Abstract
As a result of natural hybridization and human selection over millennia, the skin colors of grapes have become greatly diversified. The color is determined by the quantity and composition of anthocyanins. Color-skinned cultivars accumulate anthocyanins in their skins, whereas white-skinned cultivars do not. Myb-related transcription-factor genes such as VvmybA1 regulate anthocyanin biosynthesis. VvMYBA2r, VlmybA1-1, VlmybA1-2, and VlmybA2, which are homologs of VvmybA1, also regulate anthocyanin biosynthesis. In this study, we isolated a novel Myb-related sequence, VlmybA1-3, from cultivars of Vitis labruscana (Vitis vinifera × Vitis labrusca) by means of inverse PCR, and confirmed by means of transient gene expression assay that the gene regulates anthocyanin biosynthesis in grape berry skin. Seedlings of V. labruscana with two functional haplotypes at a region of berry color loci accumulated more anthocyanins than seedlings with a single functional haplotype. In addition, we investigated the haplotypes at the region in 35 cultivars (both V. vinifera and V. labruscana), and found certain typical characteristics. These findings will contribute to the selection of seedlings with high anthocyanin quantities in breeding programs for wine and table grapes, and will help elucidate the origin and evolution of Vitis species.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Azuma A, Kobayashi S, Yakushiji H, Yamada M, Mitani N, Sato A (2007) VvmybA1 genotype determines grape skin color. Vitis 46(3):154–155
Borovsky Y, Oren-Shamir M, Ovadia R, De Jong W, Paran I (2004) The A locus that controls anthocyanin accumulation in pepper encodes a MYB transcription factor homologous to anthocyanin2 of petunia. Theor Appl Genet 109:23–29
Boss PK, Davies C, Robinson SP (1996a) Expression of anthocyanin biosynthesis pathway genes in red and white grapes. Plant Mol Biol 32:565–569
Boss PK, Davies C, Robinson SP (1996b) Anthocyanin composition and anthocyanin pathway gene expression in grapevine sports differing in berry skin colour. Aust J Grape Wine R 2:163–170
Castellarin SD, Matthews MA, Gaspere GD, Gambetta GA (2007) Water deficits accelerate ripening and induce changes in gene expression regulating flavonoid biosynthesis in grape berries. Planta 227(1):101–112
Doligez A, Bouquet A, Danglot Y, Lahogue F, Riaz S, Meredith CP, Edwards KJ (2002) Genetic mapping of grapevine (Vitis vinifera L.) applied to the detection of QTLs for seedlessness and berry weight. Theor Appl Genet 105:780–795
Fischer BM, Salakhutdinov I, Akkurt M, Eibach R, Edwards KJ, Topfer R, Zyprian EM (2004) Quantitative trait locus analysis of fungal disease resistance factors on a molecular map of grapevine. Theor Appl Genet 108:501–515
Geekiyanage S, Takase T, Ogura Y, Kiyosue T (2007) Anthocyanin production by over-expression of grape transcription factor gene VlmybA2 in transgenic tobacco and Arabidopsis. Plant Biotechnol Rep 1(1):11–18
Goto-Yamamoto N, Mouri H, Azumi M, Edwards KJ (2006) Development of grape microsatellite markers and microsatellite analysis including oriental cultivars. Am J Enol Vitic 57(1):105–108
Hiratsuka S, Onodera H, Kawai Y, Kubo T, Itoh H, Wada R (2001) ABA and sugar effects on anthocyanin formation in grape berry cultured in vitro. Sci Hort 90:121–130
Holton T, Cornish E (1995) Genetics and biochemistry of anthocyanin biosynthesis. Plant Cell 7:1071–1083
Jeong ST, Goto-Yamamoto N, Kobayashi S, Esaka M (2004) Effects of plant hormones and shading on the accumulation of anthocyanins and the expression of anthocyanin biosynthetic genes in grape berry skins. Plant Sci 167:247–252
Kataoka I, Kubo Y, Sugiura A, Tomana T (1984) Effects of temperature, cluster shading and some growth regulators on L-phenylalanine ammonia-lyase activity and anthocyanin accumulation on black grapes. Mem Coll Agric Kyoto Univ 124:35–44
Kataoka I, Sugiyama A, Beppu K (2003) Role of ultraviolet radiation in accumulation of anthocyanin in berries of ‘Gros Colman’ grapes (Vitis vinifera L.). J Jpn Soc Hort Sci 72(1):1–6
Kliewer WM, Torres RE (1972) Effect of controlled day and night temperatures on grape coloration. Am J Enol Vitic 23(2):71–77
Kobayashi S, Ishimaru M, Hiraoka K, Honda C (2002) Myb-related genes of the Kyoho grape (Vitis labruscana) regulate anthocyanin biosynthesis. Planta 215:924–933
Kobayashi S, Goto-Yamamoto N, Hirochika H (2004) Retrotransposon-induced mutations in grape skin color. Science 304:982
Kobayashi S, Goto-Yamamoto N, Hirochika H (2005) Association of VvmybA1 gene expression with anthocyanin production in grape (Vitis vinifera) skin-color mutants. J Jpn Soc Hort Sci 74(3):196–203
Koshita Y, Kobayashi S, Ishimaru M, Funamoto Y, Shiraishi M, Azuma A, Yakushiji H, Nakayama M (2008) An anthocyanin regulator from grapes, VlmybA1–2, produces reddish-purple plants. J Jpn Soc Hort Sci 77(1):33–37
