Abstract
Main conclusion
MYBPA1-type R2R3 MYB transcription factor shows down-regulation in white mutant berries of Vaccinium uliginosum deficient in anthocyanins but not proanthocyanidins suggesting a role in the regulation of anthocyanin biosynthesis.
Berries of the genus Vaccinium are among the best natural sources of flavonoids. In this study, the expression of structural and regulatory flavonoid biosynthetic genes and the accumulation of flavonoids in white mutant and blue-colored wild-type bog bilberry (V. uliginosum) fruits were measured at different stages of berry development. In contrast to high contents of anthocyanins in ripe blue-colored berries, only traces were detected by HPLC–ESI–MS in ripe white mutant berries. However, similar profile and high levels of flavonol glycosides and proanthocyanidins were quantified in both ripe white and ripe wild-type berries. Analysis with qRT-PCR showed strong down-regulation of structural genes chalcone synthase (VuCHS), dihydroflavonol 4-reductase (VuDFR) and anthocyanidin synthase (VuANS) as well as MYBPA1-type transcription factor VuMYBPA1 in white berries during ripening compared to wild-type berries. The profiles of transcript accumulation of chalcone isomerase (VuCHI), anthocyanidin reductase (VuANR), leucoanthocyanidin reductase (VuLAR) and flavonoid 3′5′ hydroxylase (VuF3′5′H) were more similar between the white and the wild-type berries during fruit development, while expression of UDP-glucose: flavonoid 3-O-glucosyltransferase (VuUFGT) showed similar trend but fourfold lower level in white mutant. VuMYBPA1, the R2R3 MYB family member, is a homologue of VmMYB2 of V. myrtillus and VcMYBPA1 of V. corymbosum and belongs to MYBPA1-type MYB family which members are shown in some species to be related with proanthocyanidin biosynthesis in fruits. Our results combined with earlier data of the role of VmMYB2 in white mutant berries of V. myrtillus suggest that the regulation of anthocyanin biosynthesis in Vaccinium species could differ from other species studied.
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Abbreviations
- ANR:
-
Anthocyanidin reductase
- ANS:
-
Anthocyanidin synthase
- CHI:
-
Chalcone isomerase
- CHS:
-
Chalcone synthase
- DFR:
-
Dihydroflavonol 4-reductase
- F3′5′H:
-
Flavonoid 3′5′ hydroxylase
- LAR:
-
Leucoanthocyanidin reductase
- UFGT:
-
UDP-glucose: flavonoid 3-O-glucosyltransferase
References
Albert NW, Lewis DH, Zhang H, Schwinn KE, Jameson PE, Davies KM (2011) Members of an R2R3-MYB transcription family in Petunia are developmentally and environmentally regulated to control complex floral and vegetative pigmentation patterning. Plant J 65:771–784
Albert NW, Davies KM, Lewis DH, Zhang H, Montefiori M, Brendolise C, Boase MR, Ngo H, Jameson PE, Schwinn KE (2014) A conserved network of transcriptional activators and repressors regulates anthocyanin pigmentation in eudicots. Plant Cell 26:962–980
Azuma A, Kobayashi S, Mitani N, Shiraishi M, Yamada M, Ueno T, Kono A, Yakushiji H, Koshita Y (2008) Genomic and genetic analysis of Myb-related genes that regulate anthocyanin biosynthesis in grape berry skin. Theor Appl Genet 117:1009–1019
Azuma A, Yakushiji H, Koshita Y, Kobayashi S (2012) Flavonoid biosynthesis-related genes in grape skin are differentially regulated by temperature and light conditions. Planta 236:1067–1080
Bogs J, Jaffe FW, Takos AM, Walker AR, Robinson AS (2007) The grapevine transcription factor VvMYBPA1 regulates proanthocyanidin synthesis during fruit development. Plant Physiol 143:1347–1361
Castellarin SD, Di Gaspero G (2007) Transcriptional control of anthocyanin biosynthetic genes in extreme phenotypes for berry pigmentation of naturally occurring grapevines. BMC Plant Biol 7:46
Gupta V, Estrada AD, Blakley I, Reid R, Patel K, Meyer MD, Andersen SU, Brown AF, Lila MA, Loraine AE (2015) RNA-Seq analysis and annotation of a draft blueberry genome assembly identifies candidate genes involved in fruit ripening, biosynthesis of bioactive compounds, and stage-specific alternative splicing. GigaScience 4:5
