نشریه علوم باغبانی
سال سی و دوم شماره 1 (پیاپی 37، بهار 1397)

The genus Nepeta with the common Persian name “Pune-Sa” is one of the most important genera of Lamiaceae family. It has aromatic plants which are endemic of Iran. Its different species are distributed almost in all parts of Iran. Some of these species are valuable in medicine and used for medicinal purposes. Nepeta glomerulosa Boiss. is one of the most medicinal species of this family. This study was conducted to examine seed germination characteristics and to evaluate cardinal temperature of seed germination of N. glomerulosa.
Seeds of N. glomerulosa were collected from Abade region of Fars province in the middle of September 2014. Treatments were including four levels of cold period (1, 2, 3 and 4 weeks at5ºC) and four levels of potassium nitrate (0, 0.1, 0.2 and 0.3 %). Factorial experiment with a completely randomized design in three replications was done, to evaluate seed germination percentage, germination rate, T50, mean germination time, root and shoot length, root and shoot fresh and dry weight. Also, the effects of 8 constant temperatures including 5, 10, 15, 20, 25, 30, 35 and 40 ˚C and 3 variable temperatures including 5/15, 10/20 and 20/30 ˚C (12h light/12h dark), was investigated to evaluate germination percentage, germination rate, mean germination time and cardinal temperatures of seed germination.
The results showed that cold period and potassium nitrate interaction was significant only on root length. Potassium nitrate had no significant effect on germination rate, but the germination rate was increased by increasing the duration of being at5 ˚C, so that the highest germination rate was observed in 3 and 4 weeks at5 ˚C (1.7 and 1.2 /day respectively). The lowest germination rate (1.0 /day) was observed in 2 weeks at5 ˚C treatment. The maximum shoot length (51.5 mm), shoot fresh weight (30 mg), shoot dry weight (1.3 mg) and root dry weight (0.6 mg) was obtained in 2 weeks at 5 ˚C treatments andpotassium nitrate 0.1% was also the best treatment for abovementioned traits. Among the treatments, 3 weeks cold period and 0.1 % potassium nitrate showed the most positive effect on seed germination characteristics of N. glomerulosa, although, had no significant effect on germination percentage.
Based on the results of the second experiment, temperature had significant effect (P≤0.01) on germination percentage, germination rate and mean germination time. The highest amount of germination percentage (76 and 77.3 %), germination rate (4.4 and 5 /day) and mean germination time (5.1 and 5.4 /day) were recorded in the range of 20 to 25 ˚C. Germination rate and germination percentage were substantially reduced by increasing temperature above 25 ˚C. The day and night variable temperatures had significant effect on seed germination percentage and germination rate of N. glomerulosa seeds. The highest amount of germination percentage (72 and 69.3 %) and germination rate (4.6 and 5.7 /day) were observed at10/20 and 20/30 ˚C. The results of the relationships between temperature and germination rate showed that base, optimum and maximum temperatures for seeds of N. glomerulosa were 15 ˚C, 22 ˚C and 46 ˚C, respectively.
The overall results showed that most chilling occurred under natural conditions during winter and early spring which caused increasing inseed germination of lots of plants. This also found that increasingin chilling period significantly improved the seed germination characteristics of N. glomerulosa. Also, treating with potassium nitrate had significant effects on germination properties of N. glomerulosa. Although applying potassium nitrate and cold period did not show interaction effects on seed germination.
The effects of different day and night temperatures on seed germination rate and percentage have been reported in many medicinal and aromatic plants as may be due to the similarity of these conditions to the natural conditions. Comparing cardinal temperatures of N. glomerulosa with the other species of this genus indicated that they have different cardinal temperatures. Many of researchers reported that germination cardinal temperatures are significantly different amongspecies and different genotypes of species. Determining of cardinal temperatures helps us to estimate temperature range of wild species in natural areas, planting time and also suitable areas for cultivation.

Iran is the third producer of Melon (Cucumismelo L.) after China and Turkey in the World.Melon is one of the main crops in Khorasan province in the term of cultivation area. The cultivar of Khatooni, among many varieties of melon, can be considered as a breeding cultivar because of its great qualitative traits including sweetness and flavor. This cultivar is also economically important and has many attractions for export. However, this cultivar is susceptible to some of plant diseases caused by different bacterial, viral and fungal pathogens which among them vascular wilt caused by Fusariumoxysporum f. sp. melonis (Leach &Currenu) Snyder & Hansen (FOM) is one of the most destructive soil-borne diseases in melon in Iran particularly in temperate and cold regions. The control of melon diseases has depended increasingly on the extensive use of toxic chemicals (pesticides). Many of these chemicals have been shown to be toxic to non-target microorganisms and animals and may be toxic to humans. Another problem with using chemicals to control plant pathogens is that a pathogen may become resistant to the chemicals.
The most promising control approaches are included conventional breeding and genetic engineering of disease-resistant plants. However, the conventional breeding method of melon is very complicated therefore; genetic engineering could be an effective and quick tool for producing new cultivars. In vitro direct regeneration is one of the most crucial step in gene transferring programs (20). In the present study, the effects of explant type and medium were considered in order to optimize the regeneration condition for melon (cv. Khatooni).
Material and The Khatoonicv.was provided by the Seed Bank of Plant Sciences Research Center from Ferdowsi University of Mashhad. The seeds were disinfected by NaClO in 1 % concentration, washed three times by distilled water and cultivated on MS medium (Murashigo and Skoog) for germination. The explant types were; lateral buds of cotyledon and the true leaves and hypocotyls. The selected media were: M1(0.1 mg.l-1 BAP (6-Benzyl amino purine)), M2 (0.25 mg.l-1 BAP), M3 (0.5 mg.l-1 BAP), M4 (0.75 mg.l-1 BAP),M5 (1 mg.l-1 BAP)and M6(including 0.5 mg.l-1 BAP and 0.01 mg.l-1 NAA (1-Naphthalene acetic acid))The experiment was repeated three times. The experimental –was conducted factorial based on completely randomized design and statistical analysis was performed using the SAS and JMP software. The corresponded graphs were drawn by Microsoft excel 2007.
The greatest amount of shoot regeneration in cotyledons was observed in M5 with 1 mg.l-1 BAP while minimum shoot regeneration was observed in M1with 0.1 mg.l-1 BAP. Cotyledon leaves showed the best regeneration efficiency among the other evaluated explants as it has also been reported by many other researchers (12, 8, 20, 21, and 2). The observations showed that in all BAP concentration, hypocotyls only formed callus tissue and did not produce any stem. In all three explants (true leaves, cotyledon and hypocotyls) rooting was observed in a treatment combination including 0.5 mg.l-1BAP and 0.01 mg.l-1 NAA 20 days after cultivation while no stem was formed at the same time. In three weeks after exposure , a weak root system was formed on the rooting mediumin without hormonetreatment . In MS medium enriched with 0.01 and 0.05 mg.l-1 NAA an appropriate root system was formed after 8 to 10 days. The medium containing 0.01mg.l-1 NAA was more appropriate for root regeneration (rooting).
