مجله تحقیقات گیاهان دارویی و معطر ایران
سال چهلم شماره 2 (پیاپی 120، تابستان 1403)

Plants have many defense systems to overcome stresses, especially heavy metals. Some heavy metals are part of pigments and enzyme compounds and are also essential elements. In concentrations higher than plants' physiological needs, they are toxic for plants, but some heavy metals such as cadmium and lead are even unnecessary. At low concentrations, they harm plants, and for this reason, heavy metals are considered stressors. Based on this, it is imperative to investigate these metals' effects on plant oxidizing enzymes' activities. In this research, in order to investigate the reaction of oxidizing enzymes and non-oxidizing factors against the stress caused by cadmium (Cd), the required artichoke after being prepared from the mountains of Kurdistan in the presence of phosphate buffer, pH 7 and PMSF 0.02 solution as Protease inhibitor was homogenized and after centrifugation at 3000 g and 15000 g, the upper clear solution was used as a crude extract and for subsequent measurements. In the performed tests, the effect of different concentrations of cadmium chloride (CdCl2) were measured on the content of proline (Pro), phenolic compounds (Ph.C) and the activity of antioxidant enzymes such as phenyl-alanine-ammonialyse (PAL), catalase CAT), superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and polyphenol oxidase (PPO) of artichoke root. This project was carried out in the form of a completely randomized design in three replications in vials containing three milliliters of artichoke root extract with seven treatments, in which the first group was treated as a control in the presence of distilled water only. The second group was exposed to 0.25 mM CdCl2, the third, fourth, fifth, sixth, and seventh groups were exposed to 0.5, 1, 2, 5, and 10 mM CdCl2, respectively, for 10 minutes. The samples were treated at room temperature of 20 to 25 degrees and 10 minutes of natural light. Then, the activity level of each antioxidant enzyme and the level of Pro and Ph.C was measured separately. In this research, at first, the distribution diagram of the data (enzyme or protein activity against different levels of Cd) was drawn, and then various linear and non-linear regression equations were used to fit the data. In the next step, according to the best type of equation and analysis of the response type of GPX, SOD, APX, protein, Pro and Ph.C was used from nonlinear regression analysis (power, hyperbolic, asymptotic exponential and asymptotic growth models) and to express the response of enzyme activity of PPO, CAT and PAL to different levels of Cd were used from the beta model.
All models indicated Cd's stimulating effect on the studied enzymes. The research results showed that the activity of all SOD, APX, CAT, PPO, GPX, and PAL enzymes as well as the content of Pro and Ph. C increased significantly after Cd treatment. Using five non-linear regression models, the highest activity of APX enzyme at a concentration of 4.6 mM (asymptotic exponential model), GPX enzyme at a concentration of 12.3 mM (asymptotic exponential and asymptotic growth models), SOD enzyme at 14.3 mM (asymptotic growth model) was interpolated. In addition, the most production of Pro and Ph. C was obtained at concentrations of 13.6 and 14.3 mM Cd using the asymptotic and asymptotic growth models, respectively. The highest activity of the PPO enzyme was at 8 mM, CAT at 4.8 mM, and the maximum activity of PAL was at 1.4 mM. The results showed that Cd, due to the induction of oxidative stress and the increase in free radical production, leads to an increase in the content of proline, phenolic compounds, and the activity of antioxidant enzymes in artichoke roots. Further changes in the activity of these enzymes during growth indicate the presence of enzymatic regulatory mechanisms in artichoke roots against heavy metals such as cadmium. Enzymatic antioxidant defense systems, Pro and Ph. C play a crucial role in the response of artichoke roots to heavy metal cadmium stress.
In general, results showed that Cd, due to the induction of oxidative stress and the increase in free radical production, leads to an increase in the content of proline, phenolic compounds, and the activity of antioxidant enzymes in artichoke roots. Further changes in the activity of these enzymes during growth indicate the presence of enzymatic regulatory mechanisms in artichoke roots against heavy metals such as cadmium. Enzymatic antioxidant defense systems and Pro and Ph. C are crucial in the response to heavy metal cadmium stress of artichoke roots.

The Chamestan region, located in Mazandaran province, boasts unique ecological and cultural diversity. In this context, the present study aimed to investigate and document the ethnopharmacological knowledge of medicinal plants in the Chamestan area. No previous ethnobotanical reports have been recorded in this region, making the identification and introduction of medicinal plants crucial for improving healthcare and treatment conditions in the area. This research can contribute significantly to the local community's well-being and medical practices.
Ethnopharmacological information about medicinal plants in this region was collected using a semi-structured questionnaire in 1402 (solar Hijri calendar). Chamestan is situated on the outskirts of the northern natural forests. The city was divided into urban and rural sections, and samples were selected from the area with local guides. Data were gathered through semi-structured interviews, group discussions, and field walks. A total of 24 participants (13 men and 11 women) took part in the study, with 20 randomly selected and 4 purposively chosen as key informants. Demographic characteristics such as education level, gender, and age group were recorded. This research aimed to access indigenous knowledge and traditional applications for identifying medicinal plant specimens. To achieve this, information was gathered from local inhabitants, herbalists, and individuals knowledgeable about traditional medicine and medicinal plants through interviews. For quantitatively analyzing ethnobotanical data of medicinal plants, a software tool (MS Excel 2021) was used to classify and employ descriptive statistical patterns, including relative frequency of quotