شاهرخ قرنجیک

Algae by consuming CO2, light, and nutrients in wastewater not only could reduce the environmental pollutants, but also could increase the productivity of biomass and value-added byproducts such as biofuels in their cells. On the other hand, molecular techniques such as Real-Time quantitative PCR (qPCR) helps to better identify the pathways of response to environmental conditions in algal strains. In this study, at the first, the properties of prospective biodiesel were evaluated for three algal strains (Chaetomorpha antennina, Ulva intestinalis, and Bryopsis pennata) grown on different light intensity (8100, 12150, and 16200 lux) and different CO2 concentrations (380, 5190, and 10000 ppm). Finally, the expression profile of three genes involved in CO2 sequestration (CA, CIA5) and light harvesting (CAB) was investigated in superior strain. The results showed that the light intensity of 12150 lux and CO2 concentration of 5190 ppm led to the highest lipid content (30 to 37%) for all three algal strains. Also, all microalgae used in this study have a good ability to produce biofuels. Moreover, it was observed that increasing CO2 concentration has led to a decrease in the expression of the CA gene, but the highest expression of the CIA5 gene was observed in 5190 ppm CO2 concentration. The highest CAB gene expression was also observed in 12150 lux. In summary, the control of expression of the involved genes could significantly determine the final lipid productivity as well as the properties of prospective biodiesel.

Introduction A wide variety of oleaginous yeast and their productions have been used as dietary supplements in food and feed productions. Fatty acid profiles and lipid of oleaginous yeast strains were highly influenced by physic-chemical conditions of medium Material and method Pichia kudriavzevii isolated from the gill of pike perch was capable enough to produce a high quantity of lipid and polyunsaturated fatty acids (PUFA).One factor at time was used to investigate the effect of various selected substrates on growth and lipid production by Pichia kudriavzevii in batch fermentation, subsequently the key substrates were optimized for lipid, biomass, and phenolic components production by using response surface methodology. Results The results of RSM indicated that maximum lipid (28 percentage of dry weight biomass) and phenolic component (2.25 mg gallic acid /g of dry biomass) were achieved in media that had 60 g/l glucose and 9 g/l protein resources. Dry weight biomass which had the highest content of lipid produced the most phenolic components with the highest antioxidant activity (50 % Inhibition).Pichia kudriavzevii’ lipid mainly consisted of palmitic acid (21.7%), oleicacid (33.2%) and linoleic acid (23.2 %), pointing that these fatty acids could be considered a reliable source of biodiesel production. Discussion and conclusion The maximum content of phenolic components and lipid production were obtained in the same medium. To minimize the lipid oxidation, yeast was stimulated for high phenolic components production which had the highest antioxidant activity to prevent the accumulated lipid from oxidation. Overall, temperature reduction by varying methods had a significant impact on fatty acids profile of Pichia kudriavzevii lipid so that it could be regarded not only as biodiesel but also PUFAs sources.

Chickpea (Cicer arietinum L.) is a low cost crop in agronomic system of semi arid to arid region overall the world. This crop due to adaptable capacity is caltivated in wide range of environmental conditions and different type of soils. Chickpea seeds contain 24-34% protein, this is 2 to 3 times more than cereals and 14 to 24% more than tuber plants. Chickpea seeds inoculation with rhizhibium cause more N2 fixation and increase chickpea yield. Mesorhizobium ciceri is one of the rhizobium genus that can have good symbiosis with this crop and fix atmospheric nitrogen up to 60-90% of crop nitrogen requirement after establishment on root crop. Therefore it could be said that biological fertilizers can increase crop production in a sustainable system. Mesorhizobiom can increase chickpea seeds and ultimately increase soil Phosphorus and Zinc concentration. Researchs showed that sulphur deficiency cause decrease seed yield. Nevertheless soil sulphur supply can help seed filling and increase economic yield. The aim of this study was evaluating effect of Mesorhizobium and Thiobacillus bacteria’s and sulfur on yield and yield components of chickpea (Cicer arietinum L.) and some soil parameters. In order to study the effect of Mesorhizobium and Thiobacillus bacteria and sulfur on yield and yield components of chickpea (Cicer arietinum L.) and some soil parameter, a field experiment was conducted at Jotai region of Khorasan Razavi province in 2013. This research was conducted as factorial based on randomized complete block design with three replications and 18 treatments. Experimental factors were three levels of sulphur (zero, 500 and 1000 kg ha-1), thiobacillus bacteria in two level (control and inoculation) and Mesoreizhobium in three levels (control, Mesorizhobium ciceri strain 190 and Mesorizhobium ciceri strain 9). Plot size was 2×4 m2 and row spacing and intra-row spacing were 60 and 20 cm, respectively. Irrigation were done at every 14 days. Weed control were done manually. Disease and pest were not recorded during experiment. After harvesting the crop, plant and soil samples from each plots were collected and transported to laboratory evaluation. Analysis of variance were done by SPSS software. Figures were prepared by Excel 2007. Mean comparison of data were done for least significant difference (LSD) at 1 and 5% level of significant. Analysis of variance of plant height showed that main effect of sulphur on plant hight were significant at p<0.01 and interaction effect of sulphur and mesorhizibium bacteria on plant hight were significant at p<0.05. These result is consistent with results of Solemani & Asgharzadeh (2010). Analysis of variance of data showed that different species of mesorizhobium bacteria had significant effect at p<0.01 on cheickpea yield. Mean comperision showed that the highest seed yield (1146.73 kg ha-1) belong to mesorizhobium ciceri 190 and lowest seed yield (971.07 kg ha-1) belong to mesorizhobium ciceri 9. Scientific reports showed that mesorizhobium inoculation with 25 kg ha-1 zinc sulphate had significantly increased the number of pods in plant and handred seed weight. Analysis of variance of root length showed that main and interaction effect of treatments were not significant on root length excep in mesorizhobium that were significant at p<0.05. Analysis of variance of seed protein showed that main effect of sulphur, mesorizhobium, sulfhur and mesorizhobium and thiobacillus with mesorizhobium were significant at p<0.01. Role of sulphur on seed protein were well understood. Analysis of variance of seed yield showed that main effect of mesorizhobium and interaction effect of sulphur and thibacillus and sulphur and mesorizhobium on seed yields were significant at p<0.01. Shrivastava et al, (2000) showed that mesorizhobium bacteria inoculation had significant effect on soyabean yield. Analysis of variance of soil sulphur showed that interaction effect of sulphur and thiobacillus had significant effect at p<0.01 on soil sulphur, phosphorus and soil pH but had no effect on soil potassium and electrical conductivity at p<0.05. Main effect of sulphur had significant effect on soil phosphorus and soil pH at p<0.01. However the main effect of mesorizhobium bacteria had significant effect on soil phosphorus at p<0.01. Main effect of thiobacillus had only significant effect at p<0.01 on soil pH. The interaction effect of sulphur and mesorizhobium had significant effect at p<0.01 on soil phosphorus, soil potassium and soil pH at p<0.01, but interaction effect of thiobacillus and mesorizhobium bacteria had only significant effect at p<0.01 on soil phosphorus. Interaction effect of all three factors had significant effect at p<0.01 on soil phosphorus and at p<0.05 on soil sulphur, EC and soil pH. Analysis of variance of data in this research showed that factors in this study had positive and significant effect on agronomic parameters of chickpea and soil physic-chemical properties.