Many studies have been shown that, the manipulation of different phytohormones ratio such as auxin and cytokinin in cultural medium is a suitable strategy to manage differentiation and organogenesis programs in plant (9). In the present study, the effects of explant type and medium were investigated in order to optimize the regeneration condition for melon (cv. Khatooni). As a result, in all BAP concentration, hypocotyls only formed callus tissue and did not produce any stem. However, by increasing of BAP hormone concentration in the cultivated media, shoot regeneration will also increase. The result of our study showed that, cotyledon leaves were more effective on shoot regeneration compared to the true leaves and hypocotyls as it has also been reported by other researchers.

Lily (L. ledebourii) is the rarestspeciesof thegenusLilium, and grows in Caucasus region. Iranis one of the important distribution areas of this endangered species. It is important as an ornamental plant due to its large and attractive white flowers that are equal to those of commercial lilies in terms of beauty.The two main constraints on growing this plant are a low multiplication rate and the high cost of bulb production. Five to ten flowers commonly appear on each plant, even specimens with up to 15 flowers have been observed. Plant tissue culture techniques are widely used in plant propagation and using these methods can effectively provide micro-propagation of this plant in large scale. High percentage ofregeneration is necessary for plant protection, using in the breeding programs and gene transfer to this plant. Therefore, the effect of plant growth regulators and abiotic stress (ultrasound) werestudied on the bulblet production and root induction of Lilium ledebourii.
The experiment was factorial based on completely randomized design with four replicattions and was carried out in tissue culture lab of University of MohagheghArdabili in 2015. For this purpose, segmentsof scale explant was treated with ultrasound and cultured on MS medium supplemented with different concentrations of NAA and BA alone and/or in combination with each other. In this experiment, different concentrations of NAA (0, 0.01, 0.1 and 1 mgl-1) and BA (0, 0.01, 0.1 and 1 mgl-1) and different Ultrasound exposure duration (0, 5, 10, 20 and 30 second) were studied. In order to remove possible contamination from the media, all media were autoclaved for 20 minutes at 121 °C. At the end of the experiment, the number of bulblet, root length, fresh weight of bulblet were recorded. The cultures were kept at 25±2°C under illumination with daylight fluorescent lamps (30 mol m-2s-1) at 16 h photoperiod. Data was subjected for analysis of variance and compare means using SPSS 16.
The results showed that ultrasound had negative effect onroot length, so that the highest root length was observed in explants without ultrasound treatment. Result also indicated that ultrasound had positive effect on bulblet production and root induction. A different effect of growth regulators was observed in similar media on the bulblet formation percentage. The 0.1 NAA concentration had a higher efficiency while increasing NAA insignificantly decreased bulblet induction. The highest total weight and number of bulblets obtained by 0.1 mgl-1 NAA. Concentrations of NAA increased rooting percentage. Different concentrations of NAA had also significant effects on some traits. So that, the highest weight of bulblets obtained by 0.01 and 0.1 mgl-1 BA and the highest number of roots obtained in control. Bulblet maximum mean weightwas in30 seconds ofultrasoundtreatment, which hada significantdifference with the control treatment (without ultrasoundtreatment). In the other hand, ultrasound increased the number and weight of bulblets.Mechanical stress and microstreaming by acoustic cavitation might be considered as the most possible cause of the various physiological effects of ultrasound on cells. The enhancement of V-ATPase transport and ATP hydrolysis activities seem to be an ultrasound-induced metabolic response of cells. High-intensity ultrasound is well known to be destructive to biological materials, disrupting the cell membranes and deactivating biological molecules such as enzymes and DNA. Low-intensity ultrasound, on the other hand, has shown a range of sub lethal biological effects that are of potential significance in biotechnology. There are several processes that take place in the presence of cells or enzymes activated by ultrasonic waves. High-energy ultrasonic waves break the cells and denature the enzymes. Low-energy ultrasound can modify cellular metabolisms or facilitate the uptake of nutrient, and make them easily through the cellular walls and membranes. In the case of enzymes, the increase in the mass transfer rate of the reagents to the active site seems to be a most important factor.
The results showedthatthebulblet production at first stages and a little root formation in tissue culture is useful for fast bulblet inductionandthenrooting. Finally, it seems that ultrasound in combination with plant growth regulators have the potential to produce the highest average number of bulblets in the scale explant.

Ornamental plants play a vital role in meeting the mental and spiritual needs of peoplethat considered significance from commercial point of view as well. Optimal production of agricultural products requires suitable soil and adequate and absorbable nutrients for plant. Organic materials are important because ofimproving soil physical properties and soil fertility. Soil fertility depends on the content oforganic matter as well as the quality, quantity and dynamics of these materials insoil. Organic acids are an important source of organic matter. One of the most abundant forms of organic matter in nature is humic compounds which can be found in all soil and water environment. They play an important role in cation exchange, nutrients release, phosphorus buffering capacity and metal and toxic organic molecules maintenance.
In order to evaluate the effect of humic substances on morphological characteristics of geranium, thisexperiment was conducted in research greenhouse of Ferdowsi University of Mashhad during the years 2014 and 2015. The experiment was carried out as factorial based on completely randomized design with two factors and three replications. The first factor hadfour levels of humic acid (0, 0.2, 0.5 and 1 g/l), and the second hadfour levels of fulvic acid (0, 0.2, 0.5 and 1 g/ml). Treatment was usedalong with irrigation. Morphological characteristics included the number of leaves, number of nodes, number of branches, plant height, root length, leaf area, fresh weight of shoot, fresh weight of leaf, fresh weight of root, dry weight of shoot, dry weight of leaf, dry weight of root, volume of shoots, volume of root, fresh weight of shoot were measured at the end of the experiment. Statistical analysis of the results was performed by using Jmp-8 software. Charts were drawn using Excel 2010 and difference among treatments means were compared with LSD test.
The results of means comparison showed that combined use of humic acid and fulvic acid hadpositive effects on growth traits such as height, internode length, root length, fresh weight of shoot, fresh weight of root, fresh weight of leaf, dry weight of shoot, dry weight of leaf, volume of shoot, volume of root and leaf area. Research had shown that the application of humic fertilizers increased nitrogen content in shoots of the plant. It wasreported that nitrogen compounds existed in humic acid are important factors affecting the growth of plants. Humic acid also increasedshoot growth by increasing the uptake of nitrogen, calcium, phosphorus, potassium, manganese, zinc, iron and copper as well as hormone-like properties. It wasalso found that humic acid enhancedplant growth by increasing the activity of the RuBisCO enzyme and the subsequent increase in photosynthetic activity. Furthermore, humic acid reduced the pH of alkaline soils and causedthe nitrogen to be absorbed to a greater extent. The use of humic substances increased the leaf area and thus photosynthesis, therefore leading to the production of more dry matter in plants. Humic fertilizers also had a significant effect on root growth. Researchers had suggested that the presence of oxygen groups in humic acid increased lateral root growth. Although humic acid increased the growth of both root and shoot, its effectiveness on the root system wasmore evident.
