بهمن بهرام نژاد

Papaver plants from the poppy family are among the famous medicinal plants that have always been the focus of the pharmaceutical and medical industries due to the production of morphine alkaloids and isoquinoline alkaloids with antimicrobial effects. A wide variety of Papaver species can be seen in the pastures and mountains of Kurdistan province. In this research, the methanolic extracts prepared from four Papaver species were investigated about the amount of morphinan alkaloids produced and the antimicrobial effects on the growth of microorganisms in laboratory conditions.
Four Papver species including P. glaucum, P. fugax, P. argemone, and P. bracteatum were collected twice from their natural habitats in Kurdistan province in spring and were identified based on morphological characteristics under the supervision of a botanist of the province's agricultural and natural resources research center. In order to extract alkaloids, aerial parts, and roots were dried and powdered in the shade. Extracts were prepared by adding methanol and chloroform solvents in a ratio of 1 to 4 to the resulting powders of plants. The amount of morphine, codeine, thebaine, and papaverine alkaloids present in each of the samples was measured separately using high performance liquid chromatography (HPLC). Also, the antimicrobial effects of extracts against Gram-positive bacteria, Bacillus subtilis, Staphylococcus aureus, Streptococcus pyogenes, and gram-negative bacteria, Pseudomonas aeruginosa, Salmonella typhi, Escherichia coli were investigated. The pathogenicity was evaluated by measuring the growth halo, determining the minimum inhibitory concentration (MIC) and the minimum lethal concentration (MBC), using the tube dilution method.
The production of morphine alkaloids in all of the studied species was confirmed by HPLC. The amount of morphine as the dominant alkaloid among the four studied species varied from 4.21 to 21.33 mg/g dry weight. The highest amount of morphine (21.33 mg/g) was observed in P. bracteatum extract and the highest amount of thebaine and codeine (8.67 and 1.8 mg/g) was observed in P. fugax extract. The observation of non-growth aura against Pseudomonas aeruginosa and Staphylococcus aureus bacteria confirmed the antimicrobial effects of the investigated plant extracts. The detected amounts of minimum inhibitory concentration against the gram-positive and gram negative bacteria varied from 25 to 100 mg/ml and minimum lethal concentration was just observed against Escherichia coli. P. bracteatum plant extract showed the highest inhibitory effect against E. coli bacteria with a minimum inhibitory concentration of 25 mg/ml and a minimum lethal concentration of 50 mg/ml.
By examining the results, it was found that the studied Papaver species produce significant amounts and ranges of morphine alkaloids, which designates these natural species' genetic and medicinal value for further studies. In addition, the extract of these plants showed antimicrobial effects against the reference strains of gram-positive and gram-negative pathogenic bacteria used in this research. Therefore, it is suggested that more studies be conducted in the field of separation and separation of the metabolites of these plants and the investigation of their antimicrobial effects in subsequent studies to achieve effective and novel antimicrobial compounds and drugs.

Exendin-4 (Ex4) is a 39-amino acid peptide isolated from the salivary secretions of the lizard Heloderma susceptum and it has similar biological function as glucan-like peptide (GLP-1) receptor. The most important physiological role of GLP-1 is to regulate the amount of glucose metabolism by reducing the entry of glucose into the cells. Transgenic plants can be used as factories for the production of recombinant proteins. In order to maximize the efficiency of transgenic plants, the expression of foreign genes can be regulated in terms of time and place under the regulation of different promoters. Considering the nutritional value of carrot tuber and its fresh consumption, in this study, CTBEx4 gene was expressed in carrot plant using a root-specific promoter. In this study, in order to specifically express the CTBEx4 gene in carrot root, the Major Latex-Like promoter (MLL) related to the specific expression in sugar beet storage root tissue was replaced by the CaMV35S promoter in the pBI121 construct and were used for the transformation of carrot explants.

Using the CTAB method, DNA was extracted from the leaves of the sugar beet plant, then the PCR reaction was performed using specific MLL primers. The obtained fragment was replaced by the CaMV35S promoter in the pBI121 construct using restriction enzymes. Both pBI121-Mll-CTBEx4 and pBI121-CamV35S-CTBEx4 constructs were used to transform carrot leaf explants using Agrobacterium tumefaciens bacteria. Transgenic explants were indirectly regenerated into plants in MS culture media in vitro and then transferred to pot. Finally, the CTBEx4 gene expression of the transgenic plants was analyzed at mRNA and protein levels using RT-PCR and ELISA.

