Cold-induced Changes of Antioxidant Enzymes Activity and Lipid Peroxidation in Two Canola (Brassica napus L.) Cultivars


1 Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

2 Department of Plant Biology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran


This study was conducted on two canola (Brassica napus) cultivars, Okapi a winter type and cold tolerant and Rgs003 a spring type and cold sensitive. Seedlings were grown in an environmentally controlled growth room with 16 h d-1 photoperiod at 22/16 °C (day/night, control). At the 4-leaf stage, half of pots were transferred to a cold growth room for 7 d at 10/3°C (day/night, cold treatment) and their leaves were harvested as required on 0, 2, 4 and 7 d as four sampling times. Superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) activity and the amount of malondialdehyde (MDA) in samples were assessed. Analysis of variance showed that winter canola had more SOD, CAT and APX activity, lower amount of either MDA or POD activities compared with the spring canola. This study indicated that antioxidant mechanisms caused more cold tolerance in the winter cultivar of canola compared with the spring cultivar.


Azooz, MM, Ismail AM and Elhamd MF, 2009. Growth, lipid peroxidation and antioxidant enzyme activities as a selection criterion for salt tolerance of maize cultivars grown under salinity stress. International Journal of Agricultural and Biological Engineering 11: 21-26.
Baek KH and Skinner DZ, 2003. Alternation of antioxidant enzyme gene expression during cold acclimation of near-isogenic wheat lines. Plant Science 165: 1221-1227.
Balestrasse KB, Tomaro, ML, Batlle A, Noriega GO, 2010. The role of 5-aminolevulinic acid in the response to cold stress in soybean plants.Phytochemistry 71:17-18.
Bian S and Jiang Y, 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. Scientia Horticulturae 120: 264-270.
Boyer JS, 1982. Plant productivity and environment. Science218: 443-448.
Cakmak I and Horst W, 1991. Effect of aluminium on lipid peroxidation, superoxide dismutase, catalase and peroxidase activities in root tip of soybean (Glysin max). Plant Physiology 83: 463-468.
Chance B and Maehly AC, 1955. Assay of catalase and peroxidase. Methods in Enzymology 2: 764-775.
Chelikani P, Fita I and Loewen PC, 2004. Diversity of structures and properties among catalases. CMLS Cellular and Molecular Life Sciences61: 192-208.
Cunhua S, Wei D, Xiangling C, Xinna X, Yahong Z, Dong S and Jianjie S, 2010. The effects of drought stress on the activity of acid phosphatase and its protective enzymes in pigweed leaves. African Journal of Biotechnology 9: 825-833.
Dat J, Vandenabeele S, Vranova E, Van Montagu M and Van Breusegem F, 2000. Dual actions of the active oxygen species during plants stress responses. CMLS Cellular and Molecular Life Sciences 57: 779-795.
De Vos CH, Schat M, De Waal R, Vooij S and Ernst W, 1991. Increased to copper-induced damage of the root plasma membrane in copper tolerant silene cucubalus. Plant Physiology82: 523-528.
Duan M, Feng HL, Wang  LY, Li D and Meng QW, 2012. Overexpression of thylakoidal ascorbate peroxidase shows enhanced resistance to chilling stress in tomato. Journal of Plant Physiology 169: 867-877.
Food and Agriculture Organization (FAO), 2011. Crop Production Statistics.
Fry JC, 1993. Biological Data Analysis: A Practical Approach, Vol. 1. Oxford University Press, UK. 418 pp.
Fry SC, 1986. Cross-linking of matrix polymers in the growing cell walls of angiosperms. Annual Review of Plant Physiology 37: 165-186.
Ghanati F, Morita A and Yokota H, 2002. Induction of suberin and increase of lignin content by excess boron in tabacco cell. Soil Science and Plant Nutrition 48: 357-364.
Giannopolitis CN and Ries SK, 1977. Superoxide dismutases: II. Purification and quantitative relationship with water-soluble protein in seedlings. Plant Physiology 59: 315-318.
Gill SS and Tuteja N, 2010. Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry48: 909-930.
Guo FQ, Okamoto M and Crawford NM, 2003. Identification of a plant nitric oxide synthase gene involved in hormonal signaling. Science 302: 100-103.
Javadian N, Karimzadeh G, Mahfoozi S and Ghanati F, 2010. Cold-induced changes of enzymes, proline, carbohydrates, and chlorophyll in wheat. Russian Journal of Plant Physiology 57: 540-547.
