Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

RcMYBPA2 of Rosa chinensis functions in proanthocyanidin biosynthesis and enhances abiotic stress tolerance in transgenic tobacco


Proanthocyanidins (PAs) are major antioxidant flavonoids that play a key role in protecting plants against adverse environmental stress, but the transcriptional regulation of PAs synthesis in rose has not been fully investigated. Here, we report the functional characterization of RcMYBPA2 from rose (Rosa chinensis). RcMYBPA2 expression was induced by wounding, methyl viologen and salicylic acid. Overexpression of RcMYBPA2 in tobacco (Nicotiana tabacum) led to increased PAs concentrations, as well as enhanced tolerance to oxidative stress in comparison with the wild type (WT). Of special note, the transgenic plants exhibited lower levels of reactive oxygen species (ROS) than the WT, accompanied by higher levels of antioxidant enzyme activity under oxidative conditions. The ROS scavenging ability of the transgenic plants was compromised by inhibitors of antioxidant enzymes. In addition, a range of several stress-responsive genes was also up-regulated in the transgenic tobacco under oxidative stresses. Taken together, it is demonstrated that RcMYBPA2 is a positive regulator of the PAs biosynthetic pathway and participates in oxidative tolerance partly via modulating ROS scavenging ability and stress-responsive genes expression.

Key message

Overexpression of RcMYBPA2 in tobacco led to increased PA content, as well as enhanced tolerance to oxidative stress via scavenging ROS and modulating expression of stress-responsive genes.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6


  1. Abrahams S, Tanner GJ, Larkin PJ, Ashton AR (2002) Identification and biochemical characterization of mutants in the proanthocyanidin pathway in Arabidopsis. Plant Physiol 130:561–576

  2. Akagi T, Ikegami A, Yonemori K (2010) DkMyb2 wound-induced transcription factor of persimmon (Diospyros kaki Thunb.), contributes to proanthocyanidin regulation. Planta 232:1045–1059

  3. An XH, Tian Y, Chen KQ, Liu XJ, Liu DD, Xie XB, Cheng CG, Cong PH, Hao YJ (2015) MdMYB9 and MdMYB11 are involved in the regulation of the JA-induced biosynthesis of anthocyanin and proanthocyanidin in apples. Plant Cell Physiol 56:650–662

  4. Andersen CL, Jensen JL, Orntoft TF (2004) Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 64:5245–5250

  5. Barbehenn RV, Constabel C (2011) Tannins in plant–herbivore interactions. Phytochemistry 72:1551–1565

  6. Bogs J, Jaffe FW, Takos AM, Walker AR, Robinson SP (2007) The grapevine transcription factor VvMYBPA1 regulates proanthocyanidin synthesis during fruit development. Plant Physiol 143:1347–1361

  7. Cos P, De BT, Hermans N, Apers S, Berghe DV, Vlietinck AJ (2004) Proanthocyanidins in health care: current and new trends. Curr Med Chem 11:1345–1359

  8. Dixon RA (2005) Engineering of plant natural product pathways. Curr Opin Plant Biol 8:329–336

  9. Dubois A, Carrere S, Raymond O, Pouvreau B, Cottret L, Roccia A, Onesto J-P, Sakr S, Atanassova R, Baudino S, Foucher F, Bris M, Gouzy J, Bendahmane M (2012) Transcriptome database resource and gene expression atlas for the rose. BMC Genomics 13(1):638

  10. Fischer TC, Mirbeth B, Rentsch J, Sutter C, Ring L, Flachowsky H, Habegger R, Hoffmann T, Hanke MV, Schwab W (2014) Premature and ectopic anthocyanin formation by silencing of anthocyanidin reductase in strawberry (Fragaria × ananassa). N Phytol 201:440–451

  11. Foyer CH, Shigeoka S (2011) Understanding oxidative stress and antioxidant functions to enhance photosynthesis. Plant Physiol 155:93–100

  12. Gong XQ, Zhang JY, Hu JB, Wang W, Wu H, Zhang QH, Liu JH (2015) FcWRKY70, a WRKY protein of Fortunella crassifolia, functions in drought tolerance and modulates putrescine synthesis by regulating arginine decarboxylase gene. Plant Cell Environ 38:2248–2262

  13. Han YP, 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 63:2437–2447

  14. Hancock KR, Collette V, Fraser K, Greig M, Xue H, Richardson K, Jones C, Rasmussen S (2012) Expression of the R2R3-MYB transcription factor TaMYB14 from Trifolium arvense activates proanthocyanidin biosynthesis in the legumes Trifolium repens and Medicago sativa. Plant Physiol 159:1204–1220

  15. Huang XS, Liu JH, Chen XJ (2010) Overexpression of PtrABF gene, a bZIP transcription factor isolated from Poncirus trifoliata, enhances dehydration and drought tolerance in tobacco via scavenging ROS and modulating expression of stress-responsive genes. BMC Plant Biol 10:230