This experiment was conducted with the aims of evaluating the effect of humic organic fertilizers, reducing the harmful effects of chemical fertilizers and improving the quality of plant growth in geranium. According to the results obtained from the present research, it can be concluded that combined application of humic acid and fulvic acid had greater effects on the majority of traits compared to the sole application of aforementioned compounds.

Insemination combined Effective Microorganisms (EM) to the plant ecosystem can improve photosynthesis and yield of fruit plants. The useful microorganisms contained in effective microorganisms (EM) mix include photosynthetic bacteria (Rhodopseudomonaspalustris, Rhodobactersphacrodes), lactic acid bacteria (Lactobacillus plantarum, Lactobacillus casei, Streptococcus), yeasts (Saccharomyces spp.), actinomycetes (Streptomyces spp.) and fermentation fungi (Aspergillus, Penicillium).When EM applied with the soil or sprayed on the plant will be expanded, the population of photosynthetic bacteria and nitrogen stabilizer. Optimal nitrogen consumption not only causes delays old age it will also change the appearance of the plant. The availability of nitrogen to the roots especially in the early stages of growth causes elongation of the aerial parts of the plant. Proper use of nitrogen has a positive effect on flower number and the beginning of flowering in pepper but high consumption of this element has caused a delay in the formation of flowers and increase plant growth. The aim of this study was to investigate the effects of Effective Microorganisms by two methods of soil application and foliar application in different nitrogen levels on yield and morphological characteristics of strawberry cv. Paros
In order to study the impact of effective microorganisms (EM) on morphological characteristics and yield of strawberry cv. Paros a split-split-plots experiment was conducted based on Randomized Complete Block Design (RCBD) with foliar and soil treatments of effective microorganisms, each in four levels (control, one, two and three percent) and nitrogen treatment in the form of urea fertilizer in three levels (50, 100 and 150 kg ha-1of the net nitrogen) with three replications in University of Mohaghegh Ardabili during 2014-2015 years. Land was prepared in late August and potassium and phosphorous fertilizers were added as 150 and 50 kgha-1, respectively. Strawberries were planted in early November 2014 in double rows on beds covered with black polyethylene foil.Nitrogen levels in the main plots, methods of application of EM in the subplots, the concentration of effective microorganisms in the sub-sub plots were placed.Half of the amount of nitrogen at planting time, 25 percent in the April and 25 percent in the May was given with irrigation water to plants.Treatments of EM, after the start of growth seedlings weekly basis a period of two months, were applied. In soil treatment with irrigation water and foliar Treatments be applied after the irrigation (control plants with distilled water). In this study were measured traits leaf and root dry weight, root length, number of runner, chlorophyll a, band total leaf and yield. Data were analyzed by SAS ver9.3 software and means were compared by LSD test (P≤0.05).
Analysis of variance showed that the effect of different levels of effective microorganisms at the level of 1% probability were observed significant difference in the characters fresh and dry weight of root, root length, number of runner, total chlorophyll, leaf area and yield and in the probability level 5% is significant on the characters fresh and dry weight of leaves, chlorophyll aand b. Based on the results the effect of different methods of application of EM and interaction effects were not significant differences in any of the studied indicators.According to the analysis of variance,using of nitrogen had significant effect on the fresh weight yield root, root length, number of runner, chlorophyll A, B and total leaf area and yield at1% probability. The treatments had also significant effect on fresh and dry weight of leaf and root traitsat 5%,probability. Based on the results of this study the concentration of 1% and 3%of effective microorganisms and 100 kgNha-1 have positive effects on the parameters compared to the control.The results indicated that 2% EM was treated with a maximum of 185 g, and control treatment with 136 g had the lowest yield per plant.Fruit yield in plant with nitrogen application at 100 kg ha-1had the highest production (188 g) among nitrogen levels.
According to the results of this review, the best result was in treatment 2% of EM and 100 kg of nitrogen per hectare. Method used of EM in this study had no significant difference; accordingly method of consumption has no impact on the traits mentioned.

Seedless barberry (Berberis vulgaris L. var. asperma) is one of the most important horticultural and medicinal crops and also valuable fruit in South Khorasan province. Fresh barberry fruit has a very short shelf life, due to the sensitive juicy fruit tissue in the ripening stage and decay during storage. Hence, most of the harvested fruits are dried traditionally or processed as juice, jam, and other products. However, the quality of both fresh and dried fruit is low due to different environmental stresses such as drought and salt stress and consequently deficiency of minerals during growth and development and also at harvest and during storage. It has been reported that pre-harvest foliar spray of calcium salts and salicylic acid can improve resistance to pathogens before harvest and reduce physiological disorders during handling and storage in many fruits. However, there is no information regarding the effect of calcium and salicylic acid spray as well as storage duration on fresh seedless barberry fruit. The objective of the current study, therefore, was to investigate the effects of the preharvest application of calcium chloride and salicylic acid on physiological attributes and postharvest quality of barberry fruits during long-term cold storage. The experiment was conducted as split-plot in time and in a randomized complete block design with five treatments and in three replicates in the Research Orchard of Agricultural Faculty of the University of Birjand during 2014. Fifteen similar and 25-years old trees were used for the experiment. Pre-harvest treatments of foliar spraying were salicylic acid (1 and 3 mM) and calcium chloride (1 and 2%) and distilled water as control. Sprays were applied at 30 and 15 days before harvest. Fruits were harvested at commercial ripening stage, packed in plastic boxes and stored at cold room at 4±0.5 °C and 85±5 RH. Physicochemical and quality evaluations including fruit fresh weight and volume, and juice volumes of 100 berries, pH, total soluble solids (TSS), titrable acidity (TA), ascorbic acid, weight loss, electrolyte leakage of fruit and calcium content of leave and fruit were measured at harvest and after 30, 60 and 90 days of storage. Analysis of data was performed using Gen Stat program (ver 12) and comparison of means was done by LSD test at 5% probability. The analysis of variance showed that the main effects of foliar spray with calcium chloride, salicylic acid and interaction of treatments were significant in storage time. The results showed that 1 mM of salicylic acid increased the fruit fresh weight, fruit volume and fruit juice, pH, total soluble solids, ascorbic acid and titrable acidity compared to other treatments. It has been reported that salicylic acid increases cell division and growth resulting in greater yield. It was also observed that application of salicylic acid at 3mM reduced electrolyte leakage significantly that can be useful to reduce chilling injury. The lowest weight loss was obtained in sprayed fruit with calcium chloride (2%) during cold storage, in agreement with the previouswork on plum. Foliar application of calcium chloride increased the calcium content of leave and fruit, although calcium application was not a significant effect in fruit tissue. Generally, using of chemical treatments increased the physicochemical characteristics of the fruit in seedless barberry, among them calcium chloride 2% and salicylic acid at 1 mM were most effective. It can be concluded that pre-harvest spraying of seedless barberry shrub with salicylic acid and calcium chloride can improve the quality of fruit as well as extend the storage life. This is because a foliar application with salicylic acid and calcium chloride significantly affected fresh fruit weight, weight loss, fruit volume, electrolyte leakage, pH, TSS, TA and ascorbic acid in treated fruit compared to the control. Salicylic acid at 3 mM significantly decreased electrolyte leakage, which can be used as a useful tool for increasing resistance to chilling injury during long-term storage. However, further studies are needed as the effect of chemical sprays on fruit quality and quantity properties depend on different factors such as tree age and types, environmental conditions during and after the spray and also chemical concentration.

