The Effect of Low Temperatures on Domestic and Wild Pistachio Rootstocks and Interspecific Hybrids Based on Physio-biochemical Indices

Document Type : Research Article


Pistachio Research Center, Horticultural Sciences Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Rafsanjan, Iran



In recent decades, spring frost considered a serious threat in pistachio production (Pistacia vera). Considering that, Iran is one of the most important centers of pistachio production and has the highest variety of pistachios in the world, the identification of domestic and wild different rootstocks and their crosses to reach cold-tolerant rootstocks could be one of the effective solutions to face the challenge of spring frost. In this study, cold tolerance was assessed in fourteen rootstocks of pistachio domestic species (Badami Zarand, Sarakhs and Qazvini), interspecific hybrids (Qazvini×Khinjuk, Qazvini×Mutica, Qazvini × Baneh Baghi, Qazvini × Atlantica, Qazvini× Integerrima), non-domesticated species (Atlantica (P. atlantica), Integerrima, Khinjuk, Sarakhs, Mutica (P. atlantica sub. mutica) and Baneh Baghi (P. mutica × P. vera)) and UCB1 hybrid rootstock at low temperatures (4, 0 and -4 °C) for 2 hours. This experiment was conducted in a factorial completely randomized design with three replications on one-year-old seedlings in greenhouse conditions. The results showed that the rate of ion leakage, malondialdehyde and injury index percentage was significant (p <0.01) and their values were decreased in the cold-tolerant rootstocks than the cold-sensitive rootstocks. The interaction between treatments showed that the temperature of - 4 °C caused a significant reduction in ion leakage and malondialdehyde in the cold-tolerant rootstocks. Proline was increased in the cold-tolerant rootstocks compare to cold-sensitive rootstocks. The results of cation leakage percentage were significant in temperature and rootstocks (p <0.01). The results showed that the highest percentage of cation leakage related to potassium and there was a positive and significant correlation between calcium and magnesium and cell membrane stability. According to the results, the most sensitive rootstock was UCB1, Integerrima, Khinjuk and Baneh Baghi, respectively. Hybrids Qazvini , Baneh Baghi, Khinjuk and Integerrima rootstocks were significantly reduced under low-temperature stress. Mutica, Qazvini×Mutica, Sarakhs, Qazvini, Qazvini×Atlantica rootstocks, improved physiological and biochemical traits and maintained cell membrane integrity and finally showed greater tolerance to frost and freezing temperatures, which can be considered in the breeding program.


Afrousheh M, Hokmabadi H, Arab H, Tajabadipour A (2018) Evaluation of frost damage tolerance in some pistachio seedling rootstocks. Journal of Nuts. 9(1), 77-83.
Afshari H, Hokmabadi H, Ebadi A, Laee G (2010) Measurement of chemical and nonchemical parameters of three native pistachio cultivars of Damghan region (Iran) for studying spring frost. Asian Journal of Chemistry. 22(3), 2356-2366.
Afshari H, Tajabadipour A, Hokmabadi H, Mohammadi Moghadam M (2009) Determining the chilling requirements of four pistachio cultivars in Semnan Province (Iran). African Journal of Agricultural Research. 4, 55-59.
Agricultural Statistics (2019, 2020) Horticultural Products, Ministry of Jihad Agriculture, Information and Communication Technology Center, 3, p 240.
Ahmad P, Prasad MNV (2012) Environmental adaptations and stress tolerance of plants in the era of climate change. New York, Dordrecht, Heidelberg, London: Springer, ISBN:1461408156. 
Ait Barka E,  Nowak J, Christophe C (2006)  Enhancement  of  chilling resistance  of  inoculated  grapevine  plantlets  with  a plant  growth  promoting rhizobacterium,  Burkholderia  phytofirmans strain  PsJN.  Applied and Environmental Microbiology. 72, 7246– 7252.
Alipour H (2018) Photosynthesis properties and ion homeostasis of different pistachio cultivar seedlings in response to salinity stress.
