Skip to main content
SpringerLink
Log in

Implications of paraquat and hydrogen peroxide-induced oxidative stress treatments on the GABA shunt pathway in Arabidopsis thaliana calmodulin mutants

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

Abstract

Arabidopsis mutants with T-DNA insertion in seven calmodulin genes (CAM) were used to determine the specific role of CAM in the tolerance of plants to oxidative stress induced by paraquat and hydrogen peroxide (H2O2) treatments. Arabidopsis calmodulin mutants (cam) were screened for seedling growth, seed germination, induced oxidative damage, and levels of γ-aminobutyric acid (GABA) shunt metabolites. Only the cam5-4 and cam6-1 mutants exhibited an increased sensitivity to paraquat and H2O2 during seed germination and seedling growth. In response to treatments with 3 μM paraquat and 1 mM H2O2, only the cam5-4, cam6-1 mutants showed significant changes in malonaldehyde (MDA) levels in root and shoot tissues, with highly increased levels of MDA. In terms of the GABA shunt metabolites, GABA was significantly elevated in root and shoot tissues in response to the paraquat treatments in comparison to alanine and glutamate, while the levels of all shunt metabolites increased in root tissue but not in the shoot tissue following the H2O2 treatments. GABA, alanine and glutamate levels were significantly increased in root and shoot of the cam1, cam4, cam5-4, and cam6-1 mutants in response to paraquat (0.5, 1 and 3 μM), while they were increased only in the root tissue of the cam1, cam4, cam5-4, and cam6-1 mutants in response to H2O2 (200 and 500 μM, 1 mM). These data show that the cam5-4 and cam6-1 mutants were sensitive to the induced oxidative stress treatments in terms of seed germination, seedling growth, and oxidative damage. The accumulation of GABA shunt metabolites as a consequence of the induced oxidative stress treatments (paraquat and H2O2 treatments) suggests that the GABA shunt pathway and the accumulation of GABA metabolites may contribute in antioxidant machinery associated with reactive oxygen species and in the acquisition of tolerance in response to induced oxidative stress in Arabidopsis seedlings.

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

Similar content being viewed by others

Abbreviations

CAM :

Calmodulin gene

Ca2+/CaM:

Calcium/calmodulin complex

CaM:

Calmodulin protein

GABA:

γ-Aminobutyric acid

GAD:

Glutamate decarboxylase

MDA:

Malonaldehyde

ROS:

Reactive oxygen species

TBARS:

Thiobarbiturate reactive substances

UV:

Ultra-violet light (UVA or UVB)

References

  • Agarwal PK, Jha B (2010) Transcription factors in plants and ABA dependent and independent abiotic stress signaling. Biol Plant 54:201–2010

    Article  CAS  Google Scholar 

  • AL-Quraan NA, Locy RD, Singh NK (2010) Expression of calmodulin genes in wild type and calmodulin mutants of Arabidopsis thaliana. Plant Physiol Biochem 48:697–702

    Article  PubMed  CAS  Google Scholar 

  • Alscher RG, Donahue JH, Cramer CL (1997) Reactive oxygen species and antioxidants: relationships in green cells. Physiol Plant 100:224–233

    Article  CAS  Google Scholar 

  • Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399

    Article  PubMed  CAS  Google Scholar 

  • Bergmeyer HU (ed) (1983) Methods of enzymatic analysis, vol 1, 2nd edn. VCH, Weinheim

  • Bohnert HJ, Nelson DE, Jensen RG (1995) Adaptation to environmental stresses. Plant Cell 7:1099–1111

    Article  PubMed  CAS  Google Scholar 

  • Bolwell GP, Bindschedler LV, Blee KA, Butt VS, Davies DR (2002) The apoplastic oxidative burst in response to biotic stress in plants: a three-component system. J Exp Bot 53:1367–1376

    Article  PubMed  CAS  Google Scholar 

  • Bouche N, Fromm H (2004) GABA in plants: just a metabolite? Trends Plant Sci 9:110–115

    Article  PubMed  CAS  Google Scholar 

  • Bouche N, Fait A, Bouchez D, Moller SG, Fromm H (2003) Mitochondrial succinic-semialdehyde dehydrogenase of the gamma-aminobutyrate shunt is required to restrict levels of reactive oxygen intermediates in plants. Proc Natl Acad Sci USA 100:6843–6848

