هوشنگ علیزاده

Cold stress is one of the major factors limiting rice (Oryza sativa L.) productivity and yield. Two rice cultivars were chosen for this study: a japonica rice, ‘Gerde’, and an indica rice, ‘Shiroodi’, representing tolerant and sensitive cultivars, respectively. In this work, the early physiological and molecular responses of these two cultivars to 4°C cold stress were investigated. We found that cold stress caused a significant decrease in the content of chlorophyll a, chlorophyll b, and carotenoids in both rice cultivars. However, the photosynthetic pigment content in the Shiroodi cultivar decreased to a greater extent, indicating its limited genetic capacity to protect its photosynthetic machinery. During cold stress, the amounts of hydrogen peroxide and malondialdehyde produced by the Shiroodi cultivar were significantly higher than those in Gerde, suggesting a lower capacity of Shiroodi to scavenge reactive oxygen species. The higher level of beta-glucosidase gene expression in the final hours of cold treatment in the Gerde cultivar indicates the involvement of this enzyme in elevating cellular levels of active abscisic acid. Moreover, the significant increase observed in the transcript level of phenylalanine-ammonia-lyase in Gerde indicates the induction of defense responses related to the phenylpropanoid pathway. Altogether, one can conclude that the higher cold tolerance of the Gerde cultivar is multifaceted, involving various factors such as its ability to protect its photosynthetic machinery, its greater capacity for cell protection against oxidative stress, and the expression modulation of genes involved in abscisic acid metabolism and secondary metabolism.

The Valerian plant (Valeriana officinalis) contains compounds that can be used to treat nerve disorders, epilepsy, and sleep disorders. Terpenes (sesquiterpenoids) are the major group of compounds in this plant, whose biosynthesis pathway is poorly understood. Due to the lack of genomic sequence, studies based on de novo transcriptome assembly are important to elucidate more genes of the terpenoid biosynthetic pathway in this valuable medicinal plant. To reconstruct de novo assembly of RNA-Seq data from previously published SRR data for V. officinalis, Trinity and rnaSPAdes were used in this study. The primary assemblies were then analyzed using additional tools, such as CAP3 and CD-HIT-EST, in an attempt to determine the most effective de novo assembly. Based on the results of quality assessment tools such as rnaQUST, SeqKit statistics, and BUSCO, the combination of CAP3 and CD-HIT tools successfully produced an optimal assembly for Trinity (T-CAP-CD). A total of 57.78% of T-CAP-CD assembly genes were annotated in three subgroups (BP, MF, CC) with the highly accurate plant-specific Hayai-Annotation Plants annotation tool. The reconstruction of the KEGG pathway revealed 30 ortholog genes in the terpenoid biosynthetic backbone pathway and 8 ortholog genes in the sesquiterpenoid biosynthesis pathway. By performing homology search of T-CAP-CD transcriptome against PlantTFDB v.5, the most frequent transcription factor families were identified as bHLH (9.5 %), NAC (7.3 %), and MYB (6.6 %), respectively.

Early vigor, stress resistance, uniformity, high yield, and breeder right protection have turned hybrid cultivar development into the most successful breeding strategy in rapeseed (Brassica napus L.). The goals of this research were to develop the first CMS based rapeseed hybrids in Iran and to evaluate their performance and fertility. The effect of different traits on yield increasing of hybrids was also studied. Seven CMS lines were crossed with R2000 (a restorer-line) and the seed yield of hybrids was tested using a randomized complete block design with three replications in 2019-2020 and 2020-2021. Three of hybrids with 4.64, 4.40, and 4.38 tons per hectare, showed considerable yield and were significantly different than Nilufar, Neptune, and Nima (3.44, 3.37, and 3.30 tons per hectare, respectively), as check cultivars. R2000×A8 demonstrated the highest heterosis (34.71%) to high-check cultivar. All hybrids obtained were normal in terms of fertility, having no male-sterile flowers. Based on the results of principal component analysis and cluster analysis, the simultaneous increase in the number of pods per plant and the thousand seed weight, together with crossing the more distant parents will lead to more high yielding hybrids. The promising hybrids with high yield and remarkable fertility could be released as new cultivars after passing adaptability and stability experiments.