The results of the expression analysis using RT-PCR and ELISA methods indicated that the expression of CTBEx4 gene in carrot root tissue was regulated by MLL promoter, while no expression was observed in leaf tissue. Also, the results showed that the CTBEx4 gene was expressed under CaMV35S promoter regulation in both root and leaf tissues, but its expression in roots was lower (30%) than that under MLL promoter regulation. The obtained results indicated the ability of the MLL promoter to specific expression the CTBEx4 gene in the roots of carrot plants.

The MLL promoter was able to express the CTBEx4 gene in the root tissue of carrot plants at the mRNA and protein level, Also, CTBEx4 protein under MLL promoter had higher expression level compared to CaMV35S promoter.

Exendin-4 (EX4) is a protein similar to glucan-like peptide (GLP-1) and has a longer half-life and greater effect in inducing insulin secretion. The fusion of EX4 to the non-toxic cholera subunit (CTB) increases its efficiency through absorption in the digestive system. In this study, CTB-EX4 gene was introduced into lettuce by Agrobacterium tumefaciens using vector pBI121. The results showed that 20.25% of the inoculated explants were successfully transformed and regenerated (T0 generation). To create T2 plants, seeds of T1 were cultivated, and 42% of the cultivated seeds germinated. T1 plants were analyzed using PCR test. The mRNA expression of the CTB-EX4 gene was examined through RT-PCR and Real-time PCR, while the protein expression was assessed using ELISA and Western Blot tests. The number of transferred copies to T1 plants was determined using Real-time PCR. This estimation was performed using both the absolute method and the standard curve, as well as the relative method, with Acitn gene as a reference gene. The results showed that 60% of the T1 plants were transgenic and the CTB-EX4 gene was expressed at the mRNA and protein levels. The results showed that the process of transferring the CTB-EX4 gene to the lettuce plant was successful and the lettuce plant was able to express the desired gene at the RNA and protein levels. Considering the subsequent processes, including the purification and evaluation of the biological activity of the produced protein, plants can be used as a suitable bioreactor for the production of recombinant proteins.

Ferula pseudalliacea is a perennial and monocarpic plant species belonging to Apiace-ae family. This species is distributed from Middle Asia to Northern Europe. The main secondary me-tabolites present in Ferula genus comprise coumarins, sesquiterpene coumarins and di-sesquiterpene coumarins. Based on phytochemicals present in this plant species, it has been extensively utilized by local people in traditional medicine for long time. Phenylpropanoid pathway is mainly responsible for biosynthesis of these valuable phytochemicals. Cynamate-4-hydrolxylase (C4H) is one of the main and key enzyme in phenylpropanoid pathway. To date, there is no genomic or transcriptomic information in Ferula pseudalliacea and as the first report, we aimed to clone and sequence C4H and its expression analysis in root, stem, leaf, inflorescence and immature seed in this valuable medicinal plant species.

Different organs of F. pseudalliacea were collected from Gazne village near Sanandaj city in June 2019. RNA was extracted from collected organs using LiCl method. cDNA was synthesized using Yekta Tajihz Azmia kit in 20 µl reactions. PCR products amplified with pri-mer designed from conserved regions of C4H among apiaceous species and it was then cloned within pTG19. The verified recombinant plasmid were send for Applied Biosystem company for sequenc-ing. To confirm and asses the expression of C4H, both Real-Time PCR and semi-quantitative PCR were employed. The phylogenetic tree of C4H was obtained using Omega software based on the Neighbor-Joining method with bootstrap 1000 for amino acid sequence.

The obtained sequences from C4H clones was aligned and verified using BlastX. In the re-sulted dendrogram, C4H sequence from F. pseudalliacea was grouped with C4H gene from other species within Apiaceae family showing their clos relationship. The highest sequence identity was observed with C4H from Daucus carota which was fallen in same cluster with C4H from F. pseudal-liacea. It is shown that C4H has 7 motifs one of this motifs belongs to conserved motif cytochrome-cysteine 450 with FGVGRRSCPG sequence.

As the seconf most important gene in phenylpropanoid biosynthetic pathway, C4H plays an undeniable role in production of phenolic phytochemicals in particular in inflorescence which had the highest C4H expression rate. Studies performed on the C4H expression in other spe-cies revealed a specific action of C4H in biosynthesis of special secondary metabolites in specific tis-sue. As the phenolic compounds encompass a diverse range of secondary metabolites in forms of volatile and extracts, further investigations are needed to understand the exact role of C4H enzyme. Moreover, the identified gene sequence can be sued in production of recombinant constituents in other species which have the corresponding precursors.