  16. Huang XS, Luo T, Fu XZ, Fan QJ, Liu JH (2011) Cloning and molecular characterization of a mitogen-activated protein kinase gene from Poncirus trifoliata whose ectopic expression confers dehydration/drought tolerance in transgenic tobacco. J Exp Bot 62:5191–5206

  17. Huang QJ, Wang Y, Li B, Chang JL, Chen MJ, Li KX, Yang GX, He GY (2015) TaNAC29, a NAC transcription factor from wheat, enhances salt and drought tolerance in transgenic Arabidopsis. BMC Plant Biol 15:268

  18. Jia LG, Sheng ZW, Xu WF, Li YX, Liu YG, Xia YJ, Zhang JH (2012) Modulation of anti-oxidation ability by proanthocyanidins during germination of Arabidopsis thaliana seeds. Mol Plant 5:472–481

  19. Jin SX, Daniell H (2014) Expression of c-tocopherol methyltransferase in chloroplasts results in massive proliferation of the inner envelope membrane and decreases susceptibility to salt and metal-induced oxidative stresses by reducing reactive oxygen species. Plant Biotechnol J 12:1274–1285

  20. Klie M, Debener T (2011) Identification of superior reference genes for data normalisation of expression studies via quantitative PCR in hybrid roses (Rosa hybrida). BMC Res Notes 4:518

  21. Liao L, Vimolmangkang S, Wei G, Zhou H, Korban SS, Han YP (2015) Molecular characterization of genes encoding leucoanthocyanidin reductase involved in proanthocyanidin biosynthesis in apple. Front Plant Sci 6:243

  22. Lin LC, Kuo YC, Chou CJ (2002) Immunomodulatory proanthocyanidins from Ecdysanthera utilis. J Nat Prod 65:505–508

  23. Liu Y, Shi Z, Maximova S, Payne MJ, Guiltinan MJ (2013) Proanthocyanidin synthesis in Theobroma cacao: genes encoding anthocyanidin synthase, anthocyanidin reductase, and leucoanthocyanidin reductase. BMC Plant Biol 13:202

  24. Liu CG, Jun JH, Dixon RA (2014) MYB5 and MYB14 play pivotal roles in seed coat polymer biosynthesis in Medicago truncatula. Plant Physiol 165:1424–1439

  25. Liu C, Wang X, Shulaev V, Dixon RA (2016) A role for leucoanthocyanidin reductase in the extension of proanthocyanidins. Nature Plants 2:16182

  26. Lu N, Roldan M, Dixon RA (2017) Characterization of two TT2-type MYB transcription factors regulating proanthocyanidin biosynthesis in tetraploid cotton, Gossypium hirsutum. Planta 246:1–13

  27. Luo P, Shen YX, Jin SX, Huang SS, Cheng X, Wang Z, Li PH, Zhao J, Bao MZ, Ning GG (2016) Overexpression of Rosa rugosa anthocyanidin reductase enhances tobacco tolerance to abiotic stress through increased ROS scavenging and modulation of ABA signaling. Plant Sci 245:35–49

  28. Mahajan M, Yadav SK (2014) Overexpression of a tea flavanone 3–hydroxylase gene confers tolerance to salt stress and Alternaria solani in transgenic tobacco. Plant Mol Biol 85:551–573

  29. Mahajan M, Joshi R, Gulati A, Yadav SK (2012) Increase in flavan-3-ols by silencing flavonol synthase mRNA affects the transcript expression and activity levels of antioxidant enzymes in tobacco. Plant Biol 14:725–733

  30. Mellway RD, Tran LT, Prouse MB, Campbell MM, Constabel CP (2009) The wound-, pathogen-, and ultraviolet B-responsive MYB134 gene encodes an R2R3 MYB transcription factor that regulates proanthocyanidin synthesis in poplar. Plant Physiol 150:924–941

  31. Meng Y, Li N, Tian J, Gao J, Zhang C (2013) Identification and validation of reference genes for gene expression studies in postharvest rose flower (Rosa hybrida). Sci Hortic 158:16–21

  32. Naing AH, Park KI, Ai TN, Chung MY, Han JS, Kang YW, Lim KB, Kim CK (2017) Overexpression of snapdragon Delila (Del) gene in tobacco enhances anthocyanin accumulation and abiotic stress tolerance. BMC Plant Biol 17:65

  33. Nakabayashi R, Yonekura-Sakakibara K, Urano K, Suzuki M, Yamada Y, Nishizawa T, Matsuda F, Kojima M, Sakakibara H, Shinozaki K (2014) Enhancement of oxidative and drought tolerance in Arabidopsis by overaccumulation of antioxidant flavonoids. Plant J 77:367

  34. 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

  35. Ning GG, Xiao X, Lv HX, Li X, Zuo Y, Bao MZ (2012) Shortening tobacco life cycle accelerates functional gene identification in genomic research. Plant Biol 14:934–943