Preparation of cell suspension is an important step in the study of plant cells which is often done by the culture of friable embryogenic callus. The primary objective of this research was to produce friable callus and to determine an optimum culture medium for the induction of friable callus in maize plant. Using two basic media, Murashige-Skoog and Chu, with different concentrations of 2, 4 Dichlorophenoxyacetic acid and Benzyl adenine hormones (overall 16 different culture media), callus induction was performed on maize mature embryo explants. Selection criteria including high friable callus induction index, high callus size index, high callus fresh weight index, and low regeneration index were measured, and the mean values were compared using analysis of variance. Considering each of these selection criteria alone, different optimum media for callus induction were introduced. In order to determine one optimum medium considering all the criteria, the analytical hierarchy method was utilized. The optimum medium in this study was MS supplemented with 1 mg/L 2, 4-D wherein the induction, weight, and size indices were high, and the regeneration index was low. In order to evaluate and verify the selected medium, the explants were cultivated in in silico- selected medium and the growth curves were plotted for callus fresh and dry weights. Several successive subculturing of the callus was performed on the selected medium wherein no regeneration was observed. The results suggested the analytical hierarchy approach can be used as a reliable decision-making method for optimum-medium selection for inducing maize friable callus.

Verticillium wilt disease is one of the most important damaging and limiting factors of cotton cultivation worldwide. This disease reduces the quantity and quality of the product. Due to the economic importance of Verticillium wilt cotton disease, the best way to control this disease is to use tolerant cultivars. This experiment was performed to determine the effect of disease on morphological and physiological characteristics of cotton cultivars in order to identify tolerant genotypes.

The experiment was conducted in 2020 in the greenhouse of Khorasan Razavi Agricultural Education and Natural Resources Research Center (Mashhad Plant Protection Department). Ten commercial Iranian cotton cultivars belonging to Gossypium hirsutum including Varamin, Sahel, Bakhtegan, Khorshid, Armaghan, Mehr, Koker, Khordad, Kashmar and Deltapine were planted in factories in a completely randomized design with three replications in the greenhouse. The first factor was cotton cultivars and the second factor was the contamination of cotton cultivars with fungal isolates.

The results of analysis of variance of the measured traits on the studied genotypes showed that the tested cultivars in terms of leaf number, crown diameter, number of branches, leaf area, chlorophyll index, disease severity, relative leaf water content and electrolyte leakage membranes were significantly different at the 1% probability level. This difference was not observed in terms of plant height. Comparison of the mean of the above traits (except plant height) in the treatment of contamination levels, indicated that the cotton plants  inoculated with the pathogenic fungus Verticillium wilt compared to control plants (no fungal infection) had lower number of leaves, number of branches , leaf area and crown diameter. Mean comparisons of the above traits were performed between genotypes. Some genotypes can be selected for field experiments in terms of superior study traits. The results of comparing the mean interaction of infection × cultivar for all traits (except plant height) were significant at 1% level. Under the conditions of infection, the highest severity of the disease was related to Varamin and Cocker-handard cultivars. While the lowest severity of the disease was related to Bakhtegan cultivar.

The results showed that Verticillium wilt disease reduces the number of leaves, number of branches, leaf area, crown diameter, chlorophyll index and relative water content. While this disease increases the electrolyte leakage of cell membrane. In this regard, a negative and significant phenotypic correlation was observed between disease severity and number of leaves, branch, crown diameter and chlorophyll index. There was a positive and significant phenotypic correlation between disease severity and membrane electrolyte leakage. Therefore, plants with more leaves, higher chlorophyll index and relative water content and lower electrolyte leakage are more resistant to infection.

To optimize tomato (Solanum Lycopersicum) plant tissue culture condition, the effect of 13 different hormonal combinations and two explants of cotyledon and hypocotyl on the regeneration rate of three Newton, Infinity and Defines cultivars were investigated. Experiments were conducted in a factorial based on a completely randomized design with three replications. The highest shoot number (11.9 shoots per explant) was obtained for Defines cultivar on MS medium containing the hormones of TDZ=0.68, NAA=0.1milligrams per liter and cotyledon explant. Plant defense responses against environmental factors are one of the inhibitor factors of explants regeneration in plant tissue culture. Calcium ion signaling is one of the primary events and, of course, the most important event in plant defense response against pathogens, Therefore, the effect of verapamil as a calcium channel blocker was investigated on the plant defense response. For this purpose, inoculation was performed using Agrobacterium tumefaciens strain LBA4404 and based on optimized tissue culture condition. The effect of verapamil in four concentrations of 0, 50, 100 and 200 milligrams per liter was investigated in a co-culture medium. Results showed that the use of verapamil in comparison with the control sample reduced the plant defense response. Also, the use of 50 milligrams per liter of verapamil increased the amount of regeneration.

