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country (Cheraghi, 2004). Salinity in soil or water is one of the most damaging abiotic stress factors limiting crop (Debez et al., 2006). High concentration of salts in the soils immediately imposes on plants the osmotic stress effect due to lower soil water potential leading to retardation of water uptake. When exposed for longer period, salinity entails ionic stress when plants absorb and accumulate toxic level of Na + and Cl - in the cytoplasm. Salinity also induced secondary stresses such as nutritional imbalance and oxidative stress (Zhu, 2002). For worldwide crop production, the detrimental effects of salinity on plant growth are associated with low osmotic potential of soil solution (water stress), nutritional imbalance, specific ion effect (salt stress), or a combination of these factors (Levit, 1980; Ashraf and Harris, 2004). All of these agents cause adverse pleiotropic effects on plant growth and development at physiological and biochemical levels (Munns, 2002) and the molecular level (Tester and Davenport, 2003). To survive in hyper-saline soil, plants have evolved complex mechanisms that contribute to the adaptation to osmotic and ionic stresses caused by high salinity. These mechanisms include osmotic adjustment that is usually established by intake of inorganic ions as well as accumulation of compatible solute (Osmoprotectants). Inorganic ions are sequestered in the vacuole, while organic solutes are compartmentalized in the cytoplasm to balance the low osmotic potential in the vacuole (Rontein et al., 2002). Furthermore, plants have to employ a wide range of biochemical and molecular mechanisms including alteration of photosynthetic pathway, change in membrane structure, stimulation of phytohormones and induction of antioxidative enzymes (Parida and Das, 2005; Mudgal et al., 2010). Difference in salt tolerance exists not only among variant genera and species, but also within the different organs of the same species (Bankar and Ranjbar, 2010). Comparing the response of cultivars of one species to salinity provides a convenient and useful tool for unveiling the fundamental mechanisms involved in salt tolerance. The objective of this investigation was to evaluate the effects of salt stress on growth, soluble sugars and ion accumulation of three pistachio cultivars, to provide a reference for the cultivation of salt-tolerant trees. MATERIALS AND METHODS Three types of pistachio seeds (Pistacia vera L. Cultivars Akbari, Ouhadi and Kalehghouchi) were used in this study. Seeds were collected from the Anar area, Kerman province, which is one of the main pistachio growing regions in Iran with saline condition. The trial was conducted at Biology Department, Islamic Azad University, Damghan Branch, Damghan, Iran during 2011. The seeds were surface-sterilized for 10 min in 10% H2O2, washed three times with tap water to remove any trace of chemicals that could impair seed germination and were placed on sterile vermiculite at 18 to 25°C to germinate. After 21 days of germination, seedlings were sown in plastic post containing sandy soil (pH 7.6, 3.8% silt, 14.6% clay, 81.6% sand, and electrical conductivity (ECe) = 0.98 ds/m). Abbaspour et al. 2469 Seedlings were transplanted in 20 ×15 cm plastic pots containing a mixture of salinity clay: sand (1:5 v/v) (four seedlings/pot). Each pot contained 3 seedlings which were sufficiently irrigated with urban water every 4 days. All pots were placed in a greenhouse condition with mean relative humidity 45 to 60%, day / night temperature 18 to 30°C and photoperiod from 14 to 16 h (photo-synthetically active radiation of approximately of 430 to 460 mol/ms). Seedlings were grown under these conditions for 4 weeks before initiation of NaCl treatments and salinity treatments were 0, 100, 200, and 300 mM of NaCl. The soil was salinized in step-wise manner to avoid subjecting plants to an osmotic shock. After 55 days of salt treatment, the plants were harvested and separated into shoots and roots. After complete washing with distilled deionized water, fresh masses of shoots and roots and the other morphological parameters such as shoot height, number of branches and leaves were measured in fresh samples. Total leaf area was calculated with an AM-200 leaf-area meter. Total chlorophyll and cartenoid were extracted and estimated from fresh leaves according to the standard method of Arnon (1949). Chlorophyll was extracted in 80% (v/v) aceton from 1 g of fresh leaf sample in the dark at room temperature. Absorbance was measured at 663 and 645 ηm in a UV/VIS spectrophotometer. Chlorophyll concentration was calculated using the equation: Chl 0. 0202 × A645 + 0.00802 × A663 Cartenoid concentration was calculated using the equation: Car = (A436× V) / 196 × b) Where V is volume of dilution, b is the length of cell (1 cm). The value 196 is coefficients of specific absorption. The remaining samples were then over-dried at 80°C for 48 h so as to record dry masses. A flame photometer was used for Na + and K + determination. Phosphorus was analyzed colorimetrically (Boltz and Lueck, 1958) and Cl content was determined by atomic absorption. Fresh roots and leaves were collected for the determination of total soluble sugars content. Soluble sugar content was determined by 0.1 ml of the alcoholic extractor reacting with 3 ml freshly prepared anthrone (2000 mg anthrone + 100 ml 72% H2So4) then was placed in a boiling water bath for 10 min as described by Irigoyen et al. (1992). After cooling, the absorbance was read to be equal to 620 nm. Statistical analysis The experiment was arranged in a completely randomized design (CRD) in a two-factor factorial arrangement with 3 replications (n = 3). All parameters were investigated by analysis of variance (ANOVA) using SPSS software. The means were compared by the Tukey’s test at a 0.05 confidence level. RESULTS Growth performances of the plants were estimated by measuring plant height, leaf area, number of leaves and branches, and total fresh and dry weights. With increase in salinity stress, dry weight of shoots decreases in the three pistachio cultivars. However, this decline was not statistically significantly different in salinity of 100 mM but was significant different in the other salinities compared to the control plant. In all salinity levels and control conditions, Akbari cultivar exhibited larger dry weight