    Article  PubMed  CAS  Google Scholar 

  • Bouche N, Yellin A, Snedden WA, Fromm H (2005) Plant-specific calmodulin-binding proteins. Annu Rev Plant Biol 56:435–466

    Article  PubMed  CAS  Google Scholar 

  • Chen YL, Huang RF, Xiao YM, Lu P, Chen J, Wang XC (2004) Extracellular calmodulin-induced stomatal closure is mediated by heterotrimeric G protein and H2O2. Plant Physiol 136:4096–4103

    Article  PubMed  CAS  Google Scholar 

  • Coleman ST, Fang TK, Rovinsky SA, Turano FJ, Moye-Rowley WS (2001) Expression of glutamate decarboxylase homologue is required for normal oxidative stress tolerance in Saccharomyces cerevisiae. J Biol Chem 276:244–250

    Article  PubMed  CAS  Google Scholar 

  • Cuin TA, Shabala S (2007) Compatible solutes reduce ROS-induced potassium efflux in Arabidopsis roots. Plant Cell Environ 30:875–885

    Article  PubMed  CAS  Google Scholar 

  • Desikan RA, Mackerness S, Hancock JT, Neill SJ (2001) Regulation of Arabidopsis transcriptome by oxidative stress. Plant Physiol 127:159–172

    Article  PubMed  CAS  Google Scholar 

  • Dröge W (2002) Free radicals in the physiological control of cell function. Physiol Rev 82:47–95

    Google Scholar 

  • Du L, Poovaiah BW (2005) Ca+2/Calmodulin is critical for brassinosteroid biosynthesis and plant growth. Nature 437:741–745

    Article  PubMed  CAS  Google Scholar 

  • Harding SA, Oh SH, Roberts DM (1997) Transgenic tobacco expressing a foreign calmodulin gene shows an enhanced production of active oxygen species. EMBO J 16:1137–1144

    Article  PubMed  CAS  Google Scholar 

  • Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. Arch Biochem Biophys 125:189–198

    Article  PubMed  CAS  Google Scholar 

  • Hendricks SB, Taylorson RB (1975) Breaking of seed dormancy by catalase inhibition. Proc Natl Acad Sci USA 72:306–309

    Article  PubMed  CAS  Google Scholar 

  • Hong ZL, Lakkineni K, Zhang ZM, Verma DPS (2000) Removal of feedback inhibition of pyrroline-5-carboxylate synthetase results in increased proline accumulation and protection of plants from osmotic stress. Plant Physiol 122:1129–1136

    Article  PubMed  CAS  Google Scholar 

  • Hu X, Jiang M, Zhang J, Zhang A, Lin F, Tan M (2007) Calcium-calmodulin is required for abscisic acid-induced antioxidant defense and functions both upstream and downstream of H2O2 production in leaves of maize (Zea mays) plants. New Phytol 173:27–38

    Article  PubMed  CAS  Google Scholar 

  • Kumutha D, Ezhimathi K, Sairam RK, Srivastava GC, Deshmukh PS, Meena RC (2010) Waterlogging induced oxidative stress and antioxidant activity in pigeon pea genotypes. Biol Plant 53:75–84

    Article  Google Scholar 

  • Kwon SI, Lee H, An CS (2007) Differential expression of three catalase genes in the small radish (Rhaphanus sativus L. var. sativus). Mol Cell 24:37–44

    CAS  Google Scholar 

  • Miller G, Suzuki N, Rizhsky L, Hegie A, Koussevitzky S, Mittler R (2007) Double mutants deficient in cytosolic and thylakoid ascorbate peroxidase reveal a complex mode of interaction between reactive oxygen species. Plant development and response to abiotic stresses. Plant Physiol 144:1777–1785

    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 

  • Mittler R, Vanderauwera S, Gollery M, Van-Breusegem F (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9:490–498

    Article  PubMed  CAS  Google Scholar 

  • Moller IM (2001) Plant mitochondria and oxidative stress: electron transport, NADPH turnover, and metabolism of reactive oxygen species. Annu Rev Plant Physiol Plant Mol Biol 52:561–591