Salinity is one of the environmental stresses that affects bread wheat grain yield in most parts of the world. One of the basic strategies to mitgiate the effect of non-biological stresses such as salinity is genetic improvement of crop plants. Identification of stress-associated genes is a prerequisite for genetic improvement. In the present study, the role of a number of genes in the aquaporin family and genes associated with the CDPK pathway in response to environmental stresses was identified as their expression profile analysis in response to salinity stress would facilitate breeding for salinity tolerance. In 2014, the expression of two genes TaNIP and TaSC was studied in two bread wheat cultivars "Bam" tolerant to salinity and "Tajan" sensitive to salinity under salinity stress (170 mM NaCl) and Non-saline conditions. The results showed that TaNIP expression in salt-tolerant cultivar (Bam) significantly increaseed in both leaf and root tissues under salinity stress conditions, but in sensitive cultivar (Tajan) the level of TaNIP expression, especially in leaf tissues, was lower and showed a decreasing trend. In cv. Bam in both leaf and root tissues, after 24 hours, TaSC expression increased in comparion to control. In cv. Tajan, the expression in leaf tissue was generally lower and after 24 hours, the expression level significantly decreased in comparion to control (non-saline). Moreover, expression analysis under the simultaneousapplication of salicylic acid andsodium chloride showed that the expression of both genes in leaf tissues of both bread wheat cultivars significantly increased and the gene expression showed an increasing trend. In conclusion, the results of this experiment indicated that the TaNIP and TaSC genes had different expression trends in tolerant and sensitive bread wheat cultivars under salinity stress conditions. Using salicylic acid as an elicitor also activated the hormonal transduction pathway, which induced the expression of both genes. These findings highlights the role of these genes in salinity tolerance of tolerant bread wheat cv. “Bam”.

Genetically improvement for salinity resistance is one of the most effective breeding methods for wheat. But understanding the mechanism, assessing and identifying salinity tolerance genes is a prerequisite for genetic modification. The role of a number of families of transcription factors has been proven in response to environmental stresses, hence, their identification and investigation facilitates the possibility of overcoming stresses. Considering the significant role of the important family of WRKY transcription factors in response to stresses, the expression of two important genes of this family including TaWRKY10 and TaWRKY53 in different tolerant and susceptible cultivars of wheat was investigated. The results showed that TaWRKY10 exhibited different expressions in susceptible wheat (Tajan) and resistant (Bam) cultivars. In resistant cultivar (Bam) under saline conditions over a period of 48 hours, TaWRKY10 showed more expression level in both leaf and root tissues than the control and also the sensitive (Tajan) cultivar. However, the expression of TaWRKY53 was different in resistant and susceptible cultivars. TaWRKY53 in susceptible cultivar (Tajan), over the period of 48 hours, showed more expression level than the control and resistant cultivar (Bam) in leaves and root tissues. Also, by using salicylic acid under salinity stress, the expressions of TaWRKY10 and TaWRKY53 in Tajan and Bam cultivars were different. In general, our results show that WRKY transcription factors can play a role in generating resistance to salinity stress in wheat and therefore can be used in genetic improvement.

Endophytes are microorganisms (bacteria or fungi or actinomycetes) that reside inside the internal tissues of plant with no negative effect on host plants. Endophytic bacteria trigger many positive effects on host plants. The aroma and flavor of strawberry fruit are important characteristics that is affected by endophytic bacteria such as Methylobacterium. Endophytic bacteria, are effective in the synthesis of carotenoids and the production of volatile and phenolic compounds. In addition, they are effective in producing the aromatic components in fruit. Endophytic bacterial frequency and genotype can be affected by host plant genotype, climatic and soil conditions. This study was carried out to isolate endophytic bacteria from two strawberry cultivars (Kurdistan and Parous) in three regions (Angouzan, Gavrood and Noshur) in Kurdistan province. For accurate identification of bacterial isolates, 16S-rRNA gene sequencing was used for identification of purified endophytic isolates and their frequency determined at different times of fruit ripening. The results revealed that the all endophytic bacterial isolates from both cultivars were belong to the genus Methylobacterium. The frequency of these bacteria was statistically different in two cultivars and in different sampling time. The frequency of Methylobacterium significantly was higher in Kurdistan cultivar compared to Parous cultivar in all three regions (p <0.05). The highest frequency of Methylobacterium was observed in the fruit ripening time. It seems that Methylobacterium and its frequency probably have an effect on the aroma and delicacy of these two varieties.

Wheat (Triticum aestivum L.) is one of the most important grain products in the world which is the main food of the people in many parts of the world and provides about 20% of the calories needed by the world population. The late stages of wheat growth in the Mediterranean regions are usually accompanied by drought stress. Supplemental irrigation is one of the appropriate strategies to reduce the damaging effects of drought stress and improve yield and sustainability. Nitrogen is the most important nutrient in rainfed farming after wheat planting is the most important tool available to increase yield. Nitrogen fertilizer application to soil at late vegetative stage may not be appropriate due to dry soil surface and reduced root activity, Therefore Nitrogen spraying can be used as a quick and efficient way to meet plant nutrition due to its advantages such as faster and more uptake by the plant.

