محمد مجدی

Papaver plants from the poppy family are among the famous medicinal plants that have always been the focus of the pharmaceutical and medical industries due to the production of morphine alkaloids and isoquinoline alkaloids with antimicrobial effects. A wide variety of Papaver species can be seen in the pastures and mountains of Kurdistan province. In this research, the methanolic extracts prepared from four Papaver species were investigated about the amount of morphinan alkaloids produced and the antimicrobial effects on the growth of microorganisms in laboratory conditions.
Four Papver species including P. glaucum, P. fugax, P. argemone, and P. bracteatum were collected twice from their natural habitats in Kurdistan province in spring and were identified based on morphological characteristics under the supervision of a botanist of the province's agricultural and natural resources research center. In order to extract alkaloids, aerial parts, and roots were dried and powdered in the shade. Extracts were prepared by adding methanol and chloroform solvents in a ratio of 1 to 4 to the resulting powders of plants. The amount of morphine, codeine, thebaine, and papaverine alkaloids present in each of the samples was measured separately using high performance liquid chromatography (HPLC). Also, the antimicrobial effects of extracts against Gram-positive bacteria, Bacillus subtilis, Staphylococcus aureus, Streptococcus pyogenes, and gram-negative bacteria, Pseudomonas aeruginosa, Salmonella typhi, Escherichia coli were investigated. The pathogenicity was evaluated by measuring the growth halo, determining the minimum inhibitory concentration (MIC) and the minimum lethal concentration (MBC), using the tube dilution method.
The production of morphine alkaloids in all of the studied species was confirmed by HPLC. The amount of morphine as the dominant alkaloid among the four studied species varied from 4.21 to 21.33 mg/g dry weight. The highest amount of morphine (21.33 mg/g) was observed in P. bracteatum extract and the highest amount of thebaine and codeine (8.67 and 1.8 mg/g) was observed in P. fugax extract. The observation of non-growth aura against Pseudomonas aeruginosa and Staphylococcus aureus bacteria confirmed the antimicrobial effects of the investigated plant extracts. The detected amounts of minimum inhibitory concentration against the gram-positive and gram negative bacteria varied from 25 to 100 mg/ml and minimum lethal concentration was just observed against Escherichia coli. P. bracteatum plant extract showed the highest inhibitory effect against E. coli bacteria with a minimum inhibitory concentration of 25 mg/ml and a minimum lethal concentration of 50 mg/ml.
By examining the results, it was found that the studied Papaver species produce significant amounts and ranges of morphine alkaloids, which designates these natural species' genetic and medicinal value for further studies. In addition, the extract of these plants showed antimicrobial effects against the reference strains of gram-positive and gram-negative pathogenic bacteria used in this research. Therefore, it is suggested that more studies be conducted in the field of separation and separation of the metabolites of these plants and the investigation of their antimicrobial effects in subsequent studies to achieve effective and novel antimicrobial compounds and drugs.

St. John's wort (Hypericum perforatum L.) is a medicinal plant that is well known for its anti-depressant properties. Hypericin and hyperforin are the two active compounds that have the greatest medicinal value. Considering the importance of these compounds, it is necessary to have solutions to increase their production through biotechnology. Therefore, as preliminary study the expression of two key polyketide synthase genes including HpPKS2 and HpPKS1 in hypericin biosynthetic pathway was studied. Methyl jasmonate was used as elicitor to stimulate gene expression in concentrations of 0, 10 and 100 micromolar before flowering stage. Leaves and stems samples were then collected at 0, 12, 24 and 48 hours after MeJA treatment. The expression level of genes was examined by semi-quantitative RT-PCR method. The results showed that the expression level of both HpPKS2 and HpPKS1 genes were increased in leaf and stem tissues due to the application of MeJA, and at the concentrations of 10 and especially 100 µm, there was a significant difference with the control (0 concentration). Moreover, the expression level of both genes showed a rising trend in the time series of 0, 12, 24 and 48 hours after spraying MeJA in both tissues, and generally the greatest increase in expression was observed at 48 hours. The results of this study showed that both polyketide synthase genes are induced under the influence of methyl jasmonate and lead to their higher expression.