International Journal of Horticultural Science and Technology. 5(1), 19-29.
Ashraf M, Foolad M (2007) Roles of glycine betaine and proline in improving plant abiotic stress resistance. Environmental and Experimental Botany. 59, 206–216.
Aslamarz AA, Vahdati K (2010) Stomatal density and ion leakage as indicators of cold hardiness in walnut. Acta Horticulturae. 861, 321-324.
Aslamarz AA, Vahdati K, Rahemi M, Hasani D (2009) Cold-hardiness evaluation of Persian walnut by thermal analysis and freezing technique. Acta Horticulturae. 861, 269-272.
Aslamarz A, Vahdati K, Hasani D, Rahemi M, Leslie CA (2010) Supercooling and cold hardiness in the acclimated and deacclimated buds and stems of Persian walnut cultivars and genotypes. HortScience. 45 (11), 1-6.
Aslamarz AA, Vahdati K, Hasani D, Rahemi M, Leslie CA (2011) Cold hardiness and its relationship with proline content in Persian walnut. European Journal of Horticultural Sciences. 76 (3), 84–90.
Azzarello E, Mugnai S, Pandolfi C, Masi E, Marone E, Mancuso S (2009) Comparing image (fractal analysis) and electrochemical (impedance spectroscopy and electrolyte leakage) techniques for the assessment of the freezing tolerance in olive. Trees. 23(1), 159-167.
Bates LS (1973) Rapid determination of free proline for water stress studies. Plant Soil. 39, 205-207.
Beringer H, Troldenier G (1980) The influence of K nutrition on the response of plants to environmental stress. Potassium Research-Review and trends, 11th Congress of the International Potash Institute. Bern, Switzerland, pp.  89-222.
Buchanan BB, Gruissem W, Jones RL (2015) Biochemistry & molecular biology of plants, 2nd Ed West Sussex, UK: Wiley Blackwell. pp. 1280.
Chen THH, Murata N (2000) Enhancement of tolerance of abiotic stress by metabolic engineering of betaines and other compatible solutes. Current Opinion in Plant Biology. 5, 250-257.
Chen YZ, Ane L (2005) The relationship between seasonal changes in anti-oxidative  system  and  freezing  tolerance  in  the  leaves  in  woody  plants. Scientia Horticulture. 73, 272-279.
 Couée I, Sulmon C, Gouesbet G, El Amrani A (2006) Involvement of soluble sugars in reactive oxygen species balance and responses to oxidative stress in plants. Journal of Experimental Botany. 57(3), 449–459.
Czek-Kwinta RB, Antonkiewicz J, Maoelak J, Oleksiewicz A (2007) The Contents of some nutrients in the aerial parts of maize and their potential role in tolerance to Stress. Acta Physiologiae Plantarum. 29(1), 33–56.
Dionne J, Castonguay Y, Nadeau P, Desjardins Y (2001) Freezing tolerance and carbohydrate changes during cold acclimation of greentype annual bluegrass (Poa annua L.) ecotypes. Journal of Crop Science. 41, 443–451.
Esenta G, Okke A, Nalbanto B (2003) Effects of salicylic acid and cold on freezing tolerance in winter wheat leaves. Plant Growth Regulation, 41, 231-236.
Fahimirad S, Karimzadeh G, Ghanati F (2013) Cold-induced changes of antioxidant enzymes activity and lipid peroxidation in two canola cultivars. Journal of Plant Physiology and Breeding. 3(1), 1–11
Ghasemi Soluklui AA, Ershadi A, Tabatabaee ZE, Fallahi E (2014) Paclobutrazol-induced biochemical changes in pomegranate (Punica granatum L. cv.‘Malas Saveh’) under freezing stress. International Journal of Horticultural Science and Technology. 1(2),181-90
Guo WL, Chen RG, Gong ZH, Yin YX, Ahmed SS, He YM (2012) Exogenous abscisic acid increases antioxidant enzymes and related gene expression in pepper (Capsicum annuum) leaves subjected to chilling stress. Genetics and Molecular Research. 11, 4063–4080.
