Skip to main content
SpringerLink
Log in

Drought stress induces oxidative stress and the antioxidant defense system in ascorbate-deficient vtc1 mutants of Arabidopsis thaliana

  • Original Paper
  • Published:
Acta Physiologiae Plantarum Aims and scope Submit manuscript

Abstract

Drought stress has a negative impact on plant cells and results in the generation of reactive oxygen species (ROS). To increase our understanding of the effects of drought stress on antioxidant processes, we investigated the response of the ascorbate-deficient Arabidopsis thaliana vtc1 mutant to drought stress. After drought stress, vtc1 mutants exhibited increases in several oxidative parameters, including H2O2 content and the production of thiobarbituric acid reactive substances. Decreases in chlorophyll content and chlorophyll fluorescence parameters were also observed. The vtc1 mutants had higher total glutathione than did wild-type (WT) plants after 48 h of drought stress. A reduced ratio of glutathione/total glutathione and an increased ratio of dehydroascorbate/total ascorbate were observed in the vtc1 mutants compared with the WT plants. In addition, the activities of enzymes that are responsible for ROS scavenging, including superoxide dismutase, catalase, and ascorbate peroxidase, were decreased in the vtc1 mutants compared with the WT plants. Similar reductions in activity in the vtc1 mutant were observed for the enzymes that are responsible for the regeneration of ascorbate and glutathione, including monodehydroascorbate reductase, dehydroascorbate reductase, and glutathione reductase. These results suggest that low intrinsic ascorbate and impaired ascorbate–glutathione cycling in the vtc1 mutant induced a decrease in the reduced form of ascorbate, which enhanced sensitivity to drought stress.

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
Fig. 6

Similar content being viewed by others

Abbreviations

AsA:

Ascorbate acid

vtc1 :

Vitamin C-deficient mutant-1

ROS:

Reactive oxygen species

SOD:

Superoxide dimutase

APX:

Ascorbate peroxidase

GR:

Glutathione reductase

AGC:

Ascorbate–glutathione cycle

MDHA:

Monodehydroascorbate

DHA:

Dehydroascorbate

DHAR:

Dehydroascorbate reductase

CAT:

Catalase

POD:

Peroxidase

ABA:

Abscisic acid

GA:

Gibberellic acid

NPQ:

Non-photochemical quenching

GMPase:

GDP-mannose pyrophosphorylase

UV-B:

Ultraviolet ray-B

WT:

Wild type

RWC:

Relative water content

Chl:

Chlorophyll

PSII:

Photosystem II

PVP:

Polyvinyl pyrrolidone

DMAB:

Dimethylamine borane

MBTH:

3-Methyl-2-benzothiazolinone hydrazone hydrochloride monohydrate

TBARS:

Thiobarbituric acid reactive substances

TCA:

Trichloroacetic acid

BHT:

Butylated hydroxytoluene

DTT:

Dithiothreitol

EDTA:

Ethylene diamine tetraacetic acid

BSA:

Bovine serum albumin

GSSG:

Oxidized glutathione

References

  • Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126

    Article  PubMed  CAS  Google Scholar 

  • Alpert P (2006) Constrains of tolerance: why are desiccation-tolerant organisms so small or rare? J Exp Biol 209:1575–1584

    Article  PubMed  Google Scholar 

  • Anderson JV, Chevone BI, Hess JL (1992) Seasonal variation in the antioxidant system of eastern white pine needles: evidence for thermal dependence. Plant Physiol 98:501–508

    Article  PubMed  CAS  Google Scholar 

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 24:1–15

    Article  PubMed  CAS  Google Scholar 

  • Arrigoni O, De Tullio MC (2002) Ascorbic acid: much more than just an antioxidant. Biochim Biophys Acta 1569:1–9

    Article  PubMed  CAS  Google Scholar 

  • Asada K (1999) The water–water cycle in chloroplast: scavenging of active oxygens and dissipation of excess photons. Ann Rev Plant Physiol Plant Mol Biol 50:601–639

    Article  CAS  Google Scholar 

  • Attolico AD, De Tullio MC (2006) Increased ascorbate content delays flowering in long-day grown Arabidopsis thaliana (L.) Heynh. Plant Physiol Biochem 44:462–466

