Enhancing Flower Color through Simultaneous Expression of the B-peru and mPAP1 Transcription Factors under Control of a Flower-Specific Promoter
Abstract
:1. Introduction
2. Results
2.1. Expression of Anthocyanin Regulators in Floral Tissue of Tobacco Results in Flower Color Changes
2.2. Measurement of Flower Color and Petal pH
2.3. Gene Expression of B-peru and mPAP1 under Control of the Tobacco ANS Promoter Was Restricted in Floral Organs
2.4. Flower Color Modification by Two Anthocyanin Regulators
2.5. Quercetin and Cyanidin Are Significantly Increased in Transgenic Tobacco Flowers
3. Discussion
4. Materials and Methods
4.1. Gene Isolation and Vector Construction
4.2. Plant Regeneration
4.3. Measurement of Flower Color and Petal pH
4.4. Total RNA Extraction and qPCR Analysis
4.5. Flavonoid Analysis
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
- Martin, C.; Butelli, E.; Petroni, K.; Tonelli, C. How can research on plants contribute to promoting human health? Plant Cell 2011, 23, 1685–1699. [Google Scholar] [CrossRef] [PubMed]
- Grotewold, E. The genetics and biochemistry of floral pigments. Annu. Rev. Plant Biol. 2006, 57, 761–780. [Google Scholar] [CrossRef] [PubMed]
- Katsumoto, Y.; Fukuchi-Mizutani, M.; Fukui, Y.; Brugliera, F.; Holton, T.A.; Karan, M.; Nakamura, N.; Yonekura-Sakakibara, K.; Togami, J.; Pigeaire, A.; et al. Engineering of the rose flavonoid biosynthetic pathway successfully generated blue-hued flowers accumulating delphinidin. Plant Cell Physiol. 2007, 48, 1589–1600. [Google Scholar] [CrossRef] [PubMed]
- Boase, M.R.; Lewis, D.H.; Davies, K.M.; Marshall, G.B.; Patel, D.; Schwinn, K.E.; Deroles, S.C. Isolation and antisense suppression of flavonoid 3′,5′-hydroxylase modifies flower pigments and colour in cyclamen. BMC Plant Biol. 2010, 10, 107. [Google Scholar] [CrossRef] [PubMed]
- Nakatsuka, T.; Mishiba, K.; Kubota, A.; Abe, Y.; Yamamura, S.; Nakamura, N.; Tanaka, Y.; Nishihara, M. Genetic engineering of novel flower colour by suppression of anthocyanin modification genes in gentian. J. Plant Physiol. 2010, 167, 231–237. [Google Scholar] [CrossRef] [PubMed]
- Hichri, I.; Barrieu, F.; Bogs, J.; Kappel, C.; Delrot, S.; Lauvergeat, V. Recent advances in the transcriptional regulation of the flavonoid biosynthetic pathway. J. Exp. Bot. 2011, 62, 2465–2483. [Google Scholar] [CrossRef] [PubMed]
- Xu, W.; Dubos, C.; Lepiniec, L. Transcriptional control of flavonoid biosynthesis by MYB-bHLH-WDR complexes. Trends Plant Sci. 2015, 20, 176–185. [Google Scholar] [CrossRef] [PubMed]
- Lim, S.H.; Kim, D.H.; Kim, J.K.; Lee, J.Y.; Ha, S.H. A radish basic helix-loop-helix transcription factor, RsTT8 acts a positive regulator for anthocyanin biosynthesis. Front. Plant Sci. 2017, 8, 1917. [Google Scholar] [CrossRef] [PubMed]
- Aharoni, A.; de Vos, C.H.R.; Wein, M.; Sun, Z.; Greco, R.; Kroon, A.; Mol, J.N.M.; O’Connell, A.P. The strawberry FaMYB1 transcription factor suppresses anthocyanin and flavonol accumulation in transgenic tobacco. Plant J. 2011, 28, 319–332. [Google Scholar] [CrossRef]