He J, Giusti M (2010) Anthocyanins: natural colorants with health-promoting properties. Annu Rev Food Sci Technol 1:163–187
Hichri I, Barrieu F, Bogs J, Kappel C, Delrot S, Lauvergeat V (2011) Recent advances in the transcriptional regulation of the flavonoid biosynthetic pathway. J Exp Bot 62:2465–2483
Hokkanen J, Mattila S, Jaakola L, Pirttilä AM, Tolonen A (2009) Identification of phenolic compounds from lingonberry (Vaccinium vitis-idaea L.), bilberry (Vaccinium myrtillus L.) and hybrid bilberry (Vaccinium × intermedium Ruthe L.) leaves. J Agric Food Chem 57:9437–9447
Huang Y-F, Vialet S, Guiraud J-L, Torregrosa L, Bertrand Y, Cheynier V, This P, Terrier N (2014) A negative MYB regulator of proanthocyanidin accumulation, identified through expression quantitative locus mapping in the grape berry. New Phytol 201:795–809
Jaakola L (2013) New insights into the regulation of anthocyanin biosynthesis in fruits. Trends Plant Sci 18:477–483
Jaakola L, Pirttilä AM, Halonen M, Hohtola A (2001) Isolation of high quality RNA from the bilberry (Vaccinium myrtillus L.) fruit. Mol Biotechnol 19:201–203
Jaakola L, Määttä K, Pirttilä AM, Törrönen R, Kärenlampi S, Hohtola A (2002) Expression of genes involved in anthocyanin biosynthesis in relation to anthocyanin, proanthocyanidin, and flavonol levels during bilberry fruit development. Plant Physiol 130:729–739
Jaakola L, Pirttilä AM, Vuosku J, Hohtola A (2004) Method based on electrophoresis and gel extraction for obtaining genomic DNA-free cDNA without DNase treatment. Biotechniques 37:744–748
Jaakola L, Poole M, Jones MO, Kämäräinen-Karppinen T, Koskimäki JJ, Hohtola A, Häggman H, Fraser PD, Manning K, King GJ, Thomson H, Seymour GB (2010) A SQUAMOSA MADS box gene involved in the regulation of anthocyanin accumulation in bilberry fruits. Plant Physiol 153:1619–1629
Kobayashi S, Goto-Yamamoto N, Hirochika H (2004) Retrotransposon-induced mutations in grape skin color. Science 304:982
Koes R, Verweij W, Quattrocchio F (2005) Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. Trends Plant Sci 10:236–242
Koponen JM, Happonen AM, Auriola S, Kontkanen H, Buchert J, Poutanen KS, Törrönen AR (2008) Characterization and fate of black currant and bilberry flavonols in enzyme-aided processing. J Agric Food Chem 56:3136–3144
Koskimäki JJ, Hokkanen J, Jaakola L, Suorsa M, Tolonen A, Mattila S, Pirttilä AM, Hohtola A (2009) Flavonoid biosynthesis and degradation play a role in early defense responses of bilberry (Vaccinium myrtillus) against biotic stress. Eur J Plant Pathol 125:629–640
Kuhn N, Guan L, Dai ZW, Wu BH, Lauvergeat V, Gomes E, Li SH, Godoy F, Arce-Johnson P, Delrot S (2014) Berry ripening: recently heard through the grapevine. J Exp Bot 65:4543–4559
Lätti AK, Riihinen KR, Kainulainen PS (2008) Analysis of anthocyanin variation in wild populations of bilberry (Vaccinium myrtillus L.) in Finland. J Agric Food Chem 56:190–196
Lätti AK, Kainulainen PS, Hayirlioglu-Ayaz S, Ayaz FA, Riihinen KR (2009) Characterization of anthocyanins in Caucasian blueberries (Vaccinium arctostaphylos L.) native to Turkey. J Agric Food Chem 57:5244–5249
Lätti AK, Jaakola L, Riihinen KR, Kainulainen PS (2010) Anthocyanin and flavonol variation in bog bilberries (Vaccinium uliginosum L.) in Finland. J Agric Food Chem 58:427–433
Lätti AK, Riihinen KR, Jaakola L (2011) Phenolic compounds in berries and flowers of a natural hybrid between bilberry and lingonberry (Vaccinium × intermedium Ruthe). Phytochemistry 72:810–815
Li X, Sun H, Pei J, Dong Y, Wang F, Chen H, Sun Y, Wang N, Li H, Li Y (2012) De novo sequencing and comparative analysis of the blueberry transcriptome to discover putative genes related to antioxidants. Gene 511:54–61
Lin-Wang K, Bolitho K, Grafton K, Kortstee A, Karunairetnam S, McGhie TK, Espley RV, Hellens RP, Allan AC (2010) An R2R3 MYB transcription factor associated with regulation of the anthocyanin biosynthetic pathway in Rosaceae. BMC Plant Biol 10:50
Määttä-Riihinen KR, Kamal-Eldin A, Mattila PH, González-Paramás AM, Törrönen AR (2004) Distribution and contents of phenolic compounds in eighteen Scandinavian berry species. J Agric Food Chem 52:4477–4486