Mulching is a useful practice with the potential of conserving moisture, reducing evaporation, modifying soil temperature, and improving aeration as well as releasing nutrients in the soil profile. Mulching involves the use of organic materials (e.g. crop residues, straw, grasses, and farmyard manure) or inorganic and synthetic materials (e.g. polyethylene sheets, and gravels). Application of mulch can notably as a soil management method influence agricultural crop production despite the limited amount of water available in arid and semi-arid areas.
Field studies were conducted in a randomized complete block design with five replications for two years in 2014 and 2015 at Feyzabad city, Iran (34° 40´ N, 58° 25´ E). The aim of the study was to investigate the effects of different mulching treatments on physiological traits of pistachio (Pistaciavera L.) in relation to soil temperature and moisture variations. The treatments were: M1 (whitepolyethylene film mulch covered with two centimeter of soil with width cut), M2 (white polyethylene film mulch covered with two centimeter of soil with circular cut), M3 (green polyethylene film mulch which was as a single layer pulled on water strip so that irrigation water passes under the coverage), M4 (woodchip mulch), M5 (barley residue mulch), and CK (control or no mulching). The measurement (soil moisture and temperature, stomatal conductance and leaf temperature) was carried out periodically on the central trees (4 trees per treatment) every 12, 24 and 36 days after irrigation between 12-15 pm from early-May to late September. The data obtained from the experiment were subjected to an analysis of variance (ANOVA) by using SAS 9.1 software. Difference between means was compared using least significant difference test (LSD) at 5% level (p ≤ 0.05).
The results showed that the mean soil moisture percentage in all mulches especially plastic mulches was higher compared to the control over two years of experiment. During the two years of experiment, treatments of M1, M2 and M3 with 35.2, 35 and 38.9 °C and treatments of M4, M5 and CK with 28.8, 29.6 and 32.8 °C indicated the highest and lowest average soil temperature at 20-30 cm soil depth, respectively. Also, similar results were observed at 50-60 cm soil layer. M3 and CK treatments with 47.7 and 41.1 mmol/m-2s-1 had the highest and lowest mean stomatal conductance, respectively. The mean values of leaf temperature during the growing period under M1, M2, M3, M4 and M5 treatments were 2.2, 1.9, 2.4, 1.7 and 1 °C in 2014, and 1.8, 2.2, 2.2, 1.5 and 1.1 °C in 2015 lower than CK (control) treatment, respectively. According to correlation results significant difference was not observed between stomatal conductance and soil moisture, but the relationship of this parameter with soil temperature and relationship between leaf temperature with soil moisture and soil temperature were significant. Thus, this results demonstrate that stomatal conductance and soil temperature weresignificantly affected by soil temperature, so that an increase in soil temperature to 40 °C, canbe improved these traits if other soil and climatic factors werenot limiting. The results also showed that M3 mulch (plastic mulch on soil surface) despite the higher soil water storage due to providing of soil high temperatures, can be leads to moisture stress in heavy cropping year.
The results of thisstudy showed that organic and inorganic mulches improved the soil moisture content throughout the two growing seasons, thus reduced negative effects of water shortage. Mulch treatments indicated smaller fluctuations in soil temperature compared to CK (control) treatment, possibly due to insulating against heat radiation. The results of our study suggested that increase in soil temperature in M3 treatment, despite higher water storage, had limited impacts on improving stomatal conductance and leaf temperature from early July in the second year. Thus, it seems that high soil temperature in this treatment in ON year that tree's water and nutrients requirement is higher than OFF year acts as a limiting factor and restricts absorption of water and nutrients, resulting in the decrease of physiological activity of trees. Generally, it is conclude that white polyethylene films covered with soil especially M2 and organic mulches due to providing optimum soil temperature, preserving soil moisture and lower cost can be better management options to be applied in pistachio orchards especially in arid and semiarid areas, where traditional irrigation methods with high evaporation level are used.

Roselle (Hibiscus sabdarjffaL.) is an annual or biennial plant cultivated in arid and semiarid regions. This plant is grown in parts of Africa, Asia, America and Australia. This plant is ecologically short day, self-pollinated and sensitive to cold and freezing stress. Studies conducted on the medicinal plants in natural ecosystems suggested that using sustainable agriculture system provides the best conditions for the production of these plants, leading to maximum qualitative and quantitative yield in such conditions. In order to analyze the growth indices, leaf area index (LAI) and total dry matter (TDM) is required. Other indices were calculated from LAI and TDM. Considering the importance ofuses andresistant to droughtstress of roselle, this plant cultivate in provinces of Kerman and Fars recently. In order to evaluate the effect of organic, chemical and biological fertilizers on thermal requirements, developmental stages and some of physiological indices of roselle by irrigation levels, was conducted in Jiroft area.
In orderto evaluate the effect of organic, chemical and biological fertilizers on thermal requirements, developmental stages and some of physiological indices of roselle by irrigation levels, an experiment was conducted at Agricultural Research Farm Jiroft of Universityin2013-2014 growing season. Theexperiment was conducted as strip plot based on a randomized complete blocks design with three replications. Three irrigation regimes (100%, 80% and 60% of crop waterrequirement) were assigned ashorizontal factor and four fertilizer resources (mycorrhiza, vermicompost, cow manure and chemical fertilizer (NPK)) were allocated as vertical factor. Plant water requirement was calculated by using salinity parameters (sensitivity coefficient, salinity threshold and gradient of yield reduction), plant characteristics (height and the maximum depth of root), vegetation coefficients (first, middle and end of growth season), characteristics and duration of growth period with AGWAT software. Then, treatments of 80% and 60% of plant water requirements were determined and irrigation water per irrigation period was recorded by the water meter. As drip irrigation system with 0.001 Sq. M. were independently used for each treatment.Traits such as LAI, TDM, LAD, CGR and RGR indices and dry sepal yield were measured. All data were subjected to analysis of variance (ANOVA) using SAS 9.3 software. When F test indicated statistical significance at P The results indicated that roselle is short-day plant and its flowering is stimulated by nearing the short days. Fertilizer resources and irrigation water levels had not significant effect on flowering of plant. Also fertilizer resources and irrigation water levels had significant effect on LAI and TDM. The highest LAD value was found in the cow manure and vermicompost treatments. The highest CGR value was obtained from 100%, 80% and 60% respectively. Growth reduction between irrigation levels 100% and 80% was not significant. Slope decline between irrigation levels (i.e., 100% and 80%) was similar. Slope decline in control and mycorrhizal morethan vermicompost, cow manure and chemical fertilizer. Similar results also reported by Gendy et al., (2012) and Samiran et al., (2010). The results showed that the interaction of Fertilizer resource and levels irrigations on the sepal yield was significant. Sepal yield at all three levels of irrigation showed increase in the plants treated with cow manure, vermicompost and chemical compared with mycorrhizal and control treatment. The highest sepal yield (1217 kg.ha-1) was obtainedfrom100% crop water requirement cow manure treatment and the lowest sepal yield (493 kg.ha-1) was obtained at 60% crop water requirement control treatments.