- Dubos, C.; le Gourrierec, J.; Baudry, A.; Huep, G.; Lanet, E.; Debeaujon, I.; Routaboul, J.M.; Alboresi, A.; Weisshaar, B.; Lepiniec, L. MYBL2 is a new regulator of flavonoid biosynthesis in Arabidopsis thaliana. Plant J. 2008, 55, 940–953. [Google Scholar] [CrossRef] [PubMed]
- Albert, N.W.; Davies, K.M.; Lewis, D.H.; Zhang, H.; Montefiori, M.; Brendolise, C.; Murray, R.V.; Hanh, N.; Paula, E.J.; Schwinn, K.E. A conserved network of transcriptional activators and repressors regulates anthocyanin pigmentation in eudicots. Plant Cell 2014, 26, 962–980. [Google Scholar] [CrossRef] [PubMed]
- Zvi, M.M.B.; Shklarman, E.; Masci, T.; Kalev, H.; Debener, T.; Shafir, S.; Marianna, O.; Vainstein, A. PAP1 transcription factor enhances production of phenylpropanoid and terpenoid scent compounds in rose flowers. New Phytol. 2012, 195, 335–345. [Google Scholar] [CrossRef] [PubMed]
- He, X.; Li, Y.; Lawson, D.; Xie, D.Y. Metabolic engineering of anthocyanins in dark tobacco varieties. Physiol. Plant 2017, 159, 2–12. [Google Scholar] [CrossRef] [PubMed]
- Schwinn, K.E.; Boase, M.R.; Bradley, J.M.; Lewis, D.H.; Deroles, S.C.; Martin, C.R.; Davies, K.M. MYB and bHLH transcription factor transgenes increase anthocyanin pigmentation in petunia and lisianthus plants, and the petunia phenotypes are strongly enhanced under field conditions. Front. Plant Sci. 2014, 5. [Google Scholar] [CrossRef] [PubMed]
- An, C.H.; Lee, K.W.; Lee, S.H.; Jeong, Y.J.; Woo, S.G.; Chun, H.; Park, Y.I.; Kwak, S.S.; Kim, C.Y. Heterologous expression of IbMYB1a by different promoters exhibits different patterns of anthocyanin accumulation in tobacco. Plant Physiol. Biochem. 2015, 89, 1–10. [Google Scholar] [CrossRef] [PubMed]
- Dasgupta, K.; Thilmony, R.; Stover, E.; Oliveira, M.L.; Thomson, J. Novel R2R3-MYB transcription factors from Prunus americana regulate differential patterns of anthocyanin accumulation in tobacco and citrus. GM Crops Food 2017, 8, 85–105. [Google Scholar] [CrossRef] [PubMed]
- Lim, S.H.; Sohn, S.H.; Kim, D.H.; Kim, J.K.; Lee, J.Y.; Kim, Y.M.; Ha, S.H. Use of an anthocyanin production phenotype as a visible selection marker system in transgenic tobacco plant. Plant Biotechnol. Rep. 2012, 6, 203–211. [Google Scholar] [CrossRef]
- Imai, A.; Takahashi, S.; Nakayama, K.; Satoh, H. The promoter of the carotenoid cleavage dioxygenase 4a-5 gene of Chrysanthemum morifolium (CmCCD4a-5) drives petal-specific transcription of a conjugated gene in the developing flower. J. Plant Physiol. 2013, 170, 1295–1299. [Google Scholar] [CrossRef] [PubMed]
- Liu, Y.; Lou, Q.; Xu, W.; Xin, Y.; Bassett, C.; Wang, Y. Characterization of a chalcone synthase (CHS) flower-specific promoter from Lilium orential ‘Sorbonne’. Plant Cell Rep. 2011, 30, 2187–2194. [Google Scholar] [CrossRef] [PubMed]
- Lim, S.H.; Kim, D.H.; Kim, J.K.; Lee, J.Y.; Kim, Y.M.; Sohn, S.H.; Kim, D.H.; Ha, S.H. Petal-specific activity of the promoter of an anthocyanin synthase gene of tobacco (Nicotiana tabacum L.). Plant Cell Tissue Organ Cult. 2013, 114, 373–383. [Google Scholar] [CrossRef]