MicroRNAs (miRNAs) are a group of short non-coding RNA molecules with a length of about 18-24 nucleotides, which control target genes expression at the post-transcriptional level. There are no reports of miRNA detection in Rheum ribes plant. Therefore, in the present study, in order to predict potential conserved miRNAs and their target genes in R.ribes roots, total RNA was extracted from the root tissue and sequenced. Then, non-coding transcripts were identified in R.ribes transcriptome and considered as miRNA precursor candidate sequences. In this study, 82047 transcripts were created from R.ribes root by de novo assembly method. By searching the homology of these transcripts with mirbas database and secondary structure prediction, 4 conserved miRNAs belonging to mir396, mir164, mir319 and mir168 families were identified. 574 transcripts from R.ribes root transcriptome were targeted by these miRNAs. In general, due to the regulatory role of identified miRNAs on a wide range of genes, these miRNAs can be used to identify genes related to secondary metabolites.

One of the most harmful pests of Zagros forests is the Tortrix viridana (Lep. Tortricidae). Genetic diversity of Tortrix viridana host plant populations in the oak forests of northwestern Iran and the northern Zagros region was investigated using 28s gene sequence. The samples were collected from the forest areas of west Azarbaijan, Lorestan, Kurdistan and Kermanshah provinces. They were in the larval stage, were kept in laboratory conditions until they turned into pupa and then into a complete insect. DNA extraction was done by CTAB method. Also, in order to amplify the 28s region, the 28s gene sequence of Tortrix genus was used from NCBI for primer design. The desired region was amplified using the PCR method and the PCR products were sequenced. 21 samples were selected to investigate genetic diversity using the 28s gene, and 18 sequences DNAs were of suitable quality for further investigations. The DNA sequences were edited using Bioedit software and aligned using MegaX software, and the phylogenetic tree was drawn by UPGMA method with 1000 sampling repetitions. The evaluation of the genetic structure of populations showed that the diversity between populations is greater than within populations. The results of the phylogenetic tree also showed that different samples of the Tortrix viridana have genetic diversity based on geographical distance. Therefore, the time of appearance of the pest, their behavior and their type of control and management are also different.

Eurygaster integriceps is one of the known pests of wheat fields in Iran and West Asia. The role of neuropeptides in the stages of insect’s growth has led to a promising perspective for the production of a new generation of bio-insecticides based specific application. Insect’s neuropeptides along with their specific receptors are one of the most diverse proteins that control physiological and behavioral activities in insects. Allatostatin is one of the important neuropeptides in insects which, by inhibiting the youth hormone, plays a role in regulating physiological processes such as feed and metabolism in some insects. In this study, using the information obtained from adult Eurygaster integriceps transcriptome, neuropeptides and specific receptors of the allatostatin family were investigated. Bioinformatics and phylogenetic studies of the data led to identify four neuropeptides A, B and C allatostatin family, as well as the neuropeptide receptors of A and B allatostatin. The results showed that the neuropeptides of the allatostatin family identified in Eurygaster integriceps are involved in various physiological processes. Considering the important role of neuropeptides in insects, these neuropeptides can be used to design specific insecticides compatible with the environment for managing control Eurygaster integriceps in future.

Since the introduction of L-dopa (3, 4-dihydroxylphenyl-L-alanine) in the 1960s, L-dopa has been recognized as a gold-standard medical therapy for Parkinson’s disease. In the present work, a newly tyrosine-transforming strain of Paenibacillus sp. CT4W was used for the biotransformation of L-tyrosine to L-DOPA. The process parameters were optimized using single- factor experiments and Taguchi design methodology under resting cell strategy. Based on the values obtained in single-factor experiments, the optimum parameters for L- DOPA production: cell mass 6g/l, Cu+2 0.045 g/l, Temperature 30º C, pH 7, agitation 150 rpm and 0.5 g/l of yeast extract as cosubstrate. The maximum yield achieved with these parameters was 0.29 g/l after 16 h incubation. A Taguchi L16 orthogonal array was subsequently employed for further optimization. The biotransformation reaction having 5 g/l cell mass, 1.5 g/l tyrosine and 0.03 g/l Cu+2 was found to be optimum for maximum L-DOPA production. Under the optimized conditions, resting cells of Paenibacillus sp. CT4W produced 0.96 g/l L-DOPA, with a molar yield of 53.5 % after 20 h incubation.

