Document Type : Research Paper


1 Assistant Professor, Department of Agronomy and Plant Breeding, Shahid Chamran University of Ahvaz, Ahvaz, Iran

2 Associate Professor, Department of Agronomy and Plant Breeding, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran


Background and Objectives
Plant cells often increase cold tolerance by reprogramming their genes expression, which results in adjusted metabolic alternations, a process enhanced under cold acclimation (CA). In this study, responses of Carbohydrate alteration and delta12 and delta15 genes expression to cold stress (CS) phases were comparatively studied in three genotypes of bread and durum wheat differing in sensitivity. Two of them (Norstar, bread wheat and Gerdish, durum wheat) were tolerant to CS and the other one, SRN (durum wheat) was sensitive to CS.
Materials and Methods
Seeds of Norstar (hexaploid, bread wheat) and the two genotypes of Gerdish and SRN (tetraploid, durum wheat) provided by Dryland Agriculture Research Institute (DARI) of Iran were soaked in distilled water and then germinated in Petri dishes on filter paper for 72 h at 25 C in a thermostat. Subsequently, the seedlings were planted in pots. The cooling regime adopted in our experiments allowed us to differentiate the examined genotypes in terms of their tolerance to CS. In our experiment, the plants were moved from control conditions immediately into the acclimated temperature of 4–5 ºC for 14 days with the same photoperiod and irradiance. Leaf samples of genotypes were harvested and analyzed after 14 days under these conditions. After 14 days of CA, the plants were placed into a climatic chamber chilled preliminary to 0 ºC. During further treatments, the temperature was lowered gradually to -5 C (at the rate of 0.5 ºC min-1 ), and the plants were incubated at this temperature for 12 and 24 h. Total cellular RNA was extracted by Biozol method (Fersion Pooyesh, Tehran, Iran) using 80 mg FM leaflets. Applying fermentase reverse transcriptase enzyme instruction, the first strand of cDNA was produced after DNase treatment. Primers were designed using primer 3 to obtain 18–21 bp length. Carbohydrate extraction and determining the carbohydrate concentration were done by 80% ethanol and the AOAC method, respectively.
These responses confirmed the existence of a wide range of genetic capacity in durum wheat to increase cold tolerance particularly in Gerdish. The findings of the present study showed that under experimental treatments, the carbohydrate content significantly changed so that cold stress in acclimated plants increased sucrose, glucose and fructose contents particularly in Norstar and Gerdish as compared to the SRN plants. Increasing expression of delta12 and delta15 genes under cold stress in Norstar and SRN genotypes in comparison with SRN indicates the capacity of cells in increasing cold stress.
The results may be a sign for associating other metabolite or enzyme activities to create relative tolerance against cold-induced oxidative stress. Also, these responses showed high genetic diversity for cold tolerance in durum. Eventually, assessing the dynamics of cell responses after CS without CA phases could profitably be a novel path in plant stress response investigations in the short run.


Main Subjects

Anchordoguy, T. J., Rdolph, A. S., Carpenter, J. F. and Crowe, J. H. (1987). Modes of interaction of cryoprotectants with membrane phospholipids during freezing. Cryobiol, 24(4), 324-331.
AOAC (Association of official analysis committee). (1995). Official methods of analysis (16th ed.). Arlington, VA, USA: Association of Analytical Chemists.
Arroyo, A., Bossi, F., Finkelstein, R. R. and Leon, P. (2003). Three genes that affect sugar sensing (Abscisic acid insensitive 4, Abscisic acid insensitive 5 and constitutive triple response 1) are differentially regulated by glucose in Arabidopsis. Plant Physiology, 133(1), 231-242.
Balibrea, M. E., Rus-Alvarez, A. M., Bolarin, M. C. and Perez-Alfocea, F. (1997). Fastchanges in soluble carbohydrates and praline contents in tomato seedlings in response to ionic and nonionic iso-osmotic stresses. Journal of Plant Physiology, 151(2), 221-226.
Boyko, E. J., Ahroni, J. H., Stensel, V., Forsberg, R. C., Davignon, D. R., Smith, D. G. (1999). A prospective
study of risk factors for diabetic foot ulcer: The Seattle Diabetic Foot Study. Diabetes Care, 22(7), 1036-1042.
Campos, P. S., Quartin, V., Ramalho, J. C., Nunes, M. A. (2003). Electrolyte leakage and lipid degradation account for cold sensitivity in leaves of Coffea sp. plant. Journal of Plant Physiology, 160(3), 283-292.
Fernandez, A., Alvarez-Ordonez, A., Lopez, M. and Bernardo, A. (2009). Effects of organic acids on thermal inactivation of acid and cold stressed Enterococcus faecium. Food Microbiology, 26(5), 497-503.
Hurry, V. M. and Huner, N. P. A. (1991). Low growth temperature effects a differential inhibition of photosynthesis in spring and winter wheat. Plant Physiology, 96(2), 491-497.
Kaur, S., Gupta, A. K. and Kaur, N. (2000). Effect of GA3, kinetin and indole acetic acid on carbohydrate metabolism in chickpea seedlings germinating under water stress. Plant Growth Regulation, 30(1), 61-70.
Lang, P., Zhang, C., Ebel, R. C., Dane, F. and Dozier, W. A. (2005). Identification of cold acclimated genes in leaves of Citrus unshiu by mRNA differential display. Gene, 359, 111-118.
Larkindale, J. and Vierling, E. (2008). Core genome responses involved in acclimation to high temperature. Plant Physiology, 146(2), 748-761.
Lotfi Jala Abadi, A.,  Siadat, S. A.,  Bakhsandeh, A., Fathi, Gh. and Alemi Saied, Kh. (2013). Effect of chemical, organic and biological fertilizers systems on yield and yield components of wheat genotypes
(T. aestivum and T. durum) in Ahvaz conditions. Plant Productions, 36(1), 103-116. [In Farsi]
Maali Amiri, R., Yur’eva, N. O., Shimshilashvili, K. R., Goldenkova-Pavlova, I. V., Pchelkin, V. P., Kuznitsova, E. I, Tsydendambaev, V. D, Trunova, T. I., Los, D. A., Salehi, G. and Nosov, A. M. (2010). Expression of acyl lipid-12-desaturase gene in prokaryotic and eukaryotic cells and its effect on cold stress tolerance of potato Journal of Integrative Plant Biology, 52(3), 289-297.
Marti, J. and Slafer, G. A. (2014). Bread and durum wheat yields under a wide range of environmental conditions. Field Crops Research, 156, 258-271.
Mullineaux, P. M. and Baker, N. R. (2010). Oxidative stress: antagonistic signaling for acclimation or cell death. Plant Physiology, 154(2), 521-525.
Nazari, M. R., Habibpour Mehraban, F., Maali-Amiri, R. and Zeinali Khaneghah, H. (2012). Change in antioxidant responses against oxidative damage in black chickpea following cold acclimation. Russian Journal of Plant Physiology, 59(2), 183-89.
Nejadsadeghi, L., Maali-Amiri, R., Zeinali, H., Ramezanpour, S. and Sadeghzade, B. (2014). Comparative analysis of physio-biochemical responses to cold stress in tetraploid and hexaploid wheat. Cell Biochemistry and Biophysics, 70(1), 399-408.
Orlova, I. V., Serebriiskaya, T. S., Popov, V., Merkulova, N., Nosov, A. M., Trunova, T. I., Tsydendambaev, V. D. and Los, D. A. (2003). Transformation of tobacco with a gene for the thermophilic acyl-lipid desaturase enhances the chilling tolerance of plants. Plant Cell Physiol, 44(4), 447-450.
Pfaffl, M. W. (2007). Relative quantification. In M.T. Dorak (Ed.), Real-time PCR (pp. 63-82.). U.K.: Taylor and Francis Group.
Pfaffl, M. W., Horgan, G. W. and Dempfle, L. (2002). Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Research, 30(3), 36-46.
Popov, V. N., Kipaikina, N. V., Astakhova, N. V. and Trunova, T. I. (2006). Specific features of oxidative stress in the chilled tobacco plants following transformation with the desC gene for acyl-lipid D9desaturase from Synechococcus vulcanus. Russian Journal of Plant Physiology, 53(4), 469-47.

Rajabi, R. and Said Pourdad, S. (2010). A study on cold resistance in safflower varieties and lines by physiological and biochemical indices. Plant Productions, 33(2), 1-14. [In Farsi]

Shao, Y. C., Ding, H. D., Wan, Y. H., NM, Q. I., Zhu, W. M. and Yang, X. F. (2004). Effects of Cd2+ and Zn2+ stress on antioxidant enzyme system of tomato seedlings. Acta Agriculturae Shanghai, 20(4), 79-82.
Steponkus, P. L. (1984). Role of the plasma membrane in freezing injury and cold acclimation. Annual Review of Plant Physiol, 35(1), 543-584.
 © 2019 by the authors. Licensee SCU, Ahvaz, Iran. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution-Non Commercial 4.0 International (CC BY-NC 4.0 license) (