Reactive Oxygen Species Acts as an Important Inducer in Low-temperature-induced Anthocyanin Biosynthesis in Begonia semperflorens

in Journal of the American Society for Horticultural Science

Leaves of Begonia semperflorens accumulate anthocyanins and turn red under low temperature (LT). In the present work, LT increased H2O2 content and superoxide anions production rate, causing significant increases in the activities of enzymes and contents of reduced components involved in the ascorbate-glutathione cycle (AsA-GSH cycle). As a result, LT-exposed seedlings increased the expression of genes involved in anthocyanin biosynthesis, and accumulated anthocyanin. Based on LT condition, application of N,N'-dimethylthiourea (DMTU) decreased reactive oxygen species (ROS) content, and unbalanced the AsA-GSH-controlled redox homeostasis. As a result, seedlings in the LT + DMTU group did not accumulate anthocyanin. Our results suggest that ROS may act as an important inducer in LT-induced anthocyanin biosynthesis.

Contributor Notes

This research was supported by the Natural Science Foundation of Henan Province (Grant No. 182300410091) and the Key Scientific Research Project of High Education in Henan Province (Grant No. 18B220004).

We are grateful to the three anonymous reviewers for their constructive comments and suggestions.

These authors contributed equally to this work.

Corresponding author. E-mail: K.M.Zhang@henau.edu.cn.

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Article Figures

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    The dose-dependent response of hydrogen peroxide (H2O2) contents (A and B), superoxide anions (O2•-) production rate (C and D), and anthocyanin contents (E and F) to exogenous N,N'-dimethylthiourea [DMTU (1, 2, 5, and 10 mm)] and methylviologen [MV (0.005, 0.05, 0.1, and 0.5 mm)] in leaves of Begonia semperflorens. Leaf samples were harvested at 0900 HR after different treatments for 0, 1, 3, 6, 9, and 15 d. Values and error bars indicate means ±sd (n = 3–6). Means denoted by the same letter do not significantly differ at P < 0.05 in a single group according to Tukey’s test.

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    Effects of low temperature [LT (15/5 °C day/night)] and N,N'-dimethylthiourea [DMTU (5 mm)] on relative transcript levels for four key genes in anthocyanin biosynthesis [BsPAL (A), BsCHS (B), BsF3H (C), and BsANS (D)], hydrogen peroxide (H2O2) contents (E), superoxide anions (O2•-) production rate (F), and anthocyanin contents (G) in leaves of Begonia semperflorens. Leaf samples were harvested on day 12. Seedlings were sprayed with distilled water and grown at 25/15 °C day/night (CK); seedlings were sprayed with distilled water and grown at 15/5 °C day/night (LT); seedlings were sprayed with DMTU (every other day, 5 mm) and grown at 25/15 °C day/night (DMTU); seedlings were sprayed with DMTU (every other day, 5 mm) and grown at 15/5 °C day/night (LT + DMTU). Values and error bars indicate means ±sd (n = 3–6). Means denoted by the same letter do not significantly differ at P < 0.05 in a single group according to Tukey’s test.

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    Effects of low temperature [LT (15/5 °C day/night)] and N,N'-dimethylthiourea [DMTU (5 mm)] on activities of superoxide dismutase [SOD (A)], catalase [CAT (B)], monodehydroascorbate reductase [MDHAR (C)], dehydroascorbate reductase [DHAR (D)], ascorbate peroxidase [APX (E)], and glutathione reductase [GR (F)] in leaves of Begonia semperflorens. Leaf samples were harvested on day 12. Seedlings were sprayed with distilled water and grown at 25/15 °C day/night (CK); seedlings were sprayed with distilled water and grown at 15/5 °C day/night (LT); seedlings were sprayed with DMTU (every other day, 5 mm) and grown at 25/15 °C day/night (DMTU); seedlings were sprayed with DMTU (every other day, 5 mm) and grown at 15/5 °C day/night (LT + DMTU). Values and error bars indicate means ±sd (n = 3–6). Means denoted by the same letter do not significantly differ at P < 0.05 in a single group according to Tukey’s test.

Article References

  • ArrigoniO.DipierroS.BorraccinoG.1981Ascorbate free radical reductase, a key enzyme of the ascorbic acid systemFEBS Lett.125242244

  • BradfordM.M.1976A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnal. Biochem.72248254

    • Search Google Scholar
    • Export Citation
  • BrennanT.FrenkelC.1977Involvement of hydrogen peroxide in the regulation of senescence in pearPlant Physiol.59411416

  • ChoudhuryS.PandaP.SahooL.PandaS.K.2014Reactive oxygen species signaling in plants under abiotic stressPlant Signal. Behav.8e23681

  • DaltonD.A.RussellS.A.HanusF.J.PascoeG.A.EvansH.J.1986Enzymatic reactions of ascorbate and glutathione that prevent peroxide damage in soybean root nodulesProc. Natl. Acad. Sci. USA8338113815

    • Search Google Scholar
    • Export Citation
  • DasP.K.GeulB.ChoiS.B.YooS.D.ParkY.I.2014Photosynthesis-dependent anthocyanin pigmentation in ArabidopsisPlant Signal. Behav.62325

  • de HaanJ.B.CristianoF.IannelloR.C.KolaI.1995Cu/Zn-superoxide dismutase and glutathione peroxidase during agingBiochem. Mol. Biol. Intl.3512811297

    • Search Google Scholar
    • Export Citation
  • DixonR.SteeleC.1999Flavonoids and isoflavonoids - A gold mine for metabolic engineeringTrends Plant Sci.4394

  • ElstnerE.F.HeupelA.1976Inhibition of nitrite formation from hydroxylammoniumchloride: A simple assay for superoxide dismutaseAnal. Biochem.70616620

    • Search Google Scholar
    • Export Citation
  • FoyerC.H.HalliwellB.1976The presence of glutathione and glutathione reductase in chloroplasts: A proposed role in ascorbic acid metabolismPlanta1332125

    • Search Google Scholar
    • Export Citation
  • GibonY.LarherF.1997Cycling assay for nicotinamide adenine dinucleotides: NaCl precipitation and ethanol solubilization of the reduced tetrazoliumAnal. Biochem.251153

    • Search Google Scholar
    • Export Citation
  • GuX.ChenY.GaoZ.QiaoY.WangX.2015Transcription factors and anthocyanin genes related to low-temperature tolerance in rd29A:RdreB1BI transgenic strawberryPlant Physiol. Biochem.893143

    • Search Google Scholar
    • Export Citation
  • HungK.T.ChengD.G.HsuY.T.KaoC.H.2008Abscisic acid-induced hydrogen peroxide is required for anthocyanin accumulation in leaves of rice seedlingsJ. Plant Physiol.16512801287

    • Search Google Scholar
    • Export Citation
  • HughesN.M.NeufeldH.S.BurkeyK.O.2005Functional role of anthocyanins in high-light winter leaves of the evergreen herb Galax urceolataNew Phytol.168575587

    • Search Google Scholar
    • Export Citation
  • KimS.HwangG.LeeS.ZhuJ.Y.PaikI.NguyenT.T.KimJ.OhE.2017High ambient temperature represses anthocyanin biosynthesis through degradation of HY5Front. Plant Sci.81787

    • Search Google Scholar
    • Export Citation
  • LawM.Y.CharlesS.A.HalliwellB.1983Glutathione and ascorbic acid in spinach (Spinacia oleracea) chloroplasts. The effect of hydrogen peroxide and of paraquatBiochem. J.210899

    • Search Google Scholar
    • Export Citation
  • LiB.XiaY.WangY.QinG.TianS.2017Characterization of genes encoding key enzymes involved in anthocyanin metabolism of kiwifruit during storage periodFront. Plant Sci.8e29526

    • Search Google Scholar
    • Export Citation
  • LiY.L.LiuY.ZhangJ.2010Advances in the research on the AsA-GSH cycle in horticultural cropsFront. Agric. China48490

  • LiaoZ.ChenM.GuoL.GongY.TangF.SunX.TangK.2004Rapid isolation of high-quality total RNA from taxus and ginkgoPrep. Biochem. Biotechnol.34209214

    • Search Google Scholar
    • Export Citation
  • LivakK.J.SchmittgenT.D.2001Analysis of relative gene expression data using Real-Time quantitative PCR and the 2−ΔΔCT methodMethods25402408

    • Search Google Scholar
    • Export Citation
  • Lo PieroA.R.2015The state of the art in biosynthesis of anthocyanins and its regulation in pigmented sweet oranges [(Citrus sinensis) L. Osbeck]J. Agr. Food Chem.6340314041

    • Search Google Scholar
    • Export Citation
  • MarutaT.NoshiM.NakamuraM.MatsudaS.TamoiM.IshikawaT.ShigeokaS.2014Ferulic acid 5-hydroxylase 1 is essential for expression of anthocyanin biosynthesis-associated genes and anthocyanin accumulation under photooxidative stress in ArabidopsisPlant Sci.219-2206168

    • Search Google Scholar
    • Export Citation
  • MitaS.MuranoN.AkaikeM.NakamuraK.1997Mutants of Arabidopsis thaliana with pleiotropic effects on the expression of the gene for β-amylase and on the accumulation of anthocyanin that are inducible by sugarsPlant J.11841851

    • Search Google Scholar
    • Export Citation
  • NakanoY.AsadaK.1981Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplastsPlant Cell Physiol.22867880

  • NoctorG.FoyerC.H.2016Intracellular redox compartmentation and ROS-related communication in regulation and signalingPlant Physiol.17115811592

    • Search Google Scholar
    • Export Citation
  • NorrisK.M.OkieW.YakaitisC.L.PazdroR.2016The anthocyanin cyanidin-3-O-β-glucoside modulates murine glutathione homeosta sis in a manner dependent on genetic backgroundRedox Biol.9254263

    • Search Google Scholar
    • Export Citation
  • PageM.SultanaN.PaszkiewiczK.FloranceH.SmirnoffN.2012The influence of ascorbate on anthocyanin accumulation during high light acclimation in Arabidopsis thaliana: Further evidence for redox control of anthocyanin synthesisPlant Cell Environ.35388404

    • Search Google Scholar
    • Export Citation
  • PaglianoC.RavioloM.Dalla VecchiaF.GabbrielliR.GonnelliC.RascioN.BarbatoR.La RoccaN.2006Evidence for PSII donor-side damage and photoinhibition induced by cadmium treatment on rice (Oryza sativa L.)J. Photochem. Photobiol. B847078

    • Search Google Scholar
    • Export Citation
  • RaoM.V.OrmrodD.P.1995Ozone exposure decreases UV-B sensitivity in a UV-B sensitive flavonoid mutant of ArabidopsisPhotochem. Photobiol.617178

    • Search Google Scholar
    • Export Citation
  • ReczekC.R.ChandelN.S.2015ROS-dependent signal transductionCurr. Opin. Cell Biol.33813

  • RheeS.G.WooH.A.KangD.2017The role of peroxiredoxins in the transduction of H2O2 signalsAntioxid. Redox Signal.28537557

  • SilvaP.FerreiraS.NunesF.M.2017Elderberry (Sambucus nigra L.) by-products a source of anthocyanins and antioxidant polyphenolsInd. Crops Prod.95227234

    • Search Google Scholar
    • Export Citation
  • SteynW.WandS.HolcroftD.JacobsG.2002Anthocyanins in vegetative tissues: A proposed unified function in photoprotectionNew Phytol.155349361

    • Search Google Scholar
    • Export Citation
  • TakosA.M.JaffeF.W.JacobS.R.BogsJ.RobinsonS.P.WalkerA.R.2006Light-induced expression of a MYB gene regulates anthocyanin biosynthesis in red applesPlant Physiol.14212161232

    • Search Google Scholar
    • Export Citation
  • TangQ.Y.FengM.G.2002DPS data processing system for practical statistics Sci. Press Beijing China

