Journal of Applied Botany and Food Quality 86, 71 - 78 (2013), DOI:10.5073/JABFQ.2013.086.011 Department of Horticulture, Faculty of Agriculture, Shiraz University, Shiraz, Iran Growth and physiological responses of grafted and non-grafted cultivars of Ziziphus spina-christi to salinity Akhtar Shekafandeh*, Shohreh Takhti (Received May 27, 2013) * Corresponding author Summary Two grafted cultivars of Z. spina-christi (‘Danehgird’ and ‘Daneh- boland’ were grafted on wild seedlings) and non-grafted seedlings were grown in soil and perlite mixture (1:1 v/v) and treated with 0 (0 dS m-1), 3.2 (5 dS m-1), 6.4 (10 dS m-1) and 12.8 (20 dS m-1) g l-1 NaCl. After 16 weeks, salt stress resulted in a substantial decrease in root length, stem, leaf surface, lateral root number, root, stem and leaf fresh and dry weight in both grafted cultivars and seedling root- stock. These reductions were predominance in seedling rootstock than in grafted cultivars. In all organs (leaf, stem and root) of grafted ‘Danehboland’ cultivar, the K+/Na+ ratio was significantly higher than non-grafted wild seedling in saline and non-saline conditions. The proline and soluble sugar also was significantly higher in the leaves of ‘Danehboland’ cultivar than non-grafted control. The re- sults imply the predominance of the scion genotype in determining salt tolerance in comparison with rootstock seedling. Introduction Widespread in tropical and subtropical regions, Ziziphus spina- christi (L.) Desf. belonging to the Rhamnaceae family is a spiny shrub or small tree (Johnston, 1963) that are naturally distributed in southern parts of Iran and locally named ‘konar’. This species is a multipurpose plant. In addition to high value of non wood products, particularly leaves and shoots saponin, tannin substrates and fruit nutrition, this species is ecologically and economically important for its tolerance to drought and salinity (sudhersan and hussain, 2003; Sohail, 2009). The fruit is a drupe (about 1 to 1.5 cm in diameter) and is consumed either fresh or dried by the rural popu- lation (Saied et al., 2008). In the south of Iran, a wide variability in yield, fruit size, peel colour was observed among the genotypes (torahi, 2011). At present, the seedlings are used as rootstock for cultivars with high fruit quality (takhti and shekafandeh, 2012). Salinity is a main problem in arid and semi-arid regions of the world (Gebauer et al., 2004). Global warming and low rainfall intensify this difficulty. The saline growth condition causes many unfavor- able effects on plant growth and development at physiological and biochemical levels, which are due to low osmotic potential of soil solution (osmotic stress), specific ion effects, nutritional imbalance or combination of these factors (Parida and Das, 2005; Arzani, 2008). Due to water scarcity and exacerbated soil salinity in some part of Iran, cultivation of other fruit trees such as citrus is limited, so, orchardists established spina-christi grafting orchards in which selected genotypes with superior edible fruit are grafted on spina- christi seedling. Despite the fact that the most documents on salt tolerance of plants reported the important role of rootstocks, for they represent the first part to control the uptake and translocation of nutrients and salts throughout the plant (Munns, 2002; Zrig et al., 2011), there are many reports that clarify the predominant role of scion genotype in plant growth and improvement of salt effect (Moya et al., 