Alpha-linolenic acid is an omega-3 fatty that is essential for maintaining a healthy body but can't produce in the human body. The oils of some oilseeds, as well as some plants are the main sources of alpha-linolenic acid but the content of alpha-linolenic acid, in important oilseed plants like soybean and rapeseed are relatively low. The aim of this study was the cloning of Delta 15 Desaturase gene to increase alpha-linolenic acid production in the oilseed plants. For this purpose, after extracting total RNA and cDNA synthesisfrom Pichia pastoris GS115, the genewas amplified using gene-specific primers containing Kozak sequence and then cloned in PTZ57R / T vector and transformed into E.coli strain DH5α. After sequencing, the recombinant fragment was isolated from TA vector and was cloned into pBluescript KS(+) vector containing Napin promoter. The designed gene construct was cloned in pBI121 binary vector and then was transformed into Agrobacterium tumefaciens strain LBA4404. Bioinformatics characterization of the target gene was investigated by servers TopPred, TMHMM, ProtParam and SOPMA. The results of the colony PCR and amplification of the 1246 bp fragment, confirmed the accuracy of the gene cloning. The amplification of a 1210 bp fragment using an internal primer of napin promoter and Delta 15 desaturase, as well as the production of the 2145 bp fragment in enzymatic digestion confirmed the correct incorporation of the gene along with Napin promoter. Nucleotide sequencing results showed that the cloned CDS include 1246 nucleotides and translated to a protein with 415 amino acids. The amino acid sequence analysis confirmed the presence of two domains and five Transmembrane helices. Also, prediction of the protein's properties, signal peptides, and the second structure, proved that this enzyme is stable transmembrane enzymes.

Oleaginous yeasts could produce lipid with high contents of poly unsaturated fatty acids (PUFA). Isolation of oleaginous species with a strong ability to produce lipid and long-chain polyunsaturated fatty acid (LCPUFA) could be considered as an effective step to the commercialization of LCPUFA production. Having isolated an oleaginous yeast species from fish, the species were identified by the molecular method. One factor at time and response surface methods was used for optimization of lipid and oleic acid production. Fermentation was being conducted in five days. The yeast specie which isolated from the gills of trout fish was Wickerhamomyces Siamensis SAKSG. Response Surface Methodology (RSM) indicated that optimal conditions for lipid production (27% of dry biomass) was obtained at the middle levels of glucose (42 g/L) and minimum amount of soy bean (5 g/L). The optimal amount of oleic acid (66.5% of lipid) was obtained at high levels of glucose (80 g/L) and soybean (15 g/L). Docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and arachidonic acid (ARA) were also produced by this new isolated yeast. Discussion and conclusion: Wickerhamomyces SiamensisSAKSG as oleaginous yeast had great ability to produce lipid with a reasonable amount of LCPUFA. Chemical condition of media highly impact on LCPUFA production. More to the point, protein had positive effect to LCPUFA production via stimulation of the yeast growth.

Membrane-bound desaturases and related enzymes play a pivotal role in the biosynthesis of unsaturated fatty acids. Delta6 desaturase is a key enzyme in the biosynthesis of the unsaturated fatty acids. Mortierella alpina is an oleaginous fungus with active Delta 6 desaturase which hasbeengreatly considered recently.
In order to isolate and clone Δ6D gene from Mortierella alpina, after extraction of total RNA and synthesis cDNA, PCR amplification has been done using gene specific primers. The amplified fragment was cloned into the pBlueScriptSK containing seed specific promoter napin. Then the recombinant plasmid was transformed into E.coli DH5a by freezing and thawing method. The confirmed gene construct was cloned into the binary vector pBI121 and transformed into Agrobacterium LBA4404 in order to transform canola plants. Bioinformatics characterization of target gene was investigated by servers TMHMM, ProtParam and Psipred.
Correctness of cloning was confirmed by PCR with specific primers, enzymatic digestion and sequencing. The proliferation of a fragment with 830 bp using internal primer of napin promoter and Delta6 desaturase primer confirmed insertion of the gene along with napin promoter. Nucleotide sequencing results showed that cloned CDS includes 1374 nucleotides that will translate to a protein with 448 amino acids. Using bioinformatics analysis, presence of cytochrome b5 domain, three His-box, secondary and spatial structures, transmembrane and conserved domains were confirmed.
Discussion and Based on the results of BLAST analysis using nucleotide and protein sequences, and also presence of functional domains in the protein, it can be predicted that cloned CDS will show proper enzyme activity after transformation into plants. Confirming these results requires expression analysis of the gene in appropriate plant system and studying its function in the enzyme level

