Abstract
Main conclusion
Two glycosyltransferase genes belonging to UGT88 family were identified to have 6ʹ-deoxychalcone 4ʹ-glucosyltransferase activity in dahlia.
Abstract
6ʹ-Deoxychalcones (isoliquiritigenin and butein) are important pigments for yellow and orange to red flower color. 6ʹ-Deoxychalcones are glucosylated at the 4ʹ-position in vivo, but the genes encoding 6ʹ-deoxychalcone 4ʹ-glucosyltransferase have not yet been identified. In our previous study, it was indicated that snapdragon (Antirrhinum majus) chalcone 4ʹ-O-glucosyltransferase (Am4ʹCGT) has isoliquiritigenin 4ʹ-glucosylation activity. Therefore, to identify genes encoding 6ʹ-deoxychalcone 4ʹ-glucosyltransferase in dahlia (Dahlia variabilis), genes expressed in ray florets that shared high homology with Am4ʹCGT were explored. As a result, c34671_g1_i1 and c35662_g1_i1 were selected as candidate genes for 6ʹ-deoxychalcone 4ʹ-glucosyltransferases in dahlia. We conducted transient co-overexpression of three genes (c34671_g1_i1 or c35662_g1_i1, dahlia aldo-keto reductase1 (DvAKR1) or soybean (Glycine max) chalcone reductase5 (GmCHR5), and chili pepper (Capsicum annuum) MYB transcription factor (CaMYBA)) in Nicotiana benthamiana by agroinfiltration. Transient overexpression of c34671_g1_i1, DvAKR1, and CaMYBA resulted in increase in the accumulation of isoliquiritigenin 4ʹ-glucosides, isoliquiritigenin 4ʹ-O-glucoside, and isoliquiritigenin 4ʹ-O-[6-O-(malonyl)-glucoside]. However, transient overexpression of c35662_g1_i1, DvAKR1, and CaMYBA did not increase accumulation of isoliquiritigenin 4ʹ-glucosides. Using GmCHR5 instead of DvAKR1 showed similar results suggesting that c34671_g1_i1 has isoliquiritigenin 4ʹ-glucosyltransferase activity. In addition, we conducted co-overexpression of four genes (c34671_g1_i1, c35662_g1_i1 or Am4ʹCGT, DvAKR1 or GmCHR5, CaMYBA, and chalcone 3-hydroxylase from dahlia). Accumulation of butein 4ʹ-O-glucoside and butein 4ʹ-O-[6-O-(malonyl)-glucoside] was detected for c35662_g1_i1, suggesting that c35662_g1_i1 has butein 4ʹ-glucosyltransferase activity. Recombinant enzyme analysis also supported butein 4ʹ-glucosyltransferases activity of c35662_g1_i1. Therefore, our results suggested that both c34671_g1_i1 and c35662_g1_i1 are 6ʹ-deoxychalcone 4ʹ-glucosyltransferases but with different substrate preference.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
Accession numbers: c34671_g1_i1/Dv4ʹCGT (LC775776), c35662_g1_i1/Dv4ʹBGT (LC775777), c37087_g1_i1/flavonoid 3-O-glucosyltransferase-like (LC775778), c37958_g1_i1/glucosyl glycoside transferase-like (LC775779), c38610_g1_i2/flavonoid C-glucosyltransferase-like (LC775780), c38674_g1_i1/tetrahydroxychalcone 2'-glucosyltransferase-like (LC775781) and c38795_g2_i3/UDP-glycosyltransferase (LC775782).
