Abstract
Tartary buckwheat is an excellent medicinal and nutrient-rich crop that contains abundant flavonoids. Flavonoids, such as anthocyanin and proanthocyanidin, can be accumulated in plants for resistance to abiotic stresses, for instance, UV-B and cold. In this study, an R2R3-MYB transcription factor gene named FtMYB15 was isolated from tartary buckwheat. A phylogenetic tree suggested that FtMYB15 potentially plays a role in flavonoid biosynthesis. Consequently, the ectopic overexpression of FtMYB15 was conducted in Arabidopsis. Transgenic lines exerted increased pigmentation in leaves and seed coats, as well as higher accumulation levels of anthocyanin and proanthocyanidin in Arabidopsis. Specifically, anthocyanin and proanthocyanidin in transgenic lines were 8.62- and 4.93 (P < 0.01)-fold higher than wild-type Arabidopsis, respectively. Meanwhile, the expressions of both early and late biosynthesis genes, including AtCHS, AtCHI, AtF3H, AtF3′H, AtDFR, AtANS, AtBAN, and AtTT12, in the anthocyanin and proanthocyanidin pathway were enhanced (P < 0.01). Particularly, AtDFR and AtANS were upregulated 4.13 and 5.11 (P < 0.01) times higher than the control, respectively, whereas AtBAN and AtTT12 were upregulated a 100-fold (P < 0.01) compared to wild-type Arabidopsis. Only AtFLS expression decreased in transgenic Arabidopsis (P < 0.01). These results laid a foundation for better understanding of flavonoid biosynthesis and its molecular regulation mechanism in tartary buckwheat.
Similar content being viewed by others
References
Albert NW (2015) Subspecialization of R2R3-MYB repressors for anthocyanin and proanthocyanidin regulation in forage legumes. Front Plant Sci 6:1165
An X-H, Tian Y, Chen K-Q, Wang X-F, Hao Y-J (2012) The apple WD40 protein MdTTG1 interacts with bHLH but not MYB proteins to regulate anthocyanin accumulation. J Plant Physiol 169:710–717
Bai YC, Li CL, Zhang JW, Li SJ, Luo XP, Yao HP, Chen H, Zhao HX, Park SU, Wu Q (2014) Characterization of two tartary buckwheat R2R3-MYB transcription factors and their regulation of proanthocyanidin biosynthesis. Physiol Plantarum 152:431–440
Bordiga M, Travaglia F, Locatelli M, Coïsson JD, Arlorio M (2011) Characterisation of polymeric skin and seed proanthocyanidins during ripening in six Vitis vinifera L. cv. Food Chem 127:180–187
Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743
Deroles S (2009) Anthocyanin biosynthesis in plant cell cultures: a potential source of natural colourants. In: Anthocyanins. Springer, New York pp 108–167
Dong HS, Choi MG, Kang CS, Park CS, Choi SB, Park YI (2016) A wheat R2R3-MYB protein PURPLE PLANT1 (TaPL1) functions as a positive regulator of anthocyanin biosynthesis. Biochem Bioph Res Com 469:686–691
Dubos C, Stracke R, Grotewold E, Weisshaar B, Martin C, Lepiniec L (2010a) MYB transcription factors in Arabidopsis. Trends Plant Sci 15:573–581
Fabjan N, Rode J, Košir IJ, Wang Z, Zhang Z, Kreft I (2003) Tartary buckwheat (Fagopyrum tataricum Gaertn.) as a source of dietary rutin and quercitrin. J Agric Food Chem 51:6452–6455
Gesell A, Yoshida K, Lan TT, Constabel CP (2014) Characterization of an apple TT2-type R2R3 MYB transcription factor functionally similar to the poplar proanthocyanidin regulator PtMYB134. Planta 240:497–511
Gonzalez A, Zhao M, Leavitt JM, Lloyd AM (2008a) Regulation of the anthocyanin biosynthetic pathway by the TTG1/bHLH/Myb transcriptional complex in Arabidopsis seedlings. Plant J 53:814–827
Gonzalez A, Brown M, Hatlestad G, Akhavan N, Smith T, Hembd A, Moore J, Montes D, Mosley T, Resendez J (2016) TTG2 controls the developmental regulation of seed coat tannins in Arabidopsis by regulating vacuolar transport steps in the proanthocyanidin pathway. Dev Biol 419:54–63
Grotewold E, Sainz MB, Tagliani L, Hernandez JM, Bowen B, Chandler VL (2001) Identification of the residues in the Myb domain of maize C1 that specify the interaction with the bHLH cofactor R. Proc Natl Acad Sci USA 97:13579–13584
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
