Biologia plantarum 62:684-692, 2018 | DOI: 10.1007/s10535-018-0802-7

Comprehensive isolation and expression analysis of the flavonoid biosynthesis-related genes in Tricyrtis spp.

M. Otani1, Y. Kanemaki1, F. Oba2, M. Shibuya2, Y. Funayama1, M. Nakano1,*
1 Graduate School of Science and Technology, Niigata University, Niigata, Japan
2 Faculty of Agriculture, Niigata University, Niigata, Japan

Tricyrtis spp., which belong to the family Liliaceae, produce unique flowers, whose tepals have many reddish-purple spots. Although elucidation of a molecular mechanism of tepal spot formation and molecular breeding for flower colour alteration are desired for Tricyrtis spp., only one flavonoid biosynthesis-related gene, TrCHS encoding chalcone synthase (CHS), has been isolated so far. In the present study, comprehensive isolation and expression analysis of the other flavonoid biosynthesis-related genes were carried out in Tricyrtis sp. Six genes (TrCHI, TrF3H, TrF3'H, TrFLS, TrDFR, and TrANS) encoding biosynthetic enzymes chalcone isomerase (CHI), flavanone-3-hydroxylase (F3H); flavonoid 3'-hydroxylase (F3'H), flavonol synthase (FLS), dihydroflavonol 4-reductase (DFR), and anthocyanin synthase (ANS) as well as three genes (TrMYB1, TrbHLH2 and TrWDR) encoding transcription factors myeloblastosis 1 (MYB1), basic helix-loop-helix (bHLH), and WD40 repeats (WDRs) were newly isolated. Phylogenetic analysis showed that each isolated gene was classified into the monocotyledonous clade. Deduced amino acid sequences of DFRs showed that TrDFR has no substrate specificity. "Early" genes in the flavonoid biosynthetic pathway (TrCHS, TrCHI, and TrF3H) were constantly expressed in tepals during flower development, whereas expression of "late" genes (TrF3'H, TrFLS, TrDFR, and TrANS) varied with the flower developmental stage. Expression patterns of the late genes were mostly correlated with those of transcriptional factor genes, indicating that the late genes may be under the control of a transcription factor complex consisted of TrMYB1, TrbHLH2, and TrWDR. Accumulation of anthocyanins in tepals occurred slightly after transcriptional upregulation of the late genes. Results obtained in the present study may be valuable for further studies on flower colour and flower colour pattern in Tricyrtis spp.

Keywords: anthocyanins; flavonoids; flower colours; flower development; transcription factors
Subjects: flavonoid biosynthesis; anthocyanins; flower development; flower colours; transcription factors; amino acid sequences; phylogenetic tree
Species: Tricyrtis

Received: November 16, 2017; Revised: February 20, 2018; Accepted: March 14, 2018; Published: August 1, 2018Show citation

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Otani, M., Kanemaki, Y., Oba, F., Shibuya, M., Funayama, Y., & Nakano, M. (2018). Comprehensive isolation and expression analysis of the flavonoid biosynthesis-related genes in Tricyrtis spp. Biologia plantarum, 62(4), 684-692. doi: 10.1007/s10535-018-0802-7.
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    References

