صدیقه فابریکی اورنگ

The aim of this study was to investigate the effect of salinity stress on changes in morpho-physiological traits of 109 accessions of Aegilops tauschii collected from different regions of the world, along with two check cultivars, Arg (salinity tolerant) and Darya (salinity sensitive). For this purpose, a factorial experiment with two levels of salinity (zero and 300 mM NaCl) as the first factor and 111 genotypes (109 accessions and two check cultivars) as the second factor was conducted using a randomized complete block design with three replications. After salt stress treatment, some morpho-physiological traits were measured and the results were analyzed. Analysis of variance results showed that the studied accessions differed significally from each other (p≤0.01) in terms of all traits (shoot and root sodium, shoot and root potassium, shoot potassium to sodium ratio, leaf chlorophyll content, shoot fresh and dry biomass, and root fresh and dry biomass), except for the root potassium to sodium ratio. The effect of salinity stress on the aforementioned traits was also significant (p ≤ 0.01) except for root potassium. In total, some accessions collected from Iran, Georgia, Kosovo, Azerbaijan, and Turkmenistan showed acceptable performance in terms of the studied traits under salt stress conditions, when compared to the resistant check cultivar Arg. In addition, the dendrogram obtained from the cluster analysis and the survey of subgroup averages showed that the accessions collected from Iran, Georgia, Kosovo, Azerbaijan, and Turkmenistan had greater resistance and tolerance to salinity stress.

Japanese barberry (Berberis thunbergii) is a perennial and woody shrub that grows in different regions of Iran and the world. The Berberidaceae family has a high potential in food and pharmaceutical industries due to its antioxidant, antimicrobial and anticancer activities. Elicitors are biotic or abiotic stimulators that can induce responses in plants. Abiotic elicitors are used to increase secondary metabolites in plants. Nanoparticles have minimum dimensions between one and 100 nanometers, which have specific physical and chemical properties. Like many nanoparticles, zinc oxide and cerium dioxide nanoparticles are toxic to living organisms. The mechanism of toxicity of nanoparticles is generally contributed to the induction of oxidative stress which lead to the formation of free radicals. Nanoparticles of zinc oxide and cerium dioxide can increase the antioxidant capacity of plants by stimulating oxidative stress. 

In order to study the changes in the physiological and biochemical characteristics of Japanese barberry under the influence of abiotic elicitors, an experiment was conducted in a completely randomized design with three replications in the greenhouse and laboratory of the Faculty of Agriculture and Natural Resources of Imam Khomeini International University in 2019. Treatments were included zinc oxide elicitors (0.1 g/L) and cerium dioxide nanoparticles (0.0002 g/L) and control (no elicitor). In this research, the amounts of protein, antioxidant enzymes and photosynthetic pigments in tissues of Japanese barberry were measured. Data analysis was done with SAS 9.1.3 statistical software. Comparison of means was done with Duncan's multiple range test and graphs were drawn using Excel software. 

The results of the data analysis of variance showed that the effect of treatment with cerium dioxide and zinc oxide nanoparticle elicitors on the amount of leaf protein, antioxidant activity and also the amount of photosynthetic pigments was significant. Comparison of the means showed that these two elicitors increased the leaf protein content by 26.2% and 12.4%, respectively, compared to the control, but they did not have a significant effect on the root protein content. Cerium dioxide decreased the activity of guaiacol peroxidase enzyme by 77.1% in the root; But in leaves, the activity of this enzyme increased by 134.8% due to treatment with zinc oxide. The highest activity of leaf superoxide dismutase enzyme (25.2% increase compared to control) was related to zinc oxide treatment. The lowest content of catalase enzyme in leaves and roots was obtained with cerium dioxide and zinc oxide. Zinc oxide increased the activity of leaf ascorbate peroxidase enzyme by 111.8%; In the root, the highest and lowest activity levels of this enzyme were obtained in the control and cerium dioxide treatments, respectively. Both elicitors significantly increased the leaf peroxidase enzyme activity compared to the control (128.6% and 157.1%, respectively); However, only the treatment with zinc oxide caused a significant increase of 25.5% in the activity of this enzyme in the root. The highest activity of glutathione reductase enzyme in leaf and root was observed in zinc oxide treatment (23.3% and 12.5% increase compared to control). Nano zinc oxide increased the amount of total chlorophyll and carotenoid in the leaves compared to the control by 93.4 and 67.5%, respectively. 

It was concluded that zinc oxide nanoparticles elicitor had the greatest effect in increasing the amount of photosynthetic pigments and enzyme activity. By using this elicitor in Japanese barberry culture, the amount of antioxidant enzymes can be increased and the extract can be used as a strong antioxidant in pharmaceutical products.
.