Lijavetzky D, Ruiz-Garcia L, Cabezas JA, de Andres MT, Bravo G, Ibanez A, Carreno J, Cabello F, Ibanez J, Martinez-Zapater JM (2006) Molecular genetics of berry colour variation in table grape. Mol Gen Genom 276(5):427–435
Mori K, Goto-Yamamoto N, Kitayama M, Hashizume K (2007) Loss of anthocyanins in red-wine grape under high temperature. J Exp Bot 58(8):1935–1945
Negrul AM (1938) Evolution of cultivated forms of grapes. Comm Russ Acad Sci URSS 18:585–588
Ozeki Y, Chikagawa Y, Kimura S, Soh H, Maeda K, Pornsiriwong W, Kato M, Akimoto H, Oyanagi M, Fukuda T, Koda T, Itoh Y, Yamada A, Davies E, Ueno H, Takeda J (2003) Putative cis-elements in the promoter region of the carrot phenylalanine ammonia-lyase gene induced during anthocyanin synthesis. J Plant Res 116:155–159
Quattrocchio F, Wing JF, Lappen HTC, Mol JNM, Koes RE (1993) Regulatory genes controlling anthocyanin pigmentation are functionally conserved among plant species and have distinct sets of target genes. Plant Cell 5:1497–1512
Ramsay N, Walker A, Mooney M, Gray J (2003) Two basic-helix-loop-helix genes (MYC-146 and GL3) from Arabidopsis can activate anthocyanin biosynthesis in a white-flowered Matthiola incana mutant. Plant Mol Biol 52:679–688
Robbins M, Paolocci F, Hughes J, Turchetti V, Allison G, Arcioni S, Morris P, Damiani F (2003) Sn, a maize bHLH gene, modulates anthocyanin and condensed tannin pathways in Lotus corniculatus. J Exp Bot 54:239–248
Sainz M, Grotewold E, Chandler V (1997) Evidence for direct activation of an anthocyanin promoter by the maize C1 protein and comparison of DNA binding by related MYB domain proteins. Plant Cell 9:611–625
Salmaso M, Malacarne G, Troggio M, Faes G, Stefanini M, Grando MS, Velasco R (2008) A grapevine (Vitis vinifera L.) genetic map integrating the position of 139 expressed genes. Theor Appl Genet 116:1129–1143
Schwinn K, Venail J, Shang Y, Mackay S, Alm V, Butelli E, Oyama R, Bailey P, Davies K, Martin C (2006) A small family of MYB-regulatory genes controls floral pigmentation intensity and patterning in the genus Antirrhinum. Plant Cell 18:831–851
Shiraishi M, Yamada M, Mitani N, Ueno T (2007) A rapid determination method for anthocyanin profiling in grape genetic resources. J Jpn Soc Hort Sci 76(1):28–35
Spelt C, Quattrocchio F, Mol J, Koes R (2000) anthocyanin1 of petunia encodes a basic helix-loop-helix protein that directly activates transcription of structural anthocyanin genes. Plant Cell 12:1619–1632
This P, Lacombe T, Cadle-Davidson M, Owens CL (2007) Wine grape (Vitis vinifera L.) color associates with allelic variation in the domestication gene VvmybA1. Theor Appl Genet 114:723–730
Triglia T, Peterson MG, Kemp DJ (1988) A procedure for in vitro amplification of DNA segments that lie outside the boundaries of known sequences. Nucleic Acids Res 16(16):8186
Uimari A, Strommer J (1998) Anthocyanin regulatory mutations in pea: effects on gene expression and complementation by R-like genes of maize. Mol Gen Genet 257:198–204
Walker AR, Lee E, Robinson SP (2006) Two new grape cultivars, bud sports of Cabernet Sauvignon bearing pale-coloured berries, are the result of deletion of two regulatory genes of the berry colour locus. Plant Mol Biol 62(4–5):623–635
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(5):772–785
Winkel-Shirley B (2001) Flavonoid biosynthesis. A colourful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiol 126:485–493
Yakushiji H, Kobayashi S, Goto-Yamamoto N, Jeong ST, Sueta T, Mitani N, Azuma A (2006) A skin color mutation of grapevine, from black-skinned ‘Pinot Noir’ to white-skinned ‘Pinot Blanc’ is caused by the deletion of the functional VvmybA1 allele. Biosci Biotechnol Biochem 70(6):1506–1508
Yamane T, Jeong ST, Goto-Yamamoto N, Koshita Y, Kobayashi S (2006) Effects of temperature on anthocyanin biosynthesis in grape berry skins. Am J Enol Vitic 57(1):54–59
Acknowledgments
We are grateful to Dr. N. Goto-Yamamoto, National Research Institute of Brewing, for providing two grape gDNA samples. We also thank Ms. T. Nakasumi, Grape and Persimmon Research Station, National Institute of Fruit Tree Science, for technical assistance. This work was supported in part by a grant from the National Agriculture and Food Research Organization (NARO), Japan.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by H. Nybom.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Azuma, A., Kobayashi, S., Mitani, N. et al. Genomic and genetic analysis of Myb-related genes that regulate anthocyanin biosynthesis in grape berry skin. Theor Appl Genet 117, 1009–1019 (2008). https://doi.org/10.1007/s00122-008-0840-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00122-008-0840-1