Abstract
Hydroponic experiments were conducted with rice seedlings (Oryza sativa L. cv. ND1) exposed to methylene blue (MB) to investigate the accumulation of hydrogen peroxide (H2O2) and activities and gene expression of antioxidant enzymes. Results showed that a linear decrease in relative growth rate and water use efficiency was observed with rice seedlings with increasing MB concentrations. MB-induced accumulation of H2O2 was evident in both roots and shoots. The activities of antioxidative enzymes were analyzed and found to be different at different MB treatment concentrations. The activities of enzymes related to the ascorbate-glutathione cycle were more sensitive to MB treatments than other antioxidative enzymes. Transcript level, by real-time quantitative PCR, of antioxidative enzymes showed that the analyzed genes were differently expressed during different MB concentrations in both roots and shoots. The isoform analysis of superoxide dismutase (SOD) gene showed that the expressions of Cu/ZnSOD and MnSOD were relatively constant, where significant upregulation of FeSOD was observed with rice seedlings exposed to high MB concentrations. Furthermore, the expression of CAT, POD, and MDHAR genes responded biphasically to MB treatments by showing negligible changes at 1.56–15.63 μM MB and significant induction at 31.36–62.52 μM MB. The expression of GR, APX, and DHAR genes showed a remarkable induction to MB. Our results suggest that on transcription level, and in accordance with enzymatic responses, enzymes of GR, APX, and DHAR play central role in the H2O2 detoxification in rice seedlings under MB exposure.
Similar content being viewed by others
References
Achery VMM, Jena S, Panda KK, Panda BB (2008) Aluminum induced oxidative stress and DNA damage in root cells of Allium cepa L. Ecotoxicol Environ Saf 70:300–310
Ahamd A, Rafatullah M, Sulaiman O, Ibrahim MH, Hashim R (2009) Scavenging behaviour of meranti sawdust in the removal of methylene blue from aqueous solution. J Hazard Mater 170:357–365
Apel K, Hirt H (2004) Reactive oxygen species: metabolism oxidative stress, and signaling transduction. Annu Rev Plant Biol 55:373–399
Asada K (1999) The water–water cycle in chloroplasts: scavenging of active oxygen and dissipation of excess photons. Annu Rev Plant Physiol Plant Mol Biol 50:601–639
Baek KH, Skinner DZ (2003) Alternation of antioxidant enzyme gene expression during cold acclimation of near-isogenic wheat line. Plant Sci 165:1221–1227
Banerjee S, Dastidar MG (2005) Use of jute processing wastes for treatment of wastewater contaminated with dye and other organics. Bioresour Technol 96:1919–1928
Bian SM, Jiang YW (2009) Reactive oxygen species, antioxidant enzyme activities and gene expression patterns in leaves and roots of Kentucky bluegrass in response to drought stress and recovery. Sci Hortic 120:264–270
Boominathan R, Doran M (2002) Ni-induced oxidative stress in roots of the Ni hyperaccumulator, Alyssum bertolonii. New Phytol 156:205–215
Bulut Y, Aydm H (2006) A kinetics and thermodynamics study of methylene blue adsorption on wheat shells. Desalination 194:259–267
Contour-Ansel D, Ttorres-franklin ML, Cruz de Carvalho MH, D’arcy-Lameta A, Zuily-Fodil Y (2006) Glutathione reductase in leaves of cowpea: cloning of two cDNAs, expression and enzymatic activity under progressive drought stress, desiccation and abscisic acid treatment. Ann Bot 98:1279–1287
Cuypers A, Smeets K, Ruytinx J, Opdenakker K, Keunen E, Remans T, Horemans N, Vanhoudt N, Van Sanden S, Van Belleghem F, Guisez Y, Colpaert J, Vangronsveld J (2011) The cellular redox state as a modulator in cadmium and copper responses in Arabidopsis thaliana seedlings. J Plant Physiol 168:309–316
Ebbs SD, Piccinin RC, Goodger JQD, Kolev SD, Woodrow IE, Baker AJM (2008) Transport of ferrocyanide by two eucalypt species and sorghum. Int J Phytorem 10:343–357
Foyer CH, Descouvrières P, Kunert KJ (1994) Protection against oxygen radicals: an important defence mechanism studied in transgenic plants. Plant Cell Environ 17:507–523
Foyer CH, Noctor G (2005) Oxidant and antioxidant signalling in plants: a re-evaluation of the concept of oxidative stress in a physiological context. Plant Cell Environ 28:1056–1071