Fusarium solani f. sp.cucurbitae races cause crown, fruit and root rots of cucurbits and economic problem for farmers. The aim of this study is identification of Fusarium solani f. sp. cucurbitae races which collected in Shahrood, Jiroft and Varamin, provinces of Iran using morphological characteristics and fungal-specific primers. Symptomatic cucurbit plants were collected from different fields in three provinces. After isolation and purification of the causal agents, pathogenicity test and DNA extraction were performed. Specific primers for TEF-1α gene were used to distinguish among different species of Fusarium and Fusarium solani. The isolates that showed a specific 658 bp amplicon on agarose gel were selected for subsequent PCR assay in order to detection of F. solani f. sp. cucurbitae race 1 and 2. PCR assays were performed with taxon-specific primers race 1 (505bp PCR product) and race 2 (425bp PCR product). Some samples amplified a 505bp amplicon which is specific for race 1 and was in accordance with morphological characteristics. In these isolates, 5 septate macroconidia were dominant that is specific to Fusarium solani f. sp.cucurbitae race 1 macroconidia. None of the isolates were amplified by race 2 taxon-specific primers. Our study showed that, race 1 of F.solani is the dominant race of the pathogen in Shahrood, Jiroft and Varamin provinces which can help to implement effective disease management strategies.

IntroductionThe excessive use of chemical fertilizers causes environmental pollution that is led to imbalance of essential elements in agricultural production system. Organic matter application as compost in the soil can improve chemical quality and biochemical properties that increase essential elements for plant nutrition. Application of organic manure can significantly increase the soil aggregate as well. Reported that application of 7 ton ha-1 of vermicompost increased number of leaves, stem dry weight, and corn plant height as compared to control and water holding capacity increases. Sulphur in plant is near to phosphorus (0.2%). Sulphur deficiency cause severe reduction in plant growth and due to participation in protein building and its deficiency cause yellowish in younger leaves. Sulphur can be applied as elemental sulphur, with ammonium and super phosphate to the soil. Iran is situated in arid and semi arid region of the world and need to reduce the pH with sulphur application due to high pH above 8 in some parts. The aim of this research was to study the effect of above factors in yield and yield components of maize and reducing environmental pollution.
Materials and MethodsThis research carried out at 2012 in Jovein Distract suberb of Sabzevar city. Before conducting the research soil sample were collected from 0-30 cm depth and physical and chemical properties of the soil were estimated. Treatments including sulphur, thiobacillus and vermicompost were applied to soil and well mixed with soil before sowing. Each plot consists of five rows with six m length by 80 cm from each other. Seeds were sown at the depth of five cm and 20 cm from each other. This research carried out as a factorial experiment on the basis of randomized complete block design. In this research three factors including elemental sulphur, vermicompost and thiobacillus were used with three replications. Elemental sulphur in three levels (control, 500 kg ha-1 and 1000 kg ha-1), Vermicompost at two levels (control and 5000 kg ha-1 and thiobacillus with two levels (control and application) were applied. All three treatments were mixed with each other and applied to soil one month before sowing that sulphur oxidation take place. At the time of sowing basal fertilizer were applied as diammonium phosphate as (P2O5) at the rate of 150 kg ha-1 and potassium sulphate at the rate of 150 kg ha-1 as (K2O) on the bases of local recommendations. Analyses of variance were done with MSTATC and SAS and means compared with least significant difference test at 5% level. Figures were done with Excel 2007.
Results and DiscussionAnalysis of variance showed that interaction effect of sulphur, vermicompost and thiobacillus were significant on soil EC at 5% level. The highest EC was observed in 1000 kg ha-1 sulphur, 5000 kg ha-1 vermicompost and thiobacillus treatments. Analysis of variance showed that interaction effect between sulphur and thiobacillus and interaction effect of sulphur and vermicompost were significant on soil pH at 1% level. Mean comparison of data showed that highest decrease of soil pH achieved in 1000 kg ha-1 sulphur. main effect of vermicompost and interaction effect of elemental sulphur with vermicompost and sulphur with thiobacillus had significant effect on soil organic carbon at 1% level. Interaction effect of sulphur and vermicompost were significant on soil nitrogen at 5% level. Mean comparison of data showed that this increase was due to using 5000 kg ha-1 and not using sulphur. Interaction effect of sulphur, vermicompost and thiobacillus had significant on available phosphorus. Main effect of three factors and their interactions on available zinc, iron and available sulphate were significant at 5% levels.
ConclusionThe results of this research showed that application of elemental sulphur were significant on soil moisture, field capacity and soil pH and caused increase in soil EC and availability of soil zinc, iron, phosphorus, and sulphate. Organic manure like vermicompost increased soil organic carbon and soil nitrogen.