In order to study the effect of supplemental irrigation and different rates of urea fertilizer and urea foliar application on dry matter remobilization, yield and yield components and Grain protein contents experimental wheat in split –split plot in randomized complete block design with three replications It was designed and implemented in the research farm of Kurdistan University in 2016-2017. Treatments included irrigation levels (no irrigation and irrigation at booting stage) as main factor, three dry wheat cultivars (Rejaw, Sardari and Azar 2) as sub factor of different rates of nitrogen fertilizer (50 kg urea in fall (A), A + 50 kg urea in the spring (B) and B + foliar solution (20 kg urea in the booting stage) were used as a sub-factor. The studied traits included leaf dry matter, stem + leaf pod and spike in flowering and ripening stages, leaf dry matter remobilization, stem + leaf pod and panicle seed, total soil remobilized dry matter, soil yield. Dry leaf remobilization, stem + leaf pod and pod straw, total remobilization efficiency, remobilization share in grain yield, spike number per square meter, spike number per grain, 1000-grain weight, biological yield, yield, harvest index and grain protein content.Leaf dry matter, stem + Sheath dry matter and spike dry matter in flowering and maturity stages, leaf dry matter remobilization, stem + Sheath dry matter, Chaff dry matter, total dry matter remobilized, leaf,, stem + sheath, chaff and total dry matter remobilization efficiency, remobilization share in grain yield, spike number per square meter, kernals per spike, 1000-grain weight, biological yield, yield, harvest index and grain protein content were measured.                                                        

Results showed that the amount of stem + sheath, dry matter at flowering and maturity, leaf, stem + sheath, spike remobilization dry matter, total remobilization, stem+ sheath remobilization efficiency, remobilization share in grain yield, weight Spike, kernals per spike, 1000-grain weight, yield and biological yield were affected by supplemental irrigation.Among cultivars in terms of stem + sheath and chaff dry matter at flowering and maturity, leaf, stem + Sheath, spike remobilized dry matter, total remobilization, leaf remobilization efficiency, remobilization share in grain yield, weight Spike, kernals per spike, 1000-grain weight, yield, biological yield, harvest index and grain protein were different. Rejaw cultivar had more grain number per spike and yield than Azar2 and Sardari.Different rates of nitrogen fertilizer had a significant effect on stem + sheath, spike and total dry matter remobilization efficiency and remobilization share on grain yield.Supplemental irrigation at the booting stage increased dry matter stored in stem + sheath and increased the amount of remobilized dry matter during grain filling.Irrigation at booting stage increased grain yield by 12.4%. Increasing urea fertilizer and urea foliar application at the end of growth stages increased plant yield and grain quality by providing more nitrogen and stay-green.

Accumulation of nitrogen in vegetative organs and its translocation to grain of wheat are important processes that determine the yield and quality. In order to investigate the effect of supplemental irrigation nitrogen fertilizer on dry matter remobilization, yield and yield components of three wheat cultivars a split-split plot experiment was conducted based on randomized complete block design with 3 replications at Research Field of Kurdistan University, Sanandaj, west of Iran, during 2016-2017. Treatments were two levels of Irrigation (rainfed and irrigation at the booting stage) as main-plots, three rainfed cultivars (Sardari, Azar2, and Rejaw) as subplot and three rates of nitrogen (50 kg/ha N (N1), 100 kg/ha N (N2) and 100 kg/ha N plus 20 kg/ha N (N3) foliar application in Heading stage) as sub-subplot. The results showed that supplementary irrigation increased the concentration of leaf chlorophyll a, chlorophyll b, proline, and glycine betaine and decreased the electrolyte leakage. Supplemental irrigation also increased leaf nitrogen, chaff and total nitrogen content at flowering stage. Rejaw cultivar had the highest leaf nitrogen remobilization efficiency and the lowest total nitrogen remobilization efficiency. N1 fertilizer treatment led to the lowest concentration of leaf chlorophyll a and b, proline, glycine betaine, and nitrogen, and stem, chaff and total nitrogen in both flowering and maturity stages. Nitrogen remobilization was affected by supplementary irrigation and different rates of nitrogen fertilizer. It seems that supplementary irrigation in the booting stage leads to an increase in nitrogen content in the vegetative organs, which is transferred to the grain during the seed filling stage.