Japanese barberry or ornamental barberry (Berberis thunbergii) belongs to the Berberidaceae family. This family have several properties, including antioxidant, antimicrobial and anticancer activities. The healing properties of barberry have been known and studied for thousands of years. In order to study the changes in the physiological-biochemical characteristics of Japanese barberry under the influence of abiotic elicitors, an experiment was conducted in the form of a completely randomized design in the greenhouse and laboratory of the Faculty of Agriculture and Natural Resources of Imam Khomeini International University in 2018-2019. The treatments included methyl jasmonate elicitors, salicylic acid, titanium dioxide and no elicitor (control) in four replications. In this research, the amount of leaf photosynthetic pigments and antioxidant enzymes of leaf and root tissues of Japanese barberry plant were extracted and measured. The results showed that the effect of the experimental treatments on the amount of photosynthetic leaf pigments and antioxidant enzymes in the two studied tissues was significant at the 1% probability level. Comparison of the means showed that salicylic acid treatment increased the amount of chlorophyll a and total leaf chlorophyll compared to the control by 20.9% and 21.3%, respectively. Salicylic acid and methyl jasmonate increased 29.9 and 19.1 percent of carotenoid content in leaves, respectively, compared to the control. The highest protein content of the root was produced with the use of salicylic acid, which increased by 38.6% compared to the control. The activity of catalase enzyme in the root increased by salicylic acid and titanium dioxide elicitors by 76.8 and 52.2%, respectively, compared to the control. In leaves, salicylic acid and methyl jasmonate elicitors increased catalase enzyme activity by 104.1% and 57.1%, respectively. From this research, it was concluded that salicylic acid had the greatest effect in increasing the amount of pigments and the activity of antioxidant enzymes of the Japanese barberry plant, which can be used for medicinal products.

Agilops is one of the most important genetic ancestors of wheat, which is a valuable genetic resource for wheat breeders due to its tolerance to various biotic and abiotic stresses. The main goal of this research was to evaluate the genetic diversity in 89 Aegilops tauschii accessions collected from different regions of Iran and the countries of Turkey, Afghanistan, Armenia, Sweden and Azerbaijan using 32 microsatellite primer pairs. The averages of polymorphic information content (PIC), Marker index (MI), Resolution (RP) indices were equal to 0.81, 1.82 and 1.88, respectively, which indicated that the primers used are suitable with high efficiency for investigating relationships among the accessions of this species. The results of principal coordinate analysis (PCoA) showed that the first three components explained 41.88% of the total genetic variance changes. Cluster analysis classified the evaluated accessions into four main groups, which was confirmed by the results of principal coordinate analysis (PCoA). According to the cluster grouping pattern, in some cases the accessions with the same geographical origin were placed in separate groups and also the presence of non-Iranian accessions in the group of Iranian accessions was observed. The results of population structure analysis confirmed the populations genetic exchanges and showed that the classification of the studied accessions is independent of their geographical origin. In general, the results showed high genetic diversity among the studied accessions of Ae. tauschii, therefore, the high genetic diversity leads to finding new allelic sources and introducing desirable alleles, which will be of great value to wheat breeders for use in breeding programs.

Sanguinarin is an important secondary metabolite of the benzyl isoquinoline alkaloids, which is widely found in Papaveraceae family plants such as Chelidonium majus. Sanguinarin has many properties such as antimicrobial, antifungal, and anti-inflammatory. The use of abiotic elicitors is one of the effective ways to induce and increase of secondary metabolites. The elicitors such as methyl-jasmonate induce the biosynthesis and accumulation of secondary metabolites by stimulating the plant's defense system. Since there is no report on the isolation of cDNA encoding the stylopine synthase in sanguinarine biosynthesis pathway in Chelidonium majus, the full length of this gene was isolated to provide an opportunity to identify and investigate the pathways of benzylisoquinoline that have not been known so far. Also, the expression pattern of this gene was surveyed under the treatment of methyl-jasmonate to improve its production.

 In this research at the first stage, the cDNA encoding the stylopine synthase gene (STS) was identified and isolated from the root of Chelidonium majus. STS is one of the genes in sanguinarin biosynthesis pathway that converts (S)-Cheilanthifoline to (S)-Stylopine in the methyloxy bridge, which belongs to CYP719 subfamily. In the next stage, the expression pattern of STS gene was examined in a completely randomized design with four replicates by applying methyl-jasmonate (concentration of 100 μM) at four treatment levels (foliar, irrigation, foliar + irrigation and without methyl-jasmonate as control) in different organs (leaf, root and stem) and different sampling times (6, 24 and 48 hours after methyl-jasmonate treatment) In a pot experiment under greenhouse condition.

 Isolation of STS gene cDNA was done in two steps. At first, the incomplete sequence (1000 bp) was isolated, and then, by designing new primers based on the information from the primary sequence, the full length (1500 bp) of the coding region of gene was isolated and successfully cloned into pTG19-T plasmid. After sequencing and necessary modifications, the obtained gene sequence was registered in the NCBI database with the accession number KY550671.2. By evaluating the sequence characteristics and studying the phylogenetic relationships, it was determined that CYP719A3, the product of the STS gene, belongs to the P450 protein family. The results of variance analysis showed the methyl-jasmonate treatment affect significantly (P≤0.01) STS gene expression at different times in different organs of Chelidonium majus. The highest level of gene expression was observed in methyl-jasmonate foliar spraying + irrigation 48 hours after treatment in the root. The mean comparison showed that the expression level of STS gene was different in the root, leaf and stem of Chelidonium majus, so it was 1.5 and 2.5 times higher in the root than leaf and stem, respectively. The high expression of the STS gene in the root of Chelidonium majus indicates the effectiveness of the plant organ in the level of gene activity, on the other hand, the use of methyl-jasmonate could play an effective role in increasing gene expression.