2002; Santa-cruz et al., 2002; Chen et al., 2003). Colla et al. (2010) expressed that grafting is an integrative mu- tual process and both scion and rootstock can influence growth and greater root to shoot ratio and lower accumulation of Na+ and/or Cl- in shoots than ungrafted or self-grafted. The proline accumulation has often been proposed as a valuable marker for the selection of salt tolerance genotypes (Zid and Grignon, 1991). In relation to spina-christi ‘Konar’, despite its highly appreciated fruits in local markets and increasing the area under cultivations, there is limited information on physiological responses and the ex- tent of salt tolerance of these grafted plants. The objective of this research was to study the effect of different levels of NaCl salinity on growth parameters and physiological responses of three selected lo- cal genotypes non-grafted wild type, ‘Danehgird’ and ‘Danehboland’ grafted on wild spina-christi seedlings. Material and methods One year-old grafted ‘Konar’ plants (‘Danehgird’ and ‘Danehboland’ cultivars were grafted on wild seedlings) and non-grafted seedlings were used in this experiment. The plants were transferred into 20 liter plastic pots without drainage and filled with a mixture of soil (laom soil) and perlite (1:1, V:V). Field capacity of soil mixture was measured using cell pressure device (Santa Barbara Calif soil Equipment Co.) and accordingly, plants were irrigated once every three days. Commercial fertilizer NPK (20:20:20) including; iron (1 g l-1), manganese (0.5 g l-1), zinc (0.5 g l-1), copper (0.04 g l-1), boron (0.2 g l-1), molybdenum (0.04 g l-1) and calcium nitrate (0.5 g l-1) was applied to pots with irrigation water each week. The pots were placed into a plastic greenhouse with natural sunlight and temperature range 32± °C and 20± °C in the day and at night respectively. After 10 weeks when plants were well established, salinity treatments were applied. For preventing salinity shock, the salts were applied in three times with irrigation water. Treatments were 0 (0 dS m-1), 3.2 (5 dS m-1), 6.4(10 dS m-1) and 12.8 (20 dS m-1) g l-1 NaCl. At the ends of experiment (after 16 weeks), the leaf number of all treatments were counted. Leaf area was measured by leaf area meter (AM 200, ADC Bio Scientific Ltd. Taiwan). All 8 plants in each replicate were individually harvested, the lateral roots were counted, stem and root length were measured with a ruler, then they were divided into stems, leaves and roots after recording their fresh weight, all plant organs were oven-dried at 80 °C for 48 h then weighed again. Ion content Dry-ashing of plant material was obtained at 500 °C for 6 hr. Sodium (Na+) and potassium (K+) contents of samples were measured after digestion process in 1 N nitric acid solution (HNO3-), using the flame emission photometry. Chloride (Cl−) content was analyzed by titra- tion with 0.1N silver nitrate (AgNO3) in the presence of potassium bichromate according to a modified colorimetric method of Mohr (Mathieu and Pieltain, 2003). 