Sahendian savory (Satureja sahendica Bornm) is one of the native species of Iran that contain valuable secondary metabolites. The high value of the active ingredients of Sahendian savory has caused the need for methods for rapid Therefore, in this study, an in vitro tissue culture method was optimized for Sahendian savory. First, explants were prepared from sterile seedlings. Plant growth regulators treatments used for callus induction in MS medium included 2,4-D at three levels with BA (Benzyl Adenine) at four levels or NAA at two levels with BA at five levels (0, 0.5, 1, 2 and 5 mgl-1). BA treatment was also used at five levels for shoot induction. Callus formation of explants showed different reactions under the influence of different hormonal compounds. The explants showed different callogenic reactions under the consequence of different plant growth regulators combinations. The combination of 1 mgl-1 2,4-D plus 1 mgl-1 BA and the combination of 0.5 mgl-1NAA with 0.5 mgl-1 BA showed the highest callus induction rate (100%). The calli produced were green and had a spherical and firm texture. Additionally, the highest number of regenerated shoots (3 shoots) from calli were related to concentrations of 4 and 5 mgl-1 BA. These results can be used as a simple and appropriate technique in the regeneration of Sahendian savory plants for in vitro micropropagation or gene transfer.

Triticum aestivum L. is an important food crop that provides more than 40% of the per capita calories and protein in the diet in many developing countries. Wheat production plays an important role in food security and the global economy. Sunn pest is one of the most serious pests of wheat in Asia, North Africa, and Eastern Europe. Yield losses caused by Sunn pest are estimated at 50-90% in wheat in case of pest out break. The use of chemical pesticides is the most common method of controlling Sunn pest. Although chemical pesticides have decreased in recent years, the damage caused by the chemicals, and how long they remain in the environment are the causes of most worrying. Thus, finding more effective alternative methods to be safer are a necessity. The Promising new technologies are attractive on many levels, mainly because they have been developed to target specific species and eradicate pest populations more gently. Many researchers worldwide have performed several studies to control various pests using new techniques such as RNA interference, genomic modification, and microbiome studies. The use of new methods may play effective roles in controlling the sunn pest of wheat and reduce some concerns.

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.

Phenological, molecular and metabolite analysis during vernalization in wheat show that there is a tight relationship between vernalizatio fulfillment/reproductive transition with decrease in the accumulation of some metabolites and expression of cold-induced genes/proteins. In the present review, we will describe a number of similar protective metabolites and cold related proteins which are present or expressed in both sensitive and tolerant wheat genotypes. Based on several studies duration of cold acclimation in winter genotypes is quite higher which is influenced by developmental genes and consequently the level and duration of expression of cold-lnduced proteins is longer when compared with spring genotypes. Also, the initial rate of cold acclimation in cold tolerant winter wheat is higher than the cold sensitive genotypes. Our results show that the expression patterns of cold related proteins and metabolites are consistent with the trend of the developmental factors that have regulatory effects on proteome/metabolome. In addition, sub-zero acclimation of plants in field conditions during autumn/winter seasons has significant effects on numbers, levels and patterns of cold associated proteins in compared with plants acclimated in controlled condition at low temperature but above zero temperature. This present work, provide a novel insight to uncover the molecular mechanisms behind cold adaptation in wheat and pave the way to breed wheat for cold tolerance.

Medicinal plants of the Asteraceae family are important genetic resources due to their high echological flexibility in diverse climates,. Yarrow (Achillea millefolium subsp. millefolium) is a self-pollinated plant belongs to the Asteraceae which produce a wide varieties of plant secondary metabolites such as terpenoids and phenylpropanoids. In the present work, gene expression patterns of genes involved in the biosynthetic pathways of terpens and phenylpropanoids were studied to understand the regulatory mechanism behind them. 1-deoxy-D-xylose-5-phosphate reductoisomerase (DXR) and geranyldiphosphate synthase (GPPS) are important genes in the biosynthesis of monoterpenes in the 2-C-methylerythritol-4-phosphate (MEP) pathway. Phenylalanine ammonia lyase (PAL) and chalcone synthase (CHS) genes are important in the biosynthesis of phenylpropanoids. In the present study, semi quantitative RT-PCR of these genes was carried out in different developmental stages, also in response to methyl jasmonate treatment in leaves and flowers. Results showed that expression of these genes were higher in flowers and methyl jasmonate treated plants, also the expression of these genes affected by developmental stages of leaves but with different intensity.

