forough sanjarian

Water stress affects the crucial processes and yield of crops. This study aimed to investigate the effect of water stress on physiological and biochemical traits of safflower genotypes.

A factorial experiment was carried out as randomized complete block design in Seed and Plant Improvement Research Institute during 2017-19. The treatments included two irrigation levels (40 and 80% depletion of available water) and Parnian, Goldasht and Kazak genotypes. Plants were sown in soil columns with a height of 150 and a diameter of 23 cm, and irrigated by a drip system. Antioxidant enzymes, malondialdehyde, proline, relative water content, chlorophyll, seed yield and fatty acid composition were measured at the end.

Water stress significantly increased the accumulation of hydrogen peroxide and malondialdehyde in safflower genotypes which led to increased catalase and peroxidase enzyme activity by 1 and 2.5 U, respectively. The proline content increased about 16 times under water stress conditions. In contrast, the relative water content showed a significant decrease, which resulted in increased canopy temperature and decreased seed yield by 70%. The Kazak genotype had the lowest increase in canopy temperature under stress conditions and showed higher yield stability. Moreover, the fatty acid composition of seed oil changed and the amount of linoleic acid decreased by 1.8% under water stress conditions.

The results showed that in addition to drought-related physiological traits, no increase in saturated/unsaturated fatty acid ratio is also an important indicator in screening superior genotypes for cultivation in water shortage.

Inulin has been shown to possess probiotic properties and industrial applications. Chicory is the main commercial source of inulin throughout the whole world. However, the composition of the culture medium to grow chicory has a significant effect on both quality and the quantity of this active ingredient in the plant. Here, several sets of experiments were conducted in a completely randomized design under hydroponic culture. in vitro condition, to investigate the effects of phosphorus and iron fertilizer starvation on plant growth traits and also, the amount of active ingredient in chicory. Phosphorus was used at three different levels: 0.5, 1.25 and 2.5 mM while, the effect of iron treatment was investigated by removing iron from the MS culture medium and then compared to the control. The results showed that with increasing phosphorous consumption, elevated fresh and dry root weight was observed, however, iron starvation reduced root growth. In this study, the amount of inulin was significantly affected by these treatments while, the stress of phosphorus deficiency and also the elimination of iron increased the amount of inulin.

The tissue culture of Nigella arvensis L. was carried out on an MS medium supplemented with various concentrations of auxins and cytokinins. Media fortified with different concentrations (0.0, 0.5, 1.0 and 1.5 mg/L) of 2, 4-D, NAA or IBA in combination with kinetin or BAP (1.5 mg/L and 2 mg/L) were used to induce and grow callus in present and absent of light. The experimental design was performed as a randomized block factorial design with three replications. Following callus culture, antioxidant enzymes activity and total phenolic contents were evaluated. The maximum callus induction (80.9%) was obtained with 1.0 mg/L 2, 4-D and 1.5 mg/L/ kinetin in the presence of light. Plant growth regulators combination and concentration, as well as photoperiod, affected the activity of antioxidant enzymes. The highest activities of catalase and peroxidase and total phenolic contents were measured in media with BAP combinations. On the contrary, the activity of superoxide dismutase was at the least in all BAP combinations. Overall, light intensified the activities of enzymes, but it had a negative effect on total phenolic contents.

In the current study, the effects of three levels of methyl jasmonate concentration were studied on the hairy root cultures of Origanum vulgare. For this purpose, A. rhizogenes strain A13 was used to create stable hairy root lines from shoot explants. In this regard, two co-cultivation media (MS and modified MS), two inoculation times (5 and 10 min) as well two concentrations of acetosyringone (0 and 100 µM) were attentively studied. Propagated hairy roots were treated with different concentrations of methyl jasmonate (0.1, 0.2, and 0.5 µM). Sampling after 24 and 96 h of methyl jasmonate application, the activities of antioxidant enzymes were evaluated. Moreover, the gene expression level of OvDXR and OvTPS2, two important genes involved in the MEP pathway, were studied using real-time PCR technique. The highest transformation rate was observed in the explants cultured modified MS which were incubated in the bacterial suspension with acetosyringone for 10 minutes. The biomass of hairy root cultures was significantly decreased by methyl jasmonate compared to control samples. In addition, the results indicated that the concentration of methyl jasmonate, harvesting time, and their interactions significantly affected the activity of catalase, peroxidase, and SOD enzymes. The expression of OvDXR significantly increased after 24 h of treatment by 0.2 µM methyl jasmonate, while in the same sample, the expression of OvTPS2 decreased noticeably. The expression of OvDXR and OvTPS2 genes was significantly increased by 0.5 µM methyl jasmonate after 96 h. The application of methyl jasmonate as an efficient elicitor in hairy root culture system of O. vulgare is suggested for the production of valuable metabolites.