Peroxidases are used in a wide range of biotechnological processes, most of which are carried out at high temperatures and high pH levels. Since most of the commonly used peroxidases are unstable and inactive in alkaline conditions and high temprature, it is necessary to find thermoalkalophilic peroxidases for practical purposes.

In this study, extracellular production of peroxidase in the native strain Bacillus tequilensis was studied. for this purpose, Enzyme activity was evaluated using two substrates 2,4-DCP and pyrogallol in bacterial liquid culture and the effect of culture time on enzyme production, as well as the effect of parameters such as pH and temperature on enzyme activity investigated. The relative purification of the enzyme was performed using ion exchange chromatography with sephadex DEAE A50 and the kinetic parameters of enzyme activity were evaluated. In this study, kinetic parameters such as Km and Vmax were calculated.

Measurement of enzyme activity at different times of culture indicated that the highest amount of peroxidase production was obtained 72 h after bacterial culture.

Oil plants store energy in the form of neutral lipids in the organelles called oil bodies. These organelles save triacylglycerol until seed germination. In recent years, the oil bodies have been considered as an oil/water emulsion in the pharmaceutical, food, and cosmetic industries. These organelles are also effective tool for purifying, stabilizing and delivery of biotechnology products. Aqueous extraction processing (AEP) is the most common method for oil body extraction. Despite all advantages compared to organic solvent extraction, the yield of AEP still needs to be optimized. Therefore, this study surveys the efficacy of two solvents, phosphate buffer and distilled water in the oil bodies' extraction from rapeseed.

Brassica napus L. seeds were obtained from seed and plant improvement institute, Iran. To compare the efficacy of solvents, 0.1 M phosphate buffer (pH 7.5) and distilled water were used for extraction. The ground rapeseed was suspended in the buffers in a ratio of 1:10 (w/v) and stirred for 12 hours at room temperature. This step was repeated three times. Then, the extract was centrifuged at 10,000 g for 15 minutes at 4 ° C. The floating layer was carefully removed and dissolved again in the initial solvent and the pH was adjusted to 8.5 to precipitate the deflated proteins. Finally, the cream layer was retrieved using centrifuges and one-tenth of the initial buffer volume was applied to the 9 M urea buffer (pH 7.5) for 10 minutes to separate non-specific proteins from oil bodies. The purified oil-bodies were monitored under light microscopy.

According to the microscopic and macroscopic results, the stability of oil particles and efficiency of extraction would be higher by phosphate buffer due to maintaining a constant alkaline pH during the extraction. Moreover, the presence of different salts in the phosphate buffer increases the purification yield up to twice times as a result of providing osmotic pressure and increasing solubility of membrane proteins. These results emphasize the importance of membrane proteins on the formation and stabilization of oil bodies.

The most common cause of diarrhea is Shigella and no vaccine has been found so far. IpaD and B subunit of Shiga toxin proteins (STxB) play an important role in invasion, infection and pathogenesis caused by Shigella. On the other hand, Co1 ligand has introduced as one of the antigen delivery systems to M-cells in mucosal layer. In this study, nasal administration of antigen in mice, used to evaluation and comparison of immunogenicity of IpaD, IpaD-STxB and IpaD-Co1 proteins. The coding sequences of proteins cloned in pET28a vector and transformed to bacterial strain of E.coli BL21(DE3). Recombinant protein expression confirmed by using of Western bloting technique. The each one of the recombinant protein purified by affinity chromatography column and used for nasal administration in four times consecutively on mice. After the second nasal administration (first booster), a week after each administration, blood samples were collected and ELISA performed on serum. The ELISA results of serum titration showed that the highest amount of IgG has been produced against IpaD-STxB protein; and IpaD-Co1 was in next step. The immunogenicity was evaluated using active toxin E. coli O157:H7. The challenge results showed that immunized mice could endure 5 times the LD50 Shiga toxin E. coli O157: H7; but after injection of 10 times the LD50, the immunized mice by IpaD, IpaD-Co1 and IpaD-STxB were death after 24, 24 and 72 hours, respectively. According to these results recombinant protein IpaD-STxB founded to be more immunogenic than IpaD-Co1 against shiga toxin in mice.

