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IntroductionPlants are exposed to various environmental stresses during growth and development under natural and agricultural conditions. Drought is one of the most severe abiotic stresses that greatly affects plant yield. Metabolic changes under stress cause the morphological and physiological changes in the plant which may eventually lead to reduced yield. Barley is one of the most important cereals which a large part of the human population in many parts of the world are dependent on it as a source of food and animal feed.Materials and methodsIn order to investigate the effects of drought stress on some morphological and physiological traits in barley as one of the most important crop, this experiment was carried out in the greenhouse of the agriculture faculty, Ferdowsi University of Mashhad in 2019 in a factorial based on a randomized complete block design with three replications. Experimental factors were included: different levels of drought stress including 100% of field capacity (control or no stress), 80, 60 and 40% of field capacity and two barley cultivars containing semi-sensitive (Fajr30) and drought tolerant (Dasht). First, the seeds of both cultivars were planted in trays filled with coco peat, perlite and sand, and after two weeks, the seedlings were transferred to pots filled with garden soil. The pots were watered daily and after one month (4 to 6 leaf stage) until the end of the growing season, they were subjected to drought stress treatments. During the vegetative stage, morphological traits such as plant height (from crown to end of plant), stem diameter, number of leaves, number of tillers, leaf area and number of stomata below and above the leaf were examined using common methods. Physiological traits including proline, soluble sugars, photosynthetic pigments and antioxidant enzymes (catalase and peroxidase) were also measured. Analysis of variance was performed using JMP statistical software version 8 and the mean of treatments was compared using LSD test at the level of 5%.Results and discussionThe results of this experiment showed that the effect of cultivar and drought stress were significant for many of the studied traits. The highest plant height, stem diameter, number of leaves and tillers and leaf area index were observed in plants without stress (100% FC), which was significant compared to other stress level. Also, the comparison of the two cultivars showed that Dasht, as a drought tolerant cultivar, has more leaves and tillers but less leaf area and stomata than the semi-sensitive cultivar Fajr30. In other words, the tolerant cultivar produces more leaves but smaller under stress condition which finally reduces the leaf area compared to the sensitive cultivar. This can be a good solution for drought resistance through reduce evaporation from the leaf surface as well as shading. In addition, osmotic regulators such as proline and soluble sugars and the activity of antioxidant enzymes including catalase and peroxidase increased under drought stress and it was higher in tolerant cultivar. In fact, it seems that tolerant cultivar can tolerate drought stress through activating their immune system by producing osmoprotectant and increasing the activity of antioxidant enzymes.ConclusionIn general, results showed that proline, soluble sugars and antioxidant enzymes play a role in the mechanism of stress tolerance and their metabolism is affected by drought stress. The results of this experiment suggest that the accumulation of these osmoprotectans and morphological changes can be part of the drought resistance mechanisms in the drought tolerant genotype of barley, which can ultimately be used in breeding programs to improve drought tolerance. In general, the results of this experiment showed that both barley cultivars responded to drought stress, but Dasht cultivar showed more tolerance in these conditions.

Background: Cell suspension culture and elicitation have been playing an important role in the synthesis of active secondary metabolites. Rosmarinic acid (RA) is one of the active compounds found in lavender essential oil that stands out due to its antioxidant and anti-inflammatory characteristics. Objective: This research was conducted to investigate the production efficiency of RA in Lavandula angustifolia suspension culture through elicitation with methyl jasmonate (MeJA) and yeast extract (YE). Methods: Cell suspension culture established in B5 liquid media supplemented with different combinations of Plant Growth Regulators (PGRs). The influence of different PGRs treatments on cell growth and accumulation of RA were analyzed. Then the effect of concentration and time course of elicitation with YE (0.1, 0.5, and 1 g/l after 1, 3 and, 6 days of elicitation) and MeJA (50, 100 and 200 µM after 1, 2 and 3 days of elicitation) separately and in combination with each other on cell growth and intracellular and extracellular content of RA were investigated. Results: HPLC analysis showed that the highest intracellular RA content (17.03 mg/g dry weight) was observed 24 hours after the addition of 50 µM MeJA in combination with 1 g/l YE, which was approximately 33% higher than that found in leaves. Furthermore, it was 9 and 11 times higher than cultures treated with MeJA and YE alone, respectively. In addition, both elicitors significantly affected the extracellular quantity of RA than control. Conclusion: Our results documented that the application of elicitors increased biomass and RA accumulation in L. angustifolia suspension cells.