  • WinglerA.2015Comparison of signaling interactions determining annual and perennial plant growth in response to low temperatureFront. Plant Sci.519

    • Search Google Scholar
    • Export Citation
  • WuK.KimJ.LeeH.J.JungY.J.KangK.K.TyagiW.KovachM.SweeneyM.McCouchS.ChoY.G.2017Functional properties of an alternative, tissue-specific promoter for rice NADPH-dependent dihydroflavonol reductasePLoS One12e0183722

    • Search Google Scholar
    • Export Citation
  • XuZ.MahmoodK.RothsteinS.J.2017ROS induces anthocyanin production via late biosynthetic genes and anthocyanin deficiency confers the hypersensitivity to ROS-generating stresses in ArabidopsisPlant Cell Physiol.5813641377

    • Search Google Scholar
    • Export Citation
  • ZhangC.JiaH.WuW.WangX.FangJ.WangC.2015bFunctional conservation analysis and expression modes of grape anthocyanin synthesis genes responsive to low temperature stressGene574168177

    • Search Google Scholar
    • Export Citation
  • ZhangJ.ChenC.ZhangD.LiH.LiP.MaF.2014Reactive oxygen species produced via plasma membrane NADPH oxidase regulate anthocyanin synthesis in apple peelPlanta24010231035

    • Search Google Scholar
    • Export Citation
  • ZhangK.M.GuoM.L.HeD.WuR.H.LiY.H.2015aThe inhibition effect and excessive carbon flux resulting from blocking anthocyanin biosynthesis under darkness in Begonia semperflorensJ. Plant Growth Regul.352230

    • Search Google Scholar
    • Export Citation
  • ZhangK.M.LiZ.LiY.LiY.H.KongD.Z.WuR.H.PicherskyE.2013Carbohydrate accumulation may be the proximate trigger of anthocyanin biosynthesis under autumn conditions in Begonia semperflorensPlant Biol.159911000

    • Search Google Scholar
    • Export Citation
  • ZhangK.M.WangJ.W.GuoM.L.DuW.L.WuR.H.WangX.2016Short-day signals are crucial for the induction of anthocyanin biosynthesis in Begonia semperflorens under low temperature conditionJ. Plant Physiol.20417

    • Search Google Scholar
    • Export Citation
  • ZhangY.ZhengS.LiuZ.WangL.BiY.2011Both HY5 and HYH are necessary regulators for low temperature-induced anthocyanin accumulation in Arabidopsis seedlingsJ. Plant Physiol.168367374

    • Search Google Scholar
    • Export Citation
  • ZhouJ.WangJ.LiX.XiaoX.J.ZhouY.H.ShiK.ChenZ.YuJ.Q.2014H2O2 mediates the crosstalk of brassinosteroid and abscisic acid in tomato responses to heat and oxidative stressesJ. Expt. Bot.6543714383

    • Search Google Scholar
    • Export Citation
  • ZhouJ.WangJ.ShiK.XiaoX.J.ZhouY.H.YuJ.Q.2012Hydrogen peroxide is involved in the cold acclimation-induced chilling tolerance of tomato plantsPlant Physiol. Biochem.60141149

    • Search Google Scholar
    • Export Citation
  • ZhouJ.XiaoX.J.ZhouY.H.ShiK.ChenZ.YuJ.Q.2015RBOH1-dependent H2O2 production and subsequent activation of MPK1/2 play an important role in acclimation-induced cross-tolerance in tomatoJ. Expt. Bot.65595607

    • Search Google Scholar
    • Export Citation
  • ZhouY.H.HuangL.F.DuY.S.YuJ.Q.2004Greenhouse and field cucumber genotypes use different mechanisms to protect against dark chillingFunct. Plant Biol.311215

    • Search Google Scholar
    • Export Citation
  • ZhuH.ZhangT.J.ZhangP.PengC.L.SchnitzlerJ.P.2016Pigment patterns and photoprotection of anthocyanins in the young leaves of four dominant subtropical forest tree species in two successional stages under contrasting light conditionsTree Physiol.3610921104

    • Search Google Scholar
    • Export Citation

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