Jami SK, Clark GB, Ayele BT, Roux SJ, Kirti P (2012) Identification and characterization of annexin gene family in rice. Plant Cell Rep 31:813–825
Jin H, Martin C (1999) Multifunctionality and diversity within the plant MYB-gene family. Plant Mol Biol 41:577–585
Jun JH, Liu C, Xiao X, Dixon RA (2015) The transcriptional repressor MYB2 regulates both spatial and temporal patterns of proanthocyandin and anthocyanin pigmentation in Medicago truncatula. Plant Cell 27:2860–2879
Kim S-L, Kim S-K, Park C-H (2004) Introduction and nutritional evaluation of buckwheat sprouts as a new vegetable. Food Res Inter 37:319–327
Kranz HD, Denekamp M, Greco R, Jin H, Leyva A, Meissner RC, Petroni K, Urzainqui A, Bevan M, Martin C (1998) Towards functional characterisation of the members of the R2R3-MYB gene family from Arabidopsis thaliana. Plant J 16:263–276
Laitinen RA, Ainasoja M, Broholm SK, Teeri TH, Elomaa P (2008) Identification of target genes for a MYB-type anthocyanin regulator in Gerbera hybrida. J Exp Bot 59:3691–3703
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2–∆∆CT method. Methods 25:402–408
Nesi N, Jond C, Debeaujon I, Caboche M, Lepiniec L (2001) The Arabidopsis TT2 gene encodes an R2R3 MYB domain protein that acts as a key determinant for proanthocyanidin accumulation in developing seed. Plant Cell 13:2099–2114
Park NI, Li X, Suzuki T, Kim S-J, Woo S-H, Park CH, Park SU (2011) Differential expression of anthocyanin biosynthetic genes and anthocyanin accumulation in tartary buckwheat cultivars ‘Hokkai T8’and ‘Hokkai T10’. J Agr Food Chem 59:2356–2361
Peel GJ, Dixon RA (2007) Detection and quantification of engineered proanthocyanidins in transgenic plants. Nat Prod Commun 2:1009–1014
Pérez-Díaz JR, Pérez-Díaz J, Madrid-Espinoza J, González-Villanueva E, Moreno Y, Ruiz-Lara S (2016) New member of the R2R3-MYB transcription factors family in grapevine suppresses the anthocyanin accumulation in the flowers of transgenic tobacco. Plant Mol Biol 90:63–76
Ramsay NA, Glover BJ (2005) MYB–bHLH–WD40 protein complex and the evolution of cellular diversity. Trends Plant Sci 10:63–70
Terrier N, Torregrosa L, Ageorges A, Vialet S, Verriès C, Cheynier V, Romieu C (2009) Ectopic expression of VvMybPA2 promotes proanthocyanidin biosynthesis in grapevine and suggests additional targets in the pathway. Plant Physiol 149:1028–1041
Tohge T, Nishiyama Y, Hirai MY, Yano M, Nakajima JI, Awazuhara M, Inoue E, Takahashi H, Goodenowe DB, Kitayama M (2005) Functional genomics by integrated analysis of metabolome and transcriptome of Arabidopsis plants over-expressing an MYB transcription factor. Plant J 42:218–235
Vimolmangkang S, Han Y, Wei G, Korban SS (2013) An apple MYB transcription factor, MdMYB3, is involved in regulation of anthocyanin biosynthesis and flower development. BMC Plant Biol 13:56–60
Vogrinčič M, Timoracka M, Melichacova S, Vollmannova A, Kreft I (2010) Degradation of rutin and polyphenols during the preparation of tartary buckwheat bread. J Agr Food Chem 58:4883–4887
Vogt T (2010) Phenylpropanoid biosynthesis. Mol Plant 3:2–20
Wei Z, Ning G, Lv H, Liao L, Bao M (2009) Single MYB-type transcription factor AtCAPRICE: a new efficient tool to engineer the production of anthocyanin in tobacco. Biochem Bioph Res Com 388:742–747
Winkel-Shirley B (2002) Biosynthesis of flavonoids and effects of stress. Curr Opin Plant Biol 5:218–223
Acknowledgements
This work was supported by Science and Technology Department of Sichuan Province, P. R. China (Grant number: 2015HH0047). We thank the team of http://webshop.elsevier.com/ for critical reading and editing of this manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Xiaopeng Luo and Haixia Zhao have contributed equally.
Rights and permissions
About this article
Cite this article
Luo, X., Zhao, H., Yao, P. et al. An R2R3-MYB Transcription Factor FtMYB15 Involved in the Synthesis of Anthocyanin and Proanthocyanidins from Tartary Buckwheat. J Plant Growth Regul 37, 76–84 (2018). https://doi.org/10.1007/s00344-017-9709-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00344-017-9709-3