Guo Y, Liu S, Yang Z, Tian S, Sui N (2016) Responses of unsaturated fatty acid in membrane lipid and antioxidant enzymes to chilling stress in sweet sorghum (Sorghum bicolor (L.) Moench) seedling. Journal of Agricultural Science. 8(9), 71.
Gusta LV, Wisniewski M, Nesbit NT, Tanino KT (2003) Factors to consider in artificial freeze tests. Acta Horticulturea. 618, 493– 507.
Gusta LV, Wisniewski, ME, Trischuk RG (2009) Patterns of freezing in plants: The influence of species, environment and experiential procedures 213 225 Gusta L.V., Wisneiwski M. E. & Trischuk R.G. Plant cold hardiness: From the laboratory to the field CAB International.
Guy CL (2003) Freezing tolerance of plants: current understanding and selected emerging concepts. Canadian Journal of Botany.81, 1216– 1223.
Habibie A, Yazdani N, Saba MK, Vahdati K (2019) Ascorbic acid incorporated with walnut green husk extract for preserving the postharvest quality of cold storage fresh walnut kernels. Scientia Horticulturae 245: 193-199.
Hanna B, Bischofa JC (2004) Direct cell injury associated with eutectic crystallization during freezing. Cryo Bio System. 48, 8-21.
Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Archives of Biochemistry and Biophysics. 125: 180-198.
Hokmabadi H, Rezaee L, Mohammadi Moghaddam M, Mortazavi A, Serfi H, Ghorbani A, Avazabadian A, Reziabadi H (2016) Investigation of cold resistance in three commercial cultivars of Damghan pistachio and three major cultivars of pistachio through ion leakage parameters. The final report of the research project of Iran Pistachio Research Institute. Damghan Pistachio Research Station. 72 P. [In Persian].
Hosseini SM (2000) Physiological study of cold in 5 cultivars of Rafsanjan pistachio. Master Thesis in Biology. Kerman Shahid Bahonar University.
Imahori Y, Takemura M, Bai J (2008) Chilling-induced oxidative stress and antioxidant responses inmume (Prunus mume) fruit during low temperature storage. Postharvest Biology and Technology. 49(1), 54–60.
Kodra E, Steinhaeuser K, Ganguly AR (2011) Persisting cold extremes under 21st-century warming scenarios. Geophysical Research Letters. 38(8), 187.
Larcher W (2004) Ecofisiologia vegetal. São Carlos: RiMa, pp. 531.
Mansour MMF (2000) Nitrogen containing compounds and adaptation of plants to salinity stress. Biology Plant. 43, 491-500.
Matysik J, AliaBhalu B, Mohanty P (2002) Molecular mechanisms of quenching of reactive oxygen species by proline under stress in plants. Current Science. 525-532.
Mavali M (2011) Pistachio rootstocks evaluation which is resistance to cold. MS thesis, Islamic Azad University of Jiroft.
Mirmohammadi Meybodi AM, Tarkesh Isfahani S (2004) Breeding and physiological aspects of cold and freezing stress in crops. Golbon Press. pp, 312. 27-31. [In Persian].
Nasibi F, Manouchehri Kalantari K, Manzari Tavakoli Z. (2020) Effects of hydrogen sulfide on cold-induced oxidative damage in Cucumis sativus L. International Journal of Horticultural Science and Technology. 7(3), 199-211.
Nikoogoftar Sedghi M, ValizadehKaji B, Karimi R (2021) Physiological and biochemical responses of 10 commercial Iranian pistachio cultivars to freezing. Biologia. 76, 329–339.
Norozi M, ValizadehKaji B, Karimi R, Nikoogoftar Sedghi M (2019) Effects of foliar application of potassium and zinc on pistachio (Pistacia vera L.) fruit yield. International Journal of Horticultural Science and Technology. 6(1), 113-23.