    1. Adachi, Y., Mori, S., Nakano, M.: Agrobacterium-mediated production of transgenic plants in Tricyrtis hirta (Liliaceae). - Acta Hort. 673: 415-419, 2005. Go to original source...
    2. Chen, K., Liu, H., Lou, Q., Liu, Y.: Ectopic expression of the grape hyacinth (Muscari armeniacum) R2R3-MYB transcription factor gene, MaAN2, induces anthocyanin accumulation in tobacco. - Front. Plant Sci. 8: 965, 2017. Go to original source...
    3. Chen, S.M., Li, C.H., Zhu, X.R., Deng, Y.M., Sun, W., Wang, L.S., Chen, F.D., Zhang, Z.: The identification of flavonoids and the expression of genes of anthocyanin biosynthesis in the chrysanthemum flowers. - Biol. Plant 56: 458-464, 2012. Go to original source...
    4. Chiou, C.Y., Yeh, K.W.: Differential expression of MYB gene (OgMYB1) determines color patterning in floral tissue of Oncidium Gower Ramsey. - Plant mol. Biol. 66: 379-388, 2008. Go to original source...
    5. Chiu, L.W., Zhou, X., Burke, S., Wu, X., Prior, R.L., Li, L.: The purple cauliflower arises from activation of a MYB transcription factor. - Plant Physiol. 154: 1470-1480, 2010. Go to original source...
    6. Cone, K. C., Cocciolone, S.M., Burr, F.A., Burr, B.: Maize anthocyanin regulatory gene pl is a duplicate of c1 that functions in the plant. - Plant Cell 5: 1795-1805, 1993.
    7. Felsenstein, J.: Confidence limits on phylogenies: an approach using the bootstrap. - Evolution 39: 783-791, 1985. Go to original source...
    8. Forkmann, G., Ruhnau, B. Distinct substrate specificity of dihydroflavonol 4-reductase from flowers of Petunia hybrid. - Z. Naturforsch C 42: 1146-1148, 1987. Go to original source...
    9. Fornalé, S., Shi, X., Chai, C., Encina, A., Irar, S., Capellades, M., Fuguet, E., Torres, J.L., Rovira, P., Puigdomenech, P., Rigau, J., Grotewold, E., Gray, J., Caparrós-Ruiz, D.: ZmMYB31 directly represses maize lignin genes and redirects the phenylpropanoid metabolic flux. - Plant J. 64: 633-644, 2010. Go to original source...
    10. Gerats, A.G., De Vlaming, P., Doodeman, M., Al, B., Schram, A.W.: Genetic control of the conversion of dihydroflavonols into flavonols and anthocyanins in flowers of Petunia hybrida. - Planta 155: 364-368, 1982. Go to original source...
    11. Guo, N., Cheng, F., Wu, J., Liu, B., Zheng, S., Liang, J., Wang, X.: Anthocyanin biosynthetic genes in Brassica rapa. - BMC Genomics 15: 426, 2014. Go to original source...
    12. 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. 62: 2465-2483, 2011. Go to original source...
    13. Holton, T.A., Cornish, E.C.: Genetics andbBiochemistry of anthocyanin biosynthesis. - Plant Cell 7: 1071-1083, 1995. Go to original source...
    14. Jaakola, L., Määttä, K., Pirttilä, A.M., Torronen, R., Karenlampi, S., Hohtola, A.: Expression of genes involved in anthocyanin biosynthesis in relation to anthocyanin, proanthocyanidin, and flavonol levels during bilberry fruit development. - Plant Physiol. 130: 729-739, 2002. Go to original source...
    15. Jiang, C., Gu, X., Peterson, T.: Identification of conserved gene structures and carboxy-terminal motifs in the Myb gene family of Arabidopsis and Oryza sativa L. ssp. indica. - Genome Biol. 5: R46, 2004. Go to original source...
    16. Johnson, E.T., Ryu, S., Yi, H., Shin, B., Cheong, H., Choi, G.: Alteration of a single amino acid changes the substrate specificity of dihydroflavonol 4-reductase. - Plant J. 25: 325-333, 2001. Go to original source...
    17. Johnson, E.T., Yi, H., Shin, B., Oh, B.J., Cheong, H., Choi, G.: Cymbidium hybrida dihydroflavonol 4-reductase does not efficiently reduce dihydrokaempferol to produce orange pelargonidin-type anthocyanins. - Plant J. 19: 81-85, 1999. Go to original source...
    18. Kamiishi, Y., Otani, M., Takagi, H., Han, D.S., Mori, S., Tatsuzawa, F., Okuhara, H., Kobayashi, H., Nakano, M.: Flower color alteration in the liliaceous ornamental Tricyrtis sp. by RNA interference-mediated suppression of the chalcone synthase gene. - Mol. Breed. 30: 671-680, 2012. Go to original source...
    19. Katsumoto, Y., Fukuchi-Mizutani, M., Fukui, Y., Brugliera, F., Holton, T.A., Karan, M., 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. - Plant Cell Physiol. 48: 1589-1600, 2007. Go to original source...