Nicotine is a highly toxic alkaloid which can be found in tobacco processing industry. Due to the serious environmental problems and therapeutic concerns which have occurred because of presence of toxic nicotine in the environment, biodegradation of this compound by microorganisms has received considerable attention. The objective of the present study was to screen the moderately halophilic bacteria which are able to degrade nicotine and to evaluate the possibility of using the bacterial biocatalysts to clean up nicotine in contaminated environments.
Screening and selection of halophilic bacteria degrading nicotine has performed in three separate steps using enrichment culture technique, nicotine as the sole carbon and nitrogen source, and tolerance patterns. The best nicotine-degrading bacterial isolate (designated as strain ND9) was characterized on the basis of morphological and biochemical phenotypes and 16S rRNA sequencing. Growth kinetics and the removal rate of nicotine were evaluated using spectrophotometry and high performance liquid chromatography (HPLC).
Findings: According to the results obtained, growing cultures of Halomonas sp. strain ND9 (accession no. KM077028) showed the highest tolerance to nicotine and also degraded over 92% of nicotine (initial concentration of 2 g/L) as the sole source of carbon and nitrogen after 96 h incubation time.
Discussion and The present work describes the potential of moderately halophilic bacteria to be used in a de-nicotination process for the first time. Given the potential of Halomonas sp. strain ND9 in the bio-removal of nicotine, isolation and characterization of moderately halophilic bacteria as green bio-catalysts for their application in the bio-degradation of nicotine has been proposed.

To investigate the effects of nano-chelate spraying of Iron, chemical -synthesis and green-synthesis silver nanoparticles on the expression of two genes, germacrene A synthase (TpGAS) and parthenolide synthase (TpPTS) genes which are involved in the parthenolide biosynthesis pathway of Tanacetum parthenium L., a factorial experiment based on randomized complete block design was conducted with three replications. The experiment took place in the greenhouse of Zabol University in Iran. At first, the total RNA from samples of leaves, and then the synthesis of cDNA were extracted. Afterward, Real Time PCR analysis was used to determine gene expression patterns. Parthenolide concentration was measured by high-performance liquid chromatography (HPLC). The analysis of the data was conducted by SAS software, and there was a significant increase in the expressions of both germacrene A synthase (TpGAS) and parthenolide synthase (TpPTS) genes of treated plants with green-synthesis silver nanoparticles in compare with the control plants, and some plants which were treated by chemical- synthesis silver nanoparticles and nano-chelate of Iron. Furthermore, the maximum expressions of both germacrene A synthase (TpGAS) and parthenolide synthase (TpPTS) genes were identified in groups of plants which were under the influence of drought stress, or were treated by green-synthesis silver nanoparticles. A positive relation was seen between the expressions of germacrene A synthase genes (TpGAS) and parthenolide synthase (TpPTS) genes with the amount of parthenolide itself. It can be concluded that green-synthesis silver nanoparticles and water deficit stress can cause an increase on the expressions of parthenolide and germacrene A synthase genes, and as result of that, cause an increase on parthenolide production of Tanacetum parthenium.

Thymus vulgaris is one of the most important medicinal plants with more than 20 types of known important chemical compositions. A gene named 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR) acts as an important control point, as this is the first committed step in the MEP pathway. γ-terpinene is a key component in essential oil of thyme which constituted up to 30% of the essential oil in the species and is biosynthesized via γ-terpinene as a penultimate precursor. Different cytochrome P450 including CYP71D178 and CYP71D182 are involved in the biosynthesis of thymol and carvacrol in thyme and oregano via the γ-terpinene as an intermediate. The aim of the present work was to study the expression patterns of genes involved in thymol and carvacrol biosynthesis including DXR, γ-terpinene synthase, CYP71D178 and CYP71D180 genes in flowers and leaves of thyme. Gene expression analysis of DXR, γ-terpinene synthase, CYP71D178 and CYP71D180 genes were carried out by semi-quantitative RT-PCR technique. The relative gene expression of flowers compared with leaves for DXR, γ-terpinene synthase, CYP71D178 and CYP71D180 genes were 1.7, 2.7, 1.9 and 2 fold higher, respectively and the gene expression intensity varied for each gene. Expression of aforementioned genes in glandular trichomes were high, hence the higher gene expression in flowers might be related to higher density of glandular trichomes. Based on these results, higher content of essential oils in flowers compared to leaves is most likely further related to the higher expression of related genes in flowers rather than leaves.