Abbreviations
- 4′CGT:
-
Chalcone 4′-O-glucosyltransferase
- AKR:
-
Aldo-keto reductase
- CH3H:
-
Chalcone 3-hydroxylase
- CHS:
-
Chalcone synthase
- GGT:
-
Glucoside glucosyltransferase
- UGT:
-
UDP-dependent glycosyltransferase
References
Bowles D, Isayenkova J, Lim EK, Poppenberger B (2005) Glycosyltransferases: managers of small molecules. Curr Opin Plant Biol 8:254–263. https://doi.org/10.1016/j.pbi.2005.03.007
Dastmalchi M, Bernards MA, Dhaubhadel S (2016) Twin anchors of the soybean isoflavonoid metabolon: evidence for tethering of the complex to the endoplasmic reticulum by IFS and C4H. Plant J 85:689–706. https://doi.org/10.1111/tpj.13137
Deguchi A, Ohno S, Hosokawa M, Tatsuzawa F, Doi M (2013) Endogenous post-transcriptional gene silencing of flavone synthase resulting in high accumulation of anthocyanins in black dahlia cultivars. Planta 237:1325–1335. https://doi.org/10.1007/s00425-013-1848-6
Deguchi A, Ohno S, Tatsuzawa F, Doi M, Hosokawa M (2016a) Identification of the yellow pigment in Saintpaulia flowers. Hort Res (japan) 15:123–128. https://doi.org/10.2503/hrj.15.123
Deguchi A, Tatsuzawa F, Hosokawa M, Doi M, Ohno S (2016b) Quantitative evaluation of the contribution of four major anthocyanins to black flower coloring of dahlia ray florets. Hort J 85:340–350. https://doi.org/10.2503/hortj.MI-121
Gatt M, Hong D, Keith H, Brian M (1998) Polyploidy and evolution in wild and cultivated dahlia species. Ann Bot 81:647–656. https://doi.org/10.1006/anbo.1998.0614
Grabherr G, Haas J, Yassour M, Levin J, Thompson D, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng Q, Chen Z, Mauceli E, Hacohen N, Gnirke A, Rhind N, Palma F, Birren B, Nusbaum C, Lindblad-Toh K, Friedman N, Regev A (2011) Full-length transcriptome assembly from RNA-Seq data without a reference genome. Nat Biotechnol 29:644–652. https://doi.org/10.1038/nbt.1883
Halbwirth H, Wimmer G, Wurst F, Forkmann G, Stich K (1997) Enzymatic glucosylation of 4-deoxyaurones and 6′-deoxychalcones with enzyme extracts of Coreopsis grandiflora. Nutt i Plant Sci 122:125–131. https://doi.org/10.1016/S0168-9452(96)04553-0
Harborne JB (1966) Comparative biochemistry of flavonoids-I.: Distribution of chalcone and aurone pigments in plants. Phytochemistry 5:110–115. https://doi.org/10.1016/S0031-9422(00)85088-8
Harborne JB, Greenham J, Eagles J (1990) Malonylated chalcone glycoside in dahlia. Phytochemistry 29:2899–2900. https://doi.org/10.1016/0031-9422(90)87101-Y
Hofer B (2016) Recent developments in the enzymatic O-glycosylation of flavonoids. Appl Microbiol Biotechnol 100:4269–4281. https://doi.org/10.1007/s00253-016-7465-0
Hoshino A, Mizuno T, Shimizu K, Mori S, Fukada-Tanaka S, Furukawa K, Ishiguro K, Tanaka Y, Iida S (2019) Generation of yellow flowers of the Japanese morning glory by engineering its flavonoid biosynthetic pathway toward aurones. Plant Cell Physiol 60:1871–1879. https://doi.org/10.1093/pcp/pcz101
Itoh Y, Higeta D, Suzuki A, Yoshida H, Ozeki Y (2002) Excision of transposable elements from the chalcone isomerase and dihydroflavonol 4-reductase genes may contribute to the variegation of the yellow-flowered carnation (Dianthus caryophyllus). Plant Cell Physiol 43:578–585. https://doi.org/10.1093/pcp/pcf065
Jones P, Vogt T (2001) Glycosyltransferases in secondary plant metabolism: tranquilizers and stimulant controllers. Planta 213:164–174. https://doi.org/10.1007/s004250000492
Kaintz C, Molitor C, Thill J, Kampatsikas I, Michael C, Halbwirth H, Rompel A (2014) Cloning and functional expression in E. coli of a polyphenol oxidase transcript from Coreopsis grandiflora involved in aurone formation. FEBS Lett 588:3417–3426. https://doi.org/10.1016/j.febslet.2014.07.034