 Methyl-jasmonate increased the expression of STS gene in the sanguinarine biosynthesis pathway so that the highest expression was observed in the root. Therefore, the sequence of the gene encoding stylopine synthase in the sanguinarine biosynthesis pathway and its expression pattern in Chelidonium majus can be used in the pathway engineering of this valuable metabolite.

The enzyme Cheilanthifoline synthase (CFS), a member of the CYP719 protein family, is involved in the enzymatic conversion of reticulin to sanguinarine in plants. Sanguinarine is a Benzophenanthridine alkaloid that is widely found in plants of the family Papaveraceae, Famariaceae and Rotaceae. In the present study, the cDNA coding CFS was isolated from the roots of chelidonium majus. In the sequence analysis, the identification of the aromatic region, the binding region to hem group, and the EXXR conserved motif in the K helix confirmed the CFS enzyme belonging to the cytochrome P450 protein family. The expression pattern of CFS gene was evaluated in roots, stems and leaves of chelidonium majus treated with cerium dioxide and titanium dioxide nanoparticles at different times and concentrations. In plants treated with cerium dioxide, the highest expression of CFS gene was observed 72 hours after foliar spray of 100 mg/l in root and the lowest expression was observed in stem. In foliar spray of plants with titanium dioxide nanoparticles, 48 ​​hours after spray of 1000 mg/l, the highest expression increase of CFS gene showed in the root and the lowest expression was observed in the stem. The results of this study indicate an increase in transcripts of CFS gene in response to titanium dioxide and sodium dioxide nanoparticles in chelidonium majus; therefore, the obtained information from gene structure and the effectiveness of used elicitors can be useful in increasing the amount of sanguinarine alkaloid through metabolic engineering.

The aim of this study was to determine the relationships and genetic diversity between and within wild species of Aegilops using EST-derived TRAP molecular markers.

In this study, genetic diversity and relationships among eight wild species of Aegilops collected from 14 provinces of Iran were evaluated using 24 TRAP primer combinations.

The mean of PIC for studied primers was 0.95 and the lowest and highest values ​​of this index were related to TRAP20 (0.92) and TRAP10 (0.97), respectively. Also, the mean of MI was 10.77 and the lowest and highest values ​​were related to TRAP3 (6.95) and TRAP17 (17.34), respectively. The molecular analysis of variance showed 60% variance between species and 40% variance within species. Estimated genetic parameters showed that the highest levels of Na (1.07), Ne (1.26), I (0.23) and uHe (0.16) belonged to Ae. truncialis and the maximum Nei gene diversity (0.14) was related to Ae truncialis, Ae. umbelulata and Ae. neglecta. Minimum values ​​of Nei gene diversity (0.09), Shannon index (0.14), No. of effective alleles (1.16) and No. of observed alleles (0.79) was observed for Ae. crassa. The least genetic similarity was observed between Ae. crassa with Ae. cylandrica and Ae. umbelulata and the most similarity between Ae. triuncialis and Ae. umbelulata. Being in the same group of Ae. umbelulata and Ae. truncialis with cluster analysis and PCOA confirmed the high genetic similarity of these species.

The results showed high genetic diversity among and within species. Molecular analysis of variance revealed higher genetic diversity between species, indicating low gene flow and Fst, as well as a high genetic differentiation between species.

Aegilops range grasses (Poaceae family) as one of the most important ancestors of wheat is a rich source of stresses tolerance genes. For this, the fingerprinting of genetic relationships in accessions belonging to eight species of Aegilops was studied using targeted genes-related CoRAP molecular markers. In designing the fixed primers, six genes CAT, MnSoD, SoS1, miR398, miR160b and miR169gR responsible for abiotic stresses tolerance were used. The lowest and highest polymorphic information contents were related to CoRAP2 and CoRAP10 primers with 0.92 and 0.96, respectively. The marker index varied from 6.89 in CoRAP7 primer to 13.49 in CoRAP9 primer. Low gene flow (Nm) and high differentiation (Gst) was observed between species. Also, the Fst index equal to 0.45 showed that the studied populations were completely separated. Due to the high values of the effective allele number and Nei genetic diversity, two species Ae. cylandrica and Ae. caudata showed high intra-specific diversity. The greatest similarity was observed between two species Ae. truncialis and Ae. umbelulata as well as Ae. neglecta and Ae. cylanrica. Cluster analysis appropriately divided species into distinct groups, and principal coordinate analysis confirmed the results of cluster analysis. Using structure analysis of the population, the mode of gene flow and genetic intermixture among species were in accordance with the grouping results of cluster analysis and PCoA biplot.

