جستجوی مقالات مرتبط با کلیدواژه « متابولیت ثانویه » در نشریات گروه « بیوتکنولوژی و ژنتیک گیاهی »

European black cumin (Carum carvi L.) is an important species of the Apiacea family that has valuable terpene compounds. The aims of this study were, to determine the effect of different hormonal treatments on the production of callus from leaf and root explants and also to select the best explant and hormonal treatment for cell culture, and then to investigate the effect of silver nanoparticles on the stimulation of the production of secondary metabolites and the expression of the Limonene synthase gene in callus was obtained from cell culture. First, the effect of 6 hormone treatments in MS culture medium on the callus produced from two explants (leaf and root) were investigated by measuring three quantitative and qualitative traits in each of the treatments. Considering the above traits, leaf explant in MS2 hormone treatment, which included IBA (0.1 mg/l), 2IP (0.5 mg/l) and BA (0.5 mg/l), were identified as superior explant and hormone treatment. To continue the studies, the calluses obtained from the superior explant on the superior hormone treatment were transferred to the cell culture medium containing MS culture medium without agar. Then the samples were placed in a shaker at 120 rpm under dark conditions at a temperature of 24°C. After multiplying the callus and reaching the desired growth, the samples were treated with silver nanoparticles in two concentration of 50 and 100 mg/l, and samples were taken from the calluses at zero, 24 and 48 hours after the treatment. The data obtained from GC and GC/MS and the results of limonene synthase gene expression showed that silver nanoparticles as a non-living stimulus by intensifying defense responses in black cumin, caused a change in the synthesis rate of secondary  metabolites such as Limonene, Carvone, Carvacrol, Thymol, p-Cymene and γ-Terpinene and also increased the expression of Limonene synthase gene.

The success of plant breeders depends on the genetic diversity amount in plants and their wild relatives. Awareness of genetic distance and similarity makes it possible to organize hereditary reserves, effective sampling of genotypes, and better use of diversity. This research was carried out to study the genetic diversity of six basil populations (Ocimum basilicum L.) and to evaluate their biochemical response to low irrigation and two densities in the greenhouse of the Agricultural and Natural Resources Research Center, Isfahan, Iran using a factorial experiment based on completely randomized design with three replications. Basil populations as Afghan (green), Ghaemieh (violet), Qahjavaristan (green), Ardestan (green), Mobarakeh (green), and Mahyar (violet)) were sown in two densities of 6.6 and 20 plants/m2 in row distance at 50 cm with plant distance of 30 cm (equal to 6.6 plants/m2) and 10 cm (equal to 20 plants/m2). Drip irrigation was performed using evaporation pan in the form of three treatments of 60, 80 and 100% of field capacity (FC). In addition, 12 pairs of AFLP primers were used to study genetic diversity. According to the results obtained in all the irrigation conditions, the Afghan population showed the highest percentage of essential oil. Among the populations, the highest amounts of catalase, peroxidase, and superoxide dismutase were belonged to the Qahjavaristan with the values of 0.62, 0.44 and 0.53 µmol.m-1/FW, respectively. The highest amounts of chlorophyll a and b (1.49 and 0.76 mg/g FW) were belonged to the Mobarakeh. Regarding dehydration stress, it was found that the application of 80 and 60%FC led to an increase of 4% and 19% of catalase, 6% and 22% of peroxidase and 5% and 20% of superoxide dismutase, respectively. In general, the Afghan population showed the highest stress-tolerance level. Based on the analysis of AFLP results, each primer produced an average of 10.51 polymorphism bands, and primer 11 had the highest band production among the primers. Based on the genetic similarity matrix, the highest genetic similarity between Mobarakeh and Ardestan populations was at the rate of 0.99. The lowest genetic similarity (0.71) was related to Ardestan and Mahyar populations. Totally, populations with the least genetic similarity (Ardestan and Mahyar) could be used as parents in breeding programs, which leads to progeny with higher genetic diversity. The Afghan population can also be considered as parent in stress-resistant breeding programs.

Enzymes are noble metabolites which extensively utilize in different industries. Currently, the production of enzymes by microorganisms is one of the best and most effective methods for the production of these enzymes. Hydrolytic enzymes are one of the most important enzymes that produced by various microorganisms, including endophytes.

In order to investigate the presence of endophytic fungi from two plants Euphorbia esula L. and Chenopodium album L, in summer of 1395, six different plants parts including flowers, leaves, seeds, roots, crowns and stems were sampled from healthy plants in Hamadan province. After fungal purification, the presence of extracellular hydrolytic enzymes such as amylase, catalase, protease, cellulase, pectinase, and xylanase were investigated by adding the corresponding substrate in the endophytic fungal culture medium.

After sterilizing the surface of collected plants and transferring samples to Potato Dextrose Agar culture media, a total of 31 endophytic fungi were isolated. The results showed that 84% of fungal isolates had amylase, 81% catalase, 71% protease, 61% cellulase, 39% pectinase and 26% xylanase activity. Morphological studies of the isolated fungi showed that some them belongs to the Fusarium, Alternaria and Trichoderma spp.

