جستجوی مقالات مرتبط با کلیدواژه « secondary metabolites » 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 plant periwinkle with the scientific name Catharanthus roseus is an important source of anticancer and antihypertensive alkaloids. Due to the high price of these metabolites and their small content in periwinkle plant, tissue culture techniques have been suggested to increase their production. Therefore, this study was carried out with the aim of evaluating the effect of titanium oxide nanoparticles (TiO2) on the expression of key genes of the biosynthetic pathway of important medicinal constituents in periwinkle.
 
Murashige and Skoog (MS) medium was used as the basic medium along with 2,4-D (1 mg/L) and BAP (0.5 mg/L) plant growth regulators for leaf cultivation, callus induction, and cell suspension production. Periwinkle suspension culture was treated with concentrations of 0, 50, and 100 mg/l TiO2 nanoparticles at the peak of cell growth. Then, the expression of STR, SGD, DAT, and PRX genes was measured 24, 48, and 72 h after the treatment by Real-Time PCR. Tetrazolium test was also used to measure cell viability.
 
Although there was no significant difference between the percentage of cell viability after 50 and 100 mg/L NP-TiO2 treatment; however, the passage of time (from 24/48 h to 72 h) caused a decrease in cell viability. The application of a higher concentration of nanoparticles increased the expression of the key genes involved in the biosynthetic pathway of indole alkaloids. This increase continued up to 48 h after treatment, but then reduced. The highest expressions of STR, SGD, DAT, and PRX genes were obtained by 290, 186, 193, and 287% increase, respectively, in 48 h after the treatment of 100 mg/l TiO2 nanoparticles (as the most effective treatment with the highest percentage of cell viability).
 
In the face of TiO2 nanoparticles, as an elicitor and stress-causing agent, the expression of the genes involved in the biosynthesis of indole alkaloids is induced to achieve defense against the stressful agent and then the expression of the genes decreases after the passage of time and reduction of stressful symptoms. In general, the concentration of 100 mg/l TiO2 and the extraction of metabolites in 48 h after the treatment can be introduced as a promising stimulus to increase the content of indole alkaloids. According to the results, it is suggested that the exact mechanism involved in this metabolite increase be further investigated.

Sweet violet (Viola odorata L.) is a perennial plant belonging to the Violaceae family that is not only known as a medicinal and ornamental plant but also widely used in the cosmetics industry. In order to investigate the effect of different concentrations of gibberellic acid (GA3) on the quantitative and qualitative properties of sweet violet flowers, an experiment was conducted in a randomized complete block design (CRD) with three replications at Islamic Azad University of Khorasgan (Isfahan). Treatments were different levels of gibberellic acid (GA3) with three concentrations of 0 (control), 200 and 400 mg/L. Results showed that the application of  gibberellic acid at 400 ppm significantly increased flower number, flower diameter, flowering duration, anthocyanin content, flavonoids, chlorophyll and UV absorptions compared to control treatment. There was no significant difference in terms of anthocyanin, chlorophyll b and carotenoids between the treatment of 200 ppm  gibberellic acid  and control. Flower diameter, flower number, flowering duration and anthocyanin content of flowers at two concentrations of hormone (200 and 400 ppm) showed similar results. Regarding the medicinal use of sweet violet flowers, it is recommended to use hormonal treatments to increase the number of flowers and secondary metabolites of this plant.

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.

It is possible to treat Coronavirus disease with perilla (Perilla ferutescens (L.) Britton), a medicinal plant. The current study's objective is to examine how Agrobacterium rhizogenes induces hairy roots in Perilla to produce secondary metabolites. Three replications of a completely randomized design were used for this factorial experiment. Explant with four levels hypocotyl, cotyledon, leaf, and internode as well as bacterial strain with two levels were among the factors examined (ATCC-15834 and A4). The findings showed that the interaction effect of the factors under investigation was significant for hairy root features such as day to induction, percentage of induction, and fresh and dry weight after two months. The treatment combination of cotyledon explant infected with ATCC18534 produced the highest percentage of hairy root induction, fresh weight, and dry weight compared to other treatments. By utilizing a PCR reaction to track a portion of the rolB gene, it was possible to determine that the hairy roots were transgenic because the corresponding band was not amplified in the normal or non- transformed roots. In general, the findings of the present study indicated that the bacterial strain and explant had an impact on the induction of hairy roots in Perilla, and these findings are a necessary prerequisite for hairy root culture studies to produce secondary metabolites.