To study the genetic and allelic diversity in the genes encoding glutenin and gliadin proteins in wheat landraces, 98 Triticum accessions belonging to four species: T. aestivum, T. boeoticum, T. durum and T. uartu collected from different provinces of Iran were used. 13 pairs of low molecular weight glutenin primers (LMW-GS), four pairs of high molecular weight glutenin primers (HMW-GS) and four pairs of gliadin primers were amplified 1327, 548 and 152 segments, respectively. The highest genetic diversity was observed in the Glu-3A gene block, which belongs to the low molecular weight glutenin, and 13 primer pairs amplified a total of 34 alleles from 180 to 1500 bp in length, of which 11 alleles were previously reported and 23 alleles were identified as possibly new alleles. In the Glu-A1 gene block, which belongs to the high molecular weight glutenin, most allelic polymorphisms were amplified by the primers Glu-A1 and Xrj5. In the Gli-A2 gene block, only one allele was amplified by two specific primers: Gli-AS6 and Gli-AS5. The highest and lowest number of effective alleles belonged to T. aestivum and T. uartu. The average of Nei diversity index was 0.19 and its highest and lowest values ​​were related to T. aestivum and T. uartu. Based on the clustering method, 98 accessions were divided into two main groups. In the first main group, the accessions related to two species T. aestivum and T. durum were placed, which shows the greatest genetic similarity between the two species. In the second main group were placed two wild species T. boeticum and T. uartu. In overall, a large allelic diversity was observed for low molecular weight glutenin subunits in the studied Triticum species, which highlights the potential of these species as a valuable germplasm for breeding programs aimed to the bakery quality improving in wheat.

This study aimed to investigate the genetic diversity of 140 barley advanced lines, using Selection Index of Ideal Genotype (SIIG) and Adjusted Selection Index of Ideal Genotype (ASIIG) indicators for introduction and cultivation of suitable lines in rainfed conditions. The highest coefficient of variation was related to the type of spike, flag leaf area, seeds per spike, peduncle length, flag leaf width, and early growth vigor. The stepwise regression showed that the early growth vigor, 1000-seeds weight, spike type, days to heading, plant height, number of seeds per spike, and flag leaf length were the most critical components, which affect the grain yield under the rainfed conditions. The results of SIIG showed that lines 4, 43, 47, 40, 70, 96, 137, 3, 57, and 45 were the best lines with the highest SIIG values. The SIIG index divided all lines into seven groups; the average grain yield of lines in groups I and II were higher than the average grain yield of control cultivars and the other groups. According to ASIIG plots, lines 47, 3, 43, 45, 137, 96, and 40 were placed in the ideal quarter. Finally, the SIIG and ASIIG were compatible in selecting the best lines. ASIIG was more suitable for separating the ideal lines, because it participated all traits commensurate with their importance in the end product quantity and quality.

This research aimed to estimate the nucleotide polymorphism and point mutation/substitutions for ITS2 region in 12 different Triticum and Aegilops species, and to demonstrate their phylogenetic relationships. The aligned ITS2 dataset showed a total of 67 SNPs and transitions (75%) predominant over transversions (25%). The C/T transitions were observed at highest level in the studied species. The highest nucleotide substitutions occurred in ITS2 sequences of T. aestivum, T. turgidum, and A. speltoides that showed similar substitutions on the base sites 76, 193, 215, 230 and 241 (TCCAG changed to ATTTA). T. boeticum and T. urartu showed similar substitutions on the base sites 85, 201, 231, 234 and 252 (GTGCC changed to AGATA). Remarkably, the nucleotide of base site 146 was C in all Triticum species, but T in all Aegilops species. The substitution rate in A. neglecta, and A. umbellulata was zero, it was concluded that their ITS2 is conserved. The secondary structures of ITS2 in the studied species was similar to that of the other flowering plants with four helices and the variation in helices length. The helix III had the longest length compared to the other helices. The length ranges of helices I, II, IV varied from 9 (T. turgidum, and T. urartu) to 10, 11 (A. cylindrica) 13 and 6 (A. speltoides) and 10 (T. boeticum) paired bases, respectively. Using pairwise ITS2 nucleotide comparisons, the closeness of A. caudata with A. cylindrica, T. boeticum with T. urartu, A. neglecta with A. umbellulata was observed. Phylogeny trees classified 12 species into four main clades. The highest point mutations occurred in ITS2 sequences related to the species grouped in the first clade.

Dracochephalum kotschyi is an endemic medicinal plant in iran belonging to Lamiaceae family known as Zarin-Giah. Under osmotic stresses, proline accumulation is an important response of plants to these conditions. Proline is a compatible osmolyte which affects many cellular and molecular aspects of a plant in both normal and stressful situations. Proline is shown to be involved in plant development in normal conditions and in conferring resistance to a plant under biotic and abiotic stresses. Δ1-pyrroline-5-carboxylate synthetase )P5CS), one of the two main enzymes in the proline biosynthesis pathway of the glutamate precursor, has been demonstrated to play a significant role in proline accumulation in plants under drought stresses. In this research, the effect of different levels of drought stress was investigated on the relative water content, chlorophyll content, proline accumulation and P5CS gene expression in Dracocephalum kotschyi at two growth stages. This experiment was conducted as a factorial experiment with two factors including drought stress and growth stages in a completely randomized design with three replications. Drought stress treatments includes severe (25% FC), moderate (50% FC), low (75%FC) drought stress and control (100%FC). Growth stage includes vegetative and flowering stages. Analysis of variance showed that drought stress significantly increased P5CS gene expression and proline content while decreased chlorophyll a, b, total and relative water content. The highest expression level of P5CS gene and maximum amount of proline were observed at flowering stage under moderate drought stress (50% FC) conditions.