It seems that among the different fertilizer resources, cow manure and vermicompost have a greater impact on soil water holding compared to other sources of fertilizers (mycorrhiza and chemical fertilizer) by increasing soil organic matter. The results of Growth indices analysis, growth degree days and phenology assessment indicated that the most leaf area index and dry matter was observed at flowering stage and the beginning of flowering rosellewas under the influence of photoperiod. Due to soil and climatic conditions of the study area, using cow manure or vermicompost with 80% of the irrigation water requirement, roselle plant seems appropriate.

Late-season Bunch Stem Necrosis (BSN) is observed as a necrosis of the cluster stem (rachis) that leads to shriveling of berries on the affected portion of the cluster. The BSN symptoms include dark, necrotic lesions on the rachis or individual pedicels that may spread and eventually girdle the affected part of the cluster rachis. Berries distal to a lesion cease normal development, and the unripe berries either abscise or remain on the cluster in a withered condition. Frequently only the cluster tip or a shoulder is affected, while the rest of the cluster develops normally. Symptomatic and non-symptomatic clusters may be borne on the same vine. BSN has been correlated with numerous factors; however, no universal cause and effect relationships have been demonstrated. No pathogens are believed to cause this condition. Instead, certain weather conditions and vine nutrition seem to be associated with its occurrence. Low temperatures and high humidity around bloom or excessive rainfall after veraison may be related to its development. Imbalances between calcium and potassium as well as low levels of nitrogen in vines are other possible causes. Unaffected portions of clusters develop normal fruit quality.ʽBidaneSefidʼ or ʽKeshmeshiʼ is an important grape cultivar in Iran and is frequently affected by bunch stem necrosis disorder around country. The purpose of this study was to determine if mineral nutrition was associated with BSN of ʽBidaneSefidʼ grape cultivar under Urmia growing conditions.
To evaluate the effects of foliar sprays of Ca, Mg and GA3 on reduction of BSN incidence on ʽBidaneSefidʼ cultivar, and to examine the relationships between specific nutrients and the incidence of BSN this field experiment was performed based on randomized complete blocks design with eight treatments and three replicates on 2011in a vineyard around Urmia city. Vines were 8-year-old and were trained as bi lateral cordon. At the end of full bloom and a week after that the vines were sprayed two and five times with 2% MgSO4 and CaCl2 fertilizers alone and combined. GA3 also was sprayed at 70% of flowering with 20 ppm and replicated at fruit set with 40 ppm. Symptoms rate of BSN on bunches and minerals nutrient content(N, K, Ca, Mg and Ca㈏) of leaves and rachis were measuredin veraison phenological stage.
Results showed that in veraison stage the most and the least BSN incidence rate was recorded in control and the vines that were sprayed two times with 2% MgSO4, respectively. Leaf Ca content in vines treated two times with 2% CaCl2 and those sprayed five times with MgSO4ঠ was more than other treatments. Maximum amount of Mg was recorded in leaves of vines sprayed by 2% MgSO4, 5 times during growing season. These vines showed the least BSN rate, too. The most contentof K/Ca were in the leaves of control and GA3 treated vines and the least contentof those was in vines treated five times with MgSO4ঠ. Rachis tissue analysis for nutrient elements in veraisonstage did not consistently reveal any relationship between N, K, Mg, Ca and Mgয and bunch stem necrosis in bunches of studied vines of ʽBidaneSefidʼ cultivar in Urmia region. There are conflicting reports regarding the association of essential nutrients and the incidence of BSN. A high ratio of potassium to magnesium and/or calcium in affected tissues, and also the application of calcium and/or magnesium fertilizers effectively reduced the incidence of BSN in Europe. In California, BSN was not reduced by applications of calcium and magnesium. An increase in the incidence of BSN was reported with application of nitrogen fertilizers. Magnesium and calcium appeared to be involved in the disorder. The results illustrate that BSN-prone vineyards should be individually examined for nutrient imbalance or other stresses that may be contributing to BSN.
Results of this study suggest thecontrol of BSN incidence by fertilizer treatments. Foliar applications ofMg and Ca fertilizers appeared to have effect on BSN control and two times foliar applications of magnesium starting just before veraison minimize the problem. No apparent relationship was found between the petiole and/or rachis K/(Mg Ca) ratio and BSN incidence. External application of GA3 reduced the BSN incidence in comparison to control.

Salinity tolerance in plants can increase the importance of it as a result of the decreasing availability of high-quality irrigation water. Saline irrigation water can have many negative effects on crops. When irrigation water has high salinity, the salt may precipitate on the leaves as the water evaporates. Thus it can result in foliar uptake and phytotoxicity. The irrigation water may also cause accumulation of salt in the substrate, which may lead to salt uptake by the plants. Salt injury occurs when too much NaCl accumulates in the substrate. When excessive concentrations of NaCl are present in the soil, water uptake may be inhibited and it causing a physiological drought stress. However, potassium is required by plants in amounts (in kg unit) of similar or greater than nitrogen (N). K Uptake by the plant is highly selective and closely coupled to metabolic activity. At all levels in plants, within individual cells, tissues and in long-distance transport via the xylem and phloem, K exists as a free ion in solution or electrostatically bound cation. Potassium takes part in many essential processes such as enzyme activation, protein synthesis, photosynthesis, phloem transport, osmoregulation, cation-anion balance, stomatal movement and light-driven nastic movements. Potassium Chloride (KCl) is used as a source of nutrients in agricultural development and also used as relieve salinity stress.
In order to study the mitigation effects of KCl on salinity (NaCl) in mustard plant (Parkland and Goldrush), an experiment was carried out at the Research Greenhouse, Faculty of Agriculture, Ferdowsi University of Mashhad, Iran. The experiment was managed as a factorial arrangement based on completely randomized design in three replications. Treatments were included NaCl (0, 30, 60 and 90 mM) and KCl (0 and 20 mM) and two cultivars.
