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Fagopyrum tataricum FtWD40 Functions as a Positive Regulator of Anthocyanin Biosynthesis in Transgenic Tobacco

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Abstract

Tartary buckwheat (Fagopyrum tataricum Gaertn.) contains high concentration of flavonoids, which are mainly represented by rutin, anthocyanins, and proanthocyanidins. WD40 transcription factors (TFs) play significant roles in the transcriptional regulation of the anthocyanin biosynthetic pathway. In this study, a WD40-repeat protein gene (designated as FtWD40) was identified and characterized from tartary buckwheat. The bioinformatics analyses showed that the putative FtWD40 shared a high level of similarity with MtWD40-1, which is a positive regulator in anthocyanin biosynthesis of Medicago truncatula. The yeast one-hybrid assay indicated that FtWD40 had transcriptional activation activities. During florescence, FtWD40 was highly expressed in flowers compared to other organs. Furthermore, its overexpression in tobacco resulted in a remarkable deepening of petal pigmentation in flowers due to a significant increase in anthocyanins accumulation. Meanwhile, the expression of dihydroflavonol-4-reductase (DFR) and anthocyanin synthase (ANS) was upregulated 1.95- and 1.56-fold, respectively. In contrast, the expression level was lower for flavonol synthase (FLS) in the transgenic lines. To the best of our knowledge, this is the first functional characterization of a WD40 transcription factor (FtWD40) from tartary buckwheat that controls the anthocyanin pathway.

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References

  • Aharoni A, Galili G (2011) Metabolic engineering of the plant primary secondary metabolism interface. Curr Opin. Biotech 22:239–244

    CAS  Google Scholar 

  • Albert NW, Lewis DH, Zhang H, Schwinn KE, Jameson PE, Davies KM (2011) Members of an R2R3 - MYB transcription factor family in Petunia are developmentally and environmentally regulated to control complex floral and vegetative pigmentation patterning. Plant J 65:771–784

    Article  CAS  PubMed  Google Scholar 

  • An XH, Tian Y, Chen KQ, Wang XF, Hao YJ (2012) The apple WD40 protein MdTTG1 interacts with bHLH but not MYB proteins to regulate anthocyanin accumulation. J Plant Physiol 169:710–717

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • Baudry A, Heim MA, Dubreucq B, Caboche M, Weisshaar B, Lepiniec L (2004) TT2, TT8, and TTG1 synergistically specify the expression of BANYULS and proanthocyanidin biosynthesis in Arabidopsis thaliana. Plant J 39:366–380

    Article  CAS  PubMed  Google Scholar 

  • Das PK, Shin DH, Choi S-B, Park Y-I (2012) Sugar-hormone cross-talk in anthocyanin biosynthesis. Mol Cells 34:501–507

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Davies KM, Schwinn KE, Deroles SC, Manson DG, Lewis DH, Bloor SJ, Bradley JM (2003) Enhancing anthocyanin production by altering competition for substrate between flavonol synthase and dihydroflavonol 4-reductase. Euphytica 131:259–268

    Article  CAS  Google Scholar 

  • Dong W, Niu L, Gu J, Gao F (2014) Isolation of a WD40-repeat gene regulating anthocyanin biosynthesis in storage roots of purple-fleshed sweet potato. Acta Physiol Plant 36:1123–1132

    Article  CAS  Google Scholar 

  • Dubos C, Le Gourrierec J, Baudry A, Huep G, Lanet E, Debeaujon I, Routaboul JM, Alboresi A, Weisshaar B, Lepiniec L (2008) MYBL2 is a new regulator of flavonoid biosynthesis in Arabidopsis thaliana. Plant J 55:940–953

    Article  CAS  PubMed  Google Scholar 

  • Dubos C, Stracke R, Grotewold E, Weisshaar B, Martin C, Lepiniec L (2010) MYB transcription factors in Arabidopsis. Trends Plant Sci 15:573–581