The main objectives of this study were to evaluate the genetic diversity in 53 Aegilops accessions belonging to Aegilops tauschii, Ae. cylindrica and Ae. crassa as well as comparison of the efficiency of start codon targeted (SCoT) and targeted region amplification polymorphism (TRAP) markers. A total of 15 SCoT and six TRAP primer pairs amplified 165 and 201 polymorphic bands, respectively. TRAP primers indicated higher values for the number of polymorphic bands, marker index and resolving power compared to SCoT primers. However, SCoT primers showed the higher PIC value than TRAPs. The results of molecular variance analysis (AMOVA) were different for each of the marker systems, so that SCoT markers showed the highest portion of genetic variance referred to intra-species, while based on TRAP markers the highest variance was observed inter-species. SCoT markers indicated the highest values for all genetic parameters compared to TRAPs and Ae. cylindrica and Ae. tauschii exhibited the highest genetic variation based on SCoTs and TRAPs, respectively. Cluster analysis based on each marker systems as well as the pooled data classified all accessions into three main groups. The clustering pattern based on SCoT primers was clearer than TRAPs. Hence, the use of SCoT markers is recommended for population genetic structure analysis and grouping of them, while the use of TRAPs can be recommended for fine mapping studies.

 Dracocephalum kotschyi is an endemic medicinal plant in Iran belonging to the Lamiaceae family. One of the most important monoterpenes identified in this plant is limonene. Drought stress is one of the factors affecting the content of monoterpene in medicinal plants.  In this study, the effect of different levels of drought stress was investigated on the expression of some genes involved in limonene biosynthetic pathway in D. kotschyi at two growth stages.
 

 This experiment was conducted as a factorial experiment with two factor included drought stress and growth stages in a completely randomized design with three replications. Drought stress consisted of severe (25% FC), moderate (50% FC), low (75% FC) stress and non-stress or control (100% FC) and growth stages consisted of vegetative and flowering. Secondary metabolites were analyzed and the amount of limonene was measured by GC / MS using Head-space method and the relative expression of key genes involved in limonene biosynthesis was measured using Real Time PCR.
 

 Analysis of secondary metabolites showed, one of the main monoterpenes was limonene whose amounts was 20.57 % and 13.41% during vegetative and flowering stages, respectively. Drought stress significantly increased HMGR and DXS gene expression during flowering stage, while during vegetative stage no significant change was observed on expression of both genes. In both growth stages, low drought stress increased HDR gene expression, although at flowering stage this increase was not significant but severe drought decreased HDR gene expression, although this decrease was not significant at vegetative stage. Drought stress decreased LS gene expression during flowering stage and increased it during vegetative stage.
 

 The results indicated that limonene content and expression of limonene biosynthesis key genes in D. kotschyi under drought stress are growth stage-specific.

MicroRNAs are small non-coding molecules that regulate the expression of genes through mRNA digestion and/or translation inhibition. By accurately regulation the expression of the genes, these molecules allow the plant to respond appropriately to the changes in growth stages or environmental conditions. In this study, the miRNA159, miRNA160, miRNA398, and the miRNA398 targeted gene (CSD) were investigated in six species belonging to the Triticum and Aegilops species, along with the susceptible (DARYA) and tolerant (SIRVAN) check cultivars under drought (FC=25%) and non-stress (FC = 100%) conditions using real-time PCR. The qRT-PCR analysis showed that all three miRNAs had a significant (P≤0.01) increase in expression under drought stress. The highest expression of miRNA159, miRNA160, miRNA398, and CSD gene was observed in the species Ae.tauschii, T.urartu, T.durum, and T.durum, respectively. By the mean comparing of species × stress interaction, the highest expressions were observed in the three species T.durum, T.urartu and Ae.tauschii for all three miRNAs under drought stress. Also, the highest increase in CSD expression under drought stress was observed in T.urartu, T.durum and Ae.tauschii equal to 6.62, 6. 21 and 0.63 fold in compared to the non-stress condition, respectively. It was concluded that the species T.urartu, T.durum and Ae.tauschii due to their superiority in terms of containing miRNAs causer tolerance, could be suitable candidates for bread wheat germplasm enrichment and breeding to drought stress tolerance.

This experiment was carried out to investigate the effect of methyl jasmonate and salicylic acid on the antioxidant production and physiological-biochemical properties of Chelidonium majus under drought stress condition. For this, an experiment was conducted as factorial based on completely randomized design with four replications in Imam Khomeini International University during 2016-2017. Three factors included drought stress in two levels (non-stress and drought stress with 100% and 50% field capacity irrigation, respectively), elicitor in three levels (control, 100 μm methyl-jasmonate and 2 mM salicylic acid) and plant organs (root, stem and leaf). Analysis of variance and mean comparisons showed that methyl-jasmonate and salicylic acid reduced the activity of CAT enzyme. Both salicylic acid and methyl-jasmonate did not affect the activity of SOD, but methyl-jasmonate increased the activity of GPX and salicylic acid reduced APX activity under drought stress. Also, methyl-jasmonate and salicylic acid reduced APX activity compared to control treatment. The highest amount of protein content was observed in root and then in leaf. Salicylic acid increased total protein content up to 34% compared to control treatment. Both Methyl-jasmonate and salicylic acid increased chlorophyll chlorophyll, carotenoids, flavonoids and anthocyanin compared to control treatment, but salicylic acid was superior in increasing of chlorophyll and carotenoids whereas methyl-jasmonate was superior in increasing of flavonoids and anthocyanin. In conclusion, methyl-jasmonate and salicylic acid have increased the ability of celandine in response to drought stress, and also, in addition to the protective effect against drought-induced oxidation, they increased photosynthetic pigments and some of metabolites.