    Article  PubMed  CAS  Google Scholar 

  • Barth C, Moeder W, Klessig DF, Conklin PL (2004) The timing of senescence and response to pathogens is altered in the ascorbate-deficient Arabidopsis mutant vitamin c-1. Plant Physiol 134:1784–1792

    Article  PubMed  CAS  Google Scholar 

  • Bilger W, Bjo¨rkman (1990) Role of the xanthophyll cycle in photoprotection elucidated by measurements of light-induced absorbance changes, fluorescence and photosynthesis in Hedera canariensis. Photosynth Res 25:173–185

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Bray EA (2004) Genes commonly regulated by water-deficit stress in Arabidopsis thaliana. J Exp Bot 55:2331–2341

    Article  PubMed  CAS  Google Scholar 

  • Brossa R, Lo'pez-Carbonell M, Jubany-Marı′ T, Alegre L (2011) Interplay between abscisic acid and jasmonic acid and its role in water-oxidative stress in wild-type, ABA-deficient, JA-deficient, and ascorbate-deficient Arabidopsis plants. J Plant Growth Regul 30:322–333

    Article  CAS  Google Scholar 

  • Chaves MM, Oliveira MM (2004) Mechanisms underlying plant resilience to water deficits: prospects for water-saving agriculture. J Exp Bot 55:2365–2384

    Article  PubMed  CAS  Google Scholar 

  • Chen Z, Gallie DR (2006) Dehydroascorbate reductase affects leaf growth, development, and function. Plant Physiol 142:775–787

    Article  PubMed  CAS  Google Scholar 

  • Chen XY, Li W, Lu QT, Wen XG, Li HW, Kuang TY, Li ZS, Lu CM (2011) The xanthophyll cycle and antioxidative defense system are enhanced in the wheat hybrid subjected to high light stress. J Plant Physiol 167:1828–1836

    Article  Google Scholar 

  • Conklin PL, Williams EH, Last RL (1996) Environmental stress sensitivity of an ascorbic acid-deficient Arabidopsis mutant. Proc Natl Acad Sci USA 93:9970–9974

    Article  PubMed  CAS  Google Scholar 

  • Conklin PL, Norris SR, Wheeler GL, Williams EH, Smirnoff N, Last RL (1999) Genetic evidence for the role of GDPmannose in plant ascorbic acid (vitamin C) biosynthesis. Proc Natl Acad Sci USA 96:4198–4203

    Article  PubMed  CAS  Google Scholar 

  • Conklin PL, Saracco SA, Norris SR, Last RL (2000) Identification of ascorbic acid-deficient Arabidopsis thaliana mutants. Genetics 154:847–856

    PubMed  CAS  Google Scholar 

  • Demmig-Adams B, Adams WW, Heber U, Neimanis S, Winter K, Kruger A et al (1990) Inhibition of zeaxanthin formation and of rapid changes in radiationless energy dissipation by dithiothreitol in spinach leaves and chloroplasts. Plant Physiol 92:293–301

    Article  PubMed  CAS  Google Scholar 

  • Draper HH, Hadley M (1990) Malondialdehyde determination as index of lipid peroxidation. Methods Enzymol 186:421–431

    Article  PubMed  CAS  Google Scholar 

  • Foyer CH, Rowell J, Walker D (1983) Measurement of the ascorbate content of spinach leaf protoplasts and chloroplasts during illumination. Planta 157:239–244

    Article  CAS  Google Scholar 

  • Gao Q, Zhang LX (2008) Ultraviolet-B-induced oxidative stress and antioxidant defense system responses in ascorbate-deficient vtc1 mutants of Arabidopsis thaliana. J Plant Physiol 165:138–148

    Article  PubMed  CAS  Google Scholar 

  • Gechev TS, Breusegem FV, Stone JM, Denev I, Laloi C (2006) Reactive oxygen species as signals that modulate plant stress responses and programmed cell death. BioEssays 28:1091–1101

    Article  PubMed  CAS  Google Scholar 

  • Genty B, Briantais JM, Baker NR (1989) The relationship between the quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochim Biophys Acta 990:87–92

