Skip to main content
SpringerLink
Log in

Drastic anthocyanin increase in response to PAP1 overexpression in fls1 knockout mutant confers enhanced osmotic stress tolerance in Arabidopsis thaliana

  • Original Article
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

Key message

pap1 - D/fls1ko double mutant plants that produce substantial amounts of anthocyanin show tolerance to abiotic stress.

Abstract

Anthocyanins are flavonoids that are abundant in various plants and have beneficial effects on both plants and humans. Many genes in flavonoid biosynthetic pathways have been identified, including those in the MYB-bHLH-WD40 (MBW) complex. The MYB gene Production of Anthocyanin Pigment 1 (PAP1) plays a particularly important role in anthocyanin accumulation. PAP1 expression in many plant systems strongly increases anthocyanin levels, resulting in a dark purple color in many plant organs. In this study, we generated double mutant plants that harbor fls1ko in the pap1-D background (i.e., pap1-D/fls1ko plants), to examine whether anthocyanins can be further enhanced by blocking flavonol biosynthesis under PAP1 overexpression. We also wanted to examine whether the increased anthocyanin levels contribute to defense against osmotic stresses. The pap1-D/fls1ko mutants accumulated higher anthocyanin levels than pap1-D plants in both control and sucrose-treated conditions. However, flavonoid biosynthesis genes were slightly down-regulated in the pap1-D/fls1ko seedlings as compared to their expression in pap1-D seedlings. We also report the performance of pap1-D/fls1ko seedlings in response to plant osmotic stresses.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Ahn D, Lee EB, Kim BJ, Lee SY, Lee TG, Ahn M, Lim HW, Cha DS, Jeon H, Kim DK (2014) Antioxidant and lifespan extending property of quercetin-3-O-dirhamnoside from Curcuma longa L. in Caenorhabditis elegans. J Korean Soc Appl Biol Chem 57:709–714

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Allan AC, Hellens RP, Laing WA (2008) MYB transcription factors that color our fruit. Trends Plant Sci 13:99–102

    Article  CAS  PubMed  Google Scholar 

  • Baek YS, Song NY, Nam TG, Kim DO, Kang HC, Kwon OK, Baek NI (2015) Flavonoids from Fragaria ananassa calyx and their antioxidant capacities. J Korean Soc Appl Biol Chem 58:787–793

    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 

  • Borevitz JO, Xia Y, Blount J, Dixon RA, Lamb C (2000) Activation tagging identifies a conserved MYB regulator of phenylpropanoid biosynthesis. Plant Cell 12:2383–2393

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Broeckling BE, Watson RA, Steinwand B, Bush DR (2016) Intronic sequence regulates sugar-dependent expression of Arabidopsis thaliana production of anthocyanin pigment-1/MYB75. PLoS One 11:e0156673. doi:10.1371/journal.pone.0156673.eCollection

    Article  PubMed  PubMed Central  Google Scholar 

  • Broun P (2005) Transcriptional control of flavonoid biosynthesis: a complex network of conserved regulators involved in multiple aspects of differentiation in Arabidopsis. Curr Opin Plant Biol 8:272–279

    Article  CAS  PubMed  Google Scholar 

  • Coombe BG, Hale CR (1973) The hormone content of ripening grape berries and the effects of growth substances treatments. Plant Physiol 51:629–634

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dixon RA, Xie DY, Sharma SB (2005) Proanthocyanidins—a final frontier in flavonoid research? New Phytol 165:9–28

    Article  CAS  PubMed  Google Scholar 

  • Gabetta B, Fuzzati N, Griffini A, Lolla E, Pace R, Ruffilli T, Peterlongo F (2000) Characterization of proanthocyanidins from grape seeds. Fitoterapia 71:162–175

    Article  CAS  PubMed  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 

  • Guo R, Yuan G, Wang Q (2011) Sucrose enhances the accumulation of anthocyanins and glucosinolates in broccoli sprouts. Food Chem 129:1080–1087

    Article  CAS  PubMed  Google Scholar 

  • He J, Giusti MM (2010) Anthocyanins: natural colorants with health-promoting properties. Annu Rev Food Sci Technol 1:163–187

    Article  CAS  PubMed  Google Scholar 

  • Jeong SW, Das PK, Jeoung SC, Song JY, Lee HK, Kim YK, Choi G (2010) Ethylene suppression of sugar-induced anthocyanin pigmentation in Arabidopsis. Plant Physiol 154:1514–1531

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jung S (2004) Effect of chlorophyll reduction in Arabidopsis thaliana by methyl jasmonate or norflurazon on antioxidant systems. Plant Physiol Biochem 42:225–231

    Article  CAS  PubMed  Google Scholar 

  • Kang YH, Kirik V, Hulskamp M, Nam KH, Hagely K, Lee MM, Schiefelbein J (2009) The MYB23 gene provides a positive feedback loop for cell fate specification in the Arabidopsis root epidermis. Plant Cell 21:1080–1094

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lee K, Jung YJ, Shin SY, Lee YH (2016) The natural flavone eupatorin induces cell cycle arrest at the G2/M phase and apoptosis in HeLa cells. Appl Biol Chem 59:193–199

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Li S (2014) Transcriptional control of flavonoid biosynthesis: fine-tuning of the MYB-bHLH-WD40 (MBW) complex. Plant Signal Behav 9:e27522. doi:10.4161/psb.27522

    Article  PubMed  PubMed Central  Google Scholar 

  • Lin S, Zhou Z, Zhang H, Yin W (2015) Phenolic glycosides from the rhizomes of Cyperus rotundus and their antidepressant activity. J Korean Soc Appl Biol Chem 58:685–691

    Article  CAS  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 10:50–66

    Article  PubMed  PubMed Central  Google Scholar 

  • Morita Y, Saitoh M, Hoshino A, Nitasaka E, Iida S (2006) Isolation of cDNAs for R2R3-MYB, bHLH and WDR transcriptional regulators and identification of c and ca mutations conferring white flowers in the Japanese morning glory. Plant Cell Physiol 47:457–470

    Article  CAS  PubMed  Google Scholar 

  • Murphy A, Peer WA, Taiz L (2000) Regulation of auxin transport by aminopeptidases and endogenous flavonoids. Planta 211:315–324

    Article  CAS  PubMed  Google Scholar 

  • Nakabayashi R, Yonekura-Sakakibara K, Urano K, Suzuki M, Yamada Y, Nishizawa T, Matsuda F, Kojima M, Sakakibara H, Shinozaki K, Michael AJ, Tohge T, Yamazaki M, Saito K (2014) Enhancement of oxidative and drought tolerance in Arabidopsis by overaccumulation of antioxidant flavonoids. Plant J 77:367–379

    Article  CAS  PubMed  Google Scholar 

  • Nguyen NH, Kim JH, Kwon J, Jeong CY, Lee W, Lee D, Hong SW, Lee H (2016) Characterization of Arabidopsis thaliana FLAVONOL SYNTHASE 1 (FLS1) -overexpression plants in response to abiotic stress. Plant Physiol Biochem 103:133–142

    Article  CAS  PubMed  Google Scholar 

  • Page M, Sultana N, Paszkiewicz K, Florance H, Smirnoff N (2012) The influence of ascorbate on anthocyanin accumulation during high light acclimation in Arabidopsis thaliana: further evidence for redox control of anthocyanin synthesis. Plant Cell Environ 35:388–404

    Article  CAS  PubMed  Google Scholar 

  • Peer WA, Brown DE, Tague BW, Muday GK, Taiz L, Murphy AS (2001) Flavonoid accumulation patterns of transparent testa mutants of Arabidopsis. Plant Physiol 126:536–548

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Petroni K, Tonelli C (2011) Recent advances on the regulation of anthocyanin synthesis in reproductive organs. Plant Sci 181:219–229

    Article  CAS  PubMed  Google Scholar 

  • Piao HL, Lim JH, Kim SJ, Cheong GW, Hwang I (2001) Constitutive over-expression of AtGSK1 induces NaCl stress responses in the absence of NaCl stress and results in enhanced NaCl tolerance in Arabidopsis. Plant J 27:305–314

    Article  CAS  PubMed  Google Scholar 

  • Pirie A, Mullins MG (1976) Changes in anthocyanin and phenolics content of grapevine leaf and fruit tissues treated with sucrose, nitrate, and abscisic acid. Plant Physiol 58:468–472

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Santos-Buelga C, Scalbert A (2000) Proanthocyanidins and tannin-like compounds—nature, occurrence, dietary intake and effects on nutrition and health. J Sci Food Agric 80:1094–1117

    Article  CAS  Google Scholar 

  • Serna L, Martin C (2006) Trichomes: different regulatory networks lead to convergent structures. Trends Plant Sci 11:274–280

    Article  CAS  PubMed  Google Scholar 

  • Sheahan JJ, Rechnitz GA (1993) Differential visualization of transparent testa mutants in Arabidopsis thaliana. Anal Chem 65:961–963

    Article  CAS  Google Scholar 

  • Solfanelli C, Poggi A, Loreti E, Alpi A, Perata P (2006) Sucrose-specific induction of the anthocyanin biosynthetic pathway in Arabidopsis. Plant Physiol 140:637–646

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Stracke R, Werber M, Weisshaar B (2001) The R2R3-MYB gene family in Arabidopsis thaliana. Curr Opin Plant Biol 4:447–456

    Article  CAS  PubMed  Google Scholar 

  • Stracke R, Ishihara H, Huep G, Barsch A, Mehrtens F, Niehaus K, Weisshaar B (2007) Differential regulation of closely related R2R3-MYB transcription factors controls flavonol accumulation in different parts of the Arabidopsis thaliana seedling. Plant J 50:660–677

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Stracke R, De Vos RC, Bartelniewoehner L, Ishihara H, Sagasser M, Martens S, Weisshaar B (2009) Metabolomic and genetic analyses of flavonol synthesis in Arabidopsis thaliana support the in vivo involvement of leucoanthocyanidin dioxygenase. Planta 229:427–445

    Article  CAS  PubMed  Google Scholar 

  • Telias A, Lin-Wang K, Stevenson DE, Cooney JM, Hellens RP, Allan AC, Hoover EE, Bradeen JM (2011) Apple skin patterning is associated with differential expression of MYB10. BMC Plant Biol 11:93–107

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Teng S, Keurentjes J, Bentsink L, Koornneef M, Smeekens S (2005) Sucrose-specific induction of anthocyanin biosynthesis in Arabidopsis requires the MYB75/PAP1 gene. Plant Physiol 139:1840–1852

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tohge T, Nishiyama Y, Hirai MY, Yano M, Nakajima JI, Awazuhara M, Inoue E, Takahashi H, Goodenowe DB, Kitayama M, Noji M, Yamazaki M, Saito K (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 

  • Weigel D, Ahn JH, Blázquez MA, Borevitz JO, Christensen SK, Fankhauser C, Ferrándiz C, Kardailsky I, Malancharuvil EJ, Neff MM, Nguyen JT, Sato S, Wang ZY, Xia Y, Dixon RA, Harrison MJ, Lamb CJ, Yanofsky MF, Chory J (2000) Activation tagging in Arabidopsis. Plant Physiol 122:1003–1014

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xu W, Grain D, Le Gourrierec J, Harscoët E, Berger A, Jauvion V, Scagnelli A, Berger N, Bidzinski P, Kelemen Z (2013a) Regulation of flavonoid biosynthesis involves an unexpected complex transcriptional regulation of TT8 expression, in Arabidopsis. New Phytol 198:59–70

    Article  CAS  PubMed  Google Scholar 

  • Xu ZY, Kim SY, Kim DH, Dong T, Park Y, Jin JB, Hwang I (2013b) The Arabidopsis NAC transcription factor ANAC096 cooperates with bZIP-type transcription factors in dehydration and osmotic stress responses. Plant Cell 25:4708–4724

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xu W, Grain D, Bobet S, Le Gourrierec J, Thévenin J, Kelemen Z, Lepiniec L, Dubos C (2014) Complexity and robustness of the flavonoid transcriptional regulatory network revealed by comprehensive analyses of MYB-bHLH-WDR complexes and their targets in Arabidopsis seed. New Phytol 202:132–144

    Article  CAS  PubMed  Google Scholar 

  • Xu W, Dubos C, Lepiniec L (2015) Transcriptional control of flavonoid biosynthesis by MYB-bHLH-WDR complexes. Trends Plant Sci 20:176–185

    Article  CAS  PubMed  Google Scholar 

  • Yurina PN, Kloppstech K (2001) Accumulation of plastid protein precursors under Norflurazon-induced carotenoid deficiency and oxidative stress in barley. Plant Physiol Biochem 39:807–814

    Article  CAS  Google Scholar 

  • Zhao M, Morohashi K, Hatlestad G, Grotewold E, Lloyd A (2008) The TTG1-bHLH-MYB complex controls trichome cell fate and patterning through direct targeting of regulatory loci. Development 135:1991–1999

    Article  CAS  PubMed  Google Scholar 

  • Zimmermann IM, Heim MA, Weisshaar B, Uhrig JF (2004) Comprehensive identification of Arabidopsis thaliana MYB transcription factors interacting with R/B-like BHLH proteins. Plant J 40:22–34

    Article  CAS  PubMed  Google Scholar 

  • Zvi MM, Shklarman E, Masci T, Kalev H, Debener T, Shafir S, Ovadis M, Vainstein A (2012) PAP1 transcription factor enhances production of phenylpropanoid and terpenoid scent compounds in rose flowers. New Phytol 195:335–345

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by a grant from the National Research Foundation of Korea (to Hojoung Lee, 2014; Grant NRF-2014R1A1A3050272 and to Suk-Whan Hong, 2015: Grant NRF-2015RIDIAIA01058810).

Author information

Authors and Affiliations

  1. Department of Biosystems and Biotechnology, College of Life Sciences and Biotechnology, Korea University, Anam-dong 5-ga, Seongbuk-gu, Seoul, 136-713, Republic of Korea

    Won Je Lee, Chan Young Jeong, Jaeyoung Kwon, Dongho Lee & Hojoung Lee

  2. Institute of Life Science and Natural Resources, Korea University, Seoul, 136-713, Republic of Korea

    Won Je Lee, Chan Young Jeong & Hojoung Lee

  3. Department of Molecular Biotechnology, College of Agriculture and Life Sciences, Bioenergy Research Center, Chonnam National University, Gwangju, Republic of Korea

    Vu Van Kien & Suk-Whan Hong

Authors
  1. Won Je Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chan Young Jeong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jaeyoung Kwon
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vu Van Kien
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Dongho Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Suk-Whan Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hojoung Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Suk-Whan Hong or Hojoung Lee.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interests.

Additional information

Communicated by J. S. Shin.

Electronic supplementary material

Below is the link to the electronic supplementary material.

299_2016_2040_MOESM1_ESM.pdf

Fig. S1 Growth performance of Col-0, fls1ko, pap1-D, and pap1-D/fls1ko in soil. Col-0, fls1ko, pap1-D, and pap1-D/fls1ko were grown on soil for 37 days for the observation of growth performance. Before being transferred to pots, plants were sown on 2 % MS medium lacking stress inducers. Four pots were used in this experiment (PDF 123 kb)

299_2016_2040_MOESM2_ESM.pdf

Fig. S2. Final period of the life cycle of Col-0 wild-type, pap1-D/fls1ko, pap1-D, and fls1ko plants. Phenotypic performance of plants after approximately 2 months’ growth in pots. Before plants were transferred to pots, they were sown on 2 % MS medium lacking stress inducers. This experiment was carried out approximately three times (PDF 69 kb)

Table S1. Primer information. These primer sequences obtained from the ‘TAIR’ site were used for qRT-PCR (PDF 18 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lee, W.J., Jeong, C.Y., Kwon, J. et al. Drastic anthocyanin increase in response to PAP1 overexpression in fls1 knockout mutant confers enhanced osmotic stress tolerance in Arabidopsis thaliana . Plant Cell Rep 35, 2369–2379 (2016). https://doi.org/10.1007/s00299-016-2040-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-016-2040-9

Keywords

Search

Navigation