Kelus (Kelussia odoratissima Mozaff.), a medicinal plant rich in active pharmaceutical ingredients with therapeutic effects, is found only in central Zagros Mountains, west of IRAN. Despite being in danger of extinction, there are no genetic evidences regarding kelus Omics as well as valuable compounds biosynthesis pathways. MicroRNAs (miRNAs) play an important role in different processes such as growth and development, cell proliferation, response to stresses and biosynthesis metabolite. As far as the bioinformatic data are concern, the genome/transcriptome of kelus has not been sequenced. The present study was performed to identify the conserved miRNAs and their target genes in the kelus leaf transcriptome. After pair-end sequencing with the Illumina HiSeq 2500 platform, clean reads were assembled. In total, 4658 unigenes were found to contain potential miRNAs sequences. Following strict filtering criteria, five miRNAs belonging to five conserved miRNA families (miR156-3P, miR408, miR169, miR171 and miR398) were identified among candidate sequences. Results of this study revealed that the target genes of the identified miRNAs were involved in various metabolic pathways, including butanoate metabolism, glyoxylate and dicarboxylate metabolism, starch and sucrose metabolism, carbon fixation in photosynthetic organisms, peroxisome degradation, and fatty acid degradation. By affecting genes associated with six metabolic pathways, miR408 was identified as the most influential conserved microRNA in the kelus leaf transcriptome. In general, given the regulatory roles of identified miRNAs on broad spectrum of gene networks and biological processes of kelus, these miRNAs can be used as candidate genes for breeding kelus quantitative and qualitative traits.

Galega (Galega officinalis L.) from the leguminous family is used to treat diabetes. Hairy root culture is a valuable system for the production of secondary metabolites on a large scale. The use of elicitors in plant tissue and cell culture is one of the most efficient biotechnological tools for inducing biosynthesis and accumulation of secondary metabolites. This study aimed to investigate the effect of iron, silver, and silicon nanoparticles on growth and galegine content in hairy roots of G. officinalis.

In this study, induction of hairy roots in leaf, cotyledon, and hypocotyl explants of G. officinalis was performed using the A4 strain of Rhizobium rhizogenes, and the effect of different concentrations of silver, silicon, and iron oxide nano-elicitors on growth and galegine, phenol, and flavonoids content of hairy roots was investigated.

The highest rate of root induction was obtained in leaf explants with an average of 5 roots per explant. The highest root growth rate belonged to the treatments with 50 and 100 mg l-1 silicon nanoparticles for 72 h. Application of all levels of silver, iron, and silicon elicitors for 36 h resulted in a significant increase in the total phenol content of hairy roots. Thus that the highest amount of total phenol was obtained in 36 h treatment of roots with a concentration of 100 mg l-1 of silicon nanoparticles without significant difference with 10 and 20 mg l-1 silver nanoparticles. Also, the highest amount of total flavonoids was obtained at 200 mg l-1 iron nanoparticles for 36 h; and the highest content of galegine was obtained in the treatment of roots with 10 and 20 mg l-1 silver, and 100 mg l-1 silicon nanoparticles for 36 h.

The appropriate type and concentration of elicitors is an effective factor in the response of hairy roots to nano-elicitors. The highest yield of galegine (17.55 mg) was obtained in the treatment of roots with 50 mg l-1 of silicon nanoparticles for 72 h. This can be attributed to both better growth of hairy roots and high galegine content at this treatment.

Saffron (Crocus sativus L.) is one of the oldest medicinal and spice plants in the world. In order to investigate the effects of plant age on secondary metabolites and some physiological factors of saffron, a research project was conducted in a completely randomized design with three replications at the research field of medicinal plants of the University of Zanjan. Three saffron farms with different biological ages of one, two and five years old were selected as experimental treatments, and then three plots were randomly selected from each farm and the desired traits were measured. Results showed that, increasing of the age of saffron plants in this experiment increased fresh weight of plant and dry weight of stigma. 5-yraes old plants showed the highest amounts of these traits. Whereas, the contents of soluble proteins and sugars and photosynthetic pigments were higher at first year and decreased by increasing of plant age. Number of secondary metabolites decreased when the age of plants increased as in one–year old saffron plants 26 metabolites and in 5-year old plants only 15 metabolites were detected. Loliolid (18%) and thymol (17%) in 1-year old plants and thymol (19%) and camphor (14%) in 5-year old plants showed the highest level among all detected secondary compounds. Generally according to these results, plant age has an important effect on the number and kinds of secondary metabolites in saffron which should be considered at the time of saffron harvesting.