    Article  CAS  Google Scholar 

  • Giannopolitis CN, Ries SK (1977) Superoxide dismutases: II. Purification and quantitative relationship with water-soluble protein in seedlings. Plant Physiol 59:315–318

    Article  PubMed  CAS  Google Scholar 

  • Harb A, Krishnan A, Ambavaram MMR, Pereira A (2010) Molecular and physiological analysis of drought stress in Arabidopsis reveals early responses leading to acclimation in plant growth. Plant Physiol 154:1254–1271

    Article  PubMed  CAS  Google Scholar 

  • Hossain MA, Asada K (1984a) Inactivation of ascorbate peroxidase in spinach chloroplasts on dark addition of hydrogen peroxide: its protection by ascorbate. Plant Cell Physiol 25:1285–1295

    CAS  Google Scholar 

  • Hossain MA, Asada K (1984b) Purification of dehydroascorbate reductase from spinach and its characterization as a thiol enzyme. Plant Cell Physiol 25:85–92

    CAS  Google Scholar 

  • Hossain MA, Nakano Y, Asada K (1984) Monodehydroascorbate reductase in spinach chloroplasts and its participation in regeneration of ascorbate for scavenging hydrogen peroxide. Plant Cell Physiol 25:385–395

    CAS  Google Scholar 

  • Huang C, He W, Guo J, Chang X, Su P, Zhang L (2005) Increased sensitivity to salt stress in an ascorbate-deficient Arabidopsis mutant. J Exp Bot 56:3041–3049

    Article  PubMed  CAS  Google Scholar 

  • Jakab G, Ton J, Flors V et al (2005) Enhancing arabidopsis salt and drought stress tolerance by chemical priming for its abscisic acid responses. Plant Physiol 139:267–274

    Article  PubMed  CAS  Google Scholar 

  • Kiddle G, Pastori GM, Bernard S, Pignocchi C, Antoniw J, Verrier PJ et al (2003) Effects of leaf ascorbate content on defense and photosynthesis gene expression in Arabidopsis thaliana. Antioxid Redox Signal 5:23–32

    Article  PubMed  CAS  Google Scholar 

  • Kitajima M, Butler WL (1975) Excitation spectra for photosystem I and photosystem II in chloroplasts and the spectral characteristics of the distributions of quanta between the two photosystems. Biochim Biophys Acta 408:297–305

    Article  PubMed  CAS  Google Scholar 

  • Kwon SY, Choi SM, Ahn YO, Lee HS, Lee HB, Park YM et al (2003) Enhanced stress-tolerance of transgenic tobacco plants expressing a human dehydroascorbate reductase gene. J Plant Physiol 160:347–353

    Article  PubMed  CAS  Google Scholar 

  • Lukowitz W, Nickle TC, Meinke DW, Last RL, Conklin PL, Somerville CR (2001) Arabidopsis cyt1 mutants are deficient in a mannose-1-phosphate guanylyltransferase and point to a requirement of N-linked glycosylation for cellulose biosynthesis. Proc Natl Acad Sci USA 98:2262–2267

    Article  PubMed  CAS  Google Scholar 

  • Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410

    Article  PubMed  CAS  Google Scholar 

  • Muller-Moule P, Conklin PL, Niyogi KK (2002) Ascorbate deficiency can limit violaxanthin de-epoxidase activity in vivo. Plant Physiol 128:970–977

    Article  PubMed  CAS  Google Scholar 

  • Muller-Moule P, Golan T, Niyogi KK (2004) Ascorbate-deficient mutants of Arabidopsis grow in high light despite chronic photooxidative stress. Plant Physiol 134:1163–1172

    Article  PubMed  Google Scholar 

  • Noctor G, Veljovic-Jovanovic S, Foyer CH (2000) Peroxide processing in photosynthesis: antioxidant coupling and redox signalling. Philos Trans R Soc Lon B Biol Sci 355:1465–1475

    Article  PubMed  CAS  Google Scholar 

  • Okuda T, Matsuda Y, Yamanaka A, Sagisaka S (1991) Abrupt increase in the level of hydrogen peroxide in leaves of winter wheat is caused by cold treatment. Plant Physiol 97:1265–1267