  36. Paolocci F, Robbins MP, Madeo L, Arcioni S, Martens S, Damiani F (2007) Ectopic expression of a basic helix-loop-helix gene transactivates parallel pathways of proanthocyanidin biosynthesis. structure, expression analysis, and genetic control of leucoanthocyanidin 4-reductase and anthocyanidin reductase genes in Lotus corniculatus. Plant Physiol 143:504–516

  37. Ravaglia D, Espley RV, Henry-Kirk RA, Andreotti C, Ziosi V, Hellens RP, Costa G, Allan AC (2013) Transcriptional regulation of flavonoid biosynthesis in nectarine (Prunus persica) by a set of R2R3 MYB transcription factors. BMC Plant Biol 13:68

  38. Raymond O, Gouzy J, Just J, Badouin H, Verdenaud M et al (2018) The Rosa genome provides new insights into the domestication of modern roses. Nat Genet 50:772–777

  39. Schaart JG, Dubos C, Romero DLFI, 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 × ananassa) fruits. N Phytol 197:454–467

  40. Sharma SB, Dixon RA (2005) Metabolic engineering of proanthocyanidins by ectopic expression of transcription factors in Arabidopsis thaliana. Plant J 44:62–75

  41. 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

  42. Wang LJ, Ran L, Hou YS, Tian QY, Li CF, Liu R, Fan D, Luo KM (2017) The transcription factor MYB115 contributes to the regulation of proanthocyanidin biosynthesis and enhances fungal resistance in poplar. N Phytol 215:351–367

  43. Xian LH, Sun PP, Hu SS, Wu J, Liu JH (2014) Molecular cloning and characterization of CrNCED1, a gene encoding 9-cis-epoxycarotenoid dioxygenase in Citrus reshni, with functions in tolerance to multiple abiotic stresses. Planta 239:61–77

  44. Xie DY, Sharma SB, Paiva NL, Ferreira D, Dixon RA (2003) Role of anthocyanidin reductase, encoded by BANYULS in plant flavonoid biosynthesis. Science 299:396–399

  45. Xie DY, Sharma SB, Wright E, Wang ZY, Dixon RA (2006) Metabolic engineering of proanthocyanidins through co-expression of anthocyanidin reductase and the PAP1 MYB transcription factor. Plant J 45:895–907

  46. Xing W, Wang Z, Wang XQ, Bao MZ, Ning GG (2014) Over-expression of an FT homolog from Prunus mume reduces juvenile phase and induces early flowering in rugosa rose. Sci Hortic-Amsterdam 72:68–72

  47. Xu W, Lepiniec L, Dubos C (2014a) New insights toward the transcriptional engineering of proanthocyanidin biosynthesis. Plant Signal Behav 9:e28736

  48. Yoshida K, Ma D, Constabel CP (2015) The MYB182 protein down-regulates proanthocyanidin and anthocyanin biosynthesis in poplar by repressing both structural and regulatory flavonoid genes. Plant Physiol 167:693–710

  49. Yuan L, Wang L, Han Z, Jiang Y, Zhao L, Liu H, Yang L, Luo K (2012) Molecular cloning and characterization of PtrLAR3, a gene encoding leucoanthocyanidin reductase from Populus trichocarpa, and its constitutive expression enhances fungal resistance in transgenic plants. J Exp Bot 63:2513–2524

  50. Zhang W, Ning GG, Lv HY, Liao L, Bao MZ (2009) Single MYB-type transcription factor AtCAPRICE: a new efficient tool to engineer the production of anthocyanin in tobacco. Biochem Biophys Res Commun 388:742–747

  51. Zhou H, Wang KL, Liao L, Gu C, Lu ZQ, Allan AC, Han YP (2015) Peach MYB7 activates transcription of the proanthocyanidin pathway gene encoding leucoanthocyanidin reductase, but not anthocyanidin reductase. Front Plant Sci 6:908

Download references


This work was supported by grants from the National Natural Science Foundation of China (Grant No. 31800598), the Natural Science Foundation of Zhejiang Province (Grant Nos. LQ18C150001, LY17C150004) and the Fundamental Research Funds for the Zhejiang A & F University (Grant No. 2016FR033). We thank all the colleagues in our lab for constructive discussion and technical support.


The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Author information

CYY and LP conceived and designed the overall experiment. LZY and CW performed the most of the experiments and data analysis. ZC, DCX and SGY participated in tobacco transformation and physiological determination; LP, LZY and CW wrote the manuscript; CYY check the manuscript.

Correspondence to Yongyi Cui or Ping Luo.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Communicated by T. Winkelmann.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 967 KB)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Li, Z., Chen, W., Zhang, C. et al. RcMYBPA2 of Rosa chinensis functions in proanthocyanidin biosynthesis and enhances abiotic stress tolerance in transgenic tobacco. Plant Cell Tiss Organ Cult 137, 441–454 (2019).

Download citation


  • Rose
  • RcMYBPA2
  • Proanthocyanidins
  • Oxidative tolerance
  • Transgenic tobacco