Määttä-Riihinen KR, Kähkonen MP, Törrönen AR, Heinonen IM (2005) Catechins and procyanidins in berries of Vaccinium species and their antioxidant activity. J Agric Food Chem 53:8485–8491
Masuoka C, Yokoi K, Komatsu H, Kinjo J, Nohara T, Ono M (2007) Two novel antioxidant ortho-benzoyloxyphenyl acetic acid derivatives from the fruit of Vaccinium uliginosum. Food Sci Technol Res 13:215–220
Medina-Puche L, Cumplido-Laso G, Amil-Ruiz F, Hoffman T, Ring L, Rodríguez-Franco A, Caballero JL, Schwab W, Muñoz-Blanco J, Blanco-Portales R (2014) MYB10 plays a major role in the regulation of flavonoid/phenylpropanoid metabolism during ripening of Fragaria x ananassa fruits. J Exp Bot 65:401–417
Polashock J, Zelzion E, Fajardo D, Zalapa J, Georgi L, Bhattacharya D, Vorsa N (2014) The American cranberry: first insights into the whole genome of a species adapted to bog habitat. BMC Plant Biol 14:165
Riihinen K, Jaakola L, Kärenlampi S, Hohtola A (2008) Organ-specific distribution of phenolic compounds in bilberry (Vaccinium myrtillus) and ‘northblue’ blueberry (Vaccinium corymbosum x V. angustifolium). Food Chem 110:156–160
Rowland LJ, Alkharouf N, Darwish O, Ogden EL, Polashock JJ, Bassil NV, Main D (2012) Generation and analysis of blueberry transcriptome sequences from leaves, developing fruit, and flower buds from cold acclimation through deacclimation. BMC Plant Biol 12:46
Schroeter H, Heiss C, Spencer JPE, Keen CL, Lupton JR, Schmitz HH (2010) Recommending flavanols and procyanidins for cardiovascular health: current knowledge and future needs. Mol Aspects Med 31:546–557
Shimada Y, Nakano-Shimada R, Ohbayashi M, Okinaka Y, Kiyokawa S, Kikuchi Y (1999) Expression of chimeric P450 genes encoding flavonoid-3′,5′-hydroxylase in transgenic tobacco and petunia plants. FEBS Lett 461:241–245
Tanaka Y, Brugliera F (2013) Flower colour and cytochromes P450. Philos Trans R Soc Lond B Biol Sci 368:20120432
Terrier N, Torregosa L, Ageorges A, Vialet S, Verriès C, Cheynier V, Romieu C (2009) Ecotopic expression of VvMybPA2 promotes proanthocyanidin biosynthesis in grapevine and suggests additional targets in the pathway. Plant Physiol 149:1028–1041
Walker AR, Lee E, Bogs J, McDavid DAJ, Thomas MR, Robinson SP (2007) White grapes arose through the mutation of two similar and adjacent regulatory genes. Plant J 49:772–785
Zifkin M, Jin A, Ozga JA, Zaharia I, Schernthaner JP, Gesell A, Abrams SR, Kennedy JA, Constabel CP (2012) Gene expression and metabolite profiling of developing highbush blueberry fruit indicates transcriptional regulation of flavonoid metabolism and activation of abscisic acid metabolism. Plant Physiol 158:200–224
Zoratti L, Karppinen K, Luengo Escobar A, Häggman H, Jaakola L (2014) Light-controlled flavonoid biosynthesis in fruits. Front Plant Sci 5:534
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This work was financially supported by the Finnish Cultural Foundation (AP), the Jenny and Antti Wihuri Foundation (AP) and the Academy of Finland (LJ, Post-doctoral researcher’s project 123471).
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Special topic: Polyphenols: biosynthesis and function in plants and ecosystems. Guest editor: Stefan Martens.
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425_2015_2363_MOESM4_ESM.pdf
Suppl. Fig. S1 The flavonoid biosynthetic pathway. CHS, chalcone synthase; CHI, chalcone isomerase; F3H, flavanone 3-hydroxylase; FLS, flavonol synthase; F3′H, flavonoid 3′-hydroxylase; F3′5′H, flavonoid 3′5′ hydroxylase; DFR, dihydroflavonol 4-reductase; ANS, anthocyanidin synthase; LAR, leucoanthocyanidin reductase; ANR, anthocyanidin reductase; UFGT, UDP-glucose: flavonoid 3-O-glucosyltransferase 4 (PDF 108 kb)
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Primetta, A.K., Karppinen, K., Riihinen, K.R. et al. Metabolic and molecular analyses of white mutant Vaccinium berries show down-regulation of MYBPA1-type R2R3 MYB regulatory factor. Planta 242, 631–643 (2015). https://doi.org/10.1007/s00425-015-2363-8
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DOI: https://doi.org/10.1007/s00425-015-2363-8