    Article  PubMed  CAS  Google Scholar 

  • Owens CW, Belcher RV (1965) A colorimetric micro-method for the determination of glutathione. Biochem J 94:705–711

    PubMed  CAS  Google Scholar 

  • Pastori GM, Kiddle G, Antoniw J, Bernard S, Veljovic-Jovanovic S, Verrier PJ et al (2003) Leaf vitamin C contents modulate plant defense transcripts and regulate genes that control development through hormone signaling. Plant Cell 15:939–951

    Article  PubMed  CAS  Google Scholar 

  • Pignocchi C, Foyer CH (2003) Apoplastic ascorbate metabolism and its role in the regulation of cell signalling. Curr Opin Plant Biol 6:379–389

    Article  PubMed  CAS  Google Scholar 

  • Pignocchi C, Kiddle G, Hernandez I, Foster SJ, Asensi A, Taybi T et al (2006) Ascorbate oxidase-dependent changes in the redox state of the apoplast modulate gene transcript accumulation leading to modified hormone signaling and orchestration of defense processes in tobacco. Plant Physiol 141:423–435

    Article  PubMed  CAS  Google Scholar 

  • Schaedle M, Bassham JA (1977) Chloroplast glutathione reductase. Plant Physiol 59:1011–1012

    Article  PubMed  CAS  Google Scholar 

  • Veljovic-Jovanovic SD, Pignocchi C, Noctor G, Foyer CH (2001) Low ascorbic acid in the vtc-1 mutant of Arabidopsis is associated with decreased growth and intracellular redistribution of the antioxidant system. Plant Physiol 127:426–435

    Article  PubMed  CAS  Google Scholar 

  • Veljovic-Jovanovic S, Noctor G, Foyer CH (2002) Are leaf hydrogen peroxide concentrations commonly overestimated? The potential influence of artefactual interference by tissue phenolics and ascorbate. Plant Physiol Biochem 40:501–507

    Article  CAS  Google Scholar 

  • Wang FZ, Wang QB, Kwon SY, Kwak SS, Su WA (2005) Enhanced drought tolerance of transgenic rice plants expressing a pea manganese superoxide dismutase. J Plant Physiol 162:465–472

    Article  PubMed  CAS  Google Scholar 

  • Willekens H, Chamnongpol S, Davey M, Schraudner M, Langebartels C, van Montagu M, Inzé D, van Camp W (1997) Catalase is a sink for H2O2 and is indispensable for stress defence in C3 plants. EMBO J 16:4806–4816

    Article  PubMed  CAS  Google Scholar 

  • Yannarelli GG, Noriega GO, Batlle A, Tomaro ML (2006) Heme oxygenase up-regulation in ultraviolet-B irradiated soybean plants involves reactive oxygen species. Planta 224:1154–1162

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by program for National Natural Science Foundation (31060063), Gansu National Science Foundation (0803RJZA051), Gansu Provincial Key Laboratory of Aridland Crop Science(GSCS-02).The National Natural Science Foundation of China (No. 30771091 and 30970234), and the Program for New Century Excellent Talents in University (NCET) (2011 to Dr. Liu) by Ministry of Education (MOE).

Author information

Authors and Affiliations

  1. Key Laboratory of Cell Activities and Stress Adaptations, Ministry of Education, School of Life Sciences, Lanzhou University, Lanzhou, 730000, Gansu, China

    Yue Niu, Heng Liu & Guochang Zheng

  2. Gansu Provincial Key Laboratory of Aridland Crop Science, Gansu Key Laboratory of Crop Improvement and Germplasm Enhancement, College of Agronomy, Gansu Agricultural University, Lanzhou, 730070, Gansu, China

    Yuping Wang, Ping Li & Feng Zhang

Authors
  1. Yue Niu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuping Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ping Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Feng Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Heng Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Guochang Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Heng Liu.

Additional information

Communicated by G. Bartosz.

Y. Niu and Y. Wang contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Niu, Y., Wang, Y., Li, P. et al. Drought stress induces oxidative stress and the antioxidant defense system in ascorbate-deficient vtc1 mutants of Arabidopsis thaliana . Acta Physiol Plant 35, 1189–1200 (2013). https://doi.org/10.1007/s11738-012-1158-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11738-012-1158-9

Keywords

Search

Navigation