- Azuma, M.; Morimoto, R.; Hirose, M.; Morita, Y.; Hoshino, A.; Iida, S.; Oshima, Y.; Mitsuda, N.; Ohme-Takagi, M.; Shiratake, K. A petal-specific InMYB1 promoter from Japanese morning glory: A useful tool for molecular breeding of floricultural crops. Plant Biotechnol. J. 2016, 14, 354–363. [Google Scholar] [CrossRef] [PubMed]
- Sierro, N.; Battey, J.N.D.; Ouadi, S.; Bakaher, N.; Bovet, L.; Willig, A.; Goepfert, S.; Peitsch, M.; Ivanov, N. The tobacco genome sequence and its comparison with those of tomato and potato. Nat. Commun. 2014, 5, 3833. [Google Scholar] [CrossRef] [PubMed]
- Zhang, Y.; Zhang, J.; Song, T.; Li, J.; Tian, J.; Jin, K.; Yao, Y. Low medium pH value enhances anthocyanin accumulation in Malus crabapple leaves. PLoS ONE 2014, 9, e97904. [Google Scholar] [CrossRef] [PubMed]
- Schmitzer, V.; Veberic, R.; Osterc, G.; Stampar, F. Color and phenolic content changes during flower development in groundcover rose. J. Am. Soc. Hortic. Sci. 2010, 135, 195–202. [Google Scholar]
- Cavallini, E.; Zenoni, S.; Finezzo, L.; Guzzo, F.; Zamboni, A.; Avesani, L.; Tornielli, G.B. Functional diversification of grapevine MYB5a and MYB5b in the control of flavonoid biosynthesis in a petunia anthocyanin regulatory mutant. Plant Cell Physiol. 2014, 55, 517–534. [Google Scholar] [CrossRef] [PubMed]
- Spelt, C.; Quattrocchio, F.; Mol, J.; Koes, R. ANTHOCYANIN1 of petunia controls pigment synthesis, vacuolar pH, and seed coat development by genetically distinct mechanisms. Plant Cell 2002, 14, 2121–2135. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Du, H.; Wu, J.; Ji, K.X.; Zeng, Q.Y.; Bhuiya, M.W.; Su, S.; Shu, Q.Y.; Ren, H.X.; Liu, Z.A.; Wang, L.S. Methylation mediated by an anthocyanin, O-methyltransferase, is involved in purple flower coloration in Paeonia. J. Exp. Bot. 2015, 66, 6563–6577. [Google Scholar] [CrossRef] [PubMed]
- Faraco, M.; Spelt, C.; Bliek, M.; Verweij, W.; Hoshino, A.; Espen, L.; Prinsi, B.; Jaarsma, R.; Tarhan, E.; de Boer, A.H.; et al. Hyperacidification of vacuoles by the combined action of two different P-ATPases in the tonoplast determines flower color. Cell Rep. 2014, 6, 32–43. [Google Scholar] [CrossRef] [PubMed]
- Fukada-Tanaka, S.; Inagaki, Y.; Yamaguchi, T.; Saito, N.; Iida, S. Colour-enhancing protein in blue petals. Nature 2000, 407, 581. [Google Scholar] [CrossRef] [PubMed]
- Lim, S.H.; Song, J.H.; Kim, D.H.; Kim, J.K.; Lee, J.Y.; Kim, Y.M.; Ha, S.H. Activation of anthocyanin biosynthesis by expression of the radish R2R3-MYB transcription factor gene RsMYB1. Plant Cell Rep. 2016, 35, 641–653. [Google Scholar] [CrossRef] [PubMed]
- Huang, Y.J.; Song, S.; Allan, A.C.; Liu, X.F.; Yin, X.R.; Xu, C.J.; Chen, K.S. Differential activation of anthocyanin biosynthesis in Arabidopsis and tobacco over-expressing an R2R3 MYB from Chinese bayberry. Plant Cell Tissue Organ Cult. 2013, 113, 491–499. [Google Scholar] [CrossRef]
- Park, S.; Kim, D.H.; Lee, J.Y.; Ha, S.H.; Lim, S.H. Comparative analysis of two flavonol synthases from different-colored onions provides insight into flavonoid biosynthesis. J. Agric. Food Chem. 2017, 65, 5287–5298. [Google Scholar] [CrossRef]