Small and powerful promoters play a key role in recombinant protein expression in different hosts including Pichia pastoris. In the present study, expression of cellulase beta endoglucanase CMC3 gene was studied under the control of the alcohol oxidase 1 (AOX1) and methanol oxidase (MOX) promoters, with the methanol feeding, in the yeast Pichia pastoris. To avoid the gene dosage effect, colonies harbored single copy of target constructs were screened by Real-Time PCR method. The results of enzymatic assay, zymogaphy and SDS-PAGE for the culture of single copy harboring transformants showed powerful, comparable expression of CMC3 under the control of both MOX and AOX1 promoters. High-level expression of recombinant CMC3 under control of the MOX promoter indicated that this small, new powerful promoter could be considered as a promising tool for recombinant protein production in the yeast Pichia pastoris.

Terminal heat stress is one of the most important environmental factors threatening wheat production in Iran and all across the world. In order to study the effectiveness of canopy temperature and Relative water content in screening wheat lines for heat tolerant, ten wheat genotypes were evaluated in three separate randomized complete block design experiments in two years (2015-16 and 2016-17) under non- stress and heat stress conditions at Gonbad and Atrak research stations. Heat stress was applied through three methods including late sowing, plastic greenhouse and field temperature. Canopy temperature, relative water content and grain yield of genotypes were measured. All genotypes showed significant reductions in grain yield under heat stress. Based on grain yield under stress and non-stress conditions, yield loss percentage and Fisher and Maurer susceptibility index, Kauz, Shiroudi and N-92-7 were shown to be tolerant, while Bam and Verinak susceptible to heat stress. Although both canopy temperature and relative water content had a significant correlation with grain yield under heat stress, canopy temperature was significantly different between tolerant and sensitive genotypes, indicating its higher efficiency in screening for heat tolerance.

The objectives of this study were to analyze genotype by environment (GE) interactions effects on the yield of barley lines and varieties, identifying salinity tolerant genotypes with stable yield and also evaluating genotype (G), environments (E) and GE interactions using different stability parameters. The research was set up as two Randomized Complete Block design with three replications at the agriculture research station of Neishabur, Khorasan razavi, during 2013-2015. Each experiment included 17 barley varieties and promising lines. According to Eberhart and Russell stability parameters, genotypes Fajre30, Nik and MBS82-4 had general adaptation. Genotypes Fajre30, Nik, Rihan, Valfajr, MBS82-5 and Mbs87-12 were selected by Simultaneous selection for yield and yield stability (YSi) method. The results of AMMI analysis for seed yield indicated that the Genotype (G) main effects, environment (E), and GE interactions as well as two first interaction principal components (IPCA1-2) were significant. AMMI biplot was able to distinguish stable genotypes and environments with high discrimination ability from low ones. According to the AMMI analysis, genotypes MBS87-12 and Fajre30, with seed yield higher than grand mean, were the most stable genotypes and with high specific adaptation to the saline environment.

Ocimum basilicum L. (2n=48) is a medicinal plant that belongs to Lamiaceae family. Eugenol is one of the most important compounds of basil essential oil which is applied against bacteria, fungi and nematodes in various industries. Phenylpropanoids such as methyl eugenol possess anti-oxidant properties against free radicals and act as protective compounds in plant responses to biotic and abiotic stresses. Methyl eugenol O-methyl transferase (EOMTs), as a key enzyme of phenylpropanoids biosynthesis pathway, converts eugenol into methyl eugenol by methylation. In this study, we isolated, characterized and function analyzed the EOMTs gene promoter from basil plant with the aim of investigating the gene expression regulation of this gene under different environmental conditions and recognition its regulatory motifs. Analysis of EOMTs gene promoter indicated that this sequence was included some important regulatory elements which responded to high temperature and drought stress such as MYB, MYC, HSE, and WRKY. It is expected that this gene indicates expression changes under biotic and abiotic stresses because there are stress-responsive regulatory sites in ObEOMTs gene promoter. Function verification of ObEMOTs promoter in Nicotiana tabacum leaves showed that this promoter including core elements is able to direct the expression of GUS gene.