Li Y, Baldauf S, Lim EK, Bowles DJ (2001) Phylogenetic analysis of the UDP-glycosyltransferase multigene family of Arabidopsis thaliana. J Biol Chem 276:4338–4343. https://doi.org/10.1074/jbc.M007447200
Liu Y, Wang X, Mo T, Yan Y, Song Y, Zhao Y, Li J, Shi S, Liu X, Tu P (2017) Identification and functional application of a new malonyltransferase NbMaT1 towards diverse aromatic glycosides from Nicotiana benthamiana. RSC Adv 7:21028–21035. https://doi.org/10.1039/C7RA01940H
Mameda R, Waki T, Kawai Y, Takahashi S, Nakayama T (2018) Involvement of chalcone reductase in the soybean isoflavone metabolon: identification of GmCHR5, which interacts with 2-hydroxyisoflavanone synthase. Plant J 96:56–74. https://doi.org/10.1111/tpj.14014
Miyajima I, Maehara T, Kage T, Fujieda K (1991) Identification of the main agent causing yellow color of yellow-flowered cyclamen mutant. J Japan Soc Hort Sci 60:409–414. https://doi.org/10.2503/jjshs.60.409
Molitor C, Mauracher SG, Pargan S, Mayer RL, Halbwirth H, Rompel A (2015) Latent and active aurone synthase from ray florets of C. grandiflora: a polyphenol oxidase with unique characteristics. Planta 242:519–537. https://doi.org/10.1007/s00425-015-2261-0
Nakagawa T, Kurose T, Hino T, Tanaka K, Kawamukai M, Niwa Y, Toyooka K, Matsuok K, Jinbo T, Kimura T (2007) Development of series of gateway binary vectors, pGWBs, for realizing efficient construction of fusion genes for plant transformation. J Biosci Bioeng 104:34–41. https://doi.org/10.1263/jbb.104.34
Nakayama T, Yonekura-Sakakibara K, Sato T, Kikuchi S, Fukui Y, Fukuchi-Mizutani M, Ueda T, Nakao M, Tanaka Y, Kusumi T, Nishio T (2000) Aureusidin synthase: a polyphenol oxidase homolog responsible for flower coloration. Science 290:1163–1166. https://doi.org/10.1126/science.290.5494.1163
Noguchi A, Saito A, Homma Y, Nakao M, Sasaki N, Nishino T, Takahashi S, Nakayama T (2007) A UDP-glucose: Isoflavone 7-O-glucosyltransferase from the roots of soybean (Glycine max) seedlings: purification, gene cloning, phylogenetics, and an implication for an alternative strategy of enzyme catalysis. J Biol Chem 282:23581–23590. https://doi.org/10.1074/jbc.M702651200
Ohno S, Hosokawa M, Hoshino A, Kitamura Y, Morita Y, Park Y, Nakashima A, Deguchi A, Tatsuzawa F, Doi M, Iida S, Yazawa S (2011a) A bHLH transcription factor, DvIVS, is involved in regulation of anthocyanin synthesis in dahlia (Dahlia variabilis). J Exp Bot 62:5105–5116. https://doi.org/10.1093/jxb/err216
Ohno S, Hosokawa M, Kojima M, Kitamura Y, Hoshino A, Tatsuzawa F, Doi M, Yazawa S (2011b) Simultaneous post-transcriptional gene silencing of two different chalcone synthase genes resulting in pure white flowers in the octoploid dahlia. Planta 234:945–958. https://doi.org/10.1007/s00425-011-1456-2
Ohno S, Deguchi A, Hosokawa M, Tatsuzawa F, Doi M (2013) A basic helix-loop-helix transcription factor DvIVS determines flower color intensity in cyanic dahlia cultivars. Planta 238:331–343. https://doi.org/10.1007/s00425-013-1897-x
Ohno S, Hori W, Hosokawa M, Tatsuzawa F, Doi M (2018a) Post-transcriptional silencing of chalcone synthase is involved in phenotypic lability in ray florets and leaves of bicolor dahlia (Dahlia variabilis) ‘Yuino.’ Planta 247:413–428. https://doi.org/10.1007/s00425-017-2796-3
Ohno S, Hori W, Hosokawa M, Tatsuzawa F, Doi M (2018b) Identification of flavonoids in leaves of a labile bicolor flowering dahlia (Dahlia variabilis) “Yuino.” Hort J 87:140–148. https://doi.org/10.2503/hortj.OKD-099
Ohno S, Ueno M, Doi M (2020) Differences in the CaMYBA genome between anthocyanin-pigmented cultivars and non-pigmented cultivars in pepper (Capsicum annuum). Hort J 89:30–36. https://doi.org/10.2503/hortj.UTD-097
