Flower Color Modification by Manipulating Flavonoid Biosynthetic Pathway

플라보노이드 대사 조절을 통한 화색 변경

  • Lim, Sun-Hyung (Functional Biomaterial Division, National Academy of Agricultural Science) ;
  • Kim, Jae-Kwang (Biosafety Division, National Academy of Agricultural Science) ;
  • Kim, Dong-Hern (Bio-crop Development Division, National Academy of Agricultural Science) ;
  • Sohn, Seong-Han (Genomics Division, National Academy of Agricultural Science) ;
  • Lee, Jong-Yeol (Functional Biomaterial Division, National Academy of Agricultural Science) ;
  • Kim, Young-Mi (Functional Biomaterial Division, National Academy of Agricultural Science) ;
  • Ha, Sun-Hwa (Functional Biomaterial Division, National Academy of Agricultural Science)
  • 임선형 (국립농업과학원 기능성물질개발과) ;
  • 김재광 (국립농업과학원 생물안전성과) ;
  • 김동헌 (국립농업과학원 신작물개발과) ;
  • 손성한 (국립농업과학원 유전자분석개발과) ;
  • 이종렬 (국립농업과학원 기능성물질개발과) ;
  • 김영미 (국립농업과학원 기능성물질개발과) ;
  • 하선화 (국립농업과학원 기능성물질개발과)
  • Received : 2011.09.05
  • Accepted : 2011.12.28
  • Published : 2011.12.31

Abstract

Flower color is one of the main target traits in the flower breeding. Recently, technological advances in genetic engineering have been successfully reported the flower colors, such as blue roses and blue carnations that are impossible to develop by traditional breeding. Accumulated knowledge-based approaches for flavonoid biosynthesis enabled to introduce novel and unique colors into flowers. These flower color modifications have been made through the regulation of flavonoid metabolic pathway - control of endogenous gene expression and introduction of foreign genes to produce novel and specific flavonoids - and the introduction of transcription factors that are known to regulate sets of genes being involving in the flavonoid biosynthetic pathway. More empirical regulation of the flavonoids metabolism requires the understanding for regulatory mechanism of intrinsic flavonoids depending on the flower crops and the very sophisticated control of flavonoid metabolic flow. In this review, we summarized successful examples of flower color modification. It might be useful to deduce the strategy for the creation of exquisite colors in flower plants.

화색은 화훼 육종의 주요한 목표형질이다. 최근 유전공학 기술의 발달로 기존의 전통육종에서는 볼 수 없었던 파란장미와 파란카네이션과 같은 새로운 화색 개발이 성공적으로 보고되었다. 플라보노이드 생합성에 관해 축적된 지식기반 연구 결과를 바탕으로 새롭고 독특한 형질의 화색을 도입하는 것이 가능하게 된 것이다. 이러한 화색변경은 플라보노이드 대사경로의 조절, 즉 내재유전자의 발현조절 및 새로운 플라보노이드 합성 또는 특정 플라보노이드 합성을 위한 외래유전자의 추가도입과 플라보노이드 대사 전체를 조절하는 전사인자의 도입을 통해서 이루어져 왔다. 그러나 보다 실증적으로 이러한 플라보노이드 대사를 조절하기 위해서는 작물별 내재 플라보노이드의 조절 기작에 대한 이해를 바탕으로 목표로 하는 플라보노이드 합성을 위해 보다 정교한 대사흐름의 조절이 요구된다. 본 총설에서는 화훼작물의 화색변경 성공 예들을 자세히 소개하고 그 요인 분석을 통해 향후 더 성공적인 화색변경의 전략을 수립하는데 도움이 되고자 한다.