Relative water content was calculated by the following formula using leaf disc obtained from a young leaf of each plant.
(DW FW/ DW TW)*100FW=fresh weight, DW=dry weight, and TW=turgid weight
Electrolytic leakage was calculated by the following formula: EL=L1/L2 where L1 is electric conduction of leaf after putting in the deionized water in 25°C and L2 is the electric conduction of the autoclaved samples.
Leaf area was measured by Leaf area meter. Shoot and root dry weights were determined after drying the samples in 75°C for 48 h.
Chlorophyll concentration was calculated by the fallowing formula: Chla (μg/ml) = 15.65A666 – 7.340 A653
Chlb (μg/mml) = 27.05A653 – 11.21 A666
Analysis of variance was calculated using MSTAT-C.1software and means were compared by LSD test at probability level of 5%.
The results showed that the treatments of NaCl, KCl and interactions with cultivars were significantly different on dry weight, leaf area, photosynthesis, stoma conductivity and chlorophyll rate. The maximum shoot dry weight (3.44 g/plant) and photosynthesis rate was obtained from T2 (20 mMKCl and without NaCl). The maximum membrane stability index was obtained in Goldrush cultivar and T2. The minimum of these traits were observed in zero mMKCl and 90 mMNaCl. High level of NaCl (60 and 90 Mm) and increasing application of KCl could not improve all traits. According to the result of the analysis of variance increasing density of sodium chloride in planting areas has a special effect on the size of leaves, weight of dried plant and each leaf and dried root. This effect shows a meaningful variation between the weight of dried leaves and its dried root and shoots. The salty areas have a lot of negative ions like Magnesium, Chlorine, sodium and sulfate. These materials are harmful by themselves or cause affective disorder in plants metabolism. Salinity treatments applied to significant influence (p≤0.01) on the characteristics of photosynthesis, stomatal conductance and number of stomata was read out by SPAD. For example, sodium and potassium competition and chlorine and nitrate competition impairs the absorption of nutrients. The result of this reaction is that the plant needs more energy for producing organic matter so it loses most of its energy to resist against salt. This situation causes a low activity of the root and the growing of shoot consequently reduces. Also, weight and length of plant would reduce too. For example, existing potassium in salty lands causes the reduction of sodium in the shoot of plants. This research was done in a pot with the same amount of salt. Potassium causes the reduction of toxicity effects of sodium. This research showed that the potassium can regulate osmotic pressure and permeability of plant cell membranes and also cause to increase plant tolerance to salinity.
In salty condition, increasing the amount of sodium causes the reduction of potassium, compared with sodium. As a matter of fact this kind of reaction causes the reduction of potassium compared with sodium. We know that potassium can cause a suitable osmotic pressure and reduce the destructive effect of oxidation. So, amount of potassium more than sodium in salty lands is known as the standard resistance. In general, increasing the salinity of sodium chloride can decrease morphological and physiological traits of mustard. The use of potassium chloride in T2 treatment showed the best result. However, Goldrush cultivar showed better results compared with Parkland cultivar in salt tolerance.

Today, due to increasing world population, food needs to be provided from different ways. The white button mushroom has also become an important constituent of a healthy diet. Their nutritional value relies on relatively high protein, minerals, vitamins, essential amino acids content and low calories. Mushroom mycelia growth and mushroom development are not only related to genetic factors but also depends on environmental, chemical, and microbiological conditions. Casing soil can protect the compost against desiccation, supporting the mushroom against pests and diseases and providing sporophores development and development and growth of mushroom by gas exchange. Casing layer provides an environmental change in which the mushroom shifts from a vegetative stage to a reproductive one due to microorganisms in the casing soil. Casing soil bacteria influence productivity, product quality and uniformity. Physical and chemical properties of a good casing should be high porosity and water holding capacity (WHC), pH range from 7.2 to8.2, low content of soluble inorganic and organic nutrients and free of disease and pests. Many materials, alone or in combination, have been used as casing both commercially and experimentally, although only a few have been shown to be practical application. Peat is generally regarded as the most suitable casing. Because of its unique water holding and structural properties, it is widely accepted as ideal for the purposes of casing. Peats has a neutral pH and because of it contains organic matter and granular structure can stay porous even after a consecutive irrigation, hold moisture, allows appropriate gaseous exchanges and supports microbial population to release hormone-like substances which are very likely involved in stimulating the initiation of fruit bodies. Peat and limestone commonly used as a casing soil, however problems associated with its use, especially viability, depletion of reserves and alteration of ecosystems, have led to the search for alternative materials.
In order to investigate the effect of different compositions of casing soil on the white button mushroom (Agaricus bisporus Longe) cultivation, an experiment was carried out in laboratory of mushroom research, Department of Industrial Fungi Biotechnology, Iranian Academic Center for Education, Culture and Research (ACECR- Khorasan Razavi), in 2015. The experiment was conducted in a completely randomized design with 8 treatments and 3 replications. The treatments were including peat soil (100%), peat soil (60%) vermicompost (40%), peat soil (60%) cocopeat (40%), peat soil (60%) activated carbon (40%), peat soil (60%) spent mushroom compost (40%), peat soil (60%) vermicompost (30%) activated carbon (10%), peat soil (60%) cocopeat (30%) activated carbon (10%), peat soil (60%) spent mushroom compost (30%) activated carbon(10%). The treatments were applied on the compost block (40 × 60 × 20 cm sizes), that inoculated with the mushroom spawn, commercial line A15. The investigated characteristics including mushrooms yield, mean of mushroom weight, number of mushrooms per square meter, mean of stipe length, mean of cape diameter, which was measured in each harvest flash, separately. The traits were measured three times from casing to first harvest flash and harvest period.
The results showed that the longest duration between casing to first flashing belonged to peat soil (60%) spent mushroom compost (40%). The lowest duration belonged to peat soil (60%) cocopeat(40%), peat soil (60%) activated carbon (40%) and peat soil (60%) cocopeat (30%) activated carbon (10%). Treatments with the lowest duration between casing to first harvest flash, produced the most yield. The results also revealed that casing soils treatments contained cocopeat(40%) and cocopeat(30%) active carbon(10%) had the highest yield, fruit number, fruiting period and shortest duration from casing to first harvest flash as a good characteristic, although the mean of mushrooms weight and diameter of cape were lower than other treatments that it affect on marketing quality. Two treatments including spent mushroom compost (40%) and spent mushroom compost (30%) active carbon(10%), had the lowest yield, fruit number, fruiting duration, the longest duration between casing to first harvest flash as an unfavorable characteristic, although these treatments had the highest mean weight and cape diameter. Overall, the results showed that spent mushroom compost utilization as casing soil is not economically cost-effective.