    Article  CAS  PubMed  Google Scholar 

  • Espley RV, Hellens RP, Putterill J, Stevenson DE, Kutty-Amma S, Allan AC (2007) Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10. Plant J 49:414–427

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gietz D, St Jean A, Woods RA, Schiestl RH (1992) Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res 20:1425

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gonzalez A, Zhao M, Leavitt JM, Lloyd AM (2008) Regulation of the anthocyanin biosynthetic pathway by the TTG1/bHLH/Myb transcriptional complex in Arabidopsis seedlings. Plant J 53:814–827

    Article  CAS  PubMed  Google Scholar 

  • Grotewold E (2006) The genetics and biochemistry of floral pigments. Annu Rev Plant Biol 57:761–780

    Article  CAS  PubMed  Google Scholar 

  • Grotewold E, Sainz MB, Tagliani L, Hernandez JM, Bowen B, Chandler VL (2000) Identification of the residues in the Myb domain of maize C1 that specify the interaction with the bHLH cofactor R. Proc.Natl Acad Sci 97:13579–13584

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Han Y, Vimolmangkang S, Soria-Guerra RE, Korban SS (2012) Introduction of apple ANR genes into tobacco inhibits expression of both CHI and DFR genes in flowers, leading to loss of anthocyanin. J Exp Bot 65:2437–2447

    Article  Google Scholar 

  • Hernandez J, Pizzirusso M, Grotewold E (2000) The maize Mp1 gene encodes a WD-repeat protein similar to An11 and TTG. Maize Genet Coop Newsl 74:24.

    Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Horsch RB, Fry J, Hoffmann NL, Eichholtz DA, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Lai B, Du LN, Liu R, Hu B, Su WB, Qin YH, Zhao JT, Wang HC, Hu GB (2016) Two LcbHLH transcription factors interacting with LcMYB1 in regulating late structural genes of anthocyanin biosynthesis in Nicotiana and Litchi chinensis during anthocyanin accumulation. Front Plant Sci 7:166

    Article  PubMed  PubMed Central  Google Scholar 

  • Lee S, Lee J, Paek KH, Kwon SY, Cho HS, Kim SJ, Park JM (2010) A novel WD40 protein, BnSWD1, is involved in salt stress in Brassica napus. Plant Biotechnol Rep 4:165–172

    Article  Google Scholar 

  • Lepiniec L, Debeaujon I, Routaboul JM, Baudry A, Pourcel L, Nesi N, Caboche M (2006) Genetics and biochemistry of seed flavonoids. Annu Rev Plant Biol 57:405–430

    Article  CAS  PubMed  Google Scholar 

  • Li SJ, Bai YC, Li CL, Yao HP, Chen H, Zhao HX, Wu Q (2015) Anthocyanins accumulate in tartary buckwheat (Fagopyrum tataricum) sprout in response to cold stress. Acta Physiol Plantarum 37: 159–167

    Article  CAS  Google Scholar 

  • Lim SH, Song JH, Kim DH, Kim JK, Lee JY, Kim YM, Ha SH (2016) Activation of anthocyanin biosynthesis by expression of the radish R2R3-MYB transcription factor gene RsMYB1. Plant Cell Rep 35:641–653

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  PubMed  PubMed Central  Google Scholar 

  • Lin-Wang K, McGhie TK, Wang M, Liu Y, Warren B, Storey R, Espley RV, Allan AC (2014) Engineering the anthocyanin regulatory complex of strawberry (Fragaria vesca). Front Plant Sci 5:651

    Article  PubMed  PubMed Central  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Matus J, Poupin M, Cañón P, Bordeu E, Alcalde J, Arce-Johnson P (2010) Isolation of WDR and bHLH genes related to flavonoid synthesis in grapevine (Vitis vinifera L.). Plant Mol Biol Rep 72:607–620

    Article  CAS  Google Scholar 

  • Mehrtens F, Kranz H, Bednarek P, Weisshaar B (2005) The Arabidopsis transcription factor MYB12 is a flavonol-specific regulator of phenylpropanoid biosynthesis. Plant Physiol 138:1083–1096