- Luo, P.; Ning, G.; Wang, Z.; Shen, Y.; Jin, H.; Li, P.; Huang, S.; Zhao, J.; Bao, M. Disequilibrium of flavonol synthase and dihydroflavonol-4-reductase expression associated tightly to white vs. red color flower formation in plants. Front. Plant Sci. 2016, 6, 1257. [Google Scholar] [CrossRef] [PubMed]
- Matsubara, K.; Kodama, H.; Kokubun, H.; Watanabe, H.; Ando, T. Two novel transposable elements in a cytochrome P450 gene govern anthocyanin biosynthesis of commercial petunias. Gene 2005, 358, 121–126. [Google Scholar] [CrossRef] [PubMed]
- Chu, Y.X.; Chen, H.R.; Wu, A.Z.; Cai, R.; Pan, J.S. Expression analysis of dihydroflavonol 4-reductase genes in Petunia hybrida. Genet. Mol. Res. 2015, 14, 5010–5021. [Google Scholar] [CrossRef] [PubMed]
- Shih, C.H.; Chu, H.; Tang, L.K.; Sakamoto, W.; Maekawa, M.; Chu, I.K.; Wang, M.; Lo, C. Functional characterization of key structural genes in rice flavonoid biosynthesis. Planta 2008, 228, 1043–1054. [Google Scholar] [CrossRef] [PubMed]
- Zhu, Q.; Sui, S.; Lei, X.; Yang, Z.; Lu, K.; Liu, G.; Liu, Y.-G.; Li, M. Ectopic expression of the coleus R2R3 MYB-type proanthocyanidin regulator gene SsMYB3 alters the flower color in transgenic tobacco. PLoS ONE 2015, 10, e0139392. [Google Scholar] [CrossRef] [PubMed]
- Lim, S.H.; Woo, H.J.; Lee, S.M.; Jin, Y.M.; Cho, H.S. D-Amino acid oxidase (DAO) gene as a novel selection marker for plant transformation. Korean J. Plant Biotechnol. 2007, 34, 31–36. [Google Scholar] [CrossRef]
- Karimi, M.; Inzé, D.; Depicker, A. GATEWAY™ vectors for Agrobacterium-mediated plant transformation. Trends Plant Sci. 2002, 7, 193–195. [Google Scholar] [CrossRef]
- Gonnet, J.F. A colorimetric look at the RHS chart—Perspectives for an instrumental determination of colour codes. J. Hortic. Sci. 1995, 70, 191–206. [Google Scholar] [CrossRef]
Line | Flower Color | L* | a* | b* | C* | pH |
---|---|---|---|---|---|---|
NT | Pink | 78.15 ± 0.81 | 21.75 ± 1.04 | −18.96 ± 0.17 | 28.85 | 5.3 ± 0.01 |
PANSBP_DP | Dark Pink | 73.36 ± 1.71 | 25.44 ± 1.71 | −18.33 ± 0.32 | 31.35 | 5.2 ± 0.01 |
PANSBP_DR | Dark Red | 38.05 ± 0.96 | 42.97 ± 1.02 | −3.19 ± 1.41 | 43.09 | 5.11 ± 0.02 |
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Kim, D.-H.; Park, S.; Lee, J.-Y.; Ha, S.-H.; Lim, S.-H. Enhancing Flower Color through Simultaneous Expression of the B-peru and mPAP1 Transcription Factors under Control of a Flower-Specific Promoter. Int. J. Mol. Sci. 2018, 19, 309. https://doi.org/10.3390/ijms19010309
Kim D-H, Park S, Lee J-Y, Ha S-H, Lim S-H. Enhancing Flower Color through Simultaneous Expression of the B-peru and mPAP1 Transcription Factors under Control of a Flower-Specific Promoter. International Journal of Molecular Sciences. 2018; 19(1):309. https://doi.org/10.3390/ijms19010309
Chicago/Turabian StyleKim, Da-Hye, Sangkyu Park, Jong-Yeol Lee, Sun-Hwa Ha, and Sun-Hyung Lim. 2018. "Enhancing Flower Color through Simultaneous Expression of the B-peru and mPAP1 Transcription Factors under Control of a Flower-Specific Promoter" International Journal of Molecular Sciences 19, no. 1: 309. https://doi.org/10.3390/ijms19010309