Spinach (Spinaciaoleracea L.) is considered as a green leaf with fresh and canned consumption, a significant source for vitamin C, vitamin A, carotenoids, flavonoids, folic acid, calcium and magnesium.According to statistics, 4479 hectares of Iran agricultural land were devoted to spinachand the average yield per hectare is equal to 42.18(FAO, 2014).Different regions of Iran havegenetic diversity of the spinach landraceand cultivation of this plant relieson the use of the landraces.Among the different spinach landraces, a landracescalled seed barbed (Spiny), has a significantimportancebecause of some desirable characteristics.Thisstudy was conductedfor comparison of five spinach landracein Isfahan, showedthat spiny seed Varamin with 47.8 tons per hectare fresh yield allocated the highest yield.The number and size of leaves and side branches are the most important components of plant yield.Due to the fact that about 70 percent of total dietary nitrate intake per person per day will be provided through the leafy vegetables,considering the amount of accumulated nitrate in spinach is very important.To evaluate the yield and yield components of eight Iranian spinachlandrace, and two varieties of leafy American and German experiment was conducted for two years.
This research was performedfor two years, in Kabootarabad Agricultural Research Station of Isfahan to evaluate the yield and yield components of eight Iranian spinachlandraces(Varamin prickly seed, Sari, Ghaemshahr, Babul, Kashan, Najaf Abad, Varamin 88 andArdestān), and two American and German leafy cultivars. Randomized complete block design with three replications was usedfor each year and the results were reported based on two-year combined data analysis. In full leaf development stage and before the harvest of each plot, 10 plants randomly were selected and measured attributes: number of leaves, leaf length, leaf width and length of the petiole. In order to assess yield, after removing two distance of half a meter from each side, two central rows of plants per plot (surface equivalent to two square meters) were analyzed. Nitrate content of samples wasanalyzed using ion analyzer. Data statistical analysiswas performed using SAS software and means were compared by Duncan’s multiple rangetest at 5% probability level.
Result and Yield, leaf length, petiole length and height (at 1% statistical probability level) and number of leaves and nitrate content (at 5% statistical probability level) were statistically significant. Spiny seed Varamin with 53.49 t ha-1 fresh yield, produced to the highest yield. In other research according to the environmental conditions and variety, spinach fresh yield have been reported from 18.6 to 44.8 t ha-1. It looks different mass spinach; produce the highest yield in specific geographic environments. In a similar study that was conducted underIsfahan weather conditions, spiny seed Varaminlandraceproduced 47 830 kg ha-1 fresh yield. Range in plant height was from 24 cm to 30.33 cm in Najaf Abad, and Sari landraces, respectively. When spinach was encountered with optimal climate conditions, achieved the maximum height that led toa positive impact on fresh yield. Ardestānlandrace produced the lowest number of leaves (10.67)among the landraces/varieties had been studied. In a similar study of 29 spinach genotypes the leaf number were varied from 12 to 23. Petiole length range between varieties/landraces was from 9.84 to 18.33 cm. Although the petiole length, is a desired trait for mechanized harvesting, but in case of improper management of nitrogen fertilizer can be considered as an undesirable trait. Leafy Varaminlandracewith 231 and 462 ppm nitrate in blade and petiole, respectively, had the lowest and German cultivar with 583 and 1182 ppm nitrate in blade and petiole, respectively, had the highest nitrate accumulation. Despite the differences between the varieties/landraces,the contentof nitrate in petioles was greater than the leaf blade (about 1.6 times) in spinach. Higher concentrations of petiole nitrate compared to the blade considered in many research, and in caseof poor management fornitrogen fertilizers, it may increase up to6 times. In any case, leafy cultivars such as German have been modified for mechanized cultivation and have long petiole with nitrate content. That's why their petioles are separated in the industrialprocessing. It is better fresh petioles of spinach isolated or sufficiently boiled to reduce nitrate levels. According to the results obtained in this study among the examined populations of spiny seed Varaminlandraces, leafy Varamin, Najaf Abad, and Sari can produce yields similar toGerman variety and so are suitable for the production of spinach in Isfahan province.

Nowadays, horticultural products especially in our country are damaged by the climate change events (rising temperatures, water shortage associated by drought and extreme temperature fluctuations). The greatest economic damage to horticultural crops are caused by winter chilling and / or spring frosts, in which has been increased by seventimes during the last 10 years. Indeed, just during 9 months of 2016-2017 year, the chilling/frost damages were reached to 140 million euro of the total damage value of 240 million euro to agriculture (about 60%). According to the Agricultural Insurance Fund, maximum amount of compensation to farmers of the country was paid to apples, walnuts, grapes, almonds, pomegranates, oranges and pistachios orchardists, respectively. Grape (Vitisvinifera), a member of the Vitaceae family, is the most important horticultural crop in Takestan region, ranking top in producing raisins in Iran. Yearly, horticultural practices in order to reduce the severity of chilling damages to crops cost a lot under orchard conditions. Some of these practices are included burying vines under soil, winter ice-watering, flood irrigation, and turning the heaters on. Nowadays, cheap and fast alternative methods such as anti-freezes spraying havebeen considered to reduce the freeze injuries under orchards. These compounds as mechanical barriers either prevent the ice crystals formation on the sensitive plant tissues or activate the freezing tolerance mechanisms in the plants. Salicylic acid (SA), a type of phenolic acid, is one of the most secondary metabolites in the grape. SA not only plays an important role on the determination of the quality, color and taste of fruit, but also influences on the plant responses into environmental stresses such as drought, chilling, salinity and heavy metals. Natural osmolites such as soluble sugars, amino-acids and ammonium compounds are accumulated inside the plants under natural conditions. Commercial anti-freeze/anti-transpiration compounds on the basis of the mentioned natural osmolites are used to increase the freezing tolerance or postponing the bud break of horticultural crops. For example, Bio-bloom, Tiofer®, Cropaid® and Fosnutren® were used as anti-freeze compounds under almond, cherry, pistachio and apple orchards in Iran. The efficiency of this method to reduce the chilling injuries in vineyards is not well known. Therefore, to consider the effects of plant basis anti-freeze compounds on the freezing tolerance of red seedless grape under Takestan climate conditions, the current experimentation was conducted. In this regard, the important freezing tolerance determining osmolites as well as some of the qualitative and quantitative characters of vines in response to these compounds were evaluated.
In order to study the likely increase in grape spring freezing tolerance using the anti-freeze compounds under the orchard, an experiment was conducted as randomized completeblock design (RCBD) with anti-freeze compounds in Agriculture and Natural Resources Research Center of Qazvin during 2012-2013. Minimum monthly temperature during the autumn and winter months was -12.2 °C. The treatments including the spray of SA, Tiofer®, Cropaid®, Bio-Bloom® and water (control) were used on vines during fourphonological stages: before plant dormancy, during bud swelling, start of clustering and during the final clustering on the three red seedless vines per plot. Quantitative and qualitative characters such as cluster weight, yield and total soluble solids (TSS) were measured. Leaves osmolites contents such as proline (Bates et al.,1973) and glycin-betain (Grieve and Grattan.,1983) as well as the status of the freezing resistance index enzymes activities such as Glutathione peroxidase (GPX) (Updhyaya et al.,), Ascorbateperoxidase (APX) (Nakano and Asada,1981) and Catalase (CAT) (Aebi, 1984) were evaluated. All statistical analyses were made using SAS software. The comparison of mean values for the different treatments was made by ANOVA, followed by the Duncan’s test at the significant level of 0.05.
