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

So far around 500 species of Artemisia have been found in different regions of the world, and among them 34 species are endemic to Iran. It has been evidenced that Artemisinin is one of the medicinal compounds found in Artemisia that has various medical properties, especially antimalarial. The main objective of the present study was to comprise phytochemical profile and expression pattern of some genes related to artemisinin biosynthesis pathway in A. fragrans and A. annua species.

In the present study, the accumulated artemisinin content in A. fragrans and A. annua species were detected using the high-performance liquid chromatography (HPLC) technique. Moreover, phytochemical compounds in leave tissue were identified using the gas chromatography–mass spectrometry (GC-MS) technique. The gene expression patterns for some artemisinin biosynthesis related genes including 4FPSF, DBR2, HMGR1, HMGR2, WIRKY, ADS, DXS, and SQS were comprised in two studied species. In each species, association between artemisinin content and the relative expression of investigated genes were determined through correlation analysis. All statistical analysis was performed based on three replication using R software.

Based on obtained results, there was observed a significant difference between two Artemisia species in terms of artemisinin content, and the highest content was recorded for A. fragrans. Using GC-MS analysis, in general 26 and 20 phytochemical compounds were identified in A. fragrans and A. annua species, respectively. Among identified compounds, eight compounds were similar in both species. Moreover, some compounds such as Comphor, 1,8-Cineole, 4-Terpineol, and Pinocarvone were most important. According to the gene expression analysis, the highest relative expression of 4FPSF, ADS, and DXS genes were recorded in A. annua, while the highest numbers of transcripts for SQS, HMGR1, HMGR2, DRB2, and WIRKY genes were estimated in A. fragrans. The results of correlation analysis for both species showed that artemisinin content significantly and positively correlated with the expression of ADS, DBR2, DXS, and HMGR1 genes. However, in A. annua species, the SQS gene was not expressed and there was no correlation between these genes and artemisinin content.

In general, the obtained results revealed a significant difference between two studied Artemisia species in terms of the artemisinin content and other phytochemical compounds. Furthermore, our results indicated that A. fragrans could be used as an ideal source for extract artemisinin and other compounds. Hence, conducting other supplementary studies on this species is recommended.

Medicinal plants are the source of more than 25% of all drugs consumed in the world, which play a special role in the supply of anti-cancer drugs. One of the more important of these drugs is Taxol. The natural source of this valuable substance is the species of Yew (Taxus baccata). Taxol production takes place in a complex enzymatic pathway that is influenced by a variety of factors, so the aim of this study was to identify the effect of methyl jasmonate on altering the expression of the Taxadiene synthase (ts) gene and Taxol production in Yew. In the following research, plant samples were treated with methyl jasmonate in different concentrations of 0, 100, 250 and 500 μM for two different hours of 48 and 72, during which time they were stored samples in phytotron. Then, ts gene expression was measured by Real Time PCR approach. In addition, the amount of Taxol production was evaluated by HPLC technique and the obtained data was analyzed. The results showed that at 72 hours, expression of ts gene at a concentration of 250 μM was 1.13 times higher than the control plant and the lowest gene expression with less than 5% of the control plant was related to 250 μM treatment in 48 hours. Chromatography analysis showed that the highest amount of Taxol produced in the treated plant with a concentration of 500 μM methyl jasmonate and at time of 72 hours was 0.307 mg/g dry weight. Also, treatment of 250 μM methyl jasmonate with 72 hours and production of 0.08 mg Taxol was the lowest. Pearson correlation test showed that there is no clear relationship between gene expression and Taxol production. This is due to the complex effects of methyl jasmonate on gene expression and Taxol production at different time. Despite many efforts to identify the biosynthetic pathway of Taxol, a clear and accurate imagine of it still is not available. Therefore, it is important to study the important genes involved in this biosynthetic pathway.