Bread wheat is one of the most important crops in the world, which is essential in terms of global food security. However, its production is extremely compromised in agricultural regions affected by water deficiency during part of the growing season and mostly in the later stages of growth. Therefore, it is promising to identify the native drought-tolerant germplasms and molecular mechanisms used to enhance drought stress resistance. The aim of this study was to investigate the expression level of eight selected genes related to drought tolerance NCED, ABF, HKT, PAL, bHLH, ABC transporter and lipoxygenase in two native germplasms of Iranian winter wheat, one is sensitive and the other is drought tolerant. For this purpose, drought treatment was applied on native germplasms in a completely randomized design with three replications and two levels of treatment in the greenhouse. Selected gene fragments were amplified, gene expression was measured by Reverse Northern Blot and quantified using total lip software. Analysis of variance of the mean relative expression of each gene compared to the internal control gene showed that drought stress had a significant effect on the expression of all genes except bHLH gene. Biplot based on the first and second components made it possible to isolate genotypes in dehydration stress based on the expression of the seven genes evaluated. This method can be used in screening and identifying tolerant genotypes in landrace population of wheat.

Zymoseptoria tritici is a dimorphic pathogenic fungus in which switching growth pattern plays a crucial role in diseases development. Therefore, Tup1 protein, the global transcriptional repressor which is a well-known regulator of dimorphism might be a qualified candidate for preliminary study in the pathogen.

Using Saccharomyces cerevisiae Tup1p as a model, the functional domains of the protein were plotted using InterPro software. Then Zt.Tup1p tertiary structure were represented by accelrys DS visualize software. Finally, the in vitro and in vivo levels of expression of Zt.Tup were evaluated by real Time RT-PCR.

Zt.Tup1p included public structures available in the other Tup1-like eukaryotic proteins, with an N-terminal domain, a poorly conserved middle region and a C-terminal domain with seven WD repeats. Also, the results showed that the two glutamine-rich elements (Q1 and Q2) contrary to which was identified in the S. cerevisiae, do not exist in Zt.Tup1p structure. The results were indicated that the expression level of Zt.tup1 up-regulate during mycelial stage. In addition, Zt.Tup1p was partly up-regulated in 14 and 24th post-inoculation.

The presence of seven conserved WD repeats in C terminal domain of Zt.Tup1p and the other related fungi indicates that these functional domains play an important role in the life of these microorganisms. According to the data of the present study, the up-regulation of Zt.tup1 gene during mycelial stage suggests that it may play a major role in mycelium development than in conidia formation. Furthermore, it’s up-regulation during necrotrophy which are characterized by the development of pycnidia shows that the gene may play a key role in the growth and pathogenicity of Z. tritici. The present study can be considered as promising new leads for starting a new comprehensive survey for clarifying this gene detailed functions during the development and pathogenicity.

Cichorium intybus is rich in inulin and has several pharmacological applications. Hairy roots culture is a valuable biotechnological tool used to produce plant secondary metabolites. Agrobacterium rhizogenes-mediated genetic transformation of chicory to hairy roots was investigated using Agrobacterium Strains A4, A13, A7, and ATCC15834. Several hairy roots were tested, from which 17 lines were selected based on their fast-growing characteristics. Results of PCR analysis revealed foreign DNA integration into the selected transgenic hairy root lines. Finally, four Adventitious roots that contained the highest ratio of total sugar to total weight (µg/gr DW), were selected. This study investigated the effects of various levels of minerals and sucrose on the production of inulin in Cichorium hairy root culture. Different levels of sucrose, phosphate (Pi) and Iron (Fe) were evaluated, separately. It was found that an increase in sucrose levels from 3 to 5% could decrease the root growth; however, 60 g/l sucrose remarkably enhanced the inulin production rate in all the examined lines. The highest biomass was achieved by applying 3.75 mM Pi but it ended in the decreasing the inulin content per unit weight. In contrast, the highest inulin accumulation and the lowest amount of biomass were observed in 0.5 mM Pi. Fe starvation caused the biomass decrease and a significant increase in inulin accumulation. Results of this study suggest a successfully optimized culture medium to initiate the induction of Cichorium intybus hairy root cells to produce inulin as a valuable medicinal secondary metabolite.