72 A. Shekafandeh, S. Takhti Chlorophyll content Fresh leaf samples were washed with deionized water prior to extraction to remove any surface contamination. One g leaf sample was squashed in 80% acetone using a pestle and mortar. The mixture was centrifuged at 4800 rpm for 20 min. The optical density of the supernant was measured at 663 and 645 nm wavelengths and chloro- phyll content was calculated using the following equations. Mg Chl g F.W. = [20.2 (OD645nm) + 8.02 (OD663nm)] × V / F.W. × 1000 Where V is final volume of solution (ml) and F.W. leaf fresh weight (mg). Proline contents To determine proline content of the leaves, 0.5 g plant material was homogenized in 10 ml of 3% aqueous solution of sulphosalicylic acid and the homogenate filtered through Whatman # 2 filter paper. Two ml of filterate was reacted with 2 ml acid ninhydrin and 2 ml of glacial acetic acid in a test tube for 1 h at 100 °C and the reaction terminated in an ice bath. The reaction mixture was extracted with 4 ml toluene, mixed for 15 ~ 20 seconds. Chromophore containing toluene was aspirated from the aqueous phase and the absorbance was read at 520 nm in a spectrophotometer. The proline concentra- tion (μ mole g-1 F.W.) was calculated based on a standard curve. Soluble sugars content Soluble sugars were extracted from 0.1 g fresh leaves in 5 ml of 80% ethanol placed in a water bath at 70 °C for 30 min. The insoluble remains were removed by centrifuging at 5000 g for 10 min. One ml of the resulted extract was mixed with 5ml of sulfuric acid and 1 ml of 5% phenol solution. The mixed solution was vortexed after cool down to room temperature. The absorbance was recorded at 490 nm using a spectrophotometer. The soluble sugar content of each sample was determined using a standard curve for glucose and expressed as mg glucose g-1 F.W. (Mccready et al., 1950; dubois et al., 1956). Starch content From previous section, the solid debris remaining in the centrifuge tube was washed, re-extracted and re-centrifuged four-times using 80% (V/V) ethanol. The starch content in the samples was deter- mined colorimetrically using anthrone method (Mccready et al., 1950). The absorbance was read at 630 nm in a digital spectro- photometer as described by lópez et al. (2002). Statistical analysis A factorial experiment 3 (two grafted and one non-grafted ‘Konars’) × 4 (4 levels of salinity) was arranged in complete randomized design (CRD) with 4 replicates and 2 plants in each replication. Data were subjected to analysis of variance using the SPSS software (ver. 13.0) SPSS Inc. Mean differences were determined by Tukeys tests at p≤0.05. Results The results showed that leaf area, stem and root length were sig- nificantly reduced by increasing salinity. However, the reduction in growth parameters varied according to cultivar and measured factor so that, in 20 dS m-1 NaCl, the highest stem reduction (58.1%) was observed in ‘Danehboland’ and lowest reduction (50.5%) in wild cultivar. In contrast, the highest root length reduction (78.6%) was observed in wild types and the lowest root reduction was observed in grafted ‘Danehboland’ by 44% (Tab. 1). The effect of salinity on lateral root number, root fresh and dry weight showed a significant genotypic variation (Tab. 2). In non- saline condition, scions ‘Danehboland’ and ‘Danehgird’ promoted significantly lateral root number in comparison with control. The highest reduction in lateral root number was found in wild type root- stock (without grafting), which reached by 62.5% at 20 dS m-1 NaCl, whereas the lowest one was obtained in ‘Danehboland’ with 50% decreased when compared to control plants. Regardless of salt treat- ments, scion ‘Danehboland’ also produced more root fresh (15.4 g) Tab. 1: Sodium chloride effect on leaf surface, stem and root length of non-grafted wild, grated ‘Danehgird’ and ‘Danehboland’ cultivars of Z. spina- chiristi. Cultivar NaCl Leaf surface Mean Stem length Mean