2470 J. Med. Plants Res. Table 1. Fresh and dry weight (g), number of leaves and branches, leaf area (cm 2 ) and plant height (cm) of pistachio plants under NaCl stress. Plant height Leaf area Number of branches Number of leaves Root weight Fresh Dry Shoot weight Fresh Dry Cultivars 27 a 91.04 a 7 a 43 a 3.19 a 4.86 a 8.2 a 5.38 a Ak 28 a 95.4 a 7 a 46 a 1.9 ce 3.08 c 7.9 a 4.9 a Ka 27 a 93.2 a 8 a 48 a 2.01 ce 3.19 c 7.4 a 5.05 a Ou 22 ab 85.44 a 6 ac 40 a 2.4 ae 3.4 ac 5.6 b 4.7 a Ak 21 ab 87.35 a 8 a 41 a 1.2 bc 2.78 c 5.4 b 3.9 a Ka 24 ab 86.2 a 7 a 44 a 1.15 bc 2.67 c 5.2 b 3.8 a Ou 19 bd 77.37 ae 4 cd 29 c 1.7 bce 2.7 c 3.9 bd 2.7b e Ak 10 c 61.30 de 3 d 21 c 0.87 bd 2.2 dc 2.6 de 1.5 c Ka 12 c 64.35 de 3 d 24 c 0.98 bd 2.1 dc 2.3 de 1.6 c Ou 17 d 65.43 e 3 d 23 c 1.2 bd 2.2 dc 3.94 bd 1.9 ce Ak 9 c 37.3 c 2 d 10 d 1.1 d 0.49 d 1.58 e 0.23 d Ka 10 c 35.4 c 3 d 13 d 1.05 d 0.45 d 1.32 e 0.19 d Ou Within each column, means superscript with different letters are significantly different (P≤ 0.05). compared to the other two cultivars. Moreover, this state was more sensible in salinities of 200 and 300 mM and was statistically significant at P≤ 0.05. No significant difference was observed in the dry weight of shoots between the two cultivars, that is, Ka and Ou in all salinity levels and control conditions. Fresh weight was significantly declined in all salinity levels compared to control plant. However, a significant difference in fresh weight was only observed between Ak cultivar and the other two cultivars in the salinity of 300 mM (Table 1). Root weight was higher in Ak cultivar than the other two cultivars under the control and salinity stress conditions. However, this increase was statistically significant only under the control conditions. The results presented in Table 1 also indicated that root weight was decreased with exceeding stress in all pistachio cultivars. This decrease was statistically significant in salinities over 200 mM. No significant contrast was observed between the two cultivars, Ou and Ka regarding the fresh and dry weights of root under the control and different salinity conditions. No significant difference in number of branches was observed between the control and salinity level of 100 mM among different pistachio cultivars. However, under stress condition at 200 and 300 mM, number of branches showed a significant decrease in all varieties. This decrease was more pronounced in the higher level of salinity and in Ou and Ka cultivars compared to Ak cultivar in different salinities. On the other hand, no significant difference was observed between Ak cultivar and the other two varieties regarding the number of leaves for the control and low or medium salinity conditions. However, number of leaves in Ak cultivar was Salinity levels (mM) 0 100 200 300 significantly higher in the high salinity level compared to the other two cultivars (Table 1). It is of interest to mention that the number of leaves was decreased in all the different pistachio cultivars as salinity stress was increased. This decrease was statistically significant only at the medium and higher salinity levels compared to the control plants. Moreover, the reduction leaves number in Ou and Ka cultivars was more intense from moderate salinities onwards (Table 1). No tangible difference was observed for plant height between the control and the low salinity conditions among different cultivars. However, Ak cultivar had a significant increase in plant height under the moderate and high salinities in comparison with the two other ones. However, the plant height of different pistachio cultivars was shortened with exceeding salinity stress; this reduction was more intense in Ou and Ka cultivars under the moderate and high salinities (Table 1). The impact of chloride-sodium on leaf area revealed no considerable difference among different cultivars under the control and low salinity conditions. However, the difference between the three cultivars was intensified with the increase in salinity stress since this difference between Ak cultivar and two other cultivars was significant in high salinity. Ak cultivar showed the maximal leaf area among all varieties although this parameter was reduced in all varieties as stress was increased and it was more noticeable for Ou and Ka cultivars under high salinity (Table 1). The effect of chloride-sodium on the chlorophyll and cartenoid content in leaves is presented in Table (2). These results indicated that no significant difference in cartenoid pigment was observed among different pistachio cultivars
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Journal of Medicinal Plants Researc
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Editors Prof. Akah Peter Achunike E
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Electronic submission of manuscript
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Fees and Charges: Authors are requi
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Table of Contents: Volume 6 Number
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Figure 1. Chuanxiong Rhizoma (http:
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Table 1. Identification of main pea
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Table 3. Pharmacokinetic parameters
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active component study, many invest
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and 1185 kg ha -1 in the first site
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Table 1. Recommendation for use fer