The results showed that the anti-freeze compounds resulted in an significant increase in the TSS, cluster weight and yield by 22%, 17% and 26%, respectively and the most effectswas observed in Bio-bloom. Our results regarding the effect of SA on the TSS was in accordance with other researchers (ShokrollahFaam, 2011; Sedighi et al., 2011; Champa et al., 2014;Marzouk and Kasm, 2011). However the effect of other anti-freeze compounds on the quantitative characteristics has not studied yet. Tiofer and Cropaid had no significant effect on the grape cluster weight. Proline content increased significantly using anti-freeze compounds compared to control during 2012-2013. But there was no significant difference between the different anti-freeze compounds for proline content. The most important content of betaine-glysine was observed in SA and Bio-bloom treatments. A decrease in air temperature during 2013 dormancy period resulted in higher amount of osmolytes contents (betaine glycine) and freezing resistance index enzymes activities compared to those of 2012. The most important contents of these enzymes activities were obtained in SA, Tiofer, Cropaid and Bio-Bloom treatments, respectively and the less important content was observed in control.
According to these results the commercial anti-freeze compounds could be useful not only for an increase in the grape spring freeze resistance but also for an improvement in the grape cluster weight, total soluble solids and yield. Osmolytes contents such as such as proline and betaine glycine and enzymes GPX, APX, CAT are significantly higher in anti-freeze treated plants compared to control. Since high levels of soluble solids, mentioned osmolytes and enzymes resulted in an increase in cold resistance in plant, the use of antifreeze as an easy and inexpensive way to increase the cold hardiness of grapes is recommended. These results were obtained with this antifreeze in 5 ppm concentration. Therefore more researches on the other varieties using different concentrations of anti-freeze compounds are recommended.

Medicinal plants contain antioxidant compounds are benefits to human health and celery as a medicinal plant, is rich in antioxidant. Celery (Apiumgraveolens L.) seed has a lot of essential oil (EO) and this EOs has long been used for fragrance industries, spice, pharmacological and cure diseases. EO for celery seeds is 1.5-3% (v/w). Although a number of methods are applied for the acquisition of essential oils, the most frequently used method is steam distillation. Among EO extraction methods, ultrasound-assisted extraction (UAE) is an inexpensive, simple, and efficient extraction technique. This is most likely due to the intensification of mass transport and to the facilitation of solvent penetration into plant tissues which are damaged by the ultrasounds. Response surface methodology (RSM) is a powerful mathematical model with a collection of statistical techniques where in, interactions between multiple process variables can be identified with fewer experimental trials. It is widely used to examine and optimize the operational variables for experiment designing, model developing and factors and condition optimization. Antioxidants deactivate free radicals before they can damage to the bimolecular cells structure. In the present study, to find effective antioxidant constitutes of EO, antioxidant activity were determined using DPPH scavenging assay and FRAP methods.
In this research, the effect of seed to water ratio (X1), ultrasonic time (X2), and extraction time (X3) on essential oil extraction (v/w) from wild celery seeds were evaluated by the following equations. Antioxidant activity ofEO was measured via two methods of FRAP, and %DPPH30min.
%EO (v/w) = 1.200000 0.075000 X1 -0.300000X2 0.875000X3 0.725000X12 – 0.825000X22 0.725000X32 0.300000X1X2 – 0.150000X1X3 0.300000X2X3
%DPPH30min = 60.4136 6.3782 X1 -22.7935X2 3.7569X3 – 8.0662X12 – 45.5443X22 – 13.5193X32- 6.6459X1X2 – 5.8430X1X3 21.7333X2X3
FRAP (µMFe�µl EO) = 1432.94 166.65 X1 166.22X2 126.29X3 – 384.48X12 – 1215.25X22 – 322.43X32- 87.85X1X2 575.35X1X3 – 219.80X2X3
For optimization of EO extraction conditions, the RSM method was used. A Box–Behnken design (BBD) with three center points was used to study the effects of three independent variables, water to seed ratio (X1), ultrasonic time (X2), and extraction time (X3) on three dependent responses (EO, %DPPH30min, and FRAP). The independent variables were transformed to three levels (−1, 0, 1), and the complete design consisted of 15 experimental runs with three replications of the center points. The following quadratic polynomial model was fitted to the predicted responses of EO, %DPPH30min, and FRAP with changing water to seed ratio (X1 = 375, 500, and 625 ml water to 35g seed); ultrasonic time (X2= 10, 20, and 30 min); and extraction time (X3=1, 2, and 3 h) in extraction samples; Where EO, %DPPH30min, and FRAP stands (Yn) for the predicted responses for X1–X3; b0 is the constant coefficient; b1, b2, and b3 are the linear coefficients; b11, b22, and b33 are the quadratic coefficients; and b12, b13, and b23 are the cross-coefficients. The accuracy of the estimated coefficient was analyzed by ANOVA method and the model accuracy was obtained using the F test at 1 and 5 % by indicating coefficient R2. Yn = b0
According to the results, the best ratio of treatments (optimized condition) was 35g seed to 625 ml water ratio, ultrasonic time of 20 min, and extraction time of 3h. At this optimization conditions, the ratio of 35g seed/625 ml water, non-using ultrasonic, and extraction time of 3h was used as a control treatment. Under optimized condition, the highest essential oil percentage (v/w) and antioxidant activity with styles of FRAP, and %DPPH30min was obtained 2.33% (v/w), 1513.6µMFe�µl EO and 48.52%, respectively. However, at the control treatment, the highest essential oil percentage (v/w) and antioxidant activity with styles of FRAP, and %DPPH30minwas revealed at 1.45 % (v/w), 1064µMFe�µl EO and 29.30%, respectively. In the study of optimization of ultrasound-assisted extraction of anthocyanins from mulberry, using response surface methodology and also, optimization of ultrasonic-assisted extraction of pomegranate seed oil were shown similarity results.
The values of adjusted R-squared (0.9972, 0.9598, 0.9472) for the equation are reasonably close to 1, indicated a high degree of correlation between the observed and predicted values. Therefore, the experimental values were agreed with those predicted by RSM models, and it suggested that using RSM method foroptimization of extraction condition is suitable. Furthermore, application of ultrasonic technology increased essential oil extraction and its antioxidant activity. In addition, results indicated that the UAE method is a powerful tool for the extraction essential oil from seeds.