Keywords

References

  1. Aida, R., K. Yoshida, T. Kondo, S. Kishimoto, and M. Sibata. 2000. Copigmentation gives bluer flowers on transgenic torenia plants with the antisense dihydroflavonol-4-redutase gene. Plant Sci. 160:49-56. https://doi.org/10.1016/S0168-9452(00)00364-2
  2. Aida, R., K. Ohira, Y. Tanaka, K. Yoshida, S. Kishimoto, M. Shibata, and A. Ohmiya. 2004. Efficient transgene expression in chrysanthemum, Dendranthema grandiflorum (Ramat.) Kitamura, by using the promoter of a gene for chrysanthemum chlorophyll-a/b-binding protein. Breeding Sci. 54:51-58. https://doi.org/10.1270/jsbbs.54.51
  3. Annadana, S., M.J. Beekwilder, G. Kuipers, P.B. Visser, N. Outchkourov, A. Pereira, M. Udayakumar, J. De Jong, and M.A. Jongsma. 2002. Cloning of the chrysanthemum UEP1 promoter and comparative expression in florets and leaves of Dendranthema grandiflora. Transgenic Res. 11:437-445. https://doi.org/10.1023/A:1016313924844
  4. Baumann, K., M. Perez-Rodriguez, D. Bradley, J. Venail, P. Bailey, H. Jin, R. Koes, K. Roberts, and C. Martin. 2007. Control of cell and petal morphogenesis by R2R3 MYB transcription factors. Development 134:1691-1701. https://doi.org/10.1242/dev.02836
  5. Boase, M.R., D.H. Lewis, K.M. Davies, G.B. Marshall, D. Patel, K.E. Schwinn, and S.C. Deroles. 2010. Isolation and antisense suppression of flavonoid 3′,5′-hydroxylase modifies flower pigments and colour in cyclamen. BMC Plant Biol. 10:107. https://doi.org/10.1186/1471-2229-10-107
  6. Butelli, E., L. Titta, M. Giorgio, H.-P. Mock, A. Matros, S. Peterek, E.G.W.M. Schijlen, R.D. Hall, A.G. Bovy, J. Luo, and C. Martin. 2008. Enrichment of tomato fruit with healthpromoting anthocyanins by expression of select transcription factors. Nat. Biotechnol. 26:1301-1308. https://doi.org/10.1038/nbt.1506
  7. Chandler, S. and Y. Tanaka. 2007. Genetic modification in floriculture. Crit. Rev. Plant Sci. 26:169-197. https://doi.org/10.1080/07352680701429381
  8. D'Auria, J.C. 2006. Acyltransferases in plants: A good time to be BAHD. Curr. Opin. Plant Biol. 9:331-340. https://doi.org/10.1016/j.pbi.2006.03.016
  9. Davies, K.M. 2009. Modifying anthocyanin production in flowers. p. 49-83. In: K. Gould, K. Davies, and C. Winefield (eds.). Anthocyanins biosynthesis functions, and applications. Springer, NY.