Ohno S, Yokota M, Yamada H, Tatsuzawa F, Doi M (2021) Identification of chalcones and their contribution to yellow coloration in dahlia (Dahlia variabilis) ray florets. Hort J 90:450–459. https://doi.org/10.2503/hortj.UTD-305
Ohno S, Yamada H, Maruyama K, Deguchi A, Kato Y, Yokota M, Tatsuzawa F, Doi M (2022) A novel aldo–keto reductase gene is involved in 6′-deoxychalcone biosynthesis in dahlia (Dahlia variabilis). Planta 256:47. https://doi.org/10.1007/s00425-022-03958-4
Ono E, Fukuchi-Mizutani M, Nakamura N, Fukui Y, Yonekura-Sakakibara K, Yamaguchi M, Nakayama T, Tanaka T, Kusumi T, Tanaka Y (2006) Yellow flowers generated by expression of the aurone biosynthetic pathway. Proc Natl Acad Sci USA 103:11075–11080. https://doi.org/10.1073/pnas.0604246103
Ross J, Li Y, Lim EK, Bowles DJ (2001) Higher plant glycosyltransferases. Genome Biol 2(reviews3004):1. https://doi.org/10.1186/gb-2001-2-2-reviews3004
Saito N, Tatsuzawa F, Hoshino A, Abe Y, Ichimura M, Yokoi M, Toki K, Morita Y, Iida S, Honda T (2011) Anthocyanin pigmentation controlled by speckled and c-1 mutations of Japanese morning glory. J Japan Soc Hort Sci 80:452–460. https://doi.org/10.2503/jjshs1.80.452
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425. https://doi.org/10.1093/oxfordjournals.molbev.a040454
Schlangen K, Miosic S, Thill J, Halbwirth H (2010) Cloning, functional expression, and characterization of a chalcone 3-hydroxylase from Cosmos sulphureus. J Exp Bot 61:3451–3459. https://doi.org/10.1093/jxb/erq169
Stich K, Halbwirth H, Wurst F, Forkmann G (1994) Formation of 6ʹ-deoxychalcone 4ʹ-glucosides by enzyme extracts from petals of Dahlia variabilis. Zeitschrift Für Naturforschung C 49:737–741. https://doi.org/10.1515/znc-1994-11-1207
Suzuki H, Nakayama T, Yonekura-Sakakibara K, Fukui Y, Nakamura N, Yamaguchi M, Tanaka Y, Kusumi T, Nishino T (2002) cDNA cloning, heterologous expressions, and functional characterization of malonyl-coenzyme A: anthocyanidin 3-O-glucoside-6′-O-malonyltransferase from dahlia flowers. Plant Physiol 130:2142–2151. https://doi.org/10.1104/pp.010447
Taguchi G, Yazawa T, Hayashida N, Okazaki M (2001) Molecular cloning and heterologous expression of novel glucosyltransferases from tobacco cultured cells that have broad substrate specificity and are induced by salicylic acid and auxin. Eur J Biochem 268:4086–4094. https://doi.org/10.1046/j.1432-1327.2001.02325.x
Tamura K, Stecher G, Kumar S (2021) MEGA11: molecular evolutionary genetics analysis version 11. Mol Biol Evol 38:3022–3027. https://doi.org/10.1093/molbev/msab120
Welle R, Grisebach H (1998) Isolation of a novel NADPH-dependent reductase which coacts with chalcone synthase in the biosynthesis of 6ʹ-deoxychalcone. FEBS Lett 236:221–225. https://doi.org/10.1016/0014-5793(88)80318-1
Acknowledgements
This study was supported by the Grant-in-Aid for Young Scientists (No. 18K14455) from the Japan Society for the Promotion of Science to SO. Computations were partially performed on the NIG supercomputer at ROIS National Institute of Genetics (Japan).
Author information
Authors and Affiliations
Contributions
SO conceived the study. SO and MD designed the experiments. KM, HY, and SO conducted the experiments. KM and SO wrote the manuscript. All the authors read and approved the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflict of interests.
Additional information
Communicated by De-Yu Xie.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Maruyama, K., Yamada, H., Doi, M. et al. Identification of two 6ʹ-deoxychalcone 4ʹ-glucosyltransferase genes in dahlia (Dahlia variabilis). Planta 259, 114 (2024). https://doi.org/10.1007/s00425-024-04395-1
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00425-024-04395-1