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • 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

    CAS  PubMed  PubMed Central  Google Scholar 

  • Park NI, Li X, Suzuki T, Kim SJ, Woo SH, 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

    Article  CAS  Google Scholar 

  • Qi T, Song S, Ren Q, Wu D, Huang H, Chen Y, Fan M, Peng W, Ren C, Xie D (2011) The Jasmonate-ZIM-domain proteins interact with the WD-Repeat/bHLH/MYB complexes to regulate jasmonate-mediated anthocyanin accumulation and trichome initiation in Arabidopsis thaliana. Plant Cell 23:1795–1814

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rabino I, Mancinelli AL (1986) Light, temperature, and anthocyanin production. Plant Physiol 81:922–924

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ramsay NA, Glover BJ (2005) MYB–bHLH–WD40 protein complex and the evolution of cellular diversity. Trends Plant Sci 10:63–70

    Article  CAS  PubMed  Google Scholar 

  • Schaart JG, Dubos C, Romero De La Fuente I, van Houwelingen AM, de Vos RC, Jonker HH, Xu W, Routaboul JM, Lepiniec L, Bovy AG (2013) Identification and characterization of MYB-bHLH-WD40 regulatory complexes controlling proanthocyanidin biosynthesis in strawberry (Fragaria x ananassa) fruits. New Phytol 197:454–467

    Article  CAS  PubMed  Google Scholar 

  • Shin DH, 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 Co 469:686–691

    Article  CAS  Google Scholar 

  • Sompornpailin K, Makita Y, Yamazaki M, Saito K (2002) A WD-repeat-containing putative regulatory protein in anthocyanin biosynthesis in Perilla frutescens. Plant Mol Biol 50:485–495

    Article  CAS  PubMed  Google Scholar 

  • Su LT, Li JW, Liu DQ, Zhai Y, Zhang HJ, Li XW, Zhang QL, Wang Y, Wang QY (2014) A novel MYB transcription factor, GmMYBJ1, from soybean confers drought and cold tolerance in Arabidopsis thaliana. Gene 538:46–55

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Tsurunaga Y, Takahashi T, Katsube T, Kudo A, Kuramitsu O, Ishiwata M, Matsumoto S (2013) Effects of UV-B irradiation on the levels of anthocyanin, rutin and radical scavenging activity of buckwheat sprouts. Food Chem 141:552–556

    Article  CAS  PubMed  Google Scholar 

  • Van Nocker S, Ludwig P (2003) The WD-repeat protein superfamily in Arabidopsis: conservation and divergence in structure and function. BMC Genomics 4:50

    Article  PubMed  PubMed Central  Google Scholar 

  • Yang W, Wu Y, Tang Y (2009) Expressing and functional analysis of GmMYBJ6 from soybean. Yi Chuan 31:645–653

    Article  CAS  PubMed  Google Scholar 

  • Zhang ZL, Zhou ML, Tang Y, Li FL, Tang YX, Shao JR, Xue WT, Wu YM (2012) Bioactive compounds in functional buckwheat food. Food Res Int 49:389–395

    Article  CAS  Google Scholar 

  • Zhou M, Wang C, Qi L, Yang X, Sun Z, Tang Y, Tang Y, Shao J, Wu Y (2015) Ectopic expression of Fagopyrum tataricum FtMYB12 improves cold tolerance in Arabidopsis thaliana. J Plant Growth Regul 34:362–371

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by the Science and Technology Department of Sichuan Province, PR China (2015HH0047).

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Correspondence to Qi Wu.

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Yao, P., Zhao, H., Luo, X. et al. Fagopyrum tataricum FtWD40 Functions as a Positive Regulator of Anthocyanin Biosynthesis in Transgenic Tobacco. J Plant Growth Regul 36, 755–765 (2017). https://doi.org/10.1007/s00344-017-9678-6

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  • DOI: https://doi.org/10.1007/s00344-017-9678-6

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