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Sadanandan AK, Hamza S (1996c). Stu
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and it has been traced to South Ame
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Table 3. Results of phytochemical a
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(a) (b) (c) Figure 1. (a) Normal ce
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emedies in the south of Brazil. J.
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known fatty acids and terpenoids we
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Table 1. Renal function tests of ra
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Al-Radahe et al. 2271 Figure 3.nnnn
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namely disruption of the vascular e
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Zayachkivska OS, Konturek SJ, Drozd
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β-carotene were purchased from Sig
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Figure 1. Effect of ethanol concent
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Table 3. Climatic and soil factors
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content of polysaccharides. Phospha
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Serge et al. 2285 Table 1. Relative
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Percentage inhibition % of inhibiti
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Table 1. Result of phytochemical sc
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Table 3. Result of thin layer chrom
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Table 2. MIC of a compound 1 from c
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Zirihi et al. 2301 Figure 2. Termin
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Figure 5. T. catappa Linn. (Combret
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Figure 8. A. fumigatus: Aspect of c
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Figure 12. Sensitivity of A. fumiga
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Noormi et al. 2311 Table 1. Callus
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Table 2. Contd. NoC=No callus. 4.0
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Figure 2. Contd. at 2.0 mg/L yellow
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Table 1. Biochemical parameters in
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Figure 3. Bcl-xL reactions in inter
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ACKNOWLEDGMENTS The authors would l
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Figure 1. The first page of the boo
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Figure 3. The most widely included
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Figure 4. Lithographic print “St.
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Table 1. List of plants included in
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Table 1. Contd. Nedelcheva 2333 Cin
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Table 1. Contd. Nedelcheva 2335 Rhe
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14 7 1 15 1 35 17 16 Imported plant
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Academy of Science Publishing House
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et al., 1996; van Wyk and Gericke,
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DPPH inhibition (%) DPPH % inhibiti
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% Mortality Concentration (µg/ml)
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information on plants used for the
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Table 1. Levels of independent vari
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Arbutin (mg/g) Co-solvent Figure 1.
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Arbutin (mg/g) Extraction pressure
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Table 3. Arbuitn content of Asian p
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Arbutin (mg/g) Extraction pressure
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Arbutin (mg/g) Extraction temperatu
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Table 1. Chemical composition of es
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Darjazi 2369 Table 2. Statistical a
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Table 3. Correlation matrix (number
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Table 1. Precision test. extraction
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Figure 2. Effect of different alcoh
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Table 6. Results of ANOVA. Nie et a
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Table 2. Plant data and MIC values
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exhibiting the polar extracts of P.
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Table 3. Contd. AcOEt ++ ++ ++ - Me
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Figure 1. Map of Ladakh region of I
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Warghat et al. 2391 Table 3. Summar
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Table 5. Inter-Population genetic i
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Excoffier L, Smouse PE, Quattro JM
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ice residue is an ideal raw materia
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DPPH• scavenging ratio(%) OD valu
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scavenging effect on free radical a
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our new CL system. Fenton reaction
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1 2 Time (s) Figure 2. Kinetic curv
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2524 J. Med. Plants Res. ACKNOWLEDG
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UPCOMING CONFERENCES 15th European
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