Halliwell B (2006) Reactive species and antioxidants. Redox biology is a fundamental theme of aerobic life. Plant Physiol 141:312–322
Hamdaoui O (2005) Batch study of liquid-phase adsorption of methylene blue using cedar sawdust and crushed brick. J Hazard Mater B135:264–273
Kumar AK, Prasad MNV, Achary VMM, Panda BB (2013) Elucidation of lead-induced oxidative stress in Talinum triangulare roots by analysis of antioxidant responses and DNA damage at cellular level. Environ Sci Pollut Res 20:4551–4561
Li HS (2006) Principles and techniques of plant physiology and biochemistry. Higher Education Press, Beijing, China (In Chinese)
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408
Luo HJ, Li HY, Zhang XZ, Fu JM (2011) Antioxidant responses and gene expression in perennial ryegrass (Lolium perenne L.) under cadmium stress. Ecotoxicology 20:770–778
Murugesan K, Dhamija A, In-Hyun N, Young-Mo K, Yoon-Seok C (2007) Decolourization of reactive black 5 by laccase: optimization by response surface methodology. Dyes Pigments 75:176–184
Nasuha N, Hameed BH, Moha-Din AT (2010) Rejected tea as a potential low-cost adsorbent for the removal of methylene blue. J Hazard Mater 175:126–132
Neill S, Desikan R, Hancock J (2002) Hydrogen peroxide signalling. Curr Opin Plant Biol 5:388–395
Panda SK, Khan MH (2003) Antioxidant efficiency in rice (Oryza sativa L.) leaves under heavy metal toxicity. J Plant Biol 30:23–29
Pawlak S, Firych A, Rymer K, Deckert J (2009) Cu, Zn-superoxide dismutase is differently regulated by cadmium and lead in roots of soybean seedlings. Acta Physiol Plant 31:741–747
Sachs L (1992) Angewandte Statistik. Springer, Berlin
Sauer T, Cesconeto Neto G, Jose HJ, Moreira RFPM (2002) Kinetics of photocatalytic degradation of reactive dyes in a TiO2 slurry reactor. J Photochem Photobiol A 149:147–154
Senthilkumaar S, Varadarajan PR, Porkodi K, Subbhuraam CV (2005a) Adsorption of methylene blue onto jute fiber carbon: kinetics and equilibrium studies. J Colloid Interface Sci 284:78–82
Senthilkumaar S, Porkodi K, Vidyalaksmi R (2005b) Photodegradation of a textile dye catalyzed by sol-gel derived nanocrystalline TiO2 via ultrasonic irradiation. J Photochem Photobiol A 170:225–232
Smeets K, Opdenakker K, Remans T, Van Sanden S, Van Bellephem F, Semane B, Horemans N, Guisez Y, Vangronsveld J, Cuypers A (2009) Oxidative stress-related responses at transcriptional and enzymatic levels after exposure to Cd or Cu in a multipollution context. J Plant Physiol 166:1982–1992
Tan IAW, Ahmad A, Hameed BH (2008) Adsorption of basic dye using activated carbon prepared from oil palm shell: batch and fixed bed studies. Desalination 225:13–28
Trapp S, Zambrano KC, Kusk KO, Karlson U (2000) A phytotoxicity test using transpiration of willows. Arch Environ Contam Toxicol 39:154–160
Vanhoudt N, Vandenhove H, Horemans N, Wannijn J, Buganic A, Vangronsveld J, Cuypers A (2010) Study of oxidative stress related responses induced in Arabidopsis thaliana following mixed exposure to uranium and cadmium. Plant Physiol Biochem 48:879–886
Yu XZ, Lu PC, Yu Z (2012) On the role of β-cyanoalanine synthase (CAS) in metabolism of free cyanide and ferri-cyanide by rice seedlings. Ecotoxicology 21:548–556
Yu XZ, Zhang FZ (2013) Effects of exogenous thiocyanate on mineral nutrients, antioxidative responses and free amino acids in rice seedlings. Ecotoxicology 22:752–760
Yu XZ, Zhang XH, Liu W (2014) Responses of free amino acids in rice seedlings during cyanide metabolism. Environ Sci Pollut Res 21:1411–1417
Acknowledgments
This work was financially supported by the research foundations from Guilin University of Technology (Grant No. GUTRC2011007), The Guangxi Key Laboratory of Theory and Technology for Environmental Pollution Control (Grant No. 1201Z029), and The Guangxi Talent Highland for Hazardous Waste Disposal Industrialization.
Conflict of interest
The authors declare that they have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Additional information
Responsible editor: Philippe Garrigues
Rights and permissions
About this article
Cite this article
Yu, XZ., Zhang, XH. & Yue, DM. Alternation of antioxidative enzyme gene expression in rice seedlings exposed to methylene blue. Environ Sci Pollut Res 21, 14014–14022 (2014). https://doi.org/10.1007/s11356-014-3306-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11356-014-3306-9