In this study, three nonpathogenic endophytic bacterial isolates recovered from Fragaria × ananassa stolon and petal and their antagonistic activity was evaluated against Colletotrichum nymphaeae, the causal agent of strawberry anthracnose under in vitro, in vivo, and greenhouse conditions. Bacterial isolates were identified as Bacillus spp. using a combination of phenotypic, biochemical properties and molecular phylogenetic analysis of the 16S rDNA gene sequence. The living cells of bacterial isolates inhibited the mycelial growth of C. nymphaeae using dual culture method and inhibition percentage was evaluated as 41.87, 50, and 54.92 % in MarG2, MarD40 and MarD35, respectively. The volatile compounds (VOCs) produced by bacterial isolates inhibited the mycelial growth of C. nymphaeae and the highest inhibition was observed in MarD35 isolate (24.7%). Moreover, the cell free culture filtrates of bacterial isolates reduced the mycelial growth and conidial germination of pathogen and the highest inhibition was observed in MarG2 (34.96%) on mycelial growth and in MarD35 (52.07%) on conidial germination. The living cells of bacterial isolates decreased anthracnose fruit rot at postharvest and the highest inhibition was observed in MarD40 (85.63%) and MarD35 (81.72%), respectively. Furthermore, disease severity of strawberry anthracnose by bacterial isolates was reduced using drenching soil and plant spraying techniques. The findings of this study showed that bacterial isolates have the ability to prevent the growth of fungal pathogen under in vitro, in vivo and greenhouse conditions.

Symbiotic bacteria associated with the root of legume plants were isolated from soil and the root nodules of alfalfa (Medicago sativaL.), chickpea (Cicer arietinum L.) and yellows alfalfa (Melilotus officinalis L.) grown in arsenic-contaminated areas in south eastern part of Kurdistan province, Iran. According to physiological and biochemical properties, isolates were identified as the members of Rhizobium and Ensifer genus. The presence of arsenic resistant system known as ars system was confirmed by amplification using primers corresponding to arsC gene. Growth rate of strains in different concentrations of arsenate was investigated in YEMB medium supplemented with 100-400 mM arsenate. Alfalfa,s isolates (AB1, AB3), chickpea,s isolates (PA2, PC2), yellow alfalfa,s isolate (YA1) and standard strain (Sinorhizobium meliloti SM117) were tolerant to 350 mM arsenate and (AB1, PA2 and YA1) isolates were moderately capable to grow at 400 mM arsenate. Among the identified strains, AB1 and PA2, were selected for greenhouse experiments. In order to evaluate the effect of arsenic contamination on legume-rhizobia symbiosis and biomass production of alfalfa and chickpea plants, an experiment was carried out based on a completely randomized design with a factorial arrangement in three replications. Experiment factors consisted of, three levels of rhizobia inoculation (with and without inoculation of AB1 and PA2) and five levels of arsenic concentrations (0. 10, 50, 75 and 100 mgkg-1 soil) under greenhouse conditions for 8 weeks. Results obtained in this study indicated that the fresh weight and dry weight of alfalfa and chickpea (shoot and root) were decreased as the arsenic concentration of the soil was increased. The results also showed that fresh and dry shoot and root weight of alfalfa and chickpea were significantly higher in rhizobia-inoculated treatments compared to non-inoculated plants.

The gene 1-deoxy-D-xylulose 5-phosphate synthase (DXS) and TcGLIP multifunctional genes are two key genes involved in pyrethrins biosynthetic pathway, which retain insecticidal properties. DXS gene in the MEP pathway produces 1-deoxy-d-xylulose 5-phosphate using pyruvate and glyceraldehyde-3-phosphate dehydrogenase. TcGLIP gene is involved at the last steps of pyrethrins biosynthesis pathway and integrates chrysanthemyl-CoA and pyrethrolone, which are produced in the MEP pathway and lipoxygenase pathways, respectively to yield pyrethrin I. Based on the significances of pyrethrins, further identification and studies are needed on these metabolites. The aim of this study was to isolate DXS gene in pyrethrum (Chrysanthemum cinerariaefolium Vis.) from the Asteraceae, as well as expression analysis of these two genes under methyl-jasmonate treatment. In the present work, the nucleotide sequences of 1-deoxy-D-xylulose 5-phosphate synthase gene of different plant species were obtained from NCBI database and subsequently aligned with ClustalW online software. Specific primers were designed from the conserved regions of aligned sequences. The polymerase chain reaction product with 715bp length showed that the specific primers had high efficiency to isolate DXS. In addition, gene expression analysis under methyl-jasmonate treatment showed that both DXS and TcGLIP were up-regulated in response to methyl jasmonate elicitor. Our data showed that methyl jasmonate treatment in seedling stage could be used as a suitable elicitor to increase pyrethrin production.

