Abstract
The antioxidant machinery in plants consists of several components with unique or overlapping functions that combat the deleterious production of reactive oxygen species (ROS) induced by stress conditions. Tocopherols are a group of powerful antioxidants having additional roles in signaling and gene expression, with α-tocopherol being the most potent form. In the present study, we used wild-type (WT) and α-tocopherol-enriched transgenic (TR) Brassica juncea plants grown under salt, heavy metal, and osmotic stress to compare their relative tolerance to these stresses and to assess the effects of increased α-tocopherol content on the other antioxidative enzymes and molecules. The oxidative damage caused by induced stress was lower in TR plants compared to WT plants as assessed by their higher relative water content and lower electrolyte leakage, malondialdehyde content as well as H2O2 accumulation. Lesser superoxide and H2O2 accumulation was also observed by histochemical staining in TR seedlings exposed to stress. Though no significant differences were evident under normal growth conditions, TR plants showed higher activities and transcript levels of antioxidant enzymes superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase than WT plants under similar stress conditions. A decrease in ascorbate and glutathione content with marginally higher reductive ratios of these compounds was also observed in TR plants under the stress conditions. Our findings implicate the role of higher α-tocopherol levels in conferring better tolerance against salt, heavy metal, and osmotic stresses and also establish the existence of interplay between this lipid-soluble antioxidant and other water-soluble components of plant antioxidant defense.
Similar content being viewed by others
References
Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126
Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399
Barr HD, Weatherley PE (1962) A re-examination of the relative turgidity technique for estimating water deficit in leaves. Aust J Biol Sci 15:413–428
Burton GW, Ingold KU (1986) Vitamin E: application of the principles of physical organic chemistry to the exploration of its structure and function. Acc Chem Res 19:194–201
Cela J, Chang C, Munné-Bosch S (2011) Accumulation of γ- rather than α-tocopherol alters ethylene signaling gene expression in the vte4 mutant of Arabidopsis thaliana. Plant Cell Physiol 52:1389–1400
Chen GX, Asada K (1989) Ascorbate peroxidase in tea leaves: occurrence of two isozymes and the differences in their enzymatic and molecular properties. Plant Cell Physiol 30:987–998
Collakova E, DellaPenna D (2003) The role of homogentisate phytyltransferase and other tocopherol pathway enzymes in the regulation of tocopherol synthesis during abiotic stress. Plant Physiol 133:930–940
Fredstrom S (2002) Nitric oxide, oxidative stress, and dietary antioxidants. Nutrition 18:537–539
Fryer MJ (1992) The antioxidant effects of thylakoid vitamin-E (α-tocopherol). Plant Cell Environ 15:381–392
Fukuzawa K, Tokumura A, Ouchi S, Tsukatani H (1982) Antioxidant activities of tocopherols on Fe2+-ascorbate-induced lipid peroxidation in lecithin liposomes. Lipids 17:511–513
Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930
Gillespie KM, Ainsworth EA (2007) Measurement of reduced, oxidized and total ascorbate content in plants. Nat Protoc 2:871–874
Griffith OW (1980) Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinyl pyridine. Anal Biochem 106:207–212
Gupta AS, Webb RP, Holaday AS, Allen RD (1993) Overexpression of superoxide dismutase protects plants from oxidative stress. Plant Physiol 103:1067–1073
Halliwell B (2006) Reactive species and antioxidants. Redox biology is a fundamental theme of aerobic life. Plant Physiol 141:312–322
Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198
Hemavathi, Upadhyaya CP, Akula N, Young KE, Chun SC, Kim DH, Park SW (2010) Enhanced ascorbic acid accumulation in transgenic potato confers tolerance to various abiotic stresses. Biotechnol Lett 32:321–330
Hemavathi, Upadhyaya CP, Young KE, Nookaraju A, Kim HS, Heung JJ, Oh OM, Chun SC, Kim DH, Park SW (2011) Biochemical analysis of enhanced tolerance in transgenic potato plants overexpressing D-galacturonic acid reductase gene in response to various abiotic stresses. Mol Breed 28:105–115
Kulas E, Ackman RG (2001) Different tocopherols and the relationship between two methods for determination of primary oxidation products in fish oil. J Agric Food Chem 49:1724–1729