Broomrape is an obligate parasite in many economically important crops such as tomato. Success in controlling broomrape is rare hence an understanding of sucrose utilization mechanisms as a critical process for parasite provides developing specific control strategies. This parasite acts as a competitive sink for sucrose, utilizes it in the tuber and converts it to glucose and fructose. Invertases are the major enzymes in sucrose degradation and osmotic adjustment in the parasite. In this paper, the activity of two invertases (PrSAI and PrCWI) involved in different developmental stages of some broomrape species (Phelipanche aegyptiaca, P. ramosa and P. nana) on tomato was studied. Results showed that activity of both mentioned enzymes were detected in all of the developmental stages in the broomrape species; although the PrSAI activity was always higher than the PrCWI activity. The activities of PrSAI and PrCWI was increased and decreased, respectively, as progressing of the developmental stages of broomrape. Enzymes activity and protein and sugar amounts in Egyptian broomrape were high at the early stages and in branched broomrape in the late stages, the first case accorded to the earlier emergence of broomrape on the host and the later one accorded to more shoots and higher biomass in broomrape. In total, results suggested that PrSAI1 and, possibly, PrCWI are suitable targets for gene silencing strategies.

The majority of rapeseed in Iran is produced in South hot-dry and North hot-humid regions making it necessary to improve heat resistant cultivars. In order to identify heat tolerant and susceptible cultivars, nine spring varieties of rapeseed including commercial cultivars and promising lines, were evaluated under normal and heat stress conditions in research field of the University of Tehran in two sets of randomized complete blocks with three replications for two years during 2015-2016. Heat stress applied by a plastic greenhouse at flowering stage equipped with electric heater. The temperature exceeding 25.1- 39.40C inside the plastic sheet, remarkably different temperature with the open field, significant yield loss of the varieties under stressed condition in both years, illustrated that plastic greenhouse could be effectively used for applying heat stress. Based on grain yield in normal and heat-stressed conditions and tolerance indices, Roodi22, Dalgan and Mahtab were semi-tolerant while Safi5 and DH13 were shown to be the most tolerant and susceptible varieties which might be considered in production and also breeding programs.

Abrin is a plant-derived glycoprotein toxin composed of two polypeptide chains, A chain and B chain. The A-chain (molecular weight 30 kDa) as a N-glycosidase is able to catalyzes the deadenylation of the 28S rRNA and thereby inactivate ribosomes. On the other hand, B-chain (molecular weight 35 kDa) is responsible for binding to the target cell and internalization of A-chain. In this study, an abrin A-chain (ABRaA) DNA was isolated from Abrus precatorius by PCR technique and then cloned into a T-vector. This isolated sequence with 753 bp in length was registered to NCBI GenBank with the accession number MF573784, as the first report of A chain abrin DNA. In continue molecular structures and biochemicalcharacteristics of ABRaA were analyzed. The secondary and three dimensional analysis of the polypeptide structure with 28.07 kDa indicated that ABRaA is similar to the RIP-II family of plant proteins. The bacterial expression of ABRaA carried out in BL21 (DE3) strain and subsequently confirmed by Western blot.

Salt stresse is one of the most important environmental stresses, which limits plant growth and development. To survive under such conditions, plants must be able to sense and respond rapidly. Molecular studies on various plants show that these events involve complex networks of gene regulation and gene expression. Barley is one of the important crop in Triticeae plants that is tolerant in saline condition. The aim of this study is comparing gene expression patterns in that response to salinity in Afzal cultivar, one of the resistant cultivar in Iran. RNA was extracted using Trizol kit from plants under treated and control condition. RNA libraries was prepared and sequencing was done using Illumina/HiSeq platform. RNA-Seq analysis was performed using systemPipeR package. Results show that in comparative differential gene expression analysis between saline and control condition 683 genes had significant changes with FDR1.5. Bioinformatics analysis show that these genes are related to different pathway including anion ion exchanger, sodium ion transport, transition metal ion transmembrane transporter activity, protein serine/threonine phosphatase activity and lots of phosphatase activity. These large amount of achieved data can be used as a valuable information in future studies in aim of genetic manipulation in order to produce salt-tolerant cultivar.