The aim of this research was to investigate the effects of different concentrations of Azadirachta indica )neem( aqueous extract on the expression of the encoding genes of the two phenylalanine ammonialysis and peroxidase enzymes in tomato (Matin and Early Urbana cultivars) in the interaction with root knot nematode caused by Meloidogyne javanica. The tomato seedlings were inoculated in the 4-6 leafed stages with different concentrations of the neem extract and then incubated with the pathogen. The plants sampling were at six time points (0, 12, 24, 96, 192 and 288 hours) after pathogen inoculation. The gene expression of the defense enzymes was investigated using the Real-time PCR techniques. In both cultivars, the gene expression of POX and PAL enzymes in the combined treatment of the pathogen nematode and the aqueous neem extract had increasing trend with significant difference with the control. In Matin cultivar, the increase of the gene expression was higher and faster than of Early Urbana cultivar, so that the expression of PAL and POX genes in this cultivar was 18.2 and 14.5 times more than the control. In the early Urbana cultivar, the highest rate of PAL gene expression was observed at 19.2 hours and was 7.5 times, and in the case of POX gene at 96 hours and 3.9 times when it was compared with control. In greenhouse experiments, the nematode pathogenicity indexes showed a significant decrease compared with the Early Urbana cultivar. According to the results, in order to protect the plant against the invasions of the patients, the expression of the defensive genes should be increased at the earliest stages after the inoculation of the patient. Early increase in expression of genes in the seedlings treated with neem extract seems to be a justifiable phenomenon in the plant resistance to the pathogenic agent.

Zymoseptoria tritici, is a species of filamentous fungus and causes the widespread wheat disease Septoria tritici blotch (STB). Salicylic acid has a key role in plant defence reactions and is also involved in the induction of systemic acquired resistance. However, the contribution of SA to the interaction of Z. tritici -wheat in STB is not entirely clear.In this study, it was shown that the Z. tritici mycelial growth and conidia germination were significantly inhibited the presence of increasing concentration of SA in both liquid and solid media. In addition, the effect of SA on pathogenicity of Z. tritici in wheat was investigated. In this study, the inhibitory effect of SA on Z. tritici at different concentrations (1 to 20 mM) in vitro, and also, the efficacy of its exogenous application in the suppression of STB in wheat under the greenhouse condition were investigated. In vitro evaluation was done on YMDA and YMDB to determine the effect of SA on the germination of conidia and growth of mycelium, respectively. Susceptible bread wheat cultivar was grown in pot and inoculated with fungus spores and SA in a three-leaf stage for green house experiments. The results showed that the germination ofconidia was completely inhibited by 4 mM SA. Furthermore, in modified YMDA plates at over 0.8 mM, the colonies diameter was reduced significantly. The result of in planta assay indicated that the foliar application of 4 mM SA can significantly reduce the disease symptoms on the wheat leaves. Discussion and conclusion: Regarding our data, it seems that SA shows more inhibitory effect in in vitro experiments than in planta. Moreover, according to the positive effects of SA on STB, the survey results can be considered as a potential approach in the management of this disease.

This study was carried out to evaluate the effect of salicylic acid on the reduction of disease severity of septoria tritici blotch (STB) in three tetraploeid and three hexaploeid wheat cultivars, under controlled conditions at seedling stage. The 10-day-old seedlings were initially treated with 0 (control), 1, 2 and 4 mM of salicylic acid with a handy sprayer. After 24 hrs, the seedlings were inoculated with fungal spore suspensions adjusted at 107 sopres/ml. The disease progress was evaluated 21 days post inoculation based on visual estimation of the percentage of leaf area with necrotic lesions bearing pycnidia. The results showed that there were significant differences on the reduction of disease severity among different concentrations of applied salicylic acid as well as among cultivar/ concentration interactions. In addition, the results showed that application of salicylic acid reduced disease severity of both tetraploid and hexaploid wheat cultivars and 1 mM salicylic acid was the most effective concentration on the reduction of disease severity. Although, the disease severity was significantly reduced by application of 2 and 4 mM salicylic acid, the effectiveness of these concentrations was less than application of 1mM. Overall, these results revealed that activation of plant defense systems through application of salicylic acid could play a significant role in systemic acquired resistance against STB disease.