Medicinal plants have a range of significant impacts on human health due to their valuable bioactive compounds with medicinal properties. Chicory (Cichorium intybus L.) belongs to the family of Astracease, is one of the important medicinal plants which produces several types of secondary metabolites such as triterpenoid saponins. Triterpenoid saponins are the most important terpene compounds, which are often in glycosylated form as valuable substances have many applications in industry and medicine. β-amyrin synthase, an important member of the oxidosqualene cyclases family of enzymes in plants, play a crucial role in triterpene saponin biosynthesis. Because of the importance of saponins in chicory plants, partial isolation and expression of the encoding gene of β-amyrin synthase was aimed. RNA isolation, cDNA synthesis, partial cloning and nucleotide sequencing were successfully performed. Sequencing and bioinformatic analysis of the partial coding regions revealed that the partial isolated β-amyrin synthase gene in chicory is very similar to the β-amyrin synthase in other plant species, particularly Aster and Artemisia. Additionally, transcript gene expression analysis with semi-quantitative RT-PCR method and using GAPDH as reference gene was performed in leaf and root tissues of two different chicory genotypes. Results illustrated that expression levels of β-amyrin in root tissues was significantly higher than those in leaf tissues. Moreover, the transcript level of β-amyrin was considerably different between the two genotypes. The results of this study can be used in a complementary basic research in chicory and also to increase saponins through metabolic engineering.

Black cumin (Nigella sativa) is an important medicinal plant belongs to the Ranunculaceae which produce large diversity of secondary metabolites including industrial and medicinal monoterpenes/ triterpenes. In the present work, relative gene expression of genes involved in biosynthetic pathway of terpenes including a monoterpene synthase (MTS), geranyldiphosphate synthase (GDS), β- amyrin synthase (βAS) and squaleneepoxidase(SQE) were investigated in black cumin plants treated with methyl jasmonate. The plants were grown in a greenhouse sprayed with 0.1 mM methyl jasmonate. Samples were collected at 0, 4, 8, 24, 48 and 72 hours after applying the treatments. RT-PCR was performed by gene-specific primers after RNA extraction and cDNA synthesis. Results of semi quantitative RT-PCR showed that the expression pattern of GDS gene differed with the results of other genes. The highest gene expression of GDS was observed at 4 h and its expression remained at the same level till 8 h then dropped at next time periods. The expression patterns of monoterpene synthase, β-amyrin synthase and squaleneepoxidase showed rising trend. Gene expression analysis of the genes at transcript level using semi-quantitative PCR in black cumin leaves treated by methyl jasmonate revealedinduced-geneexpression which might show the roles of terpenes in defense mechanisms and signal transduction. Consequently, methyl jasmonate treatment can be considered to induce the terpene biosynthetic pathway and to study the enzymes/ metabolites associated with terpenoid metabolism.

The purpose of this research was to transform two economically important cultivars of strawberry with P5CS gene، which encodes Δ1-pyrroline-5-carboxylate synthetase (P5CS)، the key enzyme in proline biosynthesis. Shoots were obtained on MS basal medium supplemented with 2% glucose and 4 mg/l TDZ for Camarosa and Kurdistan cultivars. For genetic transformation، a binary vector PBI121 containing P5CS gene under control of the 35SCaMV promoter was used. Transformed cells (explants) were regenerated on the selective regeneration medium containing 75 mg/l kanamycin and 500 mg/l cefotaxime after five days of pre-incubation and 72 h of co-cultivation with Agrobacterium، while control explants failed to grow in the same medium. The presence of the transgene in the plant genome was confirmed by PCR. The morphology of the transgenic plants was normal as controls. Drought stress was applied using polyethylene glycol (PEG) 6000 at concentrations equal to 0، -5 and -7 Bar، respectively. Proline content was four times higher in the transformed leaves compared to that of the untransformed plants while the proline content in the roots was similar in both transgenic and wild-type plants. Overproduction of P5CS also increased chlorophyll content، shoot length، shoot fresh and dry weight in the transgenic plants under drought-stress conditions.

Black cumin (Nigella sativa) is an important medicinal plant belonging to the Ranunculaceae which produces large diversity of secondary metabolites especially terpenes. Mono (C10) and tri-terpenes (C30) are two major structural classes of terpene compounds. Several genes are involved in biosynthetic pathway of the monoterpenes known as MEP pathway، located in plastids. In the MEP pathway، monoterpene synthases are rate- limiting enzymes for monoterpene biosynthesis. In the biosynthetic pathway of monoterpenes geranyl diphosphate synthase catalyzing geranyl diphosphate (GDP) biosynthesis as universal precursor of monoterpenes. Oxidosqualenecyclases is of the most important. In this study gene expression patterns of a monoterpene synthase، GDP synthase، beta amirin synthase and squalene oxidase were evaluated under salicylic acid treatment. Gene expression analysis of the genes at trasnscript level using semi-quantitative PCR in response to salicylic acid treatment revealed their differential expression.

