Comprehensive isolation and expression analysis of the flavonoid biosynthesis-related genes in Tricyrtis spp.
- 1 Downloads
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.
Additional key wordsanthocyanins flavonoids flower colours flower development transcription factors
high performance liquid chromatography
rapid amplification of cDNA ends
reverse transcription-polymerase chain reaction
transparent testa glabra 1
Unable to display preview. Download preview PDF.
- Forkmann, G., Ruhnau, B. Distinct substrate specificity of dihydroflavonol 4-reductase from flowers of Petunia hybrid. — Z. Naturforsch C 42: 1146–1148, 1987.Google Scholar
- 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.CrossRefPubMedGoogle Scholar
- 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.CrossRefPubMedPubMedCentralGoogle Scholar
- 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.CrossRefGoogle Scholar
- 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.CrossRefPubMedGoogle Scholar
- 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.CrossRefPubMedGoogle Scholar
- 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.Google Scholar
- 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.CrossRefGoogle Scholar
- 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.CrossRefPubMedGoogle Scholar
- 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.CrossRefPubMedPubMedCentralGoogle Scholar
- 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.CrossRefPubMedPubMedCentralGoogle Scholar
- 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.CrossRefPubMedPubMedCentralGoogle Scholar