Taxol is one of the most important secondary metabolites that plays an important role in the treatment of various cancers. This valuable substance is produced through a complex pathway. Due to the low production of Taxol and the risk of extinction of yew, increasing demand of Taxol production has become very important. In this research, plant samples were treated with 0, 100, 250 and 500 μM methyl jasmonate and kept in phytotron for 48 and 72 hours. After treatment with methyl jasmonate, the expression of the stem bapt gene was investigated by Real Time PCR method and the amount of the produced Taxol was measured using high performance liquid chromatography (HPLC). The obtained data were analyzed statistically. The results showed that the expression of bapt gene at a concentration of 100 μM in 48 hours was 5.13 times higher than that of the control. In addition, the highest amount of Taxol was observed in the plant treated with 500 μM methyl jasmonate in 72 hours with 0.315 mg/g dry weight. Pearson’s correlation test showed that there was no relationship between gene expression and Taxol production. It seems to be due to the time difference between gene expression and Taxol production under methyl jasmonate. Considering the importance of Taxol in the pharmaceutical industry, it was recommended using methyl jasmonate with concentration of higher than 500 μM with periods longer than three days to examine the possible increasing of Taxol production

Plant flavonoids show positive responses to geographical changes, especially altitude, and considering that the medicinal plant of capper (Capparis spinosa) is ecologically distributed in different habitats, so in this research the effect of altitude on the content of anti-cancer flavonoids such as rutin and quercetin were studied in capper which sampled in summer from three mountainous areas of Amol (Nemarestaq, Delarestaq and Behrestaq) at four altitudes with difference heights of 150 meters from each other (minimum 850 and maximum 1650 meters) and with three replications in a completely randomized design. Total flavonoids, rutin and quercetin were measured and analyzed by spectrophotometry and HPLC. The results showed a significant difference between measured traits in different regions (p≤0.01 and p≤0.05), so that the highest amount of flavonoids, rutin and quercetin were obtained at an altitude of 1650 m in the third region (Behrestaq) with 4.51±17 μmol, 24.62±0.28 and 4.91±0.18 mg/g fresh weight, respectively. Therefore, zone and altitude had a positive effect on the amount of medicinal metabolites of capper. Also, the equation of regression lines and explanatory coefficients (r² = 0.91, r² = 0.74 and r² = 0.78) showed that the amount of flavonoids, rutin and quercetin at different altitudes in Nemarestaq, Delarestaq and Behrestaq increased with increasing altitude and the slope of the regression line for all traits was positive. Therefore, nature is the best model for obtaining more concentrated and desirable anti-cancer flavonoids in plants.

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.

Rosmarinic acid, as one of the most valuable phenolic compounds in different plant species, has severalbiological properties, including antivirus, antibacterial, antitumor and anti-inflammatory. Elicitors, byproducing messages, can stimulate the production of secondary metabolites in plants. Nitric oxide (abioticelicitor) is a stable gaseous radical, which acts as a signaling molecule in plants and in many physiologicalprocesses, development and environmental stresses. Due to the lack of information on the amount of rosmaricacid and the use of SNP as abiotic elicitor in Nepeta nuda L., the effect of SNP on production of rosmarinicacid was investigated in four concentrations (0, 25, 50 and 100 μM) in vitro shoot culture of catmint hairless.The amount of rosmarinic acid was measured by HPLC after drying samples at room temperature. The resultsof the samples analysis showed that the content of rosmarinic acid increased 37.7% in treatment of nitric oxide(25 μM) compared to the control sample (without application of nitric oxide). Also, amount of rosmarinic acidreduced with increasing nitric oxide concentration in the culture medium. So that the amount of this compoundreached from 54.73 mg.g-1 DW in the control sample to 14.61 mg.g-1 DW in treatment of nitric oxide (100μM). The results of this study indicated catmint hairless plant has the high level of rosmarinic acid and theamount of this valuable drug combination improved with the use of abiotic elicitor nitric oxide at lowconcentration.

