of Chromatography A, 1398, 57-65.

[23]   Nakatsuka, T., Mishiba, K.I., Kubota, A., Abe, Y., Yamamura, S., Nakamura, N., Tanaka, Y. and Nishihara, M. (2010) Genetic Engineering of Novel Flower Colour by Suppression of Anthocyanin Modification Genes in Gentian. Journal of Plant Physiology, 167, 231-237.

[24]   Chandler, S.F. and Sanchez, C. (2012) Genetic Modification; the Development of Transgenic Ornamental Plant Varieties. Plant Biotechnology Journal, 10, 891-903.

[25]   Napoli, C., Lemieux, C. and Jorgensen, R. (1990) Induction of a Chimer Chalcone Synthase Gene into Petunia Results in Reversible Co-Suppression of Homologous Genes in trans. Plant Cell, 2, 279-289.

[26]   van der Krol, A., Mur, L.A., Beld, M., Mol, J.N.M. and Stuitje, A.R. (1990) Flavonoid Genes in Petunia: Addition of a Limited Number of Gene Copies May Lead to a Suppression of Gene Expression. Plant Cell, 2, 291-299.

[27]   Vaucheret, H., Béclin, C., Elmayan, T., Feuerbach, F., Godon, C., Morel, J.-B., Mourrain, P., Palauqui, J.-C. and Vernhettes, S. (1998) Transgene-Induced Gene Silencing in Plants. Plant Journal, 16, 651-659.

[28]   Eamens, A., Wang, M.B., Smith, N.A. and Waterhouse, P.M. (2008) RNA Silencing in Plants: Yesterday, Today, and Tomorrow. Plant Physiology, 147, 456-468.

[29]   Han, Y., Vimolmangkang, S., Soria-Guerra, R.E. and Korban, S.S. (2012) Induction of Apple ANR Genes into Tobacco Inhibits Expression of both CHI and DFR Genes in Flowers, Leading to Loss of Anthocyanin. Journal of Experimental Botany, 63, 2437-2447.

[30]   Peltier, D., Farcy, E., Dulieu, H. and Bervillé, A. (1994) Origin, Distribution and Mapping of RAPD Markers from Wild Petunia Species in Petunia hybrida Hort Lines. Theoretical and Applied Genetics, 88, 637-645.

[31]   Dietz-Pfeilstetter, A. (2010) Stability of Transgene Expression as a Challenge for Genetic Engineering. Plant Science, 179, 164-167.

[32]   Hauser, M.T., Aufsatz, W., Jonak, C. and Luschnig, C. (2011) Transgenerational Epigenetic Inheritance in Plants. Biochimica et Biophysica Acta, 1809, 459-468.

[33]   Tsai, Y.T., Chen, P.Y. and To, K.Y. (2012) Plant Regeneration and Stable Transformation in the Floricultural Plant Cleome spinosa, a C3 Plant Closely Related to the C4 Plant C. gynandra. Plant Cell Reports, 31, 1189-1198.