Diabetes is a disease in which the body does not produce or properly utilize insulin. Although insulin is predominantly made in bacteria or yeast for commercial production, there is a burgeoning need for alternative systems in the production of insulin. In this regard, edible oyster mushroom was studied as host for the production of insulin. Therefore, a binary vector pCAMBIACTB-INS was constructed containing the cholera toxin B subunit fused with human proinsulin gene under the control of glyceraldehyde-3-phosphate dehydrogenase as homologous promoter. Then it introduced into gill tissue of edible oyster mushroom, Pleurotus ostreatus isolate Iran1649c, via Agrobacterium Tumefaciens strain GV3101 (OD600nm) and co-cultivated with P. ostreatus at 25 °C on selective medium containing hygromycin and cefotaxime. After confirming Putative transformants by PCR using gene specific primers, stable integration of the CTB-proinsulin fusion gene into P. ostreatus genome was confirmed by Southern blotting analysis. Also, the expression of transgene was confirmed by RT-PCR, ELISA and immunoblotting methods and in transgenic mycelia the recombinant CTB-proinsulin showed an expression level by 0.04% of total soluble protein. In conclusion successful expression of foreign genes in transgenic mushrooms is promising for development of biopharmaceutical in edible mushrooms.
medium containing hygromycin and cefotaxime. After confirming Putative transformants using PCR with specific primers, stable integration of the CTB-proinsulin fusion gene into P. ostreatus genome was confirmed by Southern blotting analysis. Also, the expression of transgene was confirmed by RT-PCR, ELISA and immunoblotting methods and in transgenic mycelia the recombinant CTB-proinsulin showed an expression level by 0.04% of total soluble protein. In conclusion successful expression of foreign genes in transgenic mushrooms is promising for development of biopharmaceutical in edible mushrooms.

Basil (Ocimum basilicum L.), a member of a Lamiaceae family, is used in traditional Iranian medicine. Essential oils of basil leaves are composed of terpenoids and phenylpropanoids which are important in treatment of diarrhea, coughs, warts, worms and kidney malfunctions. The valuable compound of methylchavicol makes up the bulk of plant volatile oils which recently raised researcher's attention due to its medicinal value and pharmacological activities, especially as a migraine prophylaxis agent. Chavicol O-methyl transferase (CVOMTs) is a key enzyme in phenylpropanoid pathway. It catalyzes the methylation of chavicol, to produce methyl chavicol. In current research, to explore the formation and transcriptional control exerted on these pathways, mRNA accumulation profiles of key gene in the biosynthesis pathway of methylchavicol were evaluated in the three chemotypes of O. basilicum under three treatments of water deficit stress (S1: 75% FC, S2: 50% FC and S3: 25% FC) using quantitative real-time PCR. In order to study key biosynthetic pathway accurately gene expression pattern methylchavicol combination under water deficit stress, isolation, characterization and functional analysis of promoter of CVOMTs (pCVOMTs) was investigated for the first time. The study profile of gene expression of CVOMTs indicated that the transcript accumulation of this gene was significantly affected by chemotypes and water stress levels. The level of transcripts of CVOMT gene was gradually up-regulated and then sharply increased to mainly detectable levels under S2 in cultivar 3. The analysis of promoter of CVOMT gene (pCVOMTs) revealed that pCVOMTs contained some important cis-acting element like high temperature and water stress- response related elements. So, in the chemotype 3, the increase of gene expression is related to the binding site of transcription factor that is excied by water deficit stress.

Bacillus anthracis as a spore-forming bacterium is the causative agent for anthrax. Three proteins from B. anthracis including protective antigen (PA), lethal factor (LF) and edema factor (EF) are causative factors of this bacteria. To produce plant-based vaccine, fusion protein of PA 4 domain and LF 1 domain via furin cleavage site joined to subunit B Vibrio cholera toxin (CTB) as an inducer of immune system. In this research we used 3’and modified 5’ untranslated regions of cowpea mosaic virus (CPMV) as translation enhancer and silencing inhibitor gene 2b of cucumber mosaic virus (CMV) to construct binary vector and transfer gene. Transient expression of tobacco and lettuce performed via agrobacterium and transient expression was confirmed at the level of transcriptome and proteome through RT-PCR and ELISA assay, respectively.

Salinization of crop lands is one of the most important reasons for reduction in productivity and threats food security. Activation of signaling process in plants is the first step during sense environmental cues and stress signals. Identification of the components of plant intracellular signaling mechanisms, pathways, and networks in response to salt stress in Aegilops tauschii (the D genome donor of bread wheat) by transcriptome analysis tools such as RNA Seq method is very worthwhile. RNA Seq method and functional annotation of data by MapMan suite is an efficient strategy to functional genomics analyses. In order to identify and evaluate some genes involved in signaling process, we performed library construction for two cDNA libraries from leaves of seedlings treated with 200 mM NaCl and control. Results of differential expression showed that out of 4506 differentially expressed genes, 603 (11/59%) BINs assigned to signaling process. It also became clear that receptor kinases are the largest number (222 transcripts) of differentially expressed genes in signaling process components. This study is a fundamental research that provide worthwhile functional sequences for gene transferring study to improve salinity tolerant in bread wheat.