In this study, we evaluated the effects of five levels of potassium phosphite (KH2PO3) on Induced Systemic Resistance (ISR), and also defense related enzymes and metabolites activities during different days in cucumber plants inoculated with Fusarium oxysporum. The experiment was designed as a split-plot in time based on a completely randomized design with three replications. The activity of defense-related enzymes including, catalase (CAT), guaiacol peroxidase (GPX), superoxide dismutase (SOD) and also the accumulation of secondary metabolites such as lipid peroxidation (MDA) and hydrogen peroxide (H2O2), were evaluated in a period of 11 days after pathogen inoculation. Results revealed that defense enzymes activities and metabolites accumulations were significantly increased in plants treated with potassium phosphite in comparison to control. The highest concentration of CAT enzyme (18.7 U mg-1 protein) was measured in plants treated with 4 gr l-1 potassium phosphite (KPhi4) and 5 days after inoculation, that showed 2.81 fold increase in comparison with control (no inoculate with the pathogen). Also the highest concentration of enzymes GPX (233 U mg-1 protein) and SOD (10.8 U mg-1 protein) were recorded at 3 days after inoculation in plants treated with KPhi4 in comparison with control plants. Results showed that accumulation of H2O2 in plant tissues and it's time was in accordance with increasing of defense enzymes CAT, GPX and SOD and it would indicate that the H2O2 production is the prerequisite for production of defense-related enzymes. It was also revealed that MDA accumulation in KPhi4 treatment was less than other treatments, indicating reduced tissue damage and alleviated disease symptoms.

Phytophthora species are considered as the major cause of several plant diseases resulting in huge yield losses in agricultural crops. Despite years of effort to develop Phytophthora resistance varieties, there is no reports of a resistant cucumber variety. In this study, the effect of concomitant application of potassium phosphite (KPhi) and chitosan on some physiological and molecular responses of Phytophthora capsici-challenged cucumber plants were investigated. Cucumber plants were treated with KPhi and/or Chitosan at different concentrations and were then inoculated with zoospores of P. capsici and leaf samples were collected at different time courses. Results showed that Guaiacol peroxidase (GPOD) enzymatic activity surged immediately at first and second days after pathogen inoculation with a peak in plants treated with 4 gL-1 KPhi 2 days after inoculation. Compared to GPOD, the highest superoxide dismutase (SOD) activity was observed in the same treatment but later at 5 days after inoculation. It was indicated that the activity of antioxidant enzymes was greatly influenced by application of either KPhi or chitosan while their activity was not remarkably enhanced in control plants. qPCR analysis revealed that the highest increase in glutathione peroxidase (gpx) gene expression was achieved in plants concomitantly treated with 4 gL-1 KPhi and 200 mgL-1 chitosan 5 days after inoculation. The findings of this study provide novel information regarding inducing mechanisms of KPhi and chitosan which may be effective in mitigating disease severity.

Chemical fertilizers have important role in increasing crop production in unit square meter, but studies showed that using these fertilizers for long period with strong salts in them cause damage to soil physical and chemical properties like soil infiltration, bulk density and decrease in plant root penetration that ultimately decrease crop production. The aim of this research was to estimate desirable proportions of chemical fertilizers and organic manure with the aim of reduction in consumption of chemical fertilizers
Methods and materials. For the study of the combined effect of different chemical fertilizers and manure (sheep) to reach suitable fertilizers combination (Thompson novel orange) an experiment was conducted at 2013 in one of the orchard at Sari district with low organic C. This research carried out on five years old citrus threes. This experiment carried out as factorial experiment on the base of complete randomized block design with 9 treatments and three replications. Treatments included three manure levels (0, 6 and 12 kg per) and three levels of macro fertilizer including potassium sulphate, ammonium sulphate and super phosphate triple (0, 30 and 60 percent on the bases of soil test).
Result and discussion. The results of analysis of variance on orange leaf nitrogen showed that simple effect of chemical fertilizers and organic manure and their interactions were significant at 5% level. The interaction between factors under studies for this trait showed that chemical fertilizers are under influence of organic manure. The results of analysis of variance showed that simple effect of chemical fertilizers and interaction effects of chemical fertilizers and organic manure on orange leaf phosphorus were significant at 1% level. Mean comparisions showed that highest phosphorus absorption were related to 60% chemical fertilizers use and 6 and 12 kg manure which were statically in same group. The results of analysis of variance showed that the simple effect of different levels of chemical fertilizers and its interactions with manure on orange leaf potassium absorption were significant at 1% level. Mean comparison of interactions showed that the highest potassium absorption were with T8 treatment. The analysis of variance for sulphur absorption showed that simple effect of chemical fertilizer and manure were significant at 1% level and their interactions were significant at 5% level. The results from analysis of variance showed that simple effect of chemical fertilizer and its interaction with manure on orange leaf calcium absorption were significant at 5% level and simple effect of manure on orange leaf absorption were significant at 1% level.
The highest nitrogen, phosphorus and potassium of Thompson leaf were obtained with application of 60% chemical fertilizer and 6 kg manure. The highest absorption of calcium, magnesium, Iron, manganese and zinc in Thompson orange leaf were obtained with 30% chemical fertilizer application and 12 kg manure.

With respect to the human need for functional food and the importance of long-chain polyunsaturated fatty acids as a functional food, identifying the rich resources of fatty acids greatly attract of interest of scientists. Based on this studies, it is found that microorganisms especially yeast species are rich resources of unsaturated fatty acids. In the present study, yeast samples of perch fish, which has the ability of oil production, were isolated and then their ribosomal DNA were used to identify the yeast species. According to the phylogenetic tree, it was known that the yeast species isolated from the gills of perch fish was Candida Orthopsilosi SAKSG. The response surface method showed that the highest oil content (26.3) was achived in media that had 50 g/L glucose and 5 g/L yeast extract. Moreover, maximum amount of biomass (1.16%) was obtained in medium with 46.16 g/L f glucose and 19.67 g/L yeast extract. It is notable that reduction in temperature treatment was led to improve unsaturated oil increment in this microbial lipid up to 24 percentages. reduction of temperature at the end of the fermentation time concide with rising in unsaturated fatty acids with long fatty acid more than 20 carbon, while when temperature was increased from the beginning of the process, structural unsaturated fatty acids such as linoleic and linolenic acid were increased.