Foxglove plants are important sources of medicinal cardiac glycosides compounds such as digoxin and digitoxin, which are used for treatment of congestive heart failure and cardiac arrhythmia. Digitalis has several valuable species but the only species native to Iran is digitalis nervosa. Considering the importance of study on native plants in order to be used in pharmaceutical industry this research was carried out. One of the key genes in the biosynthesis of digitalis cardenolide in plants is 3β-HSD, which encodes the enzyme 3β-HSD with important role in early stage of the cardenolide biosynthesis pathway. Purpose of the present study was to determine nucleotide sequence of 3β-HSD, phylogenetic analyses and its expression under influence of salicylic acid and methyl jasmonate elicitors in different tissues. Results revealed that a sequence encoding the enzyme show very close affinity to Digitalis lanata. In addition expression pattern of 3β-HSD in the transcript level revealed that a significant expression level was detected in leaf tissue, and also under treatment of methyl jasmonate and salicylic acid the expression level raised. Findings of our study could be used to increase cardenolide through metabolic engineering and genetic manipulation.

The gene 1-deoxy-D-xylulose 5-phosphate synthase (DXS) and TcGLIP multifunctional genes are two key genes involved in pyrethrins biosynthetic pathway, which retain insecticidal properties. DXS gene in the MEP pathway produces 1-deoxy-d-xylulose 5-phosphate using pyruvate and glyceraldehyde-3-phosphate dehydrogenase. TcGLIP gene is involved at the last steps of pyrethrins biosynthesis pathway and integrates chrysanthemyl-CoA and pyrethrolone, which are produced in the MEP pathway and lipoxygenase pathways, respectively to yield pyrethrin I. Based on the significances of pyrethrins, further identification and studies are needed on these metabolites. The aim of this study was to isolate DXS gene in pyrethrum (Chrysanthemum cinerariaefolium Vis.) from the Asteraceae, as well as expression analysis of these two genes under methyl-jasmonate treatment. In the present work, the nucleotide sequences of 1-deoxy-D-xylulose 5-phosphate synthase gene of different plant species were obtained from NCBI database and subsequently aligned with ClustalW online software. Specific primers were designed from the conserved regions of aligned sequences. The polymerase chain reaction product with 715bp length showed that the specific primers had high efficiency to isolate DXS. In addition, gene expression analysis under methyl-jasmonate treatment showed that both DXS and TcGLIP were up-regulated in response to methyl jasmonate elicitor. Our data showed that methyl jasmonate treatment in seedling stage could be used as a suitable elicitor to increase pyrethrin production.

Aim: The aim of this study was to investigate the expression of key genes, 1-deoxy-D-xylulose-5-phosphate reductoisomerase )DXR( and gamma-terpinene synthase (GTS), involve in thymol and carvacrol biosynthesis pathway in Summer savory (Satureja hortensis). This species is one of the important medicinal plants of the Lamiaceae family, and consider as an important source of the mentioned compounds.
Material and Plants were treated with salicylic acid, methyl jasmonate and UV-B rays. RNAs were extracted from control and treated plants and cDNAs were synthesized. Primers were designed for gene isolation and expression studies. Transcript expression analyses for the DXR and GTS were performed using semi-quantitative RT-PCR method.
A partial segment for DXR and GTS genes was sequenced. The relative gene expression analyses showed differential expression of both genes at transcript level in different tissues (roots, stems, leaves and inflorescence), with higher levels of expression in leaf and inflorescence. The expression of both genes under the effect of abiotic elicitors including salicylic acid, methyl jasmonate and UV-B rays exhibited significant alteration.
Under controlled conditions, using abiotic elicitors such as: salicylic acid, methyl jasmonate and UV-B radiation could elevate the level of gene expression and possibly increase the production of secondary metabolite such as: thymol and carvacrol.

Black cumin (Nigella sativa) is one of the most important medicinal plants belongs to Ranunculaceae. In spite of its significance in pharmaceutical industry, tissue culture and micropropagation of the species is not well studied yet. Hence, based on the issue we aimed to study its callogenesis and regeneration on leave explants of the species. In vitro provided explants of leave were cultured on MS medium supplemented with NAA (0.5, 1, 1.5 mgL-1) and BAP (0.2, 0.5, 1 mgL-1). Half MS medium supplemented with 0.5, 1 and 1.5 mgL-1NAA and IBA were used for rooting the regenerated shoots. Results showed that higher percentage of callogenesis was observed on MS medium supplemented with 0.5 mgL-1NAA and 0.5 mgL-1 BAP. Shooting was observed on MS medium (1mgL-1BAP) and rooting on half MS (1 mgL-1NAA), respectively. Results indicated that callogenesis, shooting and rooting of black cumin are highly affected by hormonal combination levels.