The aim of this study was to examine the effect of salicylic acid on the gene expression pattern of four enzymes; Phenylalanine ammonialyase, Polyphenol oxidase, Peroxidase and Catalase in wheat-Mycosphaerella graminicola pathosystem. For this reason, Salicylic Acid (2mm) were sprayed on wheat in two-leaf stage before inoculation with the fungal pathogen. Sampling of the plants was done at five time points (0, 3, 6, 12 and 24 h) after inoculation. The scanning of genes expression pattern of encoding enzymes were carried out by reverse northern dot blotting method. The results showed that within 24 hours post inoculation the gene expression of these enzymes significantly increased in this tolerant cultivar. SA enhanced the expression of Phenylalanine ammonialyase and Peroxidase genes in all time points. The expression of Polyphenol oxidase was increased by SA after 12h. On the other hand, increasing the expression level of these genes directly increases the activity of the enzymes which indicates direct role of these gens in plant defense system. SA caused a rapid rise in expression of Catalase gene, but this effect was not continued for 24h.

Every year thousands of people die because of stomach cancer and brain tumors. N. sativa plant has thymoquinone and this compound may prevent cancer cell line growth in many types of cancer. The purpose of this study was to examine the possibility of inhibiting cell growth in human stomach gastric adenocarcinoma (AGS) and astrocytoma cell lines (AST) by N. sativa and N. arvensis extract and to compare lethal effect of these two extracts on cancer cell line.
AGS and AST cell lines were cultured and seeded for 7 hours. After this time these cells treated with various concentration of N. sativa and N. arvensis extracts for 24 hours. Consequently, the cytotoxicity effect of extract on cell line was investigated by MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) assay.
Based on our data the extract of N. sativa has cytotoxic effects on both AGS and AST cell lines. Although in case of astrocytoma cell line this effect is not dose dependent, in AGS cell line the results do not follow the same pattern and at lower three doses the cytotoxicity effect was dose dependent. The cytotoxicity effect of N. arvensis extract on astrocytoma cell line was dose independent, but in AGS cell line this effect was dose dependent.
According to the results of this study extracts of both N. sativa and N. arvensis showed cytotoxicity effect on examined cancer cell lines.

Mortierella alpina is one of the most important fungi in food industry because of having ability of synthesizing unsaturated fatty acids, particularly Arashidonic Acid. This is a precursor of Eicosanoidregulate-lipoprotein metabolism which is involved in blood rheology, platelet activation and leukocyte-function, and the functional characteristics of the cell membrane.
In this study genetic transformation of M. alpina CBS754.68 fungus was evaluated via Agrobacterium tumefaciens and Agrobacterium rhizogenes. Agrobacteriums containing pBI121 vector were used for transformation of three days of old mycelia. Three days old hyphae were exposed to the bacteria with three level of time (one, two and three hours) in the present of acetosyringone. Mitotic stability of the third generation of transgenic (T2) was confirmed by GUS assay and amplification of CaMV 35S promoter by polymerase chain reaction.
The highest percentage of transformation and mitotic stability were obtained by using A. tumefaciens and A. rhizogenese, respectively.
Discussion and The results showed that to obtain more efficient and more stable transformation, the fundamental factor is the use of suitable species of Agrobacterium. It is the first report for transformation of autothroph strain of M. alpine via Agrobacterium.

The most important abiotic stresses is salinity, the high levels of sodium which is toxic to most varieties of plants species, and more importantly affect plant growth and limit the plants yield capacity. Hyssopus officinalis L. has been known as a culinary and medicinal herb for hundreds of years, it is cultivated globally. Although Hyssopus officinalis L. is one of valuable medicinal plants and its cultivation is continuously being extended in the world, no information is available on the responses of this plant to salinity. Since the growth stages are very sensitive to stress, in this research the effects of salinity on growth parameters of Hyssopus officinalis L. in seedling stage were studied. The treatments were six levels (0, 2, 4, 6, 8 and 10 dS m-1 of saline water). The results showed that with increasing the salinity stress, protein, cartenoids, oligosaccharides and reducing sugar, MDA and H2O2 contents significantly increased however total chlorophylls, chl a and chl b contents and total sugar and polysaccharides significantly decreased. It seems that increasing the amount of protein, cartenoids and carbohydrates are important risk factors for resistance to salt Hyssopus officinalis L.