Posmyk MM, Bailly C, Szafrańska K, Janas KM, Corbineau F (2005) Antioxidant enzymes and iso-flavonoids in chilled soybean (Glycine max (L.) Merr.) Seedlings. Journal of Plant Physiology. 162, 403–412.
Rahemi M, Yazdani F, Sedaghat S (2016) Evaluation of freezing tolerance in olive cultivars by stomatal density and freezing stress. International Journal of Horticultural Science and Technology. 3(2),145-53.
Ruelland E, Vaultier MN, Zachowski A, Hurry V (2009) Cold signaling and cold acclimation in plants. Advances in Botanical Research. 49(8), 36–150.
Sairam RK (1994) Effect of moisture stress on physiological activities of two contrasting wheat genotypes. Indian Journal Experimental Biology. 32, 584-593.
Sajadian H, Shamili M, Hokmabadi H, Tajabadipour A, Hasheminasab H (2019) Physiological Responses of Some Rootstocks and Interspecific Hybrids of Pistachio to Cold Stress under Greenhouse Conditions. Journal of Nuts. 10(2), 139-151. 
Sayyari M, Babalar M, Kalantari S, Serrano M, Valero D (2009) Effect of salicylic acid treatment on reducing chilling injury in stored pomegranates. Postharvest Biology and Technology. 53, 152-154.
Shamshiri MH, Hasani MR (2015) Synergistic accumulative effects between exogenous salicylic acid and arbuscular mycorrhizal fungus in pistachio (Pistacia vera cv. Abareqi) seedlings under drought stress.2(2), 151-160.
Shang HT, Cao SF, Yang ZF, Cai YT, Zheng YH (2011) Effect of exogenous aminobutyric acid treatment on proline accumulation and chilling injury in peach fruit after longterm cold storage. Journal of Agricultural and Food Chemistry. 59(4), 1264–1268.
Sharifkhah M, Bakhshi D, Pourghayoumi M, Abdi S, Hokmabadi H (2020) Effect of pollination time on yield and antioxidant properties of some pistachio cultivars. International Journal of Horticultural Science and Technology. 7(1), 51-8.
Singer SM, El-Tohamy WA, Hadid AF, Makhart AH (1996) Chilling and water stress injury in bean (Phaseolus vulgaris L.) seedlings reduced by pretreatment with CaCl2, mefluidide, KCl and MgCl2. Egyptian Journal of Horticulture Research. 23, 77-87.
Sofo A, Dichio B, Xiloyannis C, Masia A (2004) Effects of different irradiance levels on some antioxidant enzymes and on malondialdehyde content during rewatering in olive tree. Plant Science. 166, 293–302.
Song Y, Zhang Z, Shao H, Guo X (2008) Relationship between calcium decoding elements and plant abiotic-stress resistance. International Journal Biology Science. 4, 116-125.
Szabados L, Savoure A (2009) Proline: a multifunctional amino acid. Trends in Plant Science. 15(2), 89-97.
Tajabadipour A, Fattahi Moghadam MR, Zamani Z, Nasibi F, Hokmabadi H (2018) Evaluation of physiological and biochemical changes of pistachio (Pistacia vera L. cv. Ahmad-Aghaii) on cold tolerant and sensitive rootstocks under freezing stress conditions. Journal of Horticultural Science. 32(3), 471-484. [In Persian].
Verslues PE, Sharma S (2010) Proline Metabolism and Its Implications for Plant-Environment Interaction. Arabidopsis Book. 8, e0140.
Wahid A, Gelani S, Ashraf M, Foolad MR (2007) Heat tolerance in plants: An overview. Environmental and Experimental Botany. 61(3), 199–223.
Wisniewski M, Willick IR, Gusta LV (2017) Freeze tolerance and avoidance in plants. In, Plant stress physiology, p 179.
Zeinali Yadegari L, Heydari R, Karaptian J (2009) Alteration of biomembrane permeability and activity of soybean antioxidant enzymes in response to low temperature. Iranian Journal of Biology. 22(2), 229-236.