Root length Mean dS m-1 (mm2) (Cm) (Cm) Non-grafted 0 14.0a-d 80.6b 19.8bc Wild 5 13.2c-f 60.1de 11.4f 10 12.6def 50.0ef 7.0g 20 11.9f 39.9fg 4.2g 12.9B 57.6B 10.6C Grafted 0 14.9ab 83.0ab 21.7ab Danehgird 5 14.2abc 64.3cd 17.6cd 10 13.3c-f 50.1ef 16.1de 20 12.3ef 35.8g 13.2ef 13.7A 58.3B 17.1B Grafted 0 15.3a 94.1a 24.4a Danehboland 5 14.2abc 74.6bc 19.8bc 10 13.4b-e 53.1de 18.1cd 20 12.4ef 39.4fg 13.9ef 13.8A 65.3A 19.1A In each column, means followed by the same letter are not significantly different at p≤0.05, using Tukey’s test. (n=8). Responses of grafted and non-grafted Z. spina-christi to salinity 73 and dry weight (8.45 g) than scion ‘Danehgird’ (13.8 and 7.55 g respectively) and wild type (4.6 and 2.6 g respectively) (Tab. 2). Leaf and stem fresh and dry weight were also significantly reduced by salinity treatments. However, this reduction is varied accor- ding to cultivar (Tab. 3). For example, in 20 dS m-1 NaCl wild type, ‘Danehboland’ and ‘Danehgird’ showed 77.5, 68 and 79% reduction in leaf dry weight respectively in comparison with control plants. Regardless of salinity levels, scion ‘Danehboland’ showed higher stem fresh (12.4 g) and dry weight (7.7 g) than scion ‘Danehgird’ (11 and 6.7 g respectively) and wild type (4.6 and 2.67 g). Data showed that grafted cultivars had different behavior in Na+ and K+ uptake in comparison to non-grafted wild type. In all grafted and non-grafted plants, with increasing salinity in culture media the Na+ concentrations in different organs (root, stem and leaf) increased, however, regardless of different level of salinity, the accumulation of Na+ was significantly lower in all organs of ‘Danehboland’ and Tab. 2: Sodium chloride effect on lateral root number, root fresh and dry weight of non-grafted wild, grafted ‘Danehgird’ and ‘Danehboland’ cultivars of Z. spina-christi. Cultivar NaCl Lateral Mean Root F.W. Mean Root D.W. Mean dS m-1 root number (g) (g) Non-grafted 0 19.8d 8.4d 5.3de Wild 5 16.2de 5.4e 3.1fg 10 12.3e 3.0f 1.6gh 20 7.4f 1.5f 0.67h 13.9C 4.6C 2.7B Grafted 0 31.7b 20.7a 11.8ab Danehgird 5 25.8c 15.9b 8.6c 10 19.9d 11.7c 6.1d 20 13.2e 6.9de 3.8ef 22.6B 13.8B 7.6A Grafted 0 38.9a 21.8a 13.1a Danehboland 5 32.4b 17.3b 10.7b 10 26.2c 12.1c 6.2d 20 18.9d 7.9d 3.8ef 29.1A 15.4A 8.5A In each column, means followed by the same letter are not significantly different at p≤0.05, using Tukey’s test. (n=8). Tab. 3: Sodium chloride effect on leaf, stem fresh and dry weight of non-grafted wild, grafted ‘Danehgird’ and ‘Danehboland’ cultivars of Z. spina-christi. Cultivar NaCl Leaf Mean Leaf Mean Stem F.W. Mean Stem D.W. Mean dS m-1 F.W. (g) D.W. (g) (g) (g) Non-grafted 0 6.2a 3.4ab 7.8ef 4.7efg Wild 5 3.3bcd 1.9cd 5.1gh 2.9ghi 10 1.8def 1.2c-f 3.5hi 2.0hi 20 1.2f 0.7ef 2.2i 1.1i 3.1AB 1.8A 4.6C 2.7C Grafted 0 4.0b 2.3bc 16.5b 10.4b Danehgird 5 3.0b-e 1.5c-f 11.2d 7.4cd 10 2.3c-f 1.1def 9.5de 5.3ef 20 1.5ef 0.7ef 6.8fg 3.6fgh 2.7B 1.4B 11.0B 6.7B Grafted 0 6.8a 4.1a 19.1a 13.3a Danehboland 5 3.7bc 1.9cde 13.8c 8.4c 10 2.3c-f 0.8def 10.3d 5.7de 20 1.8def 0.6f 6.4fg 3.5fgh 3.6A 1.8A 12.4A 7.7A In each column, means followed by the same letter are not significantly different at p≤0.05, using Tukey’s test. (n=8). 