Lepidium draba L. is a perennial weed belongs to the Brassicaceae family, contains considerably amount of glucoraphanin from glucosinolates group. Hydrolysis of this metabolite via myrosinase activity produce an isothiocyanate named sulforaphane that has various anticancer properties. It has been shown that environmental conditions such as temperature and altitude affect the biosynthesis level of secondary metabolites. In the present study, mature plants of L. draba was collected from different ecotypes of Kerman province [Mahan, Baft, Jiroft (Sardooiyeh)] and then sulforaphane content of each parts (root, leaf, flower and fruit) was measured using High Performance Liquid Chromatography. The results showed that fruits have the maximum amount of this compound, followed by flowers, leaves and roots. Also, the sulforaphane content in all organs of the plants where collected from Mahan was higher than two other cities. Based on the higher temperature and lower altitude of Mahan compared to the other cities, it deduced that these factors have significant effects on biosynthesis level of this active ingredient.

Artemisia annua is particularly important for the production of artemisinin, a bioproduct which can be used to combat the causal agent of malaria, treat some kind of cancers and in other activities. The low artemisinin content in the plant has caused this compound to be among the more expensive medicines. Several attempts have been made to increase artemisinin production, for example by using different elicitors, but none of the approaches has been cost effective. In this study, the expression levels of two important genes in the artemisinin biosynthetic pathway, SQS and DBR2 and artemisinin content were investigated in Artemisia cell suspension cultures. SQS and DBR2 genes have essential roles in the regulation of artemisinin pathway. For this purpose, nano-cobalt particles in concentrations of 0.25, 2.5 and 5 mg/L were used for cell culture treatment and samples were collected after 8, 24, 48 and 72 h. The highest artemisinin content was observed 24 h after 5 mg/L nanocobalt treatment. In this case, artemisinin production was 2.25 times (113.35 mg/g d.wt) higher than that of the control. Our results showed a negative and significant correlation between SQS and DBR2 gene expression and artemisinin content at different levels of nano cobalt treatments. Results also showed an increase in nano cobalt concentration after 72 hour and an increase in nano chitosan after 4h hour caused a significant decrease in the expression of SQS and DBR2 genes. In conclusion, it appears that the content of artemisinin was increased by high concentrations of the nano cobalt particles because of a decrease in the expression of SQS and DBR2 genes.

Artemisia annua belongs to the Asteraceae family and is a medicinal annual herb that contains an anti-malaria compound called Artemisinin. In the present study, the variation of Artemisinin content was investigated in seven populations of A.annua collected from different regions in the north of Iran. The leaves of different populations were shade dried and then extracts were obtained from 1g of each leave sample. Artemisinin content in extracts was measured using HPLC-UV and the standard sample. The analysis of the amount of Artemisinin content obtained from different populations indicated that the highest and lowest amounts respectively belonged to the populations collected from Noshahr with 0/195% of dried leaf and Havigh with 0/053%. The results demonstrated a relatively low variation in the amount of Artemisinin content in populations of A.annua. The amount of Artemisinin content measured in this study is significant in comparison to the samples obtained from the other parts of the world.

The aim of this study was to evaluate the pattern of antimicrobial metabolites that are capable of inhibiting the growth of Lactobacillus indigenous intestinal bacteria are pathogenic. For this study, Lactobacillus strains were isolated from 50 samples of vegetables from the production of antimicrobial metabolites were evaluated. The anti-microbial isolates producing metabolites, phenotypic and molecular identification (16srRNA). The production phase was analyzed for each isolate. Among these strains, 2 Lactobacillus species able to produce antimicrobial metabolites were much more powerful than others. Continuing this research chemical structure of antimicrobial metabolites by HPLC and SDS-PAGE methods were evaluated.The results showed that two samples of Lactobacillus plantarum and Lactobacillus rhamnosus 20205DSM 1136JCM, are able to produce antimicrobial metabolites. These isolates the bacteria Salmonella typhimurium, Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, and fungi Aspergillus Niger and Candida albicans showed growth inhibitory effects.The results of HPLC and SDS-PAGE indicated a similar structure Lactacins and organic acid compounds produced by strains that evidence is to introduce these strains as probiotics.