Investigation of Catalase, Proxidase and Total Protein Level in Some Cold Treated Grapevine Cultivars Cold Stress Response

Document Type : Research Article


1 Tehran University

2 National Institute of Genetic Engineering and Biotechnology


Chilling is an important environmental stress that influences the yield and quality of many agricultural crops. Different plants use different systems to endure this stress and minimize its effects. One of these systems is enzymatic reaction. To find out more about responses of different grapevine species and cultivars to the low temperature conditions, their enzymatic changes were evaluated in a factorial experiment based on randomized complete design with 3 replication during different periods after chilling stress. Leaf samples of plants under cold stress had been taken and maintained in -80 °C until enzyme extraction. Low temperature around 4 °C is sufficient to induce genes that produce chilling acclimatization proteins. In the present study, leaf samples were collected from the plants that were kept at 4 °C during different time intervals, and then total proteins as well as two main antioxidant enzymes (catalase and guaiacolperoxidase) activities were measured. Results showed that as temperature decreased, enzymatic activities were increased in six Iranian grapevine cultivars (‘Atabaki’, ‘Khalili-Danedar’, ‘Shahroodi’, ‘Rajabi-Siah’, ‘Askari’ and ‘Bidane-Sefid’) as well as ‘Riparia’, an American species. The highest enzymatic activities of catalase and ceroxidase were recorded in ‘Khalili-Danedar’ and ‘Riparia’. However,the lowest activities were recorded in ‘Rajabi-Siah’, ‘Bidane-Sefid’ and ‘Shahroodi’. For all studied cultivars, peroxidase showed its highest activity at 12 h after chilling stress, then remained constant, while, the highest activity of catalase were recorded at 8 h. In addition, cold stress increased the total protein content for all studied cultivars, in which ‘Khalili-Danedar’ had the highest protein content amongstudied cultivars. Also, the highest proteins content were recorded at 12 h after exposing plants to cold.


1- افشار محمدیان م.، رضایی ش. و رمضانی ملک رودی م. 1391. بررسی مقاومت دو رقم زیتون به تنش سرما. مجله فرآیند و کارکرد گیاهی.1 (2) :1-11
2- نظری م.، معالی ا.ر. و رمضانپور س.س. 1390. بررسی پاسخ آنزیمی و بیان ژن‌های کاتالاز و پراکسیداز به تنش سرما در ژنوتیپ‌های ایرانی نخود. ژنتیک در هزاره سوم. سال نهم. 1: 2299-2290.
3- ونایی س.، سی و سه مرده ع.، و حیدری غ.1390. اثرات تنش سرما در مرحله جوانه زنی و گیاهچه ای بر فعالیت آنزیم‌های آنتی اکسیدان و برخی صفات فیزیولوژیکی در نخود. نشریه پژوهشهای زراعی ایران.9(3): 524-514.
4- Aebi H. 1984. Catalase in vitro. Methods in Enzymology. Academic press, Orlando, 105: 121-126.
5- Allen R.D., Webb R.P and Schake S.A. 1997. Use of transgenic plants to study antioxidant defenses. Free Radical Biology and Medicine 23:473-479.
6- Anderson M.D., Prasad T.K., and Stewart C.R.1995. Changes in izozyme profiles of catalase, peroxidase, and glutathione reductase during acclimation to chilling in mesocotyls of maize seedlings, Plant Physiology, 109: 1247-1257.
7- Aono M., Saji H., Sakamoto A., Tanaka K., Kondo N., and Tanaka K. 1995. Paraquat tolerance of transgenic Nicotianatabacum with enhanced activities of glutathione reductase and superoxide dismutase. Plant and Cell Physiology 36:1687-1691.
8- Apostolova P., and Yaneva I. 2006. Antioxidativedefence in winter wheat plants during early cold acclimation. General and Applied Plant Physiology, Special issue: 101-108.
9- Baker C.J., and Orlandi E.W., 1995. Active oxygen in plant pathogenesis, Annual Review Phytopathology, 33: 299–321.
10- Bradford M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72: 248-254.
11- Chance B., and Maehly A.C. 1955. Assay of catalases and peroxidases. In: Colowick, S.P., Kaplan, N.O. (Eds.), Methods in Enzymology. Academic Press, New York, 764–775.
12- Dalton T.H., Shertzer, A., and Puga, A.1999. Regulation of gene expression by reactive oxygen, Annual Review of Pharmacology and Toxicology, 39: 67-101.
13- Desikan R., Cheung M., Bright J., Henson D., Hancok J., and Neill S. 2004. ABA hydrogen peroxide and nitric oxide signalling in stomatal guard cells. Journal of Experimental Botany55(395): 205-212.
14- Foyer C. H., Lelandais M., and Kunert K. J. 1994. Oxidative stress in plants, Physiology Plant., 92: 696–717.
15- Francesca S., Luca S., and Claudio V. 1998. Changes in activity of antioxidative enzymes in wheat(Triticum aestivum) seedlings under cold acclimation. Physiologiaplantarum 104: 747-752.
16- Kukreja S., Nandwal A. S., Kumar N., Sharma S.K., Unvi V., and Sharma P. K. 2005. Plant water status, H2O2 scavenging enzymes, ethylene evolution and membrane integrity of Cicerarietinum roots as affected by salinity, Biologia Plantarum 49: 305-308.
17- Lučić B., Jovanović Ž., Radović S., and Maksimović V. 2009. Cold-induced response of buckwheat (Fagopyrum esculemtum moench) seedlings. Archives of Biological Sciences 61(3) 3-4.
18- Lukatkin A. S. 2002. Contribution of Oxidative Stress to the Development of Cold-Induced Damage to Leaves of Chilling-Sensitive Plants: 2. The Activity of Antioxidant Enzymes during Plant Chilling. Russian Journal of Plant Physiology. 49(6): 878-885.
19- Mhamdi A., Queval G., Chaouch S., Vanderauwera S., Breusegem F.V., and Noctor G. 2010. Catalase function in plants: a focus on Arabidopsis mutants as stress-mimic models. Journal of Experimental Botany, 61:4107-4320.
20- Millard P. 1988. The accumulation and storage of nitrogen by herbaceous plants. Plant Cell and Environment 11:18.
21- Mittler R., 2002. Oxidative stress, antioxidants and stress tolerance, Trends in Plant Science, 7: 405-410.
22- Mundree S.G., Baker B., Mowla S., Peters S., Marais S., Willigen C.V., Govender K., Maredza A., Muyanga S., Farrant J.M., and Thomson J.A. 2002 Physiological and molecular insights into drought tolerance. African Journal of Biotechnoly 1: 2838.
23- Pastori G.M., and Foyer C.H. 2002. Common components, networks, and pathways of crosstolerance to stress. The central role of “redox” and abscisic acid-mediated controls, Plant Physiology, 129: 460-468.
24- Prasad T.G. 1996. Mechanisms of chilling-induced oxidative stress injury and tolerance in developing maize seedling: changes in antioxidant system, oxidation of proteins and lipids, and protease activities. Plant Journal, 10: 1017–1026.
25- Sattler U., Calson P., Boiteux s., and Salles B. 2000. Detection of oxidative base DNA damage by a new biochemical assay. Archives of Biochemistry and Biophysics, 376: 26-33
26- Sharma P., and Dubey R.S. 2005. Modulation of nitrate reductase activity in rice seedlings under aluminum toxicity and water stress: role of osmolytes as enzyme protectant, Journal of Plant Physiology, 162: 854-864.
27- Sheng C.X., Yong P.L., Jin H., Ya J.G., Wen G.M., Yun Y.Z., and Shui J.Z. 2010. Responses of Antioxidant enzymes to chilling stress in tobacco seedlings, Agricultural Sciences in China, 11: 1594-1601.
28- Shin R., and Schachtman D. 2004. Hydrogen peroxide mediates plant root response to nutrient deprivation. Proceeding of the National Academy of Sciences of United States of America. 101: 8827-8832.
29- Tasgin E., Atici O., Nalbantoglu B., and Petrova L. 2006. Effects of salicylic acid and cold treatment on protein levels and on the activities of antioxidant enzymes in the apoplast of winter wheat leaves. Journal of Phytochemistry, 67: 710-715.
30- Tomashow M.F. 1999. Plant cold acclimation: freezing tolerance gene and regulatory mechanisms. Annual Review of Plant Physiology and Plant Molecular Biology50: 571-599
31- Tuteja N. 2007. Mechanisms of high salinity tolerance in plants, Methods of Enzymology, 428: 419-438.
32- Tuteja N. 2009. Cold, Salinity, and Drought Stress, Plant Stress Biology, Hirt, H. WILEYVCH Verlag GmbH &Co. KGaA, Weinheim, 137-159.
33- Wang W.B., Kim Y.H., Lee H., Yong Kim S., Deng X., and Kwak S. 2009. Analysis of antioxidant ezyme activity during germination of alfalfa under salt and drought stresses. Journal of Plant Physiology and Biochemistry, 47: 570-577.
34- 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 C3plants. EMBO Journal 16: 4806–4816.
35- Yong Z., Hao-Ru T., and Ya L. 2008. Variation in antioxidant enzyme activities of two strawbreey cultivars with short-term low temperature stress. Journal of Agricultural Sciences, 4: 456-462.