Li Y, Zhou Y, Wang Z, Sun X, Tang K (2010) Engineering tocopherol biosynthetic pathway in Arabidopsis leaves and its effect on antioxidant metabolism. Plant Sci 178:312–320
Mène-Saffrané L, DellaPenna D (2010) Biosynthesis, regulation and functions of tocochromanols in plants. Plant Physiol Biochem 48:301–309
Miller G, Suzuki N, Ciftci-Yilmaz S, Mittler R (2010) Reactive oxygen species homeostasis and signaling during drought and salinity stress. Plant Cell Environ 33:453–467
Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410
Munné-Bosch S, Alegre L (2002) The function of tocopherols and tocotrienols in plants. Crit Rev Plant Sci 21:31–57
Rathinasabapathi B, Kaur R (2006) Metabolic engineering for stress tolerance. In: Madhava Rao KV, Raghavendra AS, Janardhan Reddy K (eds) Physiology and molecular biology of stress tolerance in plants. Springer, The Netherlands, pp 255–299
Sairam RK, Rao KV, Srivastava GC (2002) Differential response of wheat genotypes to long term salinity stress in relation to oxidative stress, antioxidant activity and osmolyte concentration. Plant Sci 163:1037–1046
Sattler SE, Gilliland LU, Magallanes-Lundback M, Pollard M, DellaPenna D (2004) Vitamin E is essential for seed longevity and for preventing lipid peroxidation during germination. Plant Cell 16:1419–1432
Sattler SE, Mène-Saffrané L, Farmer EE, Krischke M, Mueller MJ, DellaPenna D (2006) Non-enzymatic lipid peroxidation reprograms gene expression and activates defense markers in Arabidopsis tocopherol-deficient mutants. Plant Cell 18:3706–3720
Scarpeci TE, Zanor MI, Carrillo N, Mueller-Roeber B, Valle EM (2008) Generation of superoxide anion in chloroplasts of Arabidopsis thaliana during active photosynthesis: a focus on rapidly induced genes. Plant Mol Biol 66:361–378
Semchuk NM, Lushchak OV, Falk J, Krupinska K, Lushchak VI (2009) Inactivation of genes, encoding tocopherol biosynthetic pathway enzymes, results in oxidative stress in outdoor grown Arabidopsis thaliana. Plant Physiol Biochem 47:384–390
Smith IK, Vierheller TL, Thorne CA (1989) Properties and functions of glutathione reductase in plants. Physiol Plant 77:449–456
Szarka A, Tomasskovics B, Bánhegyi G (2012) The ascorbate–glutathione–α-tocopherol triad in abiotic stress response. Int J Mol Sci 13:4458–4483
Upadhyaya CP, Venkatesh J, Gururani MA, Asnin L, Sharma K, Ajappala H, Park SW (2011) Transgenic potato overproducing L-ascorbic acid resisted an increase in methylglyoxal under salinity stress via maintaining higher reduced glutathione level and glyoxalase enzyme activity. Biotechnol Lett 33:2297–2307
Velikova V, Edreva A, Loreto F (2004) Endogenous isoprene protects Phragmites australis leaves against singlet oxygen. Physiol Plant 122:219–225
Yoshimura K, Miyao K, Gaber A, Takeda T, Kanaboshi H, Miyasaka H, Shigeoka S (2004) Enhancement of stress tolerance in transgenic tobacco plants overexpressing Chlamydomonas glutathione peroxidase in chloroplasts or cytosol. Plant J 37:21–33
Yusuf MA, Kumar D, Rajwanshi R, Strasser RJ, Tsimilli-Michael M, Govindjee, Sarin NB (2010) Overexpression of gamma-tocopherol methyl transferase gene in transgenic Brassica juncea plants alleviates abiotic stress: physiological and chlorophyll a fluorescence measurements. Biochim Biophys Acta 1797:1428–1438
Yusuf MA, Sarin NB (2007) Antioxidant value addition in human diets: genetic transformation of Brassica juncea with γ-TMT gene for increased α-tocopherol content. Transgenic Res 16:109–113
Acknowledgments
This work was partially funded by the Council of Scientific and Industrial Research (CSIR) Grant No. 38/1126/EMR-II. DK and PS acknowledge the financial support from CSIR and University Grants Commission (UGC). MAY is a UGC-Dr. D.S. Kothari Postdoctoral Fellow. Research in the laboratory of NBS is supported by U.G.C.-C.A.S., U.G.C.-R.N.W., Department of Science and Technology (D.S.T.)-F.I.S.T., and D.S.T.-PURSE.
Conflict of interest
The authors declare that they have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Additional information
Handling Editor: Bhumi Nath Tripathi
Rights and permissions
About this article
Cite this article
Kumar, D., Yusuf, M.A., Singh, P. et al. Modulation of antioxidant machinery in α-tocopherol-enriched transgenic Brassica juncea plants tolerant to abiotic stress conditions. Protoplasma 250, 1079–1089 (2013). https://doi.org/10.1007/s00709-013-0484-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00709-013-0484-0