Black cumin (Nigella sativa) is an important medicinal plant belongs to the Ranunculaceae which produce large diversity of secondary metabolites including industrial and medicinal monoterpenes/ triterpenes. In the present work, relative gene expression of genes involved in biosynthetic pathway of terpenes including a monoterpene synthase (MTS), geranyldiphosphate synthase (GDS), β- amyrin synthase (βAS) and squaleneepoxidase(SQE) were investigated in black cumin plants treated with methyl jasmonate. The plants were grown in a greenhouse sprayed with 0.1 mM methyl jasmonate. Samples were collected at 0, 4, 8, 24, 48 and 72 hours after applying the treatments. RT-PCR was performed by gene-specific primers after RNA extraction and cDNA synthesis. Results of semi quantitative RT-PCR showed that the expression pattern of GDS gene differed with the results of other genes. The highest gene expression of GDS was observed at 4 h and its expression remained at the same level till 8 h then dropped at next time periods. The expression patterns of monoterpene synthase, β-amyrin synthase and squaleneepoxidase showed rising trend. Gene expression analysis of the genes at transcript level using semi-quantitative PCR in black cumin leaves treated by methyl jasmonate revealedinduced-geneexpression which might show the roles of terpenes in defense mechanisms and signal transduction. Consequently, methyl jasmonate treatment can be considered to induce the terpene biosynthetic pathway and to study the enzymes/ metabolites associated with terpenoid metabolism.

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.

Black cumin (Nigella sativa) is an important medicinal plant belonging to the Ranunculaceae which produces large diversity of secondary metabolites especially terpenes. Mono (C10) and tri-terpenes (C30) are two major structural classes of terpene compounds. Several genes are involved in biosynthetic pathway of the monoterpenes known as MEP pathway، located in plastids. In the MEP pathway، monoterpene synthases are rate- limiting enzymes for monoterpene biosynthesis. In the biosynthetic pathway of monoterpenes geranyl diphosphate synthase catalyzing geranyl diphosphate (GDP) biosynthesis as universal precursor of monoterpenes. Oxidosqualenecyclases is of the most important. In this study gene expression patterns of a monoterpene synthase، GDP synthase، beta amirin synthase and squalene oxidase were evaluated under salicylic acid treatment. Gene expression analysis of the genes at trasnscript level using semi-quantitative PCR in response to salicylic acid treatment revealed their differential expression.

Feverfew (Tanacetum partheniumL. Schulz Bip.) is a medicinal herb belonging to the Asteraceae which recently raised researcher’s attention due to its medicinal value and pharmacological activities¡ especially as a migraine prophylaxis agent and also for treatment of cancer. Parthenolide has a sesquiterpene lactone structure which is most likely synthesized through the mevalonic acid (MVA) pathway. Recently¡ it has been shown that there is cross a talk between the MVA and the MEP pathways through IDP (Iosopentenyl diphosphate) exchange as a precursor for the biosynthesis of different terpenes; hence¡ parthenolide biosynthesis could be affected by the MEP pathway as well. In the present work¡ the relative gene expression of two key genes of the MVA pathway¡ including 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) and germacrene A synthase (GAS) and two key genes of the MEP pathway including 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) and hydroxy-2-methyl-2-(E)-butenyl4-diphosphate reductase (HDR) were examined in leaves derived from the vegetative or generative phases¡ also in flowers using real time PCR. The results of our study showed that expression of these genes depend on the growth stage and genotype. Moreover¡ in which in the flowers and leaves derived from vegetative phase in different genotypes only the TpGAS gene expression showed a significant difference¡ while in the leaves derived of generative phase the relative gene expression showed a significant difference for TpHMGR¡ TpGAS and TpDXR.

Monoterpenes are one of the major groups of terpenes which are present in black cumin (Nigella sativa L.) as limonene، carvacrol، thymoquinone etc. One of the key genes in monoterpene biosynthesis is geranyl diphosphate synthase (GPPS) that produces a monoterpene precursor. The aim of this study was to investigate the expression of the genes in black cumin. RNA was extracted، cDNA was synthesized and a partial sequence of GPPS gene was isolated using degenerate primers and sequenced. Moreover، RNA was extracted from different tissues of the species and semi-quantitative RT-PCR using specific primers of GPPS gene and GAPDH as housekeeping gene in order to study the pattern of GPPS gene expression. Results showed that transcript expression of geranyl diphosphate synthase in stem and leaf is significantly higher than that of root، capsule and seed.