One of the most important proteins in gene silencing and creation of small RNA، are argonaute proteins. Most of these proteins have an endonucleolytic activity so argonaute proteins play a key role in creation of defense mechanism against certain pathogenic viruses. In current study we studied the putative argonaute genes in six medicinal plants using bioinformatic tools. Sequences of argonaute genes of six medicinal plants were retrieved from target database using Arabidopsis Argonaut blast. Sequence alignment، 3D structure، phylogenetic tree and conserved domain were generated for six medicinal plants argonautes. Results showed that studied medicinal plants have 6 to 18 argonaute proteins in the genome. There were three classes of Argonaute proteins similar to that of Arabidopsis، and each varies in length from 846 to 1187 AA، with a average of 970 AA and 108 kDa. six medicinal plants argonaute proteins had PAZ، PIWI and MID conserved domains. Our results showed that six medicinal plants have three classes of argonautes، This indicates is that the conservation of argonaute proteins in different plants.

Total of nineteen isolates of Botrytis sp. were collected from infected strawberry plants of hydroponic cultivated greenhouse and fields surround kurdistan province. According to the morphological characteristics، isolates were identified as Botryits cinerea species. For confirming the species identification by molecular method specific primers C729 were used and PCR test repeated twice. The results showed that these isolates amplified the 700 bp band which is specific for this species. Genetic diversity of existing isolates was investigated by RAPD analysis. According to the results obtained، isolates were clustered into four groups. Considering this clustering mode، in first glance، most of the greenhouse isolates were grouped together and separated from the group of field isolates، but some of them didn’t follow the pattern and were divided differently. Likewise، study among field isolates collected surround the province and their relationships، showed that there is no correlation between molecular clustering and geographical origin of these isolatesو but also it was clear that some of the geographically further isolates were more similar to each other rather than isolates from closer geographical regions.

To investigate drought stress effects on grain yield and its component in chickpea genotypes and its relation with physiological characteristics, nineteen chickpea genotypes were evaluated in field and pot experiments at the Faculty of Agriculture, University of Kurdistan, Sanandaj, Iran, in 2010-11 cropping season. Field experiment was carried out as split plot arrangement in randomized complete blocks design (RCBD) with three replications. Irrigation and rainfed conditions were assigned to main plots and chickpea genotypes were randomized in subplots. Grain yield, some yield components and drought resistance and susceptible indices were measured and recorded. Pot experiment was conducted as factorial arrangement using RCBD with two factors including irrigation treatments (irrigation at -3 (control) and -12 bar (stress) of soil water potential) and 19 chickpea genotypes as second factor with three replications. In this experiment, stomatal conductance at the vegetative stage and polyphenol oxidase activity at the vegetative and reproductive stages were studied. Chickpea genotypes divided in high and low yield potential and drought tolerant and susceptible genotypes based on seed yield under drought and control conditions. Results showed that drought stress reduced pod and grain per square meter by 43% and 44%, respectively and caused 43% reduction of grain yield, however, it did not have significant effect on 100 grain weight. In pot experiment, drought stress reduced stomatal conductance by 78% and drought tolerant genotypes had 33% higher stomatal conductance in comparison to drought susceptible genotypes. There were highly significant positive correlation (r = 0.52**) between polyphenol oxidase activity and stomatal conductance. It can be conclude that polyphenol oxidase activity and stomatal conductance would be useful physiological traits for improvement of drought tolerant genotypes in chickpea breeding programs.

Among proteins involved in gene silencing and creation of small RNA, argonaute proteins play an important role that has been shown to possess endonucleolytic activity. In the present study, we aim to use bioinformatics tools for analyzing putative argonaute genes in forest strawberry. A BLAST analysis was performed against the available strawberry genomic sequence draft to identify argonaute sequences using previously reported argonaute sequences of Arabidopsis. Sequence alignment, 3D structure, phylogenetic tree and conserved domains were generated for strawberry argonautes. Results showed that 13 putative argonaute genes are spread on different chromosomes of forest strawberry. There were three classes of argonaute proteins similar to that of Arabidopsis, and each varies in length from 845 to 1956 AA, with an average of 1076 AA and 120 kDa. Forest strawberry argonaute proteins have PAZ, PIWI and MID conserved domains. argonaute 6 contained a new domain GT1-Sucrose-synthase which has not been reported for plants and animals previously. Our results showed that forest strawberry have three classes of argonautes and a new argonaute that needs more studies.