IntroductionLegumes are very important source of protein after cereals. These plants are capable of biological nitrogen fixation and increasing soil fertility as cover crop or including in crop rotation and reducing soil erosion which improve sustainable agriculture. Increasing use of chemical fertilizer which has negative effect on soil structure which cause unbalance on physical and chemical propertied of soil which ultimately reduce nutrient element absorption. Nowadays due to environmental problems use of biological fertilizers are good substitutions. On the other hand biological fertilizer use improve soil fertility. Protein quantity and quality are basic nutritional problem. According to use of more than 85000 tones chemical fertilizers in legume cultivation in Iran. It is necessary to conduct correct and efficient inoculation for each legume crop in the country including bean, which is one of the important crop for human consumption. In most cases biological fertilizers are complementary of chemical fertilizers in a sustainable agriculture. The aim of this study was to compare the effect of interaction effect of mycorrhiza vesicular, Psodomonas bacteria and humic acid on yield and yield components of bean.
Material & MethodsMycorrhiza inoculation was prepared in soil science Lab in Tabriz University in sterile sandy loam. Bacterial inoculation Pseudomonas putida prepared in soil biology of Tabriz University. Humic acid extraction was done with method of Kay et al., (2004). For Seed bean Phaseolus vulgaris variety Cos-16 from agronomy and plant breeding department of Tabriz University which is resistant to drought. Soils for this experiment were collected from Khalat Poshan research station from 0 to 25 cm depth and passed through 4mm sieve after sun sterile method. Soil physical and chemical properties were estimated with standard methods. Each pot was filled with three kilogram sterile soil. Seventy gram of mycorrhiza inoculation was spread in 5 cm of top soil and then bacteria inoculation was added to soil just below the seeds. Humic acid extracted from vermicompost and applied to soil in the powder form in three levels, control (H0), 200 mg kg-1(H1), and 400 mg kg-1 (H2). Pots were place in green house at 25C. After treatment application 5 seeds were sown in each pots and pot moisture kept at 0.9 FC. Pots were irrigated with distilled water whenever needed. After 4 months plant roots and shoot properties and plant yield and yield components were estimated. This experiment was conducted in the form of factorial on the base of complete randomize block design in Tarbiat Moalem University of Tabriz. First and second factor were in two levels, control and applied for mycorrhiza (M0 and M1) and bacteria P. putida (S0 and S1) and humic acid in three levels control (H0), 200 mg kg-1 (H1) and 400 mg kg-1 (H2) in three replications. Data analysis was done with MSTATC and SPSS and mean comparison were done with LSD test for significance and figures drown with Excel 2007.
Results & DiscussionAnalysis of variance of data for plant height showed that interaction effects of humic acid and bacteria were significant at 1% level and mycorrhiza was significant at 5% level. Mean comparison of results showed that least plant height mean from interaction of humic acid and bacteria with S0H1 treatment with (28.58 cm) and the highest was (30.35cm) with S1H1 treatment. Analysis of variance showed that main effect of mycorrhiza were significant at 1% level and interaction effect of humic acid and bacteria at 5% level on shoot wet weight of bean. Analysis of variance for main effect of humic acid and mycorrhiza were significant at 1% level for shoot dry weight and main effect of bacteria and interaction effect of bacteria and mycorrhiza were significant at 5% level for shoot dry weight of bean. Analysis of variance showed that main effect of mycorrhiza and humic acid were significant at 1% level and interaction effect of mycorrhiza and humic acid at 5% level on root dry weight. Analysis of variance of Mycorrhiza and bacteria main effect on ratio of root dry weight to shoot were significant at 1% level and main effect of humic acid and interaction effect of humic acid and mycorrhiza were significant at 5% level. Analysis of variance showed that main effect of mycorrhiza and humic acid were significant at 1% level and main effect of bacteria and interaction effect of mycorrhiza and bacteria were significant at 5% level for seed numbers of bean. Analysis of variance showed that main effect of bacteria and mycorrhiza and interaction effect of bacteria and mycorrhiza were significant at 1% level and interaction effect of bacteria and mycorrhiza were significant at 5% level on stem length. Analysis of variance showed that main effect of humic acid and mycorrhiza were significant at 1% level and main effect of bacteria and interaction effect of humic acid and bacteria were significant at 5% level for stem diameter. Analysis of variance showed the main effect of mycorrhiza was significant at 1% level and main effect of humic acid and interaction effect of bacteria and mycorrhiza and interaction effect of bacteria and humic acid and main effect of humic acid were significant at 5% level for the seed yield.
ConclusionAnalysis of variance of experiment treatments in this research showed that treatment applied had positive effect on agronomic properties of bean and humic acid could have direct and positive effect on plant root and shoot growth.

The hairy roots induced by Agrobacterium rhizogenes due to the stability and mass production in hormone free culture provide an appropriate tissue for secondary metabolite production, and the content of metabolites produced could be affected by bacterial strain. In the current study, the effect of different strains of A. rhizogenes including A4, A13 and 15834 on the percentage of hairy root induction of Echinacea purpurea (L.) moench. in the leaf and hypocotyl explants were studied. In another experiment, total phenolics and polysaccharides content of hairy roots were measured compared to the control (untransformed) roots. The results showed that the percentage of hairy root induction was significantly affected by bacterial strains, so that in E. purpurea leaves incubated by strains A13, 15834 and A4, the hairy root induction was 60%, 40% and 0, and in hypocotyl explants was 85%, 45% and 70%, respectively. Maximum total phenolic compounds were observed in A4 strain. As well, in comparison to control, the polysaccharide content in hairy root induced by strains 15834 and A13 increased and by A4 strain decreased significantly. Differences observed in this study about hairy root induction and secondary metabolites production could be due to different ability of A. rhizogenes strains in T-DNA transferring to plant cells.