Stem reserves of barely can be an important contributor to grain filling particularly under condition is limited such as salinity stress. The aim of this research was to study dry matter accumulation and remobilization ability in different internodes of some Iranian Barley varieties and promising lines under salinity and normal conditions. The research was set up as two Randomized Complete Block design with three replications at the agriculture research station of Neishabur, Khorasan Razavi, during 2013-2014. The amount of dry matter accumulation and remobilization were determined using method. The relationship between length and specific weight of internodes with dry matter accumulation and Remobilization was investigated. The results showed that lower internodes had maximum dry weight and specific weight in both of experimental conditions. Maximum remobilization was obtained from penultimate and lower internodes in normal and salinity conditions, respectively. A positively significant correlation was observed between seed weight per spike and the amount of Remobilization from all of internodes in saline condition. Results indicated that dry matter accumulation and remobilization ability are two different mechanisms that inherent from parents to progeny. Also in this study it was found that new tolerant promising lines compared to old tolerant varieties, had internodes specific weight, dry matter accumulation and remobilization ability.

Currently, millions of people all over the world for escaping the lethal effects of diabetes require persistent injections of insulin. Thus, to develop a system with low cost and high production of insulin that can fit into the consumers, is essential. With evolving genetic engineering techniques, production of human insulin in bacteria and yeasts were feasible, but due to drawbacks of producing recombinant proteins in those organisms including high cost, the possibility of contamination with toxic proteins and costly purification steps, most research has focused on the production of recombinant proteins in plants. Therefore, this investigation was done to express transiently the human proinsulin in alfalfa leaves by two different constructs. Construct pLPC as a fusion gene B subunit heat liable toxin of E.coli - proinsulin - penetratin and constructs pEPC incorporation into a peptide containing extensin leader - Proinsulin - penetratin. These constructs were transferred into Agrobacterium strain LBA4404 and agroinfiltration. Gene expression in alfalfa leaves was attested by RT-PCR and specific primers. The production of active proinsulin was confirmed by medical diagnostic mono-bind kits. The amount of production of active proinsulin in agroinfiltrated alfalfa leaves with constructs pLPC and pEPC were estimated 76 and 71.6 nano-grams per gram of fresh alfalfa leaves respectively.

Artemisia annua is an annual herb native to Asia most probably China produce one of the most important sesquiterpene namely artemisinin which has antimalarial and anticancer activity. Three transcription factors belonging to WRKY family, two transcription factors belonging to MYB family and two enzymes DBR2 and OPR3 were isolated from A. annua leaves and first inserted into pJET1.2 vector then sub cloned into cowpea mosaic virus (CPMV) based viral vector (pEAQ-HT). Thegenes were transiently expressed in A. annua young leaves of two different chemotypes ANAMED and Iranian wild type under the control of a constitutive cauliflower mosaic virus (CaMV) 35S promoter as vacuum infiltration. An agroinfitration method was carried out for transient expression of different genes. The analysis of metabolites by GC-MS method showed that the transcription factors which have expressed transiently, could effect on the rate of DHAA and AA metabolites. The results show that the TFs (WRKY and MYB) are involved in artemisinin biosynthesis pathway.

Germination is one the most important growth phases in plants that injury from drought stress extremely. Germination divided to 3 phases although before third phases، many activity (radicle protrusion) initiated. Many metabolically activity occurred specially in first phase since seed embryo is a live organ، although differences in protein patterns arised in this phase. This study was carried out to understand the effect of drought stress on protein patterns in the first phase of germination. In order to analysis the effects of drought stress (-12 bar) on wheat embryo proteome (Sardari and Qods respectively tolerant and susceptiblecultivar)، factorial experiment was conducted in a Randomized completely block design with three replications. Drought stress diminished germination percentage and caused adverse effects on the proteins pattern. Two-dimensional gel electrophoresis analysis of wheat embryo axis showed 1000 spots were reproducible that 25 spots between treatments indicated significant differences on 5درصد level. In control condition، in comparison tolerant to susceptiblecultivar، 22 and 6 spots and in stress condition، 21 and 10 spots were down and up regulated respectively. The results indicated that frequency of protein under drought stress depends on severity and amount of stress and imbibition time.