In order to investigate the responses of Sardari wheat ecotypes to drought stress, 12 spikes of Sardari wheat were selected from different regions of Kurdistan province and planted as head to row. In the second year, a split- plot factorial field experiment was conducted. In this experiment water stress and irrigation were considered as the levels of main factor and 12 Sardari wheat ecotypes allocated to subplots. In the third year, the harvested wheat ecotypes in the second year were grown under control and drought conditions to evaluate grain yield and stress susceptibility index (SSI). Results showed that photosynthesis rate, rate of leaf water loss, protein and chlorophyll content, mesophyll conductance, photosynthetically water use efficiency and sub stomatal CO2 concentration were affected significantly by water stress in all wheat landraces. The results revealed that ecotypes with higher photosynthetic rate showed higher transpiration. These ecotypes exhibited better mesophyll conductance and lower SSI. Leaf protein patterns of ecotypes by electrophoresis were similar to that of soluble protein content under drought stress. The results showed that under drought stress, the new bands of protein were identified which correlated with pattern of photosynthesis of ecotypes in this condition.

Cyclotides-like proteins are a family of small disulfide-rich and plant-derived peptides made up of ~30 amino acids. They contain a unique structural motif that consists of a head-to-tail cyclic backbone and knotted arrangement of three disulfide bonds, together referred to as the cyclic cystine knot (CCK). The cyclotides are antimicrobial circular peptides that have a variety of biological activities such as anti-HIV, anti-tumor, cytotoxic, hemolytic activity and etc. In current study, we have cloned new members of the cyclotide-like gene family from Zea Mays leaf cDNA using 3'RACE-PCR and specefic primers based on sequences found in a continuous region of homology present in EST database. Clones were randomly picked and sequenced. Nine cyclotide-like genes were identified. Phylogenetic analysis divided them in two separate classes; first class with length 300-500nt showed similarity with classical cyclotide-like genes in databases. In contrast the other class was very different with length 90-500nt and showed variations in number of disulfide bands. The cloning of these genes in maize showed that they are very diverse and may have important role in plant response to different stresses.

In order to investigate of the effects of water stress on seed yield and gas exchange and to evaluate the relationship between source and sink in sunflower Azargol variety, a split-plot factorial field experiment was conducted in 2008. In this experiment, water stress (no irrigation from 30 days after sowing till ripening) and irrigation (irrigation at -3 bar soil water potential) were considered as main factors and leave removal at five levels (0% as control, 25%, 50%, 75%, and 100% defoliation) as sub factors in flowering period (R5) and end of seed setting (R7). For sink limitation, after pollination, one half of seeds in the head were removed. The results showed that defoliation and drought stress reduced grain yield through biomass, number of filled seed and harvest index reduction. Drought stress showed no significant effects on seed oil content, while decreased protein content. Results indicated that water stress decreased photosynthesis rate, stomatal conductance and increased water use efficiency, but defoliation increased photosynthesis rate. However it can be concluded that at water stress conditions, both source and sink restricted grain yield whereas at irrigation conditions, the sink limitation determined final grain yield. The reduction of seed oil and increasing source limitation under drought stress conditions showed that use of varieties with lower source limitation associated with increased seed oil content. Under irrigated conditions, varieties with high number of grains per head can eliminate sink limitation and lead to improved grain yield.

In order to study the effects of different levels of water stress on growth and some biochemical traits of three pear genotypes (Nazkeh, Lasoreah and Kevilah), a pot experiment was established. Three pear genotype seedlings from Kurdistan province were exposed to four water supply regimes. Control seedlings were kept well watered everyday. Water stress treatments were irrigated when soil water potential reached – 0.4, -0.8 and -1.2 MPa. Plant height, stomatal density, leaf chlorophyll content (chlorophyll a, b as well as total) water were assessed following the stress period, but proline and Total Soluble Sugar (TSS) were determined both during water stress and after the plants being rewatered. The results of statistical analysis revealed that stomatal density and leaf chlorophyll contents (a, b and total) were not affected by stress treatments. Water stress exerted significant effects on growth and biochemical characteristics. Plant height decreased during water stress. Plant growth stopped in -0.8 and -1.2 treatments in Nazkeh and Lasoreh but plant height increment was 9.4 % of control in Kevilah in -0.8 MPa treatment. Kevilah had a higher growth rate in -0.4 MPa treatment as compared with the other genotypes. Proline and total soluble sugars increased during water stress period but reache the control levels 2 days past rewatering in all water stress treatments. The free proline and TSS content of leaves in Kevilah genotype increased at a higher rate when in water stress conditions. The highest contents of proline and TSS were observed in Kevilah genotype in -1.2 MPa treatment. The results indicated that Kevileh genotype was more resistant to drought stress.