74 A. Shekafandeh, S. Takhti ‘Danehgird’ cultivars in comparison to wild type (Tab. 4, Tab. 5 and Tab. 6). For example, leaf Na+ concentration in ‘Danehboland’ and ‘Danehgird’ was 11.4 and 12.4 mg g-1 D.W. respectively which was significantly lower than wild type (17.5 mg g-1 D.W.). In 5, 10 and 20 dS m-1 NaCl, leaves of all genotypes showed a significant decrease in K+ concentration (Tab. 6). The same trend observed for accumu- lation of K+ in stem, and root. Regardless of salt concentration, all organs (leaf, stem and root) of grafted cultivars maintained a higher ratio of K+/Na+ than wild type. (Tab. 4, Tab. 5 and Tab. 6). With increasing salinity, the Cl- content in all plant parts (leaf, stem and root) of grafted and non-grafted cultivars increased. But this accumulation is more pronounced in roots than stems and leaves Tab. 4: Sodium chloride effect on root Na+, K+ (mg g-1 D.W.) and K+/Na+ ratio of non-grafted wild, grafted ‘Danehgird’ and ‘Danehboland’ cultivars of Z. spina-christi. Cultivar NaCl Root Na+ mg Mean Root K+ mg Mean Root Mean dS m-1 g-1 D.W. g-1 D.W. K+/ Na+ Non-grafted 0 24.2de 20.3cd 0.8bcd Wild 5 28.8c 17.2ef 0.6d 10 34.3b 16.1fg 0.5d 20 37.9a 14.7g 0.4d 31.3A 17.1C 0.6C Grafted 0 14.3g 21.7bc 1.5bc Danehgird 5 18.1f 18.7de 1.0bcd 10 22.4e 17.3ef 0.8cd 20 25.4d 15.3fg 0.6d 20.1B 18.2B 1.0B Grafted 0 9.0h 30.1a 3.5a Danehboland 5 14.4g 23.1b 1.6b 10 18.8f 20.3cd 1.1bcd 20 22.9de 16.4efg 0.7cd 16.3C 22.5A 1.7A In each column, means followed by the same letter are not significantly different at p≤0.05, using Tukey’s test. (n=8). Tab. 5: Sodium chloride effect on stem Na+, K+ (mg g-1 D.W.) and K+/Na+ ratio of non-grafted wild, grafted ‘Danehgird’ and ‘Danehboland’ cultivars of Z. spina-christi. Cultivar NaCl Stem Na+ Mean Stem K+ Mean Stem K+/Na+ Mean dS m-1 mg g-1 D.W. mg g-1 D.W. Non-grafted 0 15.2e 28.8c 1.9cd Wild 5 21.7cd 23.2d 1.2def 10 26.4b 18.3e 0.7ef 20 31.0a 15.6f 0.5f 23.6A 21.5C 1.1C Grafted 0 10.0fg 31.6ab 3.2b Danehgird 5 15.4e 27.7c 1.8cde 10 19.8d 22.3d 1.1def 20 23.2c 17.0ef 0.7ef 17.1B 24.6B 1.7B Grafted 0 7.7g 34.0 4.5a Danehboland 5 11.1f 29.3bc 2.7bc 10 16.4e 23.0d 1.4def 20 21.8d 17.8ef 0.8def 14.2C 26.0A 2.4A In each column, means followed by the same letter are not significantly different at p≤0.05, using Tukey’s test. (n=8). Responses of grafted and non-grafted Z. spina-christi to salinity 75 (Tab. 7), which means the plants limited the translocation of Cl- to aerial parts. In all cultivars, with increasing NaCl the amount of sugar increased and the amount of starch decreased and in all levels of salinity, ‘Danehboland’ leaves showed significantly higher sugar and starch contents in comparison with non-grafted wild type and grafted ‘Danehgird’ (Fig. 1). The leaf proline content increased with increasing the salinity in all cultivars and in contrast the amount of chlorophyll decreased. In all levels of salinity, scion-stock relation- ship had no effect on chlorophyll content in comparison with non- grafted wild type, but in 3.2 and 6.4 g l-1 NaCl scion ‘Danehboland’ cultivar showed higher proline concentration in comparison with others (Fig. 2). Tab. 6: Sodium chloride effect on leaf Na+, K+ (mg g-1 D.W.) and K+/Na+ ratio of non-grafted wild, grafted ‘Danehgird’ and ‘Danehboland’ cultivars of Z. spina-christi. Cultivar NaCl Leaf Na+ Mean Leaf K+ Mean Leaf K+/Na+ Mean dS m-1 mg g-1 D.W. mg g-1 D.W. Non-grafted 0 12.3ef 30.7b 2.5cde Wild 5 15.7cd 25.7c 1.7ef 10 18.6b 22.2de 1.2ef 20 23.6a 18.1g 0.8f 17.5A 24.2C 1.5B Grafted 0 8.3gh 35.6a 4.3ab Danehgird 5 10.1fg 30.3b 3.0bcd 10 13.4de 24.2cd 1.8def 20 17.7bc 18.4fg 1.1f 12.4B 27.1B 2.5A Grafted 0 7.4h 36.8a 5.0a Danehboland 5 9.1gh 32.4b 3.6abc 10 12.4ef 26.2c 2.1def 20 16.7bc 21.2ef 1.3ef 11.4B 29.2A 3.0A In each column, means