Stored product pests are a major problem in the storage of agricultural products that cause damage from harvest until consumption. Greater Wax Moth (Galleria mellonella L.) is one of the most important pests of stored products and beehives. The most common method to control this pest in many countries is use of chemical compounds. However, these compounds have disadvantages such as pesticide residues in wax, the development of resistance in pest and irreversible effects on the environment and humans. The use of insect pathogenic fungi due to its low risk on mammals and natural enemies can be a good alternative to conventional chemical pesticides. Response surface methodology (RSM) is a statistical technique that is employed to optimize processes that are affected by several variables. This technique uses regression analysis to obtain optimal equations to estimate the values. Using this method, while maintaining the quality in the experiments, the number of those could be reduced. Therefore, this study was aimed to evaluate response surface methodology to determine the effect of optimum lethal level of concentration of B. bassiana conidia, temperature as well as humidity variables on the mortality of fifth instar larvae of greater wax moth.
Wax moth-eating insects were raised in plastic containers containing artificial food and old black wax at 30 ± 1 ° C and a relative humidity of 85 ± 1 % and photoperiod of 14:10 h (L: D). Isolation of insect pathogenic fungus B. bassiana was done by using Galleria Bait Method (GBM). For this purpose, after preparation of the fungus suspension from the infected larvae, 1 ml volume of the suspension was transferred to the water-agar 1.2% and then sealed petri dishes incubated at 30 ° C for three days. After identifying the single colony and formation of pure isolates, microscopic slides were prepared and eventually recovered isolates were recognized as B. bassiana. The bioassay was performed by determining the lethal concentrations of the B.bassiana that cause 20% to 80% casualties with a lot of concentration by immersion method for 10 seconds. Concentrations 1×106 and 1×108 conidia/ml were identified as high and low lethality ranges, respectively. In this study, the central composite design and response surface methodology with three independent variables including temperature (25-35°C), humidity (70-80 percent) and concentration (1×106-1×108 conidia/ml) and six replications in the central point of the design (to calculate the repeatability of the process) were used to evaluate the increase in mortality. The number of experiments was twenty and the dependent variable (response) was the mortality of the fifth instar larvae of greater wax moth. For each experiment, 10 last instar larvae were randomly selected and then 10 sterile petri dishes containing sterile wax to feed insect were prepared. Larvae were immersed for 10 seconds in a solution containing the fungus and then were placed in containers.
Analysis of variance (ANOVA) for the quadratic response surface model to factor mortality of the fifth instar larvae of greater wax moth showed that quadratic model is statistically significant (P≤0.001). Also high R2 (R2 = 0.9430) and coordination of adjusted R2 (Adj R2 = 0.9211) indicates the strength of the model to predict. According to tests, the optimal conditions for achieving maximum mortality of the fifth instar larvae of greater wax moth is 25 ° C temperature, 75% humidity and 1×108 conidia/ml concentration, respectively. Settings applied to the optimization process, was including maximum mortality. The effect of temperature on mortality of the fifth instar larvae of this insect showed that the mortality rate decreased with increasing temperature. Cause of mortality reduction as increasing the temperature is probably related to the characteristics of this fungus that could be affected by temperature, so that the impact of this fungus increases with decreasing temperature. The impact of the concentration on mortality rates showed that by gradually increasing of concentrations, the amount of mortality increases. This is because at the high concentrations of conidia, many more conidia could contact with the body of fifth instar larvae of this insect and could infect and destroy the larvae very quickly.
The results of current research indicate the efficiency of response surface method to optimize the use of insect pathogenic fungi. Among the conditions that were imposed on mortality, it was found that the increase in mortality is influenced by the quadratic response surface model of concentration and temperature, so that increasing the concentration caused increase in the mortality. Also increasing the temperature caused a decline in mortality rate.

Determination of genetic diversity in plant material is the initial step to identify and preservation of genetic resources and also is important in conducting of breeding programs. In order to study genetic variation among 25 grapevine cultivars of Shahrood and Sisakht regions, nine retrotransposonal primer combinations including 3 IRAP primers and 6 REMAP primers combination were used. Totally, 87 bands produced that 79 bands were polymorphic among cultivars. Numbers of polymorphic bands varied from 4 (Tvv1Fa㋍) to 12 (Vin1Fa) bands per primer. Expected heterozygosis ranged from 0.29 (Tvv1Fa Ms8) to 0.45 (Vine1Fa Ms3) with an average of 0.35. Average of polymorphism percentage among evaluated cultivars was 95.83%. Principle coordinates analysis (PCoA) showed that first five components explained 68.5% of total variance. Cluster analysis using UPGMA method based on Jaccard similarity coefficient with cophenetic correlation coefficient of 0.89, divided grapevine cultivars into 6 different groups. In the first to sixth group, 13, 4, 2, 3, 2 and 1 cultivars were assigned respectively. According to this grouping, cultivars named Syah-Daneh Boland, Syah-Daneh Gerd and Asgari from SySakht city were assigned in three different groups that showed genetic differences among these cultivars. The results of this study indicated that IRAP and REMAP markers developed based on active retrotransposons in the grapevine genome can be used as efficient molecular markers to accelerate grapevine breeding programs.