Juhnke H, 1996. Mutants that show increased sensitivity to hydrogen peroxide reveal an important role for the pentose phosphate pathway in protection of yeast against oxidative stress. Molecular and General Genetics 252: 456-464.
Kazemi Shahandashti SS, Maali Amiri R, Zeinali H and Ramezanpour SS, 2012. Change in membrane fatty acid compositions and cold-induced responses in chickpea. Molecular Biology Reports DOI 10.1007/s11033-012-2130-x.
Kwon SY, Lee HS and Kwak SS, 2001. Development of environmental stress-tolerant plants by gene manipulation of antioxidant enzymes. Plant Pathology 17: 88-93.
Liu X and Huang B, 2000. Heat stress injury in relation to membrane lipid peroxidation in creeping bentgrass. Crop Science 40: 503-510.
Lopez-Huertas E, 2000. Stress induces peroxisome biogenesis genes. European Molecular Biology Organization Journal 19: 6770-6777.
Luo Y, Tang H and Zhang Y, 2011. Production of reactive oxygen species and antioxidant metabolism about strawberry leaves to low temperature. Journal of Agricultural Science 3:89-96.
Mittler R, 2002. Oxidative stress, antioxidants and stress tolerance. Trends in Plant Science 7: 405-410.
Moussa H and Abdel-Aziz SM, 2008. Comparative response of drought tolerant and drought sensitive maize genotypes to water stress.Australian Journal of Crop Science 1: 31-36.
Nokano Y and Asada K, 1981. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiology 22: 867-880.
Nordin-Henriksson K and Trewavas AJ, 2003. The effect of short-term low-temperature treatments on gene expression in Arabidopsis correlates with changes in intracellular Ca2+ levels. Plant Cell Environment 26: 485-496.
Omidi H, 2010. Changes of proline content and activity of antioxidative enzymes in two canola genotype under drought stress. American Journal of Plant Physiology 5: 338-349.
Ozkur O, Ozdemir F, Bor M and Turkan I, 2009. Physiochemical and antioxidant responses of the perennial xerophyte Capparis ovata Desf. to drought. Environmental and Experimental Botany 66:487-492.
Parvanova D, Popova A, Zaharieva I, Lambrev P, Konstantinova T, Taneva S, Atanassov A, Goltsev V and Djilianov D, 2004. Low temperature tolerance of tobacco plants transformed to accumulate proline, fructans, or glycine betaine. Variable chlorophyll fluorescence evidence. Photosynthetica 42: 179-185.
Passardi F, Longe F, Penel D and Dunand C, 2004. The class III peroxidase multigenic family in rice and its evolution in land plants. Phytochemistry 65: 1879-1893.
Prasad TK, 1996. Mechanisms of chilling-induced oxidative stress injury and tolerance in developing maize seedlings: changes in antioxidant system, oxidation of proteins and lipids, and protease activities. Plant Journal 10: 1017-1026.
Radyuk MS, Domanskaya IN, Shcherbakov RA and Shalygo NV, 2010. Effect of low above-zero temperature on the content of low-molecular antioxidants and activities of antioxidant enzymes in green barley leaves. Russian Journal of Plant Physiology 56: 175-180.
Rajabi S, Karimzadeh G, Ghanati F and Asilan KS, 2012. Salt-induced changes of antioxidant enzymes activity in winter canola (Brassica napus) cultivars in growth chamber. Journal of Plant Physiology and Breeding 2: 11-21.
Ryan B and Joiner BL, 2001. Minitab Handbook, 4th edn. Duxbury Press, California, USA.
Uemura, M, Tominaga, Y, Nakagawara, C, Shigematsu, S, Minami, A and Kawamura, Y 2006. Responses of the plasma membrane to low temperatures. Physiologia Plantarum 126:81-89.
Wang-Hao Z, Yu-Zhen C and Cun-Fu L, 2007. Differences in biochemical responses to cold stress in two contrasting varieties of rapeseed (Brassica napus L.). Forestry Studies inChina 9: 142-146.
Willekens H, Chamnongpol S, Davey M, Schraudner M, Langebartels C, Van Montagu M, Inze D and Van Camp W, 1997. Catalase is a sink for H2O2 and is indispensable for stress defense in C3 plants. European Molecular Biology Organization Journal 16: 4806-4816.
Xu J, Zhang Y, Guan Z, Wei W, Han L and Chai T, 2008. Expression and function of two dehydrins under environmental stresses in Brassica juncea L. Molecular Breeding 21: 431-438.
Zhang WP, Jiang B, Lou LN, Lu MH, Yang M and Chen JF, 2011. Impact of salicylic acid on the antioxidant enzyme system and hydrogen peroxide production in Cucumis sativus under chilling stress. Zeitschrift für Naturforschung C66: 413-422.
Zhu J, Dong CH and Zhu JK, 2007. Interplay between cold-responsive gene regulation, metabolism and RNA processing during plant cold acclimation. Current Opinion in Plant Biology 10: 290-295.