followed by the same letter are not significantly different at p≤0.05, using Tukey’s test. (n=8). Tab. 7: Sodium chloride effect on leaf, stem and root Cl− (%) of non-grafted wild, grafted ‘Danehgird’ and ‘Danehboland’ cultivars of Z. spina-christi. Cultivar NaCl Root Cl− Mean Stem Cl− Mean Leaf Cl− Mean dS m-1 % % % Non-grafted 0 5.1h 2.9h 3.0g Wild 5 7.6fg 3.9gh 4.4fg 10 8.9ef 6.7cde 6.6de 20 11.6cd 8.1bc 6.8de 8.3B 5.4C 5.2C Grafted 0 5.5gh 3.6h 4.5fg Danehgird 5 7.9f 5.4efg 5.5ef 10 9.1ef 7.2bc 7.2cd 20 13.1c 8.8b 7.3bcd 8.9B 6.2B 6.1B Grafted 0 10.2de 4.4fgh 7.1cd Danehboland 5 12.9c 5.8def 8.5abc 10 19.5b 7.8bc 8.9ab 20 22.7a 9.9a 9.2a 16.3A 6.9A 8.4A In each column, means followed by the same letter are not significantly different at p≤0.05, using Tukey’s test. (n=8). 76 A. Shekafandeh, S. Takhti Discussion Salt stress resulted in a considerable decrease in leaf surface, shoot and root length, lateral root number, root, stem an leaf fresh and dry weight in both grafted cultivars and seedling rootstock of spina- christi ‘Konar’. These reductions were predominant in seedling root- stock than in grafted cultivars (Tab. 1, Tab. 2 and Tab. 3), which means that different cultivars grafted on the same seedling root- stock improved the growth behavior of the root system. In respect to some traits such as lateral root number, shoot and root length, grafted ‘Danehboland’ cultivar even achieved better than grafted ‘Danehgird’. Hence, the scion genotype played a major role in estab- lishing the growth rate of grafted ‘konars’, which is in accordance with the results obtained in salt stress citrus (banuls and priMo- Millo, 1995; Chen et al., 2003) and tomato (santa-cru, 2002). In grape vines, sivritepe et al. (2010) explained the preference of the scion genotype by determining variation in leaf-levels physiological characters of grafted vines. The most important parts of the plant exposed with soil-related stress factors such as salinity are root systems. The enhanced salt tolerance of grafted ‘konar’ cultivars may be associated with the root system. In fact, in non-saline condition, grafting significantly increased lateral root and root dry weight in comparison with non- grafted seedling rootstock. In salt stress condition, two of the above factors decreased, but the reduction in grafted plants was signifi- cantly smaller than in non-grafted seedlings. In tomatoes, He et al. (2009) also observed that root dry mass declined at high NaCl in comparison with non-saline conditions, but the decrease was smaller in rootstock-grafted plants. In non-saline condition, the Na+ ion content in root of the grafted ‘konar’ cultivars were significantly lower than Na+ ion content in root of seedling rootstock, that means, scion genotypes limited the Na+ uptake from root zone. In the same trend, in salt stress condi- tion (5, 10, 20 dS m-1 NaCl) the accumulation on Na+ in root of grafted plants were significantly less than non-grafted controls. In addition, Na+ content in stems and leaves of rootstock seedlings and in grafted ‘konar’ cultivars were less than their roots in all levels of salt treatments conditions. The preservation of Na+ in the roots of two grafted ‘Konars’ aid to decline its concentration in the leaves that may help to avoid affecting plant metabolism processes. In this respect, tabatabaie (2010) previously reported similar results with olive tree cultivars subjected to high salt stress, where salt tolerance Fig. 2: Sodium chloride effect on leaf total