Black cumin (Nigella sativa) is a medicinal plant of the Ranunculacea family which raised attention due to its pharmaceutical properties. Medical significance of N. sativa mainly attributed to its oxygenated monoterpenes which are biosynthesized via the methyl erythritol phosphate (MEP) pathway located in plastids. In this study, the essential oil components of leaves, flowers, and developmental stages of seed including half black seeds, soft black seeds, and hard black seeds were analyzed in N. sativa. Whereas no terpene was detected in flowers and leaves, seeds were found to be the major site of biosynthesis and accumulation of terpenes, and the amount of terpene compounds changed during seed maturation. The essential oil consists of monoterpenes (more than 99%) and sesquiterpenes (less than 1%). In order to improve our understanding of monoterpene metabolism, the partial sequence of a hypothetical monoterpene synthase (NsTPS2) was isolated from N. sativa plant using RACE-PCR technique. This monoterpene synthase was identified from RNA sequencing data from soft black seeds. Except of the highly conserved DDXXD motif in NsTPS2 which is necessary to validate monoterpene synthases, no other conserved regions of other identified monoterpene synthases were observed. Dendrogram analysis revealed that NsTPS2 had the highest homology with a terpene synthase (72.89%) from Aconitum carmichaelii and these two sequences were grouped in the same group. Nigella sativa and Aconitum carmichaelii both belong to the Ranunculacea family. This indicates that the genetic information of plants of the Ranunculacea family can be used to isolate different monoterpene synthase. The results of this research can be useful in genetic manipulation and metabolic engineering of Nigella sativa.

Active substances or secondary metabolites in medicinal plants are regularly produced in response to biotic and abiotic stresses and determine the ability of plants to adapt during growth and development. In order to increase the quantity and quality of the effective substance of the green cumin medicinal plant, with the scientific name Cuminum cyminum L from the Chetrian family, we need to identify the germplasm and useful genes in domestic and wild populations of this plant. In the current research, five selected genes in the biosynthesis pathway of flavonoids in cumin, obtained from previous research based on NGS method, which had the highest expression changes in drought stress, were validated. DNA from young leaves of selected ecotypes was extracted and quality measured with agarose gel. Considering the PCR test and the observation of a single band in all cases and the lack of difference in the length of the amplified fragment among the selected ecotypes; The length polymorphism of the fragments resulting from the amplification was rejected due to the presence of deletion and addition regions between different genotypes. This experiment could not confirm the placement of the two studied ecotypes of Sadouq and Rafsanjan in terms of seed yield, essential oil and extract content, in two different groups in terms of the studied genes. Also, he could not confirm the difference in the expression of two genes (with sequence numbers DN1196 and DN32640) in drought stress and normal irrigation in the previous research

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.

Extraction of genomic DNA of high quality and quantity from some medicinal plants is difficult. It can be attributed to the presence of high amounts of specialized metabolites biosynthesized into their cells. On the other hand, molecular biological studies require successful extraction of high-quality DNA. Hence, the present study aimed to compare four modified procedures of DNA extraction including Murry and Thompson (1980), Dellaporta et al. (1983), Doyle and Doyle (1990) and Ziegenhagen et al. (1993) from five genera of Lamiaceae. Fresh and dried leaves of Rosmarinus officinalis, Lavandula officinalis, Salvia hydrangea, Dracocephalum kotschyi and Ziziphora tenuior were used for the procedures. Following measurement of the extracted DNA quantity and quality, the DNA preparations were tested by PCR amplification using two inter-simple sequence repeats (ISSR) primers. All of the procedures were capable of extracting DNA from five genera. The modified Murray and Thompson method was found to be the most efficient DNA extraction method, indicating high DNA concentration (570.57ng/µl), good-quality DNA (1.80), affordable cost and less time. The results also showed that the quality and yield of extracted DNA from fresh leaves were relatively higher than those of dried leaves. In addition, banding pattern obtained using 1% agarose gel and PCR showed the same results. According to the data, Murray and Thompson method is recommended for DNA extraction from fresh and dried leaves of five studied medicinal plants.