    Article  PubMed  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:472–497

    Article  Google Scholar 

  • Neill SJ, Desikan RA, Hancock JT (2002) Hydrogen peroxide signaling. Curr Opin Plant Biol 5:388–395

    Article  PubMed  CAS  Google Scholar 

  • Niyogi KK (1999) Photoprotection revisited: genetic and molecular approaches. Annu Rev Plant Physiol Plant Mol Biol 50:333–359

    Article  PubMed  CAS  Google Scholar 

  • Oracz K, El-Maarouf BH, Farrant JM, Cooper K, Belghazi M, Job C, Job D, Corbineau F, Bailly C (2007) ROS production and protein oxidation as a novel mechanism for seed dormancy alleviation. Plant J 50:452–465

    Article  PubMed  CAS  Google Scholar 

  • Prasad TK, Anderson MD, Martin BA, Stewart CR (1994) Evidence for chilling-induced oxidative stress in maize seedlings and a regulatory role for hydrogen peroxide. Plant Cell 6:65–74

    Article  PubMed  CAS  Google Scholar 

  • Rentel MC, Knight MR (2004) Oxidative stress-induced calcium signaling in Arabidopsis. Plant Physiol 135:1471–1479

    Article  PubMed  CAS  Google Scholar 

  • Rogozhin VV, Kuriliuk TT, Filippova NP (2000) Change in the reaction of the antioxidant system of wheat sprouts after UV-irradiation of seeds. Biofiz 45:730–736

    CAS  Google Scholar 

  • Scandalios JG, Guan LM, Polidoros A (1997) Catalases in plants: gene structure, properties, regulation and expression. In: Scandalios JG et al (eds) Oxidative stress and the molecular biology of antioxidant defenses Cold Spring Harbor Laboratory Press, Plainview, pp 343–406

  • Shelp BJ, Bown AW, McLean MD (1999) Metabolism and functions of gamma-aminobutyric acid. Trends Plant Sci 4:446–452

    Article  PubMed  Google Scholar 

  • Simontacchi M, Caro A, Fraga CG, Puntarulo S (1993) Oxidative stress affects [alpha]-tocopherol content in soybean embryonic axes upon imbibition and following germination. Plant Physiol 103:949–953

    PubMed  CAS  Google Scholar 

  • Smirnoff N, Cumbes QJ (1989) Hydroxyl radical scavenging activity of compatible solutes. Phytochemistry 28:1057–1060

    Article  CAS  Google Scholar 

  • Vranova E, Inze D, Van-Breusegem F (2002) Signal transduction during oxidative stress. J Exp Bot 53:1227–1236

    Article  PubMed  CAS  Google Scholar 

  • Weber H, Chetelat A, Reymond P, Farmer EE (2004) Selective and powerful stress gene expression in Arabidopsis in response to malondialdehyde. Plant J 37:877–888

    Article  PubMed  CAS  Google Scholar 

  • Yang T, Poovaiah BW (2002) Hydrogen peroxide homeostasis: activation of plant catalase by calcium/calmodulin. Proc Natl Acad Sci USA 99:4097–4102

    Article  PubMed  CAS  Google Scholar 

  • Zhang G, Bown AW (1997) The rapid determination of gamma-aminobutyric acid. Phytochemistry 44:1007–1009

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biotechnology and Genetic Engineering, Faculty of Science and Arts, Jordan University of Science and Technology, Irbid, 22110, Jordan

    Nisreen A. AL-Quraan

  2. Department of Biological Sciences, Auburn University, Auburn, AL, 36849, USA

    Robert D. Locy & Narendra K. Singh

Authors
  1. Nisreen A. AL-Quraan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Robert D. Locy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Narendra K. Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nisreen A. AL-Quraan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

AL-Quraan, N.A., Locy, R.D. & Singh, N.K. Implications of paraquat and hydrogen peroxide-induced oxidative stress treatments on the GABA shunt pathway in Arabidopsis thaliana calmodulin mutants. Plant Biotechnol Rep 5, 225–234 (2011). https://doi.org/10.1007/s11816-011-0174-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11816-011-0174-3

Keywords

Search

Navigation