chlorophyll (mg g-1 F.W.) and leaf proline content (µmole g-1 F.W.) of non-grafted wild, grafted ‘Danehgird’ and ‘Danehboland’ cultivars of Z. spina-christi. In each measured factor (chlorophyll, proline), means followed by the same letter are not significantly different at p≤0.05, using Tukey’s test. The data are the means±SE (n=8). Fig. 1: Sodium chloride effect on leaf sugar, starch content mg g-1 D.W. of non-grafted wild, grafted ‘Danehgird’ and ‘Danehboland’ cultivars of Z. spina- christi. In each measured factor (sugar, starch), columns followed by the same letter are not significantly different at p≤0.05, using Tukey’s test. The data are the means±SE ( n=8). Responses of grafted and non-grafted Z. spina-christi to salinity 77 in olive trees was hypothesized to be associated with the ability to reduce the uptake and/or transport of saline ions. In both grafted cultivars and seedling rootstock, with increasing salinity, the leaf K+/Na+ ratio decreased, which can be attributed to a failure in the K+/Na+ selectivity mechanisms. In the leaves of two grafted cultivars the K+/Na+ ratio was recorded above 2 in all salt concentrations, except in 20 dS m-1 NaCl for grafted ‘Danehboland’ and in 10 dS m-1 and 20 dS m-1 in grafted ‘Danehgird’ (Tab. 6). A high K+/Na+ ratio in the leaves is often considered as a salt-tolerance marker (chartzoulakis et al., 2002; dasgan et al., 2002; Meena et al., 2003). So, scion genotypes had an ameliorating effect on K+/ Na+ ratio which helps the grafted plants do better their metabolic functions. The finding from chloride analysis showed that both grafted ‘konars’ and seedling rootstock exhibited a significant variation at all salinity levels and in all plant parts (leaf, stem and root). Grafted and ungrafted ‘konar’ plants limited the translocation of Cl- to the aerial parts; which is an important mechanism to avoid the delete- rious effects of salinity in plants (chartzoulakis, 2005), however, grafted ‘Danehboland’ cultivar accumulated significantly more Cl- in their different parts than non-grafted seedlings. It is interesting to note that this grafted cultivar had higher root, stem and leaf fresh and dry weight and also accumulated more k+ than non-grafted root- stock. It seems that this cultivar uses Cl- to balance electrical charge and pH of their cells (roussos et al., 2006). K+ is the major cation within plants which counter balance negative charge of anions and stabilizes the pH, osmotic potential and turgor pressure within cells. It also plays a crucial role in the activation of the enzyme involved in the metabolism (chelli-chaabouni et al., 2010). In the present study, a positive correlation was recorded between proline and NaCl concentration in the leaves of non-grafted wild seedlings and two grafted cultivars. The same trend was observed between soluble sugar and salt concentration. It has been shown that these compatible solutes play a major role in osmoregulation (Hare et al., 1998; parida and das, 2005; reJskovà et al., 2007). In this regard, grafted ‘Danehboland’ achieved better than grafted ‘Danehgird’ and non-grafted wild type. The results are in agreement with those previously reported by hokMabadi et al. 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Address of the authors: Dr. Akhtar Shekafandeh and Eng Shohreh Takhti, Department of Horti- culture, Faculty of Agriculture, Shiraz University, Shiraz, Iran, P.O. Box 7144465186. E-mail; shekafan@shirazu.ac.ir E-mail; shoreh_g_t@yahoo.com