Background: Datura innoxia Mill. is one of the medicinal plant which can produce tropan alkaloids such as hyoscyamine and scopolamine with analgesic and antiasthmatic activities. Objective: In this research, the effect of various factors such as culture media and bacteria strain were studied on hairy roots induction and secondary metabolites production in Datura innoxia. Methods: Strains A4, A13, R1000, MSU, 15834, 2656 and 11325 of Agrobacterium rhizogenes with two different concentrations (OD = 0.5, 1) used for co-cultivation of leaf and stem explants prepared from 1-3-month-old and 5-7-month-old plants to optimize the production of hairy roots and then 6 different media were used for establishment and growth of hairy roots. Results: The results of data analysis showed that A4 strain with 86.6 % hairy root production was the best strain than others. Also, dark conditions, younger explants and higher concentration of bacteria caused the highest amount of induction of hairy roots. ½ MS medium was recognized as the best medium for the growth of hairy roots. The results of HPLC analysis also showed that the level of hyoscyamine in hairy roots was lower than that of normal roots which, of course, can be compensated by the higher growth of hairy roots. Conclusion: The appearance and growth of hairy roots depends on various factors such as bacterial strain, culture medium, explant and its age and other factors that need to be optimized at the beginning of any research.

Background: Datura species produce a wide range of tropane alkaloids with medicinal values. D. innoxia is a species of the Datura genus which is poisonous and has an unpleasant odor due to the presence of tropane alkaloids. Objective: The study aimed to induction of polyploidy by colchicine treatment on downy thorn-apple and the effect of it on appearance and hyoscyamine content. Methods: In order to induce tetraploidy in downy thorn-apple, a factorial experiment was carried out with two factors: colchicine concentrations (0.05, 0.1, and 0.2 %) and exposure time (48 and 72 h) with 3 plants for each treatment with using a cotton ball included colchicine on shoot tip. Morphological changes were observed and the plants were analyzed by flow cytometry in near to the flowering time. Results: The results showed that the concentration and exposure time of colchicine, and their interaction affected the percentage of tetraploidy. The maximum percent of tetraploidy was observed in concentration of 0.2 % and 72 h. On the other hand, the application of different concentrations of colchicine and exposure time didn’t have a significant effect on plant survival. The number of stomatal guard cells were decreased but no significant differences were observed in the content of total chlorophyll and carotenoids. Also, the hyoscyamine content in tetraploid plants was decreased to half in comparison to diploid ones. Conclusion: In general, induction of tetraploidy in this plant could change some phenotypic traits but has a negative effect on hyoscyamine content.

Background: Thalictrum minus L. is a medicinal plant contains berberine, which is used for complementary treating of some diseases. Excessive harvesting of this species from rangelands to extract the alkaloid has endangered its survival. Objective: This study was aimed for in vitro propagation of this species and berberine production. Methods: The seeds were germinated at in vitro culture conditions. Then, the leaves were used as explants for callus induction in two MS and LS basal media supplemented with BA (0, 0.1, 0.5 and 1 mg/L) in combination with different concentrations of auxins, NAA, 2,4-D or IAA. Results: The results showed that MS medium with the plant growth regulator combination of 0.5 mg/L BA and 0.5 mg/L NAA had the highest callus formation and 100 % regeneration. Some calli were transferred to three liquid culture media (MS, LS, or ½ MS) with 1 mg/L BA + 0.1 mg/L NAA or without plant growth regulator to produce the berberine. The HPLC results identified 6 % of berberine in the liquid medium. This amount of berberine can be easily released into the culture medium, therefore, it will be easier to extract. Conclusion: This method can be successfully used for propagation and extraction of berberine without plant destruction.