The results obtained in this study showed that endophytic fungi isolated from Euphorbia esula L. and Chenopodium album L, plants are capable of in vitro production of various enzymes. Among them, the presence of important enzymes like cellulase, pectinase and protease in some isolates, emphasis the significance of results. Based on the vast application and magnitudes of these enzymes in different industries, the importance of these fungi and demands for more experiments on enzyme activity arespecified.

In this study for the first time, different strains of Agrobacterium rhizogenes, co-cultivation times, and concentrations of acetosyringon in leaf explants of Echium khuzistanicum were examined to produce hairy roots.

The leaves of 21-day-old plant, different concentrations of acetosyringone (0,100,200 µM) and 3 strains of Agrobacterium (A4, ATCC15834 and 11325) were used for hairy root induction. In each explant, surficial wounds were created by a syringe contaminated with the bacterium and put for 10 minutes in bacterial suspension. After infection, the explants were transferred in the ½ MS solid medium supplemented with ascorbic acid (100 mg/L) to a light free growth chamber with 25 ° C temperature for 48 or 72 hours. PCR technique was used to confirm transformation by gene amplification with the rolBprimers. HPLC analysis was conducted on a high performance liquid chromatography system for shikonin measurement.  

The results of this study showed that, the first hairy roots emerged from wounded places after 14-21 days. The presence of A. rhizogenes T-DNA in the hairy root genome was confirmed by PCR using specific primers for rolB gene. Strain ATCC15834 had the highest rate of hairy root induction at 100 and 200 µM concentrations of acetosyringone indicating that the phenolic compound, acetosyringone, was effective in increasing hairy root induction. Generally, High concentrations of acetosyringone and more co-cultivation time together resulted in a significant increase in the hairy root induction in three strains of Agrobacterium rhizogenesis. Line 1 of hairy root produced 254.4±0.56 µg/g FW of shikonin.  

For the first time the results of this research showed that transformation and hairy root induction in E.khuzistanicum is possible by Agrobacterium rhizogenes and it can be used in the gene transfer and hairy root culture in order to shikonin production.

Trigonella foenum-graecum is annual plant and dicotyldones of the fabaceae family. Root, leaf and seed have important medicinal compounds. Manipulation of cell culture media with elicitors is one of the important strategies for induction of secondary metabolism and production of valuable metabolites. Salicylic acid as a non-biological elicitor is effective in increasing the production of pharmaceutical metabolites. The aim of this study was to investigate the effect of salicylic acid on growth, some physiological and trigonelline production in in vitro culture of fenugreek. Fenugreek cell culture was obtained using cuticle extract of TN-47-155 fenugreek genotype in MS medium containing 0.35 mg / l TDZ and 0.05 mg / l IBA. Salicylic acid at concentrations of 12.5, 25, 50 mg / L were used. The cells were then treated with triglyceride and other physiological parameters after one week of treatment. The results of HPLC showed that cell growth and viability of the cells decreased as compared to the control. The amount of hydrogen peroxide and membrane lipid peroxidation in the cells increased compared to the control (without hormone) treatment. All treatments increased the production of TG and total phenol. Treatment of cells with 50 mg / l (75%) salicylic acid increased triglyceride levels twice as much as control (35%). Salicylic acid can be used as a stimulant in fenugreek cell culture and induces higher triglyceride production.

Hairy root cultures are an effective method for production of secondary metabolites, because the hairy roots are genetically and biologically stable and they are able to produce metabolite within a short time without needs to hormone. Chicory (Cichorium intybus L.) is one of the important medicinal plants that contains a number of important medicinal compounds. In this research, hairy root induction was established through the mediation of the ATCC11325 strain of Agrobacterium rhizogenes. In first experiment, the effects of type and age of explant and co-culture times on the efficiency of hairy root induction were investigated. In the second experiment, we studied the effect of different hairy root lines on growth rate. In third experiment, the effects of various concentration of NAA (0, 0.5, 1 and 1.5 mg/l) and sucrose (3, 4, 5 and 6%) on biomass accumulation were investigated. Results showed that maximum hairy root induction (75.55 percent) and number of roots (7.26 roots per explant) obtained from 5-day-old cotyledons. The results revealed that 1.5 mg l-1 NAA in combination with 3 and 4% sucrose were superior for highest fresh (1.4 and 1.3 g) and dry weight (0.107 and 0.100 g) productivity and growth index (21.42 and 19.96). The highest total phenolic content (6.76 mg g-1 DW) and flavonoid content in 270 nm wavelengths was observed in hairy roots that were grown in medium supplemented with 1.5 mg l-1 NAA and 4% sucrose while the maximum flavonoid content in 300 and 330 wavelengths was achieved in 1.5 mg l-1 NAA and 5% sucrose.