Ajowan (Trachyspermum ammi L.) is an annual plant in Apiaceae family which is used in traditional medicine and medical science. This study was carried out to determine the quantitative expression pattern of some genes on monoterpenoid biosynthesis pathway and essential quality in Ajowan treated by methyl jasmonate. Ajowan essential oil compounds and the amount of MECPS, DXR,  and GDs genes expression as main genes related to Thymol biosynthesis pathway in response to methyl jasmonate in 4 different times were analyzed.  

After flowering, the 0.1 mM methyl jasmonate hormone was sprayed onto the plants. Samples were taken in 4 times (after 8, 24, and 48 treatment and control treatment). RNA extraction, cDNA synthesis, and Real-Time PCR have done from flowers and the date from these gene expressions were analyzed. Essential chemical compounds were studied by GC-GC/MS. Generally, in 4 samples (samples were taken after 8, 24, and 48 treatment and control treatment), 10 different compounds were distinguished.  

Based on results, the study of the essential oils of Ajowan showed that the changes of the amount of secondary compounds in the essential oil, the amount of γ-Terpinene (precursor of monoterpenoids like Thymol) went up after 24 hours methyl jasmonate treatment. Due to Real-Time PCR dates, the quantity of all 3 studied genes increased after 24 hours methyl jasmonate treatment. The increasing of these genes expression can be causes the incline level of γ-Terpinene in essential oil.  

Increasing the amount of 3 mentioned genes expression and also a high amount of γ-Terpinene in essential oils after 24 hours in flowers show that this hormone can affect the gene expression in MEP pathway which causes monoterpenoids production increase after one day of using the hormone.

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.

Induced hairy roots by Agrobacterium rhizogenes are a suitable tissue for the production of secondary metabolites, due to the stability and high production of roots in hormonal-free conditions. Trigonella foenum – graecum L. is one of the medical plants in the family Fabaceae. Many useful components in Trigonella foenum L. leafs are Calcium, Iron, Phosphorus, Carotene, Ascorbic acid, Protein, Thiamine, Riboflavin. Also, this plant widely used as anti-diabetes, blood glucose, cholesterol- lowering, Anti-cancer, Antibacterial, and food stuffing. In this study, hairy roots were induced using A. rhizogenes strains A4, ATCC11325 and ATCC15834 by injection method in complete seedling, cotyledon and hypocotyl explants. The effect of strain type and explant type, were investigated on the effectiveness of hairy roots. The explants were cultured in B5 medium with 4 replications. Hairy roots appeared in bacterial inoculation after 2 to 3 weeks. The highest amount of transgenic (93%), hairy roots number (10.43 number) and the maximum root length (6.64 cm) were observed in complete seedling explant of 10 days and A4 strain. In the hypocotyl explants, the hairy roots were not observed.

Savory (Satureja khuzistanica) is one of the nine endemic species which have phenolic compounds such as carvacrole and thymol in its essential oil and rosmarinic and other phenolic acids in its extract, has strong antioxidant and antimicrobial properties and remarkable effects on several diseases. Drought stress was induced by stopping irrigation at flowering stage. Samples were taken five times with three interval days and several physiological traits were measured. For oil extraction, Clevenger apparatus was used. For determination of essential oil’s components and methanolic extract compounds, GC, GC/MS and HPTLC were utilized. Profiling of volatiles using GC/MS, showed an increasing-decreasing trend at major phenolic and terpenes compounds such as thymol, γ- Terpinene, p- Cymene, Rosmarinic acid and Caffeic acid. Drought stress also led to a significant increase in oil yield, soluble sugars and proline as well as a significant reduction in leaf water potential, relative water content and pigments. Metabolite profiling of Satureja khuzistanica represents the strategies employed by savory in generating different biochemical phenotype. Applicable results of this study for increasing product quality is effective application of drought stress before harvesting. The results verified that drought stress affected physiological characteristics and secondary metabolism of savory which were useful for future work by using metabolic engineering focusing on important species to increase main compounds, such as carvacrol and rosmarinic acid and caffeic acid.

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.