The WRKY gene family encodes a large group of transcription factors that regulate genes involved in plant response to biotic and abiotic stresses. Sorghum is a notable grain and forage crop in semi-arid regions because of its unusual tolerance against hot and dry environments. In this study, we performed a genome-wide analysis of WRKYs using a genome assembly of sorghum bicolor. First, all possible WRKY gene sequences as well as all possible WRKY protein sequences in the Sorghum bicolor genome database were identified using the NCBI website. A set of 85 WRKY genes was identified in the S. bicolor genome and classified into three groups (I–III). Among the members of the group I, the SbWRKY13 had a different and novel zinc finger motif as compared to other members of this group. It was also found that mutations occurred at R, K, Y and Q in the conserved WRKYGQK sequence. No complete gene duplication was found in gene copy number investigation, suggesting that the expansion of SbWRKY genes was not necessarily based on the gene duplication events or duplication of SbWRKY genes had probably happened in the past times. Gene cluster analysis showed that the number of genes on chromosomes were positively correlated with the number of clusters. The study of amino acid composition revealed that totally in all groups, Alanine and Proline were the most frequent residues while Cysteine had the lowest frequency. The study of introns in the SbWRKY domain showed that the majority of SbWRKY genes had two types of introns in their WRKY domains (phase 0 and phase 2). Also, investigation of conserved known motifs revealed that there were six, two and one known motifs outside of the region of the SbWRKY domain for groups IIa, IIb and IIc, respectively. The results describe evolution and functional differentiations of WRKY transcription factors in Sorghum bicolor.

Plant organs and latex of greater celandine are rich in isoquinolin alkaloids that sanguinarine, a member of benzofenanthridine, is one of the most important metabolites, which its accumulation mainly reported in the roots. Sanguinarine biosynthesis requires seven enzymatic steps from the pre-substance s-reticulin using berberine bridge enzyme (BBE), cheilanthifoline synthase (CFS), stylopine synthase (STS), tetrahydroprotoberberine cis- N-methyltransferase (TNMT), protopine 6- hydroxylase (P6H), S-cis-N-methylstylopine 14-hydroxylase (MSH) and dihydro benzophenanthridine oxidase (DBOX). The study of biosynthetic pathways and the identification of factors affecting the production of secondary metabolites are important steps in metabolite engineering. Nanoparticles are a new category of elicitors that act as inducers for the production of secondary metabolites. For this purpose, the effect of titanium dioxide nanoparticles was investigated on the expression changes of the celandine sanguinarine biosynthetic pathway genes in a factorial experiment with three factors included titanium dioxide concentrations, spraying period and post-induction time. Analysis of variance for the expression of STS, P6H, BBE and MSH genes revealed significant differences in titanium dioxide concentrations, post-induction times, spraying period, different organs and their interactions at P≤0.05, 0.01 levels of probability. The accumulation of STS and P6H transcripts in the roots was 2-3 times higher than the leaves and stems, and also the transcripts of the BBE and MSH genes was four times higher in the root than the stem. Totally, the concentration of 500 mg/L titanium dioxide increased the expression of STS, P6H, BBE and MSH genes by 63, 66, 500 and 91%, respectively, relative to control treatment. The 24 and 48 hours post-induction time resulted in maximum expression in all four studied genes in comparison to 72 hours post-induction time, and 500 mg/L titanium dioxide showed the highest expression for STS, P6H and BBE genes, and 1000 mg/L for MSH gene. From the results it can be concluded that titanium dioxide nanoparticles are effective in increasing the production of celandine sanguinarine through the regulation of its biosynthetic pathway genes.

The olive is considered as one of the most important horticultural crops, with more abundant number of cultivars. Thus, the aim of this research was to study the genetic diversity in a set of olive cultivars in Taroum station of Tran. In this research, 25 CBDP markers were used to study the genetic diversity and relationships of 20 promising and commercial Iranian and foreign varieties of olive. The 25 used primers amplified a total of 755 polymorph alleles, and the mean of PIC and MI indices were 0.941 and 5.64, respectively, indicating the high efficiency and differentiation power of CBDP markers. The genetic parameters of Nei and Shannon indices were higher in foreign cultivars than those of Iranian cultivars. The high gene flow (4.46) was observed among olive cultivars, which confirmed the low values of the inter-population differentiation (Gst) and FsT indices. Analysis of molecular variance (AMOVA) differentiated the total genetic variation into inter-group (Iranian and foreign cultivars) (17%) and within-group (83%) diversity. The cluster analysis using Dice coefficient and UPGMA method divided the Iranian and foreign cultivars into five groups, which the promising cultivar T24 and Koroneiki with the maximum difference with others were placed in separate groups. The Principal Coordinate Analysis (PCoA) was able to distinguish promising Iranian cultivars from the other commercial cultivars. The population structure analysis confirmed the results of the cluster and PCoA analysis and clearly separated the two groups of Iranian and foreign cultivars. In conclusion, it was observed that the promising cultivar T24 with having a genetic similarity to both groups of Iranian and foreign cultivars can be a suitable candidate for commercialization with complementary characteristics of both groups. Also, it is recommended to use this marker to study the genetic diversity of other olive varieties.