Desaturase enzymes (EC: 1.14.19.3) catalyze dehydrogenation reactions and creates a double bond in the fatty acid chains. These enzymes classified into two groups, soluble and membrane each with different consensus motifs. Despite the Delta-6 desaturase is a key enzyme in the biosynthesis of unsaturated fatty acids but due to lack of structural information of membrane-bound desaturase, it's three-dimensional and crystal structure has not been determined. Mortierella alpina is a rich source for production of unsaturated fatty acid arachidonic and has an active Delta-6 desaturases enzyme. The aim of this study was obtain encoding sequences of Delta-6 desaturase gene, and determination of functional domains using comparative analysis and molecular model and propose a membrane topology model for this enzyme. Thus, after extraction of total RNA and cDNA synthesis using gene specific primers, PCR product was cloned into pBlueScriptSK vector and then sequenced. Based on bioinformatics analysis by Swissmodel, Predict and Phobius servers presence of functional domains cytochrome b5 carrying motif (HPGG), three His-box motifs and transmembrane regions were identified. Then we proposed a membrane topology model for the Delta-6 desaturase enzyme.

Soil microorganisms with potential for alleviation of salt stress in combination with plant growth promotion would be a promising approach in sustainable agriculture. In the present study, interaction of two varieties of bread wheat (Triticum aestivum L.), Pishtaz and Zarrin and three salt tolerant ectoines producing bacteria including S. Cellulosae, S. rimosus C-2012 and Streptomyces Strain S2 was undertaken in normal and salt conditions. Diversity and distribution of 16S rRNA gene of rhizospheric bacteria in soil inoculated with Streptomyces in salt and normal conditions were studied by denaturing gradient gel electrophoresis (PCR-DGGE) method. Results showed that bacterial cell free extract of all strains reduced root length but S2 and C-2012 increased plant dry weight related to control. Soil treatment with strain C-2012 increased Pishtaz root and shoot fresh and dry weight and Zarrin root fresh and dry weight in normal conditions. At salt conditions, plant growth promotion of C-2012 limited to increase Pishtaz root fresh and dry weight and Zarrin leaf area. Root extract of wheat plantlets increased C-2012 population (cfu) and decreased mycellial aggregation. The effect of Pishtaz root extract on bacterial cfu was more than Zarrin. Based on PCR-DGGE data, the diversity of 16S rRNA gene in rhizosphere changed when Streptomyces or salt was added to soil. PCR-DGGE profiles of 16S rRNA gene in rhizosphere of wheat varieties were also different. Our observations certify that beneficial role of salt tolerant Streptomyces for wheat growth promotion at normal or saline conditions is plant variety dependent.

In vitro propagation of three Iranian apricot cultivars, “Ghavami”, “RajabAli” and “Khiveei” was studied by direct micropropagation technique. To optimize sterile manipulation, shoot nods of one year-old dormant vegetative shoots and current growing vegetative shoot were cultured in WPM medium after treatment by HgCl2, citric acid, ethanol and NaClO (10, 15, 20 min). Establishment of buds was tested in the MS and WPM mediums. Proliferation of sprouting buds was evaluated in WPM medium supplemented with three concentrations of (0.5, 1 and 2 mg/l) BAP and 0.05 mg/l of IBA. A half-Strength MS medium supplemented with three concentrations (0.5, 1 and 2 mg/l) of IBA was used as a rooting medium. The greatest number of contamination-free shoots in HgCl2 0.01% and citric acid 0.07% treatments were obtained. No significant difference was found between two different mediums in terms of bud vegetative growth. The shoot number and length were significantly affected by apricot varieties and BAP concentrations. The greatest numbers of proliferated shoot were observed in Rajabali and Ghavami varieties at 1 mg/l of BAP, whereas maximum number of shoot in Khiveei cultivar was observed in 0.5 mg/l of BAP. The maximum shoot length in Rajabali and Ghavami cultivars was obtained in 2 mg/l of BAP.

Among the various fungal strains screened for citric acid production, Aspergillus niger is known to produce considerable amounts of citric acid and other organic acids when cultivated in submerge fermentation. In this study, optimization of the medium components was carried out using "one-factor-at-a-time" and response surface methods (RSM). One-factor-at-a-time indicated that the amount of citric acid production was increased along with the increasing of agitation speed (from 150 to 200 rpm), raising the incubation temperature (from 17 to 32) and decreasing pH value (to 2). Moreover, in comparison with the other groups, citric acid production in the treatment containing soy bean powder and minerals revealed a significant increase (up to 25 g/l). The results showed that application of ultrasonic wave during the growth phase could remarkably enhance the production of citric acid. Based on the results of one-factor-at-a-time, sucrose and soy bean powder were the selected additives to test their effect on citric acid production using RSM. The two variables were identified to have significant effects on citric acid production and the maximum citric acid production of 58 g/l was resulted from the combination of 230.87 g/l sucrose and 200.81 g/l soy bean powder. In this study, the significant increase in the production of citric acid in the optimal conditions indicated the using of appropriate statistical methods and also the correct levels of selected variables.

In this research, genetic variation among 22 grapevine cultivars of Shahrood region and 3 cultivars from Sisakht city were evaluated using 6 ISSR primers. The used primers produced totally 69 bands, which 65 bands showed polymorphism among evaluated cultivars. Primers UBC825 with 16 bands and UBC857 with 6 bands produced the highest and lowest numbers of polymorphic bands, respectively. Polymorphism percentage using these markers was 94.1%. Mean of expected heterozygosis (He) and mean of Shannon index were 0.37 and 0.54, respectively. The highest (0.61) and lowest (0.48) amount of Shannon index was belong to the primers UBC 825 and UBC849, respectively. Also primer UBC825 showed the highest amount of the expected heterozygosity (0.42), highest number of effective allele (1.75) and highest number of different alleles (2). Cluster analysis using Jaccard similarity coefficients and UPGMA algorithm put the 25 studied cultivars in three different groups. The results of this study showed that ISSR markers can be used as effective tools to evaluate the genetic variation and to accelerate grapevine breeding programs.