  10. Di Stilio, V.S., C. Martin, A.F. Schulfer, and C.F. Connelly. 2009. An ortholog of MIXTA-like2 controls epidermal cell shape in flowers of Thalictrum. New Phytol. 183:718-728. https://doi.org/10.1111/j.1469-8137.2009.02945.x
  11. Dubos, C., J. Le Gourrierec, A. Baudry, G. Huep, E. Lanet, I. Debeaujon, J.M. Routaboul, A. Alboresi, B. Weisshaar, and L. Lepiniec. 2008. MYBL2 is a new regulator of flavonoid biosynthesis in Arabidopsis thaliana. Plant J. 55:940-953. https://doi.org/10.1111/j.1365-313X.2008.03564.x
  12. Espley, R.V., R.P. Hellens, J. Putterill, D.E. Stevenson, S. Kutty-Amma, and A.C. Allan. 2007. Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10. Plant J. 49:414-427. https://doi.org/10.1111/j.1365-313X.2006.02964.x
  13. Fukusaki, E., K. Kawasaki, S. Kajiyama, C.I. An, K. Suzuki, Y. Tanaka, and A. Kobayashi. 2004. Flower color modulations of Torenia hybrida by downregulation of chalcone synthase genes with RNA interference. J. Biotechnol. 111:229-240. https://doi.org/10.1016/j.jbiotec.2004.02.019
  14. Gachon, C.M., M. Langlois-Meurinne, and P. Saindrenan. 2005. Plant secondary metabolism glycosyltransferases: The emerging functional analysis. Trends Plant Sci. 10:542-549. https://doi.org/10.1016/j.tplants.2005.09.007
  15. Grotewold, E. 2005. Plant metabolic diversity: A regulatory perspective. Trends Plant Sci. 10:57-62. https://doi.org/10.1016/j.tplants.2004.12.009
  16. Grotewold, E. 2006. The genetics and biochemistry of floral pigments. Annu. Rev. Plant Biol. 57:761-780. https://doi.org/10.1146/annurev.arplant.57.032905.105248
  17. Han, Y.J., Y.M. Kim, J.Y. Lee, S.J. Kim, K.C. Cho, T. Chandrasekhar, P.S. Song, Y.M. Woo, and J.I. Kim. 2009. Production of purple-colored creeping bentgrass using maize transcription factor genes Pl and Lc through Agrobacteriummediated transformation. Plant Cell Rep. 28:397-406. https://doi.org/10.1007/s00299-008-0648-0
  18. Hanumappa, M., G. Choi, S. Ryu, and G. Choi. 2007. Modulation of flower colour by rationally designed dominant-negative chalcone synthase. J. Exp. Bot. 58:2471-2478. https://doi.org/10.1093/jxb/erm104
  19. Hiratsu, K., K. Matsui, T. Koyama, and M. Ohme-Takagi. 2003. Dominant repression of target genes by chimeric repressors that include the EAR motif, a repression domain, in Arabidopsis. Plant J. 34:733-739. https://doi.org/10.1046/j.1365-313X.2003.01759.x
  20. Jeong, S.W., P. Kumar, S.C. Jeoung , J.Y. Song, H.Y. Lee, Y.K. Kim, W.J. Kim, Y.I. Park, S.D. Yoo, S.B. Choi, G. Choi, and Y.I. Park. 2010. Ethylene suppression of sugar-induced anthocyanin pigmentation in Arabidopsis. Plant Physiol. 154:1514-1531. https://doi.org/10.1104/pp.110.161869
  21. Katsumoto, Y., M. Fukuchi-Mizutani, Y. Fukui, F. Brugliera, T.A. Holton, M. Karan, N. Nakamura, K. Yonekura-Sakakibara, J. Togami, A. Pigeaire, G.Q. Tao, N.S. Nehra, C.Y. Lu, B.K. Dyson, S. Tsuda, T. Ashikari, T. Kusumi, J.G. Mason, and Y. Tanaka. 2007. Engineering of the rose flavonoid biosynthetic pathway successfully generated blue-hued flowers accumulating delphinidin. Plant Cell Physiol. 48:1589-1600. https://doi.org/10.1093/pcp/pcm131
  22. Koes, R., W. Verweij, and F. Quattrocchio. 2005. Flavonoids: A colorful model for the regulation and evolution of biochemical pathways. Trends Plant Sci. 10:236-242. https://doi.org/10.1016/j.tplants.2005.03.002
  23. Li, S.J., X.M. Deng, H.Z. Mao, and Y. Hong. 2005. Enhanced anthocyanin synthesis in foliage plant Caladium bicolor. Plant Cell Rep. 23:716-720. https://doi.org/10.1007/s00299-004-0871-2
  24. Lim, S.-H., S.H. Ha, J.K. Kim, S.H. Sohn, D.H. Kim, J.S. Kim, Y.M. Kim, and J.R. Lee. 2010. Expression of phenylpropanoid biosynthetic genes in transgenic tobacco plants with maize B-peru Gene. Kor. J. Intl. Agri. 22:72-77.