Engineered plants as bioreactors for production of heterologous proteins are appropriate replacement for conventional expression systems such as mammalian cell culture and transgenic animals. For efficient production of recombinant proteins, the type of host species is very important and its choice depends on the type of recombinant proteins, plant life cycle and its performance, and maintenance or production costs. The most important benefits of plant-based expression systems could be included: cost effective of the plant systems, no contamination of recombinant proteins derived from plants with human pathogenic microorganisms due to host-less of plants to human pathogens, easy and low cost of extraction and purification of proteins, more post-translational changes required for the stability and activity of protein by plants. For development of plant-based production of recombinant proteins, optimization of protein expression level is essential. In this regard; various factors controlling the expression of target gene at levels of transcription, translation, post-translational modifications and recombinant protein accumulation are involved. On the other hand, lower amounts of expressed recombinant proteins are serious problem limiting commercial exploitation of plants bioreactors. Thus improving the accumulation of recombinant proteins in transgenic plants is an important factor in the desirability of an expression system. In this regard, the accession of signals in C-terminal of target proteins leads to increased retention and accumulation of proteins in the endoplasmic reticulum. After production of recombinant proteins, the use of affinity tag for purification of proteins are very effective and useful, so protein purification will possible without prior notification of their biochemical properties.

This experiment was conducted to investigate the effect of drought stress on photosynthetic pigments contents, biomass of roots and shoots, total protein content and antioxidant enzymes activity and non-enzymatic antioxidant of six species of Aegilops genus along with two drought resistant and susceptible wheat cultivars as checks in a factorial based on randomized complete block design with three replications. Drought stress treatments were applied at three levels of non-stress (FC=100%), moderate stress (FC=50%) and severe stress (FC=25%) conditions. Ae. caudata, Ae. triuncialis and Ae. cylindrica were ranked higher than resistance cultivar, Sirvan, as check in term of root fresh and dry weights under stresses. The highest total chlorophyll content was related to Ae. cylindrica, Ae. umbellulata and Ae. caudata in stresses. Carotenoid showed decreasing trend in all studied species except for Ae. umbellulata, and Ae cylindrica had the highest amount at both stress and non- stress conditions. The highest SOD under severe stress was observed in Ae. crassa and Sirvan as resistant cultivar. Maximum APX under severe stress was observed in Ae. triuncialis and Ae. caudata and the lowest activity was in Ae. neglecta. The highest level of GPX was observed in Ae. triuncialis, Ae. caudata and Sirvan cultivar under drought stress. Also the maximum CAT enzyme under severe stress was observed in Ae. triuncialis, Ae. caudata and Ae. crassa, respectively, and Ae. umbellulata and Darya cultivar had the lowest amount of CAT enzyme. In conclusion, because the three species Ae. caudata, Ae. triuncialis and Ae. cylindrica had the highest amounts for the traits associated with increased drought tolerance, including root volume, carotenoid, CAT, GPX and APX enzymes, and also these species were same in having CC genome. Therefore, it seems that the CC genome could have a high potential for future wheat breeding programs in increasing of drought tolerance.

Perception of the genetic diversity in domesticated and wild relatives of a plant species is important for their use in breeding programs. Considering that a large part of wheat cropping fields in Iran is located in arid and semi-arid areas, and that plants adapt to environmental stresses with different strategies, such as changes in antioxidant enzymes. Therefore, it is necessary to study more about the resistant species and the recognition of their mechanism of tolerance. In order to investigate the effect of drought stress on the content of photosynthetic pigments, fresh and dry biomass of roots and shoots and antioxidant enzymes activity in five ancestral and cultivated species of Aegilops and Triticum (Aegiliops tauschii ،Aegiliops speltoides ،Triticum aestivum ، Triticum urartu و Triticum durum), a factorial experiment was conducted based on randomized complete block design with three replications at three levels of normal irrigation (100% FC), medium stress (FC 50%) and severe stress (FC% 25). By mean comparing, the highest activity of CAT, SOD, APX and GPX was observed in severe stress compared to non-stress condition, with an increase of 80, 34, 84 and 29%, respectively. The results showed that the species T. aestivum and T. urartu in terms of SOD, Ae. tauschii for CAT, T. durum for APX and Ae. Speltoides in terms of GPX are considered as candidate species for drought tolerance breeding due to the highest activity of mentioned enzymes in both stresses. Total chlorophyll decreased in moderate and severe stresses by 11. 5 and 66%, respectively, however, the two species Ae. speltoides and T. durum showed the lowest reduction compared to normal condition. The carotenoid, which is a non-enzymatic antioxidant, decreased with increasing of drought severity. Not only this reduction was not occurred in T. durum, but also it increased with drought severity. Positive correlation (0. 60) was observed between the amount of carotenoid and GPX activity under stress conditions in contrast to non-stress condition. Root dry weight showed a positive correlation (0. 81) with APX enzyme under stress conditions. However, in non-stress condition, this correlation was not significant. The highest activity of studied enzymes was observed in severe stress compared to non-stress condition. T. aestivum and T. urartu in terms of SOD, Ae. tauschii for CAT, T. durum for APX and Ae. Speltoides in terms of GPX are considered as candidate species for drought tolerance breeding due to the highest activity of mentioned enzymes in both stresses. Total chlorophyll decreased in moderate and severe stresses by 11. 5 and 66%, respectively, however, the two species Ae. speltoides and T. durum showed the lowest reduction compared to normal condition. The carotenoid, which is a non-enzymatic antioxidant, decreased with increasing of drought severity. Not only this reduction was not occurred in T. durum, but also it increased with drought severity.