Betulinic, oleanolic and ursolic acid are highly valuable pentacyclic triterpenoids because of their wide spectrum of biological activities such as anti-inflammatory, hepatoprotective, antitumor, anti-HIV, antimicrobial, antifungal, anti-ulcer, gastroprotective, hypoglycemic, and antihyperlipidemic. Salvia sahendica Boiss. & Buhse is an aromatic, endemic perennial herb which grows in the restricted regions of Northwest of Iran. In the present study, callus induction and cell suspension culture establishment of the plant for the production of triterpenoids were studied. Callus induction was conducted from in vitro-grown leaf explant cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of 2,4-D (0.5, 1.0, 1.5 and 2.0 mg l−1) and BA (0.5, 1.0 and 1.5 mg l−1) base on factorial complete random design experiment. Maximum callus induction (100%) was obtained in MS medium supplemented with 1 mg l−1 2,4-D and 0.5 mg l−1 BA. The study of cell growth curve showed that the highest fresh weight (13.2g) and dry weight (2.2g) obtained after 4 week of culture. High content of ursolic acid (173.81 mg/100g DW), betulinic acid (162.2 mg/100g DW) and oleanolic acid (174.32 81 mg/100g DW) were obtained at third week of culture. Results showed that the content of betulinic acid and ursolic acid at third week of culture was 10.4 and 1.5-fold than the aerial part of wild growing plant. These results can be considered for scaling-up of triterpenoids production in bioreactors

Papaverace family includes valuable alkaloids with commercial importance and Papaver bracteatum belongs to this family. Elicitors are useful tool to enhance the production of secondary metabolites and benzophenanthridine in the poppy plants. In this study, two different elicitors, Salicylic acid and Methyl jasmonate were evaluated on the root’s explants, in order to increase the production of alkaloids in Persian poppy. Time had a great effect on the treatment of both elicitors. Overall, treatment with Methyl jasmonate has increased the amount of morphine alkaloid more than salicylic acid in cell culture. Moreover, the highest concentration of morphine was obtained by increasing the expression of TYDC and COR genes under both elicitor treatments. Having codeine and morphine in the cell culture may be attributed to the expression of TYDC as specific gene in the root. The results of this study showed that the use of biotic elicitors can motivate biosynthetic cycle of the production and accumulation of morphine alkaloid as secondary metabolite in the plant and high levels of this alkaloid can be obtained comparing to control plants.

Cuminum cyminum is a drug plant of Apiaceae and is full of secondary metabolite such as flavonoids and anthocyanin compounds. Salicylic acid belongs to a group of phenolic compounds acts as a key secondary messenger in activating specific defensive response of plant and also is used as inducer in provement of biosynthesis of secondary metabolites. In this research the effect of salicylic acid on gene expression of flavone synthase and flavonoid and anthocyanins compounds was studied. In addition the effect of salicylic acid on malondialdehyde , hydrogen peroxide and antioxidant enzymes activity of catalase, peroxidase and superoxid dismutase are studied. Seedlings of cuminum cyminum in vegetative growth stage (21 days) were treated by 1.5 mM salicylic acid and was harvested on 1, 2, 3 and 4 days after treatment. The results showed that gene expression of flavone synthase, flavonoid and anthocyanin compounds in comparison with the control increased significantly. The most increase was observed in third day which were about twice more than control. But in fourth day was decreased but still was greater than control. In fact according to the research the positive correlation between gene expression of flavone synthase and flavonoids and anthosyanins compounds was observed. Comparing with control, the results showed that malondialdehyde, hydrogen peroxide and antioxidant enzymes activity are increasedin different harvest stages under treatment with salicylic acid. So salicylic acid as an inducer increased antioxidant system of plant and increased flavonoid and anthocyanin compounds of cuminum cyminum by increase of flavone synthase gene expression.

Alecost (Tanacetum balsamita L.), belongs to the Asteraseae family, is a pharmacologically important species rich in important secondary metabolites including flavones, sesquiterpene lactones, phenylpropane compounds and derivatives, tannins and essential oils. Alecost has been used both fresh or dried as flavouring or food additive. Additionally, it has medicinal properties and is applied in aromatic products. In addition to traditional farming, in vitro hairy root culture has been found to be suitable for the production of secondary metabolites. Therefore, in order to establish a protocol for hairy root culture of alecost, root induction by co-cultivation with Agrobacterium rhizogenes (strain A4) was assessed in this study. Different explants (cotyledon, young leaf, stem and root) showed different responses to hairy root induction by Agrobacterium rhizogenes. Moreover, the frequencies of hairy root induction for different types of explants were considerably different. The induced roots were shown to be transformed by PCR using primers specific for rolB. This is the first report of hairy root induction in alecost and the results may be useful in genetic manipulation of Tanacetum balsamita and use of hairy root culture to produce high-value secondary metabolites.

Psoralen is one of the important secondary metabolites in Psoralea corylifolia. The important aspect of medicinal value of psoralen can be indicated to anticancer activity against leukemia, lung and breast cancers. In this research due to the pharmaceutical importance of psoralen, pathway of psoralen synthesis was studied. For the first time we isolate and characterized the gene coding psoralen key enzyme from the leaves of the Psoralea corylifolia by the different molecular techniques. Result of molecular analysis revealed that the length of this gene was 1237 bp. Also, the sequence analysis of nucleotides by using available bioinformatics data in NCBI found that psoralen synthase gene has 93% similarity with this gene in Ammi magus.