Fusarium head blight (FHB), is the most destructive disease of wheat, producing the mycotoxin deoxynivalenol, a protein synthesis inhibitor, which is harmful to humans and livestock. dsRNAmycoviruses-infected-isolates of Fusariumgraminearum, showed changes in morphological and pathogenicity phenotypes including reduced virulence towards wheat and decreased production of trichothecene mycotoxin (deoxynivalenol: DON). Previous studies indicated that over expression of yeast acetyl transferase gene (ScAYT1) encoding a 3-O trichothecene acetyl transferase that converts deoxynivalenol to a less toxic acetylated form, leads to suppression of the deoxynivalenol sensitivity in pdr5 yeast mutants. To identify whether ScAYT1 over-expression in transgenic tobacco plants can deal with mycotoxin (deoxynivalenol) in fungal extract and studying the effect of dsRNA contamination on detoxification and resistance level, we have treated T1 AYT1 transgenic tobacco seedlings with complete extraction of normal F. graminearum isolate carrying dsRNA metabolites. First, we introduced AYT1into the model tobacco plants through Agrobacterium-mediated transformation in an attempt to detoxify deoxynivalenol. In vitro tests with extraction of dsRNA carrying and cured isolates of F. graminearum and 10 ppm of deoxynivalenol indicated variable resistance levels in transgenic plants.Discussion and The results of this study indicate that the transgene expression AYT1 and Fusarium infection to dsRNA can induce tolerance to deoxynivalenol, followed by increased resistance to Fusarium head blight disease of wheat.

Fusarium graminearum is causal agent of economically catastrophic disease of cereal Fusarium Head Blight (FHB) around the world. In addition to causing a loss of yield, this fungus causes serious threats to humans and animals due to the contamination of grain with the trichothecene mycotoxin. TRI101 gene, a Fusarium spp. gene, encodes an enzyme that transfers an acetyl group to the C3 hydroxyl of trichothecenes. We introduced TRI101 gene from F. graminearum to tobacco to test its usefulness for decontamination of mycotoxins. The acetyltransferase activity of this gene was detected in transgenic plants. The growth pattern of T1 seedling was examined at present of deoxynivalenol (DON) and crude extract of mycotoxin. A significant difference in growth rate was seen in plants expressing TRI101 as compared to wild type in DON assay. There was no significant difference in growth rate of the wild type and transgenic plants in presence of a mixture of mycotoxins. According to the results, it seems although FgTRI101 enzyme is suitable for detoxification of DON, more studies are required to evaluate its performance in detoxification of mixture of Fusarium toxins.

One of the important technical obstacles in the study of many filamentous fungi is the development of efficient transformation system. Transformation of filamentous fungi is difficult because they have a cell wall and for most frequently used approaches preparing of protoplast is necessary. Protoplast preparation is batch-dependent and frequently frustrating work. In this study, a modified method was introduced for gene transfer to the plant pathogenic fungus, Fusarium graminearum, the major causal agent of Fusarium Head Blight disease in small grains. This protocol was based on protoplast-PEG method. Age of mycelia, enzymes exposure time and mycelium/enzyme ratio were optimized for the purpose of protoplast preparation. The outcome showed that the best result for protoplast preparation was obtained when 1.5 × 105 spores germinated for 6 h then exposed to 10 ml of enzyme solution for 3 h. The effect of other parameters that might enhance transformation yields including PEG concentration, DNA quantity and number of protoplasts was also examined. The most efficient condition of transformation involved the use of 106 protoplasts, 20 µg DNA and 30% PEG (w/v). In the course of this study, a simple and appropriate modified protocol for transformation of F. graminearum was established. The method introduced is also more economical and faster than other current methods.

Fusarium graminearum is causal agent of economically catastrophic disease of cereal Fusarium Head Blight (FHB) around the world. In addition to causing a loss of yield, this fungus causes serious threats to humans and animals due to the contamination of grain with the trichothecene mycotoxin. TRI101 gene, a Fusarium spp. gene, encodes an enzyme that transfers an acetyl group to the C3 hydroxyl of trichothecenes. We introduced TRI101 gene from F. graminearum to tobacco to test its usefulness for decontamination of mycotoxins. The acetyltransferase activity of this gene was detected in transgenic plants. The growth pattern of T1 seedling was examined at present of deoxynivalenol (DON) and crude extract of mycotoxin. A significant difference in growth rate was seen in plants expressing TRI101 as compared to wild type in DON assay. There was no significant difference in growth rate of the wild type and transgenic plants in presence of a mixture of mycotoxins. According to the results, it seems although FgTRI101 enzyme is suitable for detoxification of DON, more studies are required to evaluate its performance in detoxification of mixture of Fusarium toxins.

The primary causal agent of FHB (F. graminearum) can produce deoxynivalenol (DON), a trichothecene mycotoxin that enhances disease severity and poses a health hazard to human and monogastric animals. To study the possible ways for reducing the effect of DON, an acetyltransferase (AYT1) gene from yeast was cloned and transformed into tobacco as a model plant. AYT1 encodes an enzyme that transfers an acetyl moiety to the C3 hydroxil group of trichothecene. The results of in vitro culture resistance tests suggested detoxification and partial protection against DON mycotoxin in transgenic plants.