Different parts of the greater celandine herb contain important alkaloids such as chelodonin, chloritin, sanguinarine, and berberine. Sanguinarine is a four-member benzofenanthridine that is widely found in Papaveraceae, Fumariaceae and Rutaceae. This biologically active alkaloid, singuainarin, has a wide range of potentially useful pharmacological properties, such as anti-microbial, anti-inflammatory, and anti-tumor. The first step in biosynthesis pathway of sanguinarine alkaloid requires the transformation of S-reticulin to S-skolorine, which catalyzes by berberine beridge enzyme encoded by the BBE gene. In order to prepare plant materials for gene isolation, and the expression of BBE gene in four drought levels including severe, moderate, weak stress and non stress (40, 55, 70 and 100% field capacity, respectively) in three organs, root, stem and leaf, an factorial experiment was carried based on completely randomized design with three replications. RNA was extracted from root, stem and leaf to isolate the cDNA encoding BBE enzyme and study the gene expression. The cDNA encoding BBE enzyme with approximate length of 1500 bp was isolated from root and successfully integrated into PTG19-T plasmid and cloned at E. coli. The sequencing results showed that the isolated gene with a length of 1441 bp could translate to a peptide chain with 483 amino acids. The predicted BBE peptide chain had NX(S/T) motif as glycosylation sites, the flavin binding domain for the binding of flavinated histidine, SGGH motif, and the Glu sequence which plays an important catalytic role in the formation of berberine bridge in flavoprotein enzymes. Based on the mean comparison results, the highest and lowest expressions of this gene were obtained on root under weak drought stress (FC 70%) and on leaf and shoot under non-stress condition (100% FC), respectively. In general, it can be concluded that the weak drought stress can increase the BBE gene expression, and possibly this could result in higher production of sanguinarine in celandine.

In order to evaluation of phytochemical and growth changes from Thymus vulgaris L. under moderate abiotic stresses, an experiment was carried out at Imam Khomeini International University in a completely randomized design with three replications during 2016. The treatments consisted of drought (50% FC), salinity (100 mM NaCl) and non-stress control (100% FC). Root fresh weight, root dry weight, root length, main branch diameter, sub-branch diameter, sub-branch number, shoot dry weight, shoot fresh weight, plant height, relative water content and chlorophyll content were measured. Also, total phenols, flavonoids and anthocyanins were assayed in leaves, roots and stems by using spectrophotometer. Statistical analysis showed that drought and salinity had significant effects on morphological traits, except for shoot dry weight and number of lateral branches. Generally, morphological traits affected by drought stress in compared with non-stress condition; so that, plant height, main branch diameter, lateral branch diameter, root dry weight, root fresh weight and shoot fresh weight decreased under drought stress compared to non-stressed plants. Analysis of variance showed that stresses had significant effects on leaf and stem flavonoids and phenol contents. By contrast, there was no significantly difference between drought and saline stresses in terms of total flavonoids and phenol of roots. Mean comparisons showed that the amount of leaf phenols and flavonoids increased under drought stress in compared with non-stress condition. In conclusion, this study showed that drought and salinity were increased secondary metabolites in thyme, but the effect of salinity in increasing of secondary metabolites was lower than drought stress.

The greater celandine (Chelidonium majus L.) contains important alkaloids such as Sanguinarine. Sanguinarine is an active alkaloid with potentially antimicrobial, anti-inflammatory and anti-tumor properties that is widely found in plants of Papaveraceae family. In this research, the isolation and sequencing of (s)-cis-N-methyl stylopine 14-hydroxylas (MSH) coding gene was performed as one of the key enzymes in Sanguinarine pathway from celandine. Then, the changes in cmMSH gene expression were investigated in roots, leave and stems at four salinity levels (0, 25, 50, and 100 mM) in a factorial experiment based on completely randomized design. In the present study, the cDNA encoding MSH enzyme was isolated from root and successfully integrated into PTG19-T plasmid and cloned at E. coli. After confirmation of recombinant clones by PCR, plasmids of recombinant bacteria were extracted and the gene segment sequenced. The sequenced segment had 784 nucleotides with an open reading frame of 261 amino acids which the derived protein with functional domains including helix K region, heme-binding region and aromatic region belonged to the Cytochrome-P450 protein family. In the phylogeny tree, the protein sequence of isolated cmMSH gene had the most similarity with methylstylopine 14-hydroxylase enzyme of poppy species. Mean comparison of gene relative expression showed that 50 mM salinity had maximal effect on increasing of cmMSH gene expression in root tissue. Also, the expression pattern showed that with increment the salinity up to 50 mM, the expression of cmMSH gene increased in leaves and roots, but with increment the salinity to 100 mM the gene expression decreased 35% in both tissues.