Beauveria spp. as entomopathogenic fungi have extremely diverse insect host ranges.Here, a comparative transcript analysis of a Bassiasin I (BSNI), a Dipeptidyl peptidase (DPP), a Cytochrom P450 monooxygenase (CYP52), a Mitogen activated protein kinase (MAPK), and a Neuronal calcium sensor 1 (NCS) in two isolates of Beauveria bassiana., with low and high virulence, were carried out. For this, induction media, containing isolated cuticles from four insects, and a check medium lacking any cuticular extract were prepared and isolates were allowed to grow on them. Interestingly, up-regulation of BSNI and DPP in induction media was noted in comparison to the check, following normalization with γ-ACTIN. CYP52 was up-regulated in synthetic medium in comparison to cuticular extract. MAPK expression was greater in most virulent isolate once compared with the low virulent isolate. The expression of MAPK was greater in the presence of Caliptamus italicus cuticles and synthetic medium as opposed to others. NCS expression remained on changed in all media tested and can be suggested as a reference gene in transcriptomics of Beauveria spp. genes in future.

Citrus are one of the important orchard fruit production that after banana is second in production at the world level and every year, Chemical fertilizers having most important role in increasing crops productions, but in long application of fertilizers cause soil destructions and polluting underground water. Also soils of dry regions are very poor in organic matter level. Nowadays in most countries, climatically condition and poor management cause poor organic matter content of soils. In Iran more than 60 percent of cultivated lands having less than 0.5 up to 1 percent organic matter. This may be due to intensive cultivation and poor managements For this reason if we have combine applications of manure and chemical fertilizers, the results would be much better. The aim of this research was to evaluate suitable ratio of manure and chemical fertilizer in order to reduce the chemical fertilizer use in citrus orchard in north of Iran. This experiment was conducted in one of the orchard at Sari district with low organic C. This research carried out on five years old citrus threes. This experiment carried out as factorial experiment on the base of complete randomized block design with 9 treatments and three replications. Treatments included three manure levels (0, 6 and 12 kg per tree) and three levels of macro fertilizer including potassium sulphate, ammonium sulphate and super phosphate triple (o, 30 and 60 percent on the bases of soil test). Total treatment were 27 plots, (each plots were includes two threes).all treatments were applied at March. All analysis was done with standard methods. This experiment was done as factorial on the bases of complete randomized block design with 9 treatments and three replications. The treatments were as follows:T1: Zero percent chemical fertilizer and zero kg manureT2: 30 percent chemical fertilizer (potassium sulphate 50 kgha-1, ammonium sulphate 30 kg ha-1 and super phosphate triple 45 kg ha-1) and zero kg manure.T3: 60 percent chemical fertilizer (potassium sulphate 100 kgha-1, ammonium sulphate 6 kg ha-1 and super phosphate triple 90 kg ha-1) and zero kg manure.T4: Zero percent chemical fertilizer and 6 kg manureT5: 30 percent chemical fertilizer (potassium sulphate 50 kgha-1, ammonium sulphate 30 kg ha-1 and super phosphate triple 45 kg ha-1) and 6 kg manure.T6: 60 percent chemical fertilizer (potassium sulphate 100 kgha-1, ammonium sulphate 6 kg ha-1 and super phosphate triple 90 kg ha-1) and 6 kg manure.T7: Zero percent chemical fertilizer and 12 kg manureT8: 30 percent chemical fertilizer (potassium sulphate 50 kgha-1, ammonium sulphate 30 kg ha-1 and super phosphate triple 45 kg ha-1) and 12 kg manure.T9: 60 percent chemical fertilizer (potassium sulphate 100 kgha-1, ammonium sulphate 6 kg ha-1 and superphosphate triple 90 kg ha-1) and 12 kg manure.All data analysis was done with MSTATC software and mean data comparison done with Duncan test levels at 1 or 5 percent levels. The results of analysis of variance showed that simple effect of manure and fertilizers and their interactions had no significant effect on TSS, but simple effect of fertilizer and their interactions between them had significant effect. The simple effect of chemical fertilizer and its interaction with manure were significant at 1 percent level on titration able pH of fruit juice and the simple effect of manure effect on fruit juice pH were significant at 5 percent level. The simple chemical fertilizer effect on C vitamin was not significant but simple effect of manure and its interaction effect with chemical fertilizer were significant at 5 percent level. The analysis of variance showed that simple effect of chemical fertilizer on fruit juice potassium were not significant, but simple effect of manure and its interaction with chemical fertilizer were significant at 5 percent level. Analysis of variance showed that simple effect of manure and fertilizer on the fruit juice calcium was significant at 1 percent level but their interactions were significant at 5 percent level. Simple effect of manure and fertilizer and their interactions on the fruit magnesium, iron manganese, zinc and copper were significant at 1 percent level. Analysis of variance showed that simple effect of manure and fertilizer and their interactions on the fruit juice magnesium, manganese, iron and copper was significant at percent level, but simple effect of manure on fruit juice zinc were significant at 5 percent level. In this study the treatment with combined application of 30 percent and 12 kg manure had highest Ca, Mg, Fe, Mn and zinc concentration in fruit, also combined application of 60 percent fertilizer and 12 kg manure had effect on pH and potassium of fruit juice.