  25. Lin-Wang, K., K. Bolitho, K. Grafton, A. Kortstee, S. Karunairetnam, T.K. McGhie, R.V. Espley, R.P. Hellens, and A.C. Allan. 2010. An R2R3 MYB transcription factor associated with regulation of the anthocyanin biosynthetic pathway in Rosaceae. BMC Plant Biol. 10:50. https://doi.org/10.1186/1471-2229-10-50
  26. Mathews, H., S.K. Clendennen, C.G. Caldwell, X.L. Liu, K. Connors, N. Matheis, D.K. Schuster, D.J. Menasco, W. Wagoner, J. Lightner, and D.R. Wagner. 2003. Activation tagging in tomato identifies a transcriptional regulator of anthocyanin biosynthesis, modification, and transport. Plant Cell 15:1689-1703. https://doi.org/10.1105/tpc.012963
  27. Matsui, K., Y. Umemura, and M. Ohme-Takagi. 2008. AtMYBL2, a protein with a single MYB domain, acts as a negative regulator of anthocyanin biosynthesis in Arabidopsis. Plant J. 55:954-967. https://doi.org/10.1111/j.1365-313X.2008.03565.x
  28. Meyer, P., I. Heidmann, G. Forkmann, and H. Saedler. 1987. A new petunia flower colour generated by transformation of a mutant with a maize gene. Nature 330:677-678. https://doi.org/10.1038/330677a0
  29. Milkowski, C. and D. Strack. 2004. Serine carboxypeptidase-like acyltransferases. Phytochemistry 65:517-524. https://doi.org/10.1016/j.phytochem.2003.12.018
  30. Mitsuda N., Y. Takiguchi, M. Shikata, K. Sage-Ono, M. Ono, K. Sasaki, H. Yamaguchi, T. Narumi, Y. Tanaka, T. Terakawa, K. Gion, R. Suzuri, Y. Tanaka, T. Nakatsuka, S. Kimura, M. Nishihara, T. Sakai, R. Endo-Onodera, K. Saitoh, K. Isuzugawa, Y. Oshima, T. Koyama, M. Ikeda, M. Narukawa, K. Matsui, M. Nakata, N. Ohtsubo, and M. Ohme-Takagi. 2011. The new FioreDB database provides comprehensive information on plant transcription factors and phenotypes induced by CRES-T in ornamental and model plants. Plant Biotechnol. 28:123-130. https://doi.org/10.5511/plantbiotechnology.11.0106a
  31. Mitsuda, N., Y. Umemura, M. Ikeda, M. Shikata, T. Koyama, K. Matsui, T. Narumi, R. Aida, K. Sasaki, T. Hiyama, Y. Higuchi, M. Ono, K. Isuzugawa, K. Saitoh, R. Endo, K. Ikeda, T. Nakatsuka, M. Nishihara, S. Yamamura, T. Yamamura, T. Terakawa, N. Ohtsubo, and M. Ohme-Takagi. 2008. FioreDB: A database of phenotypic information induced by the chimeric repressor silencing technology (CRES-T) in Arabidopsis and floricultural plants. Plant Biotechnol. 25:37-44. https://doi.org/10.5511/plantbiotechnology.25.37
  32. Momonoi, K., K. Yoshida, S. Mano, H. Takahashi, C. Nakamori, K. Shoji, A. Nitta, and M. Nishimura. 2009. A vacuolar iron transporter in tulip, TgVit1, is responsible for blue coloration in petal cells through iron accumulation. Plant J. 59:437-447. https://doi.org/10.1111/j.1365-313X.2009.03879.x
  33. Morita, Y., M. Saitoh, A. Hoshino, E. Nitasaka, and S. Iida. 2006. Isolation of cDNAs for R2R3-MYB, bHLH and WDR transcriptional regulators and identification of c and ca mutations conferring white flowers in the Japanese morning glory. Plant Cell Physiol. 47:457-470. https://doi.org/10.1093/pcp/pcj012
  34. Nakamura, N., M. Fukuchi-Mizutani, K. Miyazaki, K. Suzuki, and Y. Tanaka. 2006. RNAi suppression of the anthocyanidin synthase gene in Torenia hybrida yields white flowers with higher frequency and better stability than antisense and sense suppression. Plant Biotechnol. 23:13-18. https://doi.org/10.5511/plantbiotechnology.23.13