    20. Koes, R., Verweij, W., Quattrocchio, F.: Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. - Trends Plant Sci. 10: 236-242, 2005. Go to original source...
    21. Kranz, H.D., Denekamp, M., Greco, R., Jin, H., Leyva, A., Meissner, R.C., Petroni, K., Urzainqui, A., Bevan, M., Martin, C., Smeekens, S., Tonelli, C., Paz-Ares, J., Weisshaar, B.: Towards functional characterisation of the members of the R2R3-MYB gene family from Arabidopsis thaliana. - Plant J. 16: 263-276, 1998. Go to original source...
    22. Kumar, S., Stecher, G., Tamura, K.: MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. - Mol. Biol. Evol. 33: 1870-1874, 2016.
    23. Kunihiro, S., Tanabe, D., Niwa, Y., Kitamura, K., Abe, J., Yamada, T.: Isolation and molecular characterization of a Lotus japonicus R2R3-MYB subgroup 7 transcription factor gene. - Plant Biotechnol. 34: 45-49, 2017. Go to original source...
    24. Lai, Y.S., Shimoyamada, Y., Nakayama, M., Yamagishi, M.: Pigment accumulation and transcription of LhMYB12 and anthocyanin biosynthesis genes during flower development in the Asiatic hybrid lily (Lilium spp.). - Plant Sci. 193-194: 136-147, 2012. Go to original source...
    25. Li, Q., Wang, J., Sun, H.Y., Shang, X.: Flower color patterning in pansy (Viola × wittrockiana Gams.) is caused by the differential expression of three genes from the anthocyanin pathway in acyanic and cyanic flower areas. - Plant Physiol. Biochem. 84: 134-141, 2014. Go to original source...
    26. Liu, C., Long, J., Zhu, K., Liu, L., Yang, W., Zhang, H., Li, L., Xu, Q., Deng, X.: Characterization of a citrus R2R3-MYB transcription factor that regulates the flavonol and hydroxycinnamic acid biosynthesis. - Sci. Rep. 6: 25352, 2016. Go to original source...
    27. Montefiori, M., Brendolise, C., Dare, A.P., Lin-Wang, K., Davies, K.M., Hellens, R.P., Allan, A.C.: In the Solanaceae, a hierarchy of bHLHs confer distinct target specificity to the anthocyanin regulatory complex. - J. exp. Bot. 66: 1427-1436, 2015. Go to original source...
    28. Mori, S., Asano, S., Kobayashi, H., Nakano, M.: Analyses of anthocyanidins and anthocyanins in flowers of Muscari spp. - Bull. Fac. Agr. Niigata Univ. 55: 13-18, 2002.
    29. Mori, S., Oka, E., Umehara, H., Kobayashi, H., Hoshi, Y., Kondo, M., Ogata, K., Nakano, M.: Stability of β-glucuronidase gene expression in transgenic Tricyrtis hirta plants after two years of cultivation. - Biol. Plant 52: 513-516, 2008. Go to original source...
    30. Nakano, M., Nomizu, T., Mizunashi, K., Suzuki, M., Mori, S., Kuwayama, S., Hayashi, M., Umehara, H., Oka, E., Kobayashi, H., Asano, M., Sugawara, S., Takagi, H., Saito, H., Nakata, M., Godo, T., Hara, Y., Amano, J.: Somaclonal variation in Tricyrtis hirta plants regenerated from 1-year-old embryogenic callus cultures. - Sci. Hort. 110: 366-371, 2006. Go to original source...
    31. Nakatsuka, A., Mizuta, D., Kii, Y., Miyajima, I., Kobayashi, N.: Isolation and expression analysis of flavonoid biosynthesis genes in evergreen azalea. - Sci. Hort. 118: 314-320, 2008a. Go to original source...
    32. Nakatsuka, T., Haruta, K.S., Pitaksutheepong, C., Abe, Y., Kakizaki, Y., Yamamoto, K., Shimada, N., Yamamura, S., Nishihara, M.: Identification and characterization of R2R3-MYB and bHLH transcription factors regulating anthocyanin biosynthesis in gentian flowers. - Plant Cell Physiol. 49: 1818-1829, 2008b.
    33. Nakatsuka, T., Nishihara, M., Mishiba, K., Yamamura, S.: Temporal expression of flavonoid biosynthesis-related genes regulates flower pigmentation in gentian plants. - Plant Sci. 168: 1309-1318, 2005. Go to original source...
    34. Noda, N., Aida, R., Kishimoto, S., Ishiguro, K., Fukuchi-Mizutani, M., Tanaka, Y., Ohmiya, A.: Genetic - Phytochemistry 50: 1181-1184, 1999.
    35. Paz-Ares, J., Ghosal, D., Wienand, U., Peterson, P.A., Saedler, H.: The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators. - EMBO J. 6: 3553-3558, 1987. Go to original source...
    36. Perfus-Barbeoch, L., Jones, A.M., Assmann, S.M.: Plant heterotrimeric G protein function: insights from Arabidopsis and rice mutants. - Curr. Opin. Plant Biol. 7: 719-731, 2004. Go to original source...
    37. Petroni, K., Tonelli, C.: Recent advances on the regulation of anthocyanin synthesis in reproductive organs. - Plant Sci. 181: 219-229, 2011. Go to original source...
    38. Quattrocchio, F., Wing, J., Van der Woude, K., Souer, E., De Vetten, N., Mol, J., Koes, R.: Molecular analysis of the anthocyanin2 gene of petunia and its role in the evolution of flower color. - Plant Cell 11: 1433-1444, 1999. Go to original source...
    39. Rabino, I., Mancinelli, A.L.: Light, temperature, and anthocyanin production. - Plant Physiol. 81: 922-924, 1986. Go to original source...
    40. Saitou, N., Nei, M.: The neighbor-joining method: A new method for reconstructing phylogenetic trees. - Mol. Biol. Evol. 4: 406-425, 1987.
    41. Schwinn, K., Venail, J., Shang, Y., Mackay, S., Alm, V., Butelli, E., Oyama, R., Bailey, P., Davies, K., Martin, C.: A Small family of MYB-regulatory genes controls floral pigmentation intensity and patterning in the genus Antirrhinum. - Plant Cell 18: 831-851, 2006. Go to original source...
    42. Schwinn, K.E., Ngo, H., Kenel, F., Brummell, D.A., Albert, N.W., McCallum, J.A., Pither-Joyce, M., Crowhurst, R.N., Eady, C., Davies, K.M.: The onion (Allium cepa L.) R2R3-MYB gene MYB1 regulates anthocyanin biosynthesis. - Front. Plant Sci. 7: 1865, 2016.
    43. Smith, T.F., Gaitatzes, C., Saxena, K., Neer, E.J.: The WD repeat: a common architecture for diverse functions. - Trends Biochem. Sci. 24: 181-185, 1999. Go to original source...
    44. Spelt, C., Quattrocchio, F., Mol, J.N., Koes, R.: Anthocyanin1 of petunia encodes a basic helix-loop-helix protein that directly activates transcription of structural anthocyanin genes. - Plant Cell 12: 1619-1632, 2000. Go to original source...
    45. Stracke, R., Ishihara, H., Huep, G., Barsch, A., Mehrtens, F., Niehaus, K., Weisshaar, B.: Differential regulation of closely related R2R3-MYB transcription factors controls flavonol accumulation in different parts of the Arabidopsis thaliana seedling. - Plant J. 50: 660-677, 2007. Go to original source...
    46. Suzuki, K., Suzuki, T., Nakatsuka, T., Dohra, H., Yamagishi, M., Matsuyama, K., Matsuura, H.: RNA-seq-based evaluation of bicolor tepal pigmentation in Asiatic hybrid lilies (Lilium spp.). - BMC Genomics 17: 611, 2016. Go to original source...
    47. Tatsuzawa, F., Saito, N., Miyoshi, K., Shinoda, K., Shigihara, A., Honda, T.: Diacylated 8-C-glucosylcyanidin 3-glucoside from the flowers of Tricyrtis formosana. - Chem. Pharm. Bull. 52: 631-633, 2004. Go to original source...
    48. Van Nocker, S., Ludwig, P.: The WD-repeat protein superfamily in Arabidopsis: conservation and divergence in structure and function. - BMC Genomics 4: 50, 2003. Go to original source...
    49. Walker, A.R., Davison, P.A., Bolognesi-Winfield, A.C., James, C.M., Srinivasan, N., Blundell, T.L., Esch, J.J., Marks, M.D., Gray, J.C.: The TRANSPARENT TESTA GLABRA1 locus, which regulates trichome differentiation and anthocyanin biosynthesis in Arabidopsis, encodes a WD40 repeat protein. - Plant Cell 11: 1337-1350, 1999. Go to original source...
    50. Wang, L.M., Zhang, J., Dong, X.Y., Fu, Z.Z., Jiang, H., Zhang, H.C.: Identification and functional analysis of anthocyanin biosynthesis genes in Phalaenopsis hybrids. - Biol. Plant. 62: 45-54, 2018. Go to original source...
    51. Winkel-Shirley, B.: Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. - Plant Physiol. 126: 485-493, 2001. Go to original source...
    52. Yamagishi, M., Shimoyamada, Y., Nakatsuka, T., Masuda, K.: 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, 2010. Go to original source...
    53. Zhao, D., Tang, W., Hao, Z., Tao, J.: Identification of flavonoids and expression of flavonoid biosynthetic genes in two coloured tree peony flowers. - Biochem. Biophys. Res. Commun. 459: 450-456, 2015a. Go to original source...
    54. Zhao, X., Yuan, Z., Feng, L., Fang, Y.: Cloning and expression of anthocyanin biosynthetic genes in red and white pomegranate. - J. Plant Res. 128: 687-696, 2015b. Go to original source...

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