Introduction Mentha is one of the most important genera of aromatic plants which belongs to the Lamiaceae family. The genus of Mentha is distributed across Asia, Africa, Australia, Europe, and North America. Mentha longifolia L., also known as wild mint, is a fast-growing aromatic perennial herb. It is widely used as herbal medicine and is beneficial for the immune system and fighting with secondary infections. The essential oil of this plant is partly responsible for the decongestant, antispasmodic and antibiotic effects. Currently, much emphasis is being laid on conserving plant germplasm as valuable bio-resources. Selection between and within accessions for a high level of herbage yield and other characters requires an effective tool to be applied by mint breeders. Achieving to cultivars that are more capable of optimum producing is a breeding goal. The objectives of this study were to analyze the diversion of the agronomical traits of Mentha longifolia L. genotypes from different regions of Iran under a similar condition in order to find the superior genotypes and introduce for the domestication of this plant. Material and MethodThis research was performed at the research field of the Ferdowsi University of Mashhad. The field is located at 36˚15' North latitude and 59˚38' East longitude, at an altitude of 985 meters. The information related to temperature and precipitation was obtained from climate station. Soil sampling was done in the depth of 0.3 meter and physical experiments on samples were done before starting the experiment. The field was fertilized by 25 kg/ha animal manure. Seeds of 20 genotypes of M. longifolia L. were prepared from Gene Bank of Research Institute of Forests and Rangelands and a view of the wild mint genotypes distribution was presented on the map. The experiment was performed in a field with 500m2 areas in 2015-2016 growing seasons. The experiment in form of compound analysis arranged in complete randomized design with three replicates of 20 wildmint's genotypes in every replication. In each furrow, fourteen bush was totally studied, in which the distance between the two plants was 20 cm. The plot was considered 1.2 by 3 m and the distance between rows was 0.50 m. The dimension in the plots for every replication area was 3.6 m2 and the distance between blocks were 1.5 and between experimental units were 0.5 meters. Picking up was done after elimination of 0.5 meters from every side of replication. The essential oil was extracted using Clevenger type apparatus and by hydrodistillation. Results and DiscussionAnalysis of variance showed that the effect of the diversity among the genotypes on morphological, agronomical and essential oil yield parameters of wild mint was significant (P≤0.01). The results of this study showed that Mentha longifolia L. herb has a high diversity under the same culture conditions among genotypes collected from 20 regions of Iran. Correlation result shows that vegetative organs have more effect on the essential oil content than the reproductive organs. This may be due to the distribution of essential oil accumulation and storage sites, which requires further research in this regard. The study of vegetative and reproductive characteristics suggests that the genotypes of the dry and semi-arid climates have different conditions, which may causes the separation of their genotypes compared with the other genotypes. The bi-plot, based on PC1 and PC2, reflects the relationships between the studied genotypes. The results of bi-plot of cluster analysis confirmed that genotypes were divided into two main groups based on agronomical and morphological traits. Conclusion Finally, plants of genotype G13 belongs to Hormozgan province and then G16 genotype belongs to Khuzestan province were selected because of more biomass, more aerial part volume and also the most essential oil performance compared to the other genotypes. Desirable traits for the aerial part such as the time of flowering, leaf length, leaf width, plant height, the highest number of leaves in the stem and internode distance are important traits that should be considered. In addition, the cultivation of these genotype in order to the further production can be the great help in the domestication of this species and, given the fact that the diversity is a precursor of breeding, this research can be an introduction for future breeding operations. However, further research is needed to confirm the phytochemical superior genotypes.

Geranium is one of the most economically important pot plants in the world and is widely used in landscape. In order to produce disease-free and homogenic plants, proliferation by tissue culture is one of the alternative methods for mass production of this valuable plant. The aim of this study was to achieve an efficient and rapid method for micropropagation of Geranium by comparing leaf discs, petiole, stem and root explants on MS and B5 media. Material and methods For callus induction, stem, leaf and root explants were cultured on MS basal medium with 11 different kinds of hormonal combination, containing auxin (NAA and IAA) and cytokinin (BAP, Kin and Zeatin) with various concentrations. For direct regeneration of stem, leaf and petiole explants, 10 different culture media were used in MS and B5. After shooting, the explants were transferred to B5 culture medium containing auxin (0.5 mg/l NAA and 0.5 mg/l IAA) for root induction. Results The best hormonal treatment for callus induction was 1mg/l NAA+10mg/l Kin by using stem and leaf explants as 92.5% and 88.75%, respectively. Also, the maximum fresh and dry weight of callus were related to stem explants on MS culture medium included 1mg/l NAA+10mg/l Kin, which were not significantly different from leaf explants on the same culture medium. The maximum percentage of direct regeneration and number of branches, respectively as 71.25% and 7.12 shoots, were related to stem explant on B5 medium containing 2mg/l BAP+1mg/l Zeatin, and the maximum shoot length, as 2.56 cm, was observed during direct regeneration in MS culture medium included 0.2 mg/l NAA+0.5 mg/l BAP+1 mg/l Zeatin. The shoots were successfully rooted and adapted to the environment. Conclusions The results of the present study were showed that the micropropagation of ornamental geranium plant is affected by the type of explant, the concentration of growth regulators and the type of culture medium and the stem explant is efficient in callus induction and direct regeneration.

Background: Papaver bracteatum Lindl. (Persian poppy) has a very long history as the major source of natural alkaloids. Among Papaver species, Persian poppy can be considered as a rich source of thebaine. Objective: The purpose of this study was to evaluate the thebaine content in different organs and different populations of Persian poppy. Methods: Seven populations of Persian poppy were collected from five different geographical regions of Iran including Damavand (Rineh, Abask), Chalous (Yoush, Pol-e Zanguleh), Mahabad, Meshgin Shahr and Zanjan (Taham) and were screened for its thebaine content. Extraction of thebaine was performed using the ultrasonic-assisted extraction (UAE) method. The parameters including pH, amount of solvent, extraction temperature and time, the repetition of extraction cycles and also the particle size were investigated and optimal conditions were determined to reach the maximum efficiency. Also, the correlation between phenology of the plant and thebaine content was studied during the developmental stages of poppy from pre-flowering to lancing. The developed UAE method followed by high performance liquid chromatography (HPLC) was used for quantitative analysis. Results: Thebaine content was significantly different in various organs and populations grew at different regions. The thebaine content was in the range of 0.9-1.4 % and 0.1-0.45 % in capsules and stems, respectively. The highest thebaine content was obtained in the capsules of Mahabad population. Conclusion: Extraction led to a high purity thebaine from Persian poppy. Eventually, a comprehensive profile of thebaine in different Persian poppy populations from different geographical regions in Iran was released.

An important equation usually used in modeling neuronal dynamics is cable equation. In this work, a numerical method for the fractional cable equation which involves two Riemann-Liouville fractional derivatives is proposed. Our computational technique is based on collocation idea where a combination of Bernoulli polynomials and Sinc functions are used to approximate the solution to this problem. The constructed approximation by our method convert the fractional cable equation into a set of algebraic equations. Also, we provide two numerical examples to confirm the accuracy and effectiveness of the present method.

Royal jelly (RJ) from queen honeybee larva as a traditional medicine agent has a variety of pharmacological benefits. In the present study, the effect of Royal jelly was investigated on the urinary bladder cancer cell line (HTB-9 5637). To determine the cell viability in different concentrations of Royal jelly, MTT assay was performed. An in vitro wound healing assay was applied to investigate the effect of RJ on cell migration. The activity and gene expression level of matrix metalloproteinase 2 and 9 was assessed by zymography and Real time PCR respectively. R.J.S at the concentration of 0.7 mg/ml had a significant effect on reducing the proliferation rate of 5637 cells after 72h (p < 0.009). R.J.S significantly decreased cell migration and induced a significant decrease in the transcriptional level of MMP9 after 72h (0.5x; P < 0.049). However R.J.S did not impose any effect on the expression level and activity of matrix metalloproteinase 2. These results indicate the potential of RJ as a promised natural anti-proliferative and anti-metastatic drug in combination with advanced therapy methods for cancer treatment. Royal jelly has the potential to be more focused as an anti-metastatic drug to control tumor growth and can be considered as a more effective alternative to the current chemotherapy drugs.

Silver nanoparticles are widely used in manufacturing of different products considering their unique physical and chemical properties. Also they have been noticed in medical diagnosis and treatment because of their antibacterial properties. Physical and chemical methods of producing nanoparticles, are expensive and are not safe enough as toxic substances may remain in the final preparations. To solve this problem, biological production of nanoparticles is considered as an efficient alternative method. In present study, synthesis of silver nanoparticles via seeds, petals, roots, and hairy root extracts of Calendula officinalis were performed. These nanoparticles were characterized by means of spectrophotometer, particle size analyzer and transmission electron microscope. Nanoparticles which were synthesized by hairy root extract showed the highest absorption at 430 nm (1/6 a.u) and the smallest size (5/3 nm), in comparison to other examined particles. Results confirmed the better performance of hairy root extracts in the synthesis of silver nanoparticles.

Introduction: Nowadays, different solvents are used to extract plant antioxidants. Each solvent has a special ability to extract antioxidants materials. Accordingly, in this research, some medicinal herbs were collected from different vegetation zones of Iran and evaluated based on the majority of useful solvents. The best solvent for the extraction of antioxidants can be identified and used to determine the best herbal medicine. Materials & Methods: In order to investigate the type of solvent extraction on the amount of antioxidant material of some indigenous medicinal plants of different regions of Iran such as Mentha asiatica, Ziziphora persica, Otostegia persica, Crocus sativus, Achillea millefolium, Mentha longifolia, Hyssopus officinalis, Thymus vulgaris, Fumaria officinalis and Teucrium polium, were done the experiment in The factorial based on a completely randomized design with three replications. DPPH was used to measure antioxidant activity. Findings: Results showed that the extract (hydro, hydroalcoholic, acetone and methanolic) and the interaction of extract and plant on antioxidant properties had a different effect on phenol and flavonoid content (P

Introduction: Anthurium is a popular genus of the Araceae (order Spathiflorae).The flower consists of a protruding spadix containing numerous florets, subtended by a brightly colored modified leaf, the spathe. Anthuriums are bisexual and protogynous.Anthuriumscherzerianum as the most important species ofAnthurium genus is a potted perennial plant. Due to having beautiful, attractive and long-life flowers, A. scherzerianum can be used for the production of pot and cut flowers. Tissue culture is suggested as the most commonly method in order to rapid propagation and removing disease in a short period of time. This method also recommended for Anthuriumbecause of problems in classical propagation method of this flower..The three basic propagation methods for Anthuriumare propagation by seed, traditional vegetative and tissue culture.Micropropagation of Anthurium is using forcommercial production. Materials and Methods: In this study, the effect of plant growth regulators and explants on indirect regeneration of A. scherzerianumdetermined in separate experiments. In the first experiment, callogenesis was done by leaf explants on MS medium containing growth regulators, BA in three concentrations (0.5, 1.25 and 2 mg/l) in combination with 2, 4-D (0.5, 1.25 and 2 mg/l) or NAA (0.5, 1.25 and 2 mg/l) and the combinations of TDZ (0.5, 1.25 and 2 mg/l) with 2, 4-D (0, 0.5 mg/l). In the second experiment, regeneration was done on MS medium containing 0.75 mg/l BA with 0.05 mg/l 2, 4-D and 0.1 mg/l NAA and also in combination with TDZ (0.75mg/l). For rooting, MS medium containing different concentrations of IBA and IAA (0, 0.2 and 1 mg/l) were used. Callus induction, regeneration and rooting experiments were done based on completely randomized design, with 12, 6 and 6 replications, respectively.Data from all the schemes used in this study were analyzed with SAS statistical software. The comparison of means using Duncan's multiple range test was evaluated at the 5% level. Results and Discussion: Analysis of variance showed that the effect of explant type and hormone combinations was significant on the percentage of callogenesis, callus volume and survival percentage. The interaction effect of explant type and combination of hormones was also significant on percentage of callogenesisand the volume of callus. Means comparisons showed that the highest callogenesis, viability and callus volume were achieved on MS medium containing 2 mg/l of BA and 0.5 mg/l of 2, 4-D. Petiole explants, also produced the highest percentage of callus (95%), survival rate (96%) and callus with dimensions of 6 mm2. Callus formation in leaf vein explants was higher than others. The effect of explant type and hormone combinations on regeneration, number of branches, number of leaves and leaf length was significant.The interaction of explant and hormone combinations on regeneration, number of branches, number of leaves and leaf length was also significant. Moreover, results of regeneration experiment indicated that the maximum number of shoots (6.9) and the maximum shoot length (5 cm), number of leaves (18) and the leaf length (2.8 mm) were achieved in 0.75 mg/l BA mg/l of and 0.05 mg/l 2, 4-D. In this study, petiole explants were also regenerated earlier than leaf explants.The effect of hormone combinations and concentrations was significant on rooting specially on the number of roots and root length.Furthermore, results of rooting experiment revealed that the highest rooting percentage (95%), the maximum number of roots (4.5 per plantlet) and the longest roots (3.5 cm) were produced in the medium containing 0.2 mg/l of IBA. Finally, the rooted plantlets were adapted (90%) in vivo condition by placing them on a mixture of cocopeat and perlite (2:1) substrate. Conclusion: In this study callugensis, regeneration and rooting of A.scherzerianum’s petiole and leaf explants were studied and different levels of plant growth regulators used for callugensis and regeneration. In this study petiole explants showed the highest callugenesis and regeneration. MS medium containing BA (2 mg/l) and 2, 4-D (0.5 mg/l), was the best for callugenesis. Also the highest percentage of regeneration was observed in medium containing BA (0.75 mg/l) and 2, 4-D (0.05 mg/l). Moreover low concentration (0.2 mg/l) of auxin has a better effect on rooting than high levels (1mg/l) so that the highest rooting percentage was produced in medium containing IBA (0.2 mg/l) and the lowest rooting percentage was produced in medium containing IAA (1 mg/l). Anthurium plantlets acclimized is cocopeat and perlite substrate (2: 1) with 90% acclimation.

Introduction Cold and freezing stress is one of the most important environmental stresses that affect the growth and yield of crops. One of the important processes in plant resistance to cold stress is known to acclimatize. In this process, the internal mechanisms of plants causes higher resistance to freezing stress after exposure to temperatures close to zero. Low temperature stress induce or inhibit the expression of certain genes. P5CS protein and transcription factor DREB (CBF) play an important role in adaptation and cold tolerance in plants. P5CS activates the enzymes in proline synthesis pathway and cause accumulation of this substance in plant that protects membranes and proteins against different effects of high concentrations of inorganic ions and high and low temperatures. Also DREB proteins with a conserved sequence bind to specific DNA sequences upstream of responsible genes in resistance to cold stress and leads to their expression. In this study the expression of P5CS and (DREB1)CBF genes that have an important role in the face of many plants to freezing and cold stress were examined in chickpea in both resistant and susceptible genotypes and used of Real time PCR method to evaluate the expression of these genes under stress and non-stress. Materials & Methods In the first experiment, the expression pattern of P5CS and CBF (DREB1) genes were evaluated at normal (23°C), acclimation (10°C) and freezing (-10°C)temperatures in two resistant (MCC426) and susceptible (MCC505) genotypes of chickpea, in two organs (leaf and stem). Briefly pots containing plants of both genotypes stored at 23°C for two weeks. After this period, plants are divided into two parts, half remained in the normal conditions and the other half were transferred to 10°C for cold acclimation. After 10 days, seedlings of normal and acclimation conditions transferred to a thermogradient freezer and after 15 minutes at -10°C, sampling was carried out from the leaves and stems. The simultaneous sampling of the control plants (normal conditions) were also done. After RNA extracting from samples and determine the concentration and quality by nanodrop and treatment with DNase I enzyme, cDNA synthesis was performed. After designing appropriate primers, the PCR process was optimized and Real time PCR was performed under optimized process conditions and gene expression levels were calculated. In the Second experiment free proline changes of seedlings in the acclimation phase were evaluated. Briefly after transferring of plants in the acclimation conditions, amount of proline at them were measured at appropriate intervals after exposure in acclimation conditions. Results & Discussion Results of analysis showed that the expression of these genes were significantly higher in resistant genotype than the susceptible one. Because of these genes are the important factors in plant resistance to stress especially cold and freezing stress, this can confirm that MCC426 genotype is resistant and MCC505 genotype is sensitive to cold. Also the interaction of genotype and organ sample in the control condition showed significant difference only for expression of P5CS gene. Since this gene is an important gene in freezing and cold stress, increased expression of this gene in MCC426 genotype that is resistant to cold not unexpected. But expression of CBF gene occurred only after cold stress induction and in the absence of stress is poor expression. In addition, it was revealed that the interaction of genotype and acclimation treatment on CBF genes in both genotypes had significant differences, but in P5CS gene only in MCC426, this trend was observed. However, changes of free proline as one of the mechanisms of cell protection in acclimation period in both genotypes showed that proline contents of MCC426 in response to acclimation treatment was gradually increased, but this increase in MCC505 genotype occurred during the early stages of acclimation, and proline has decreased over time. Conclusion According to the results, it generally seems, one of the main reason for tolerance to chilling is the high expression levels of the mentioned genes during chilling and freezing stress and therefore it can be used for inducing freezing tolerance into sensitive plants.

Variegation in flower color is commonly observed in many plant species and also occurs on petunia (Petunia hybrida) as an ornamental plant. Variegated plants are highly valuable in the floricultural market. To gain a global perspective on genes differentially expressed in variegated petunia flowers, we investigated the expression of chalcone synthase (chs) and chalcone isomerase (chi) as two essential genes in biosynthesis pathway of pigment production. Also, we measured the concentration level of total flavonoids, naringenin chalcone and naringenin to evaluate the probably relationship between the expression profile of chs and chi genes and the concentration of mentioned pigments. The results indicated that chalcone synthase and chalcone isomerase expression had different profile in different petal color of Petunia hybrida. Because red flower color in petunia is related to the synthesis of pelargonidin-based (orange to red) pigments, our results suggest that the low chalcone synthase and chalcone isomerase expression levels in white petals reduce dihydrokaempferol formation, thereby inhibiting pelargonidin production. In contrast, the high expression levels of these genes observed in red petals ensure sufficient anthocyanin yields to make flowers red.

Among abiotic stresses, salinity has been increasing over the time for many reasons like using chemical fertilizers, global warming and rising sea levels. Under salinity stress, the loss of water availability, toxicity of Na+ and ion imbalance directly reduces carbon fixation and biomass production in plants. K+ is a major agent that can counteract Na+ stresses, thus the potential of plants to tolerate salinity is strongly dependent on their potassium nutrition. HKTs (High-affinity K+ Transporters) are a family of transporters that mediate Na+-specific or Na+-K+ transport and play a key role in the regulation of ion homeostasis. In this study, we intended to focus on Electrolyte Leakage, ratio of K+/Na+, transcriptomic responses of a subclass two HKT in the roots of Aeluropus littoralis under salt stress. We investigated a noticeably different expression pattern over studied time points and found a snappy increase of AlHKT and rebalance of K+ concentration. It can be suggested that the early and high response of a Na+-K+ coupled transporter acted as a part of A. littoralis salt tolerance.

Foot and Mouth Disease (FMD) is a very dangerous livestock disease which causes a serious loss in the production of milk and meat. Therefore, producing an effective recombinant subunit vaccine virus this disease is of great importance. Transient gene expression is a valuable tool to reach rapid and acceptable recombinant vaccine. An Agrobacterium-mediated transient gene expression assay was carried out in spinach (Spinacia oleracea) leaves for expression of a chimeric gene encoding a part of capsid protein of Foot and Mouth Disease virus called VP1. The plant leaves were transformed via agroinfiltration procedure. The presence of foreign gene and its expression in transformed plants were confirmed through polymerase chain reaction (PCR), real time PCR, protein Dot blot and ELISA. The results obtained in this examination showed quite a high level of gene expression in spinach leaves, showing that transient gene expression can be applied as an effective and time-saving procedure for the production of recombinant proteins. The procedures for transformation, detection of recombinant protein and its application for molecular experiments are described in the study.

Induced resistance to pathogen is one of the ways which plants use against biotic stresses. Pathogenesis-related proteins(PR), including chitinase and β-1,3-glucanase synthesized by the plant in response to pathogenic infection. In this investigation, the expression of two isoforms: acidic and basic of chitinase and β-1,3-glucanasegenes were analyzed at 0, 6, 12, 24, 48 and 72 hours after fungal inoculation of chickpea genotypes (MCC496 as resistant and MCC403 as sensitive) with race 6 of Ascochyta rabiei by semi-quantitative RT-PCR. The results showed that the expression of both acidic isoforms of chitinase and β-1,3-glucanase genes were increased at 24 and 48 hours, respectively after fungal inoculation in genotype MCC496, but the expression level in sensitive genotype increased in early time after inoculation and then decreased. The basic isoform of chitinase was expressed in sensitive genotype more than the resistant genotype. The basic isoform of β-1,3-glucanase was not expressed in these genotypes. Totally, acidic isoforms of these enzymes specifically chitinase would play roles in plant defense against Ascochyta blight

Pod borer is one of the main causes for yield loss of chickpea. Therefore, breeding of chickpea for resistance to this pest is important. The use of Cry toxin from Bacillus thuringiensis is an effective strategy for producing of transgenic resistant chickpea to this pest. These toxins are able to become active in the midgut of larvae and disrupt the insect's digestive system. We studied the stability and expression of cry1Ac gene obtained as T2 transgenic chickpea with cry1Ac gene and nptII gene with binary T-DNA in T3 and T4 generations of transgenic plants and observed the transgenic lines with cry1Ac gene and no nptII gene suggesting that separation between cry1Ac and nptII genes was occurred by recombination between two T-DNAs. In T3, PCR results showed that 6 of 25 putative transgenic plants had cry1Ac gene but all of them showed the nptII gene. From six samples with positive PCR in cry1Ac gene, five of them had positive results in RT-PCR reaction, confirming the expression of cry1Ac gene in transgenic lines. PCR results in T4 plants showed that 73 of 94 plants had cry1Ac gene and 81 of 94 samples included the nptII gene. In 10 samples cry1Ac gene separated from nptII gene. According ELISA results, in all samples tested, Cry1Ac protein was expressed in different concentrations. Bioassay tests showed that all pod borer larvae fed by leaves of transgenic plants, were dead, but all survived when the larvae were fed with leaves of non-transgenic plants. So, expected phenotype was observed successfully.

Saffron (Crocus sativus L.) is the most valuable food additive in the world which little production and high price of it caused some adulterations such as plant and chemical material similar to saffron. There are several methods for detecting these fraudulent based on morphological and chemical tests but they are not effective in some cases. In this research a novel molecular method based on ITS-2 marker is introduced. A common forward primer based on 5.8s rDNA for all plant frauds such as safflower, corn stigmas, pomegranate, turmeric and capsicum slices was designed then specific reverse primers based on ITS-2 for any frauds have been designed for polymerase chain reaction. Related ITS-2 bands were amplified in any adulterations in saffron. Specific primer for camel and cow meet fibers was designed based on cytochrome b gene and could amplified the related bands. Multiplex PCR with all of these primers could amplify all of the bands related to any adulterations. Furthermore, using 20% polyacrylamide gel lead to good segregation of bands. This method can be used successfully for detection of low percentage (1%) of fraudulent in saffron. So this marker can be used efficiently for detection of these frauds in commercial saffron.

Expressed sequence tags simple sequence repeats (EST-SSRs) are important sources for investigation of genetic diversity and molecular marker development. Similar to genomic SSRs, the EST-SSRs are useful markers for many applications in genetics and plant breeding such as genetic diversity analysis, molecular mapping and cross-transferability across related species and genera. In spite of low polymorphism, these markers show variation in the expressed part of the genome. In this study, Medicago truncatula EST-SSRs were used for investigation of transferability between M. truncatula and some chickpea (Cicer arietinum L.) genotypes, also genetic diversity between used chickpea genotypes was determined. In this research, 650 M. truncatula ESTs were searched to find simple sequence repeats (SSRs). A total of 131 EST-SSRs were contained di-and trinucleotide motif SSRs. In this study, thirty pairs of primers were designed to amplify over 10 chickpea genotypes. Thirteen primer pairs (43%) generated reproducible bands in at least one chickpea genotype that eight bands (61.5%) were polymorphic in the chickpea genotypes. A total of 24 alleles were amplified with an average of 3 alleles per primer. The average of polymorphism information content (PIC) was 78.75% and transferability across M. truncatula and C. arietinum was 43.32%. The results indicate that the developed EST-SSR markers from M. truncatula as a model plant are valuable genetic markers for legume species such as chickpea. In addition to suitability of EST-SSR markers for genetic diversity analysis, their broad range of transferability also proved their potential for comparative genomics studies.

In this paper, the effects of a number of growth regulators as well as supplements to the Murashige and Skoog (MS) basal medium were evaluated on the regeneration of Alstroemeria rhizome explants. In the first experiment the effects of three cytokinins (BA, TDZ and 2IP each at 0.5, 1 and 2 mg/l) in combination with NAA (0.2 mg/l), followed by another PGR combination of 2IP (at 0.5, 1 and 2 mg/l) with NAA (0 and 0.2 mg/l), on regeneration of rhizome-derived explants, was investigated. Through the second experiment, the effects of a number of supplements, including glucose (30 g/l as the alternative for sucrose), casein hydrolysate (1 g/l), asparagine and glutamine, (each at 30 mg/l) added to MS medium, containing 1 mg/l BA and 0.2 mg/l NAA, was examined on rhizome explants� regeneration. Among the tested cytokinins, BA induced better regeneration of rhizome explants, resulting in a higher number of shoots compared to the other cytokinins. A medium supplemented with 1 mg/l BA and 0.2 mg/l NAA proved to be the most effective, with an average of 4.16 regenerated shoots per explant. In the second PGR combination, addition of NAA at 0.2 mg/l improved regeneration, compared to NAA-free treatments. In the second experiment, glucose substitution for sucrose improved regeneration with an average of 5.10 regenerated shoots per explant, compared to 4.16 shoots in sucrose-containing medium; whereas glutamine and asparagine (with 2.66 shoots) and casein hydrolysate (with 3.80 shoots) showed a negative influence on rhizome explants� regeneration.

Saffron (Crocus sativus) is the most valuable and indigenous crop in Iran. The stigmas of flower are used as a popular natural flavouring, colouring and medicinal agent. However, the market suffers from frauds in this plant such as mixing with safflower petals due to high profit. Identification of these frauds with conventional and biochemical methods is difficult and low sensitive. Therefore, application of molecular markers such as random amplified polymorphic DNA (RAPD)/sequence characterized amplified regions (SCAR) is being considered as an alternative. In this study, DNA was extracted from dry stigmas of 5 Saffron accessions and dry petals of 7 safflower cultivars. RAPD reactions with ten 15-mer random primers resulted in two specific monomorphic bands (500 and 700 bp) for safflower, while they were absent in saffron accessions. PCR analysis with specific SCAR primers amplified two specific bands (414 and 589 bp) for safflowers in different combinations of saffron stigmas and safflower petals. This was the case with very low rates or 1% of safflower. Therefore, this method seems to be suitable for fraud identification of safflower petals in commercial saffron samples.

Saffron (Crocus sativus L.) is an indigenous and the most valuable and marginal plant in Iran. However, limited knowledge exists on its molecular biology. The importance of this plant is due to the color, flavor and medicinal properties of its red stigmas. Saffron stigmas contain a high amount of carotenoids such as crocetin and crocin. Two genes, bch and pds, have essential role in carotenoid production. In this study, the effect of four different irrigation regimes was evaluated on the expression of bch and pds genes. Semi-quantitative RTPCR showed no significant difference in the expression levels of genes of interest related to the internal standard (18S rRNA). Results of Real-Time PCR assays showed that the expression of bch and pds genes were affected by irrigation treatments as their expression decreased in irrigated plants in comparison to non irrigated ones, exept for one irrigation treatment (one irrigation in September) where the pds gene expression showed higher level. However, the expression profile of the genes was almost the same in all treatments. The comparison between results of two techniques indicated that the Real-Time PCR is more accurate for determination of the level of transcript in the Iranian saffron. It was interesting that by decreasing of irrigation, the expression level of these two genes increased indicating that abiotic stress and drought can affect on the gene expression relating to the saffron color.

Pod borer is one of the main factors for yield decrease of chickpea. Therefore, breeding ofchickpea for resistance to this pest is important. Modified cry1Ac gene of Bacillus thuringiensis inpCry1Ac-nptII plasmid containing twin T-DNA for cry1Ac and nptII genes have been used fortransformation of chickpea. At last 38 plants had nptII gene that 36 plants of them had cry1Ac geneand two plants had only nptII gene. Western blotting analysis showed that 30 plants had Cry1Acprotein expression. Transformation percentage was 0.37 in this experiment. Only 7 plants produced T1seeds. Regarding to western blotting and PCR analysis for T0 and T1, plants were devided to somegroups with different expressions. Bioassay with neonate of pod borer showed high mortality in lineswith high Cry1Ac protein expression.Key words: Bt, Chickpea, cry1Ac, pod borer, transformation