  35. Nakamura, N., M. Fukuchi-Mizutani, Y. Fukui, K. Ishiguro, K. Suzuki, H. Suzuki, K. Okazaki, D. Shibata, and Y. Tanaka. 2010. Generation of pink flower varieties from blue Torenia hybrida by redirecting the flavonoid biosynthetic pathway from delphinidin to pelargonidin. Plant Biotechnol. 27:375-383. https://doi.org/10.5511/plantbiotechnology.10.0610a
  36. Nakatsuka, T. and M. Nishihara. 2010. UDP-glucose:3-deoxyanthocyanidin 5-O-glucosyltransferase from Sinningia cardinalis. Planta 232:383-392. https://doi.org/10.1007/s00425-010-1175-0
  37. Nakatsuka, T., C. Pitaksutheepong, S. Yamamura, and M. Nishihara. 2007a. Induction of differential flower pigmentation patterns by RNAi using promoters with distinct tissue-specific activity. Plant Biotech. Rep. 1:251-257. https://doi.org/10.1007/s11816-007-0039-y
  38. Nakatsuka, T., K. Mishiba, A. Kubota, Y. Abe, S. Yamamura, N. Nakamura, Y. Tanaka, and M. Nishihara. 2010. Genetic engineering of novel flower colour by suppression of anthocyanin modification genes in gentian. J. Plant Physiol. 167:231-237. https://doi.org/10.1016/j.jplph.2009.08.007
  39. Nakatsuka, T., K. Mishiba, Y. Abe, A. Kubota, Y. Kakizaki, S. Yamamura, and M. Nishihara. 2008a. Flower color modification of gentian plants by RNAi-mediated gene silencing. Plant Biotechnol. 25:61-68. https://doi.org/10.5511/plantbiotechnology.25.61
  40. Nakatsuka, T., K. Sato, H. Takahashi, S. Yamamura, M. Nishihara. 2008b. Cloning and characterization of the UDP-glucose: anthocyanin 5-O-glucosyltransferase gene from blue-flowered gentian. J. Exp. Bot. 59:1241-1252. https://doi.org/10.1093/jxb/ern031
  41. Nakatsuka, T., K.S. Haruta, C. Pitaksutheepong, Y. Abe, Y. Kakizaki, K. Yamamoto, N. Shimada, S. Yamamura, and M. Nishihara. 2008c. Identification and characterization of R2R3- MYB and bHLH transcription factors regulating anthocyanin biosynthesis in gentian flowers. Plant Cell Physiol. 49:1818-1829. https://doi.org/10.1093/pcp/pcn163
  42. Nakatsuka, T., M. Nishihara, K. Mishiba, and S. Yamamura. 2006. Heterologous expression of two gentian cytochrome P450 genes can modulate the intensity of flower pigmentation in transgenic tobacco plants. Mol. Breeding 17:91-99. https://doi.org/10.1007/s11032-005-2520-z
  43. Nakatsuka, T., Y. Abe, Y. Kakizaki, S. Yamamura, and M. Nishihara. 2007b. Production of red-flowered plants by genetic engineering of multiple flavonoid biosynthetic genes. Plant Cell Rep. 26:1951-1959. https://doi.org/10.1007/s00299-007-0401-0
  44. Nakayama, T., K. Yonekura-Sakakibara, T. Sato, S. Kikuchi, Y. Fukui, M. Fukuchi-Mizutani, T. Ueda, M. Nakao, Y. Tanaka, T. Kusumi, and T. Nishino. 2000. Aureusidin synthase: A polyphenol oxidase homolog responsible for flower coloration. Science 290:1163-1166. https://doi.org/10.1126/science.290.5494.1163
  45. Nielsen, K., S.C. Deroles, K.R. Markham, J.M. Bradley, E. Podivinsky, and D. Manson. 2002. Antisense flavonol synthase alters copigmentation and flower color in lisianthus. Mol. Breed. 9:217-229. https://doi.org/10.1023/A:1020320809654
  46. Nishihara, M. and T. Nakatsuka. 2010. Genetic engineering of novel flower colors in floricultural plants: Recent advances via transgenic approaches. Methods Mol. Biol. 589:325-347.
  47. Nishihara, M., T. Nakatsuka, and S. Yamamura. 2005. Flavonoid components and flower color change in transgenic tobacco plants by suppression of chalcone isomerase gene. FEBS Lett. 579:6074-6078. https://doi.org/10.1016/j.febslet.2005.09.073
  48. Nishihara, M., T. Nakatsuka, K. Hosokawa, T. Yokoi, Y. Abe, K. Mishiba, and S. Yamamura. 2006. Dominant inheritance of white-flowered and herbicide-resistant traits in trangenic gentian plants. Plant Biotechnol. 23:25-31. https://doi.org/10.5511/plantbiotechnology.23.25
  49. Ono, E., M. Fukuchi-Mizutani, N. Nakamura, Y. Fukui, K. Yonekura-Sakakibara, M. Yamaguchi, T. Nakayama, T. Tanaka, T. Kusumi, and Y. Tanaka. 2006. Yellow flowers generated by expression of the aurone biosynthetic pathway. Proc. Nat. Acad. Sci. USA 103:11075-11080. https://doi.org/10.1073/pnas.0604246103
  50. Pattanaik, S., Q. Kong, D. Zaitlin, J.R. Werkman, C.H. Xie, B. Patra, and L. Yuan. 2010. Isolation and functional characterization of a floral tissue-specific R2R3 MYB regulator from tobacco. Planta 231:1061-1076. https://doi.org/10.1007/s00425-010-1108-y
  51. Quattrocchio, F., A. Baudry, L. Lepiniec, and E. Grotewold. 2006. The regulation of flavonoid biosynthesis. p. 97-122. In: E. Grotewold (ed.). The science of flavonoids. Springer, NY.
  52. Quattrocchio, F., J. Wing, K. van der Woude, E. Souer, N. de Vetten, J. Mol, and R. Koes. 1999. Molecular analysis of the anthocyanin2 gene of petunia and its role in the evolution of flower color. Plant Cell 11:1433-1444.
  53. Schwinn, K., J. Venail, Y. Shang, S. Mackay, V. Alm, E. Butelli, R. Oyama, P. Bailey, K. Davies, and C. Martin. 2006. A small family of MYB-regulatory genes controls floral pigmentation intensity and patterning in the genus Antirrhinum. Plant Cell 18:831-851. https://doi.org/10.1105/tpc.105.039255
  54. Seitz, C., M. Vitten, P. Steinbach, S. Hartl, J. Hirsche, W. Rathje, D. Treutter, and G. Forkmann. 2007. Redirection of anthocyanin synthesis in Osteospermum hybrida by a two-enzyme manipulation strategy. Phytochemistry 68:824-833. https://doi.org/10.1016/j.phytochem.2006.12.012
  55. Shimada, N., T. Nakatsuka, M. Nishihara, S. Yamamura, S. Ayabe, and T. Aoki. 2006. Isolation and characterization of a cDNA encoding polyketide reductase in Lotus japonicus. Plant Biotechnol. 23:509-513. https://doi.org/10.5511/plantbiotechnology.23.509
  56. Tanaka, Y., F. Brugliera, and S. Chandler. 2009. Recent progress of flower colour modification by biotechnology. Int. J. Mol. Sci. 10:5350-5369. https://doi.org/10.3390/ijms10125350
  57. Tanaka, Y., Y. Katsumoto, F. Brugliera, and J. Mason. 2005. Genetic engineering in floriculture. Plant Cell Tiss. Org. Cult. 80:1-24. https://doi.org/10.1007/s11240-004-0739-8
  58. Ueyama, Y., K. Suzuki, M. Fukuchi-Mizutani, Y. Fukui, K. Miyazaki, H. Ohkawa, T. Kusumi, and Y. Tanaka. 2002. Molecular and biochemical characterization of torenia flavonoid 3′-hydroxylase and flavone synthase II and modification of flower color by modulating the expression of these genes. Plant Sci. 163:253-263. https://doi.org/10.1016/S0168-9452(02)00098-5
  59. Ueyama, Y., Y. Katsumoto, Y. Fukui, M. Fukuchi-Mizutani, H. Ohkawa, T. Kusumi, T. Iwashita, and Y. Tanaka. 2006. Molecular characterization of the flavonoid biosynthetic pathway and flower color modification of Nierembergia sp. Plant Biotechnol. 23:19-24. https://doi.org/10.5511/plantbiotechnology.23.19
  60. Umehara, M., A. Hanada, S. Yoshida, K. Akiyama, T. Arite, N. Takeda-Kamiya, H. Magome, Y. Kamiya, K. Shirasu, K. Yoneyama, J. Kyozuka, and S. Yamaguchi. 2008. Inhibition of shoot branching by new terpenoid plant hormones. Nature 455:195-200. https://doi.org/10.1038/nature07272
  61. van der Krol, A.R., P.E. Lenting, J. Veenstra, I.M. van der Meer, R.E. Koes, A.G.M. Gerats, J.N.M. Mol, and A.R. Stuitje. 1988. An anti-sense chalcone synthase gene in transgenic plants inhibits flower pigmentation. Nature 333:866-869. https://doi.org/10.1038/333866a0
  62. Verweij, W., C. Spelt, G.P. Di Sansebastiano, J. Vermeer, L. Reale, F. Ferranti, R. Koes, and F. Quattrocchio. 2008. An $H^{+}$ P-ATPase on the tonoplast determines vacuolar pH and flower colour. Nat. Cell Biol. 10:1456-1462. https://doi.org/10.1038/ncb1805
  63. Winefield, C., H. David, E. Swinny, H. Zhang, H. Arathoon, T. Fischer, H. Halbwirth, K. Stich, C. Gosch, G. Forkmann, and K. Davies. 2005. Investigation of the biosynthesis of 3-deoxyanthocyanins in Sinningia cardinalis. Physiologia Plantarum 124:419-430. https://doi.org/10.1111/j.1399-3054.2005.00531.x
  64. Yamagishi, M., Y. Shimoyamada, T. Nakatsuka, and K. Masuda. 2010. Two R2R3-MYB genes, homologs of petunia AN2, regulate anthocyanin biosyntheses in flower tepals, tepal spots and leaves of Asiatic hybrid lily. Plant Cell Physiol. 51:463-474. https://doi.org/10.1093/pcp/pcq011
  65. Yamasaki, M., M. Oda, N. Koizumi, K. Mitsukuri, M. Johkan, T. Nakatsuka, M. Nishihara, and K. Mishiba. 2011. De novo DNA methylation of the 35S enhancer revealed by highresolution methylation analysis of an entire T-DNA segment in transgenic gentian. Plant Biotechnol. 28:223-230. https://doi.org/10.5511/plantbiotechnology.10.1222a
  66. Yoshida, K., M. Mori, and T. Kondo. 2009. Blue flower color development by anthocyanins: From chemical structure to cell physiology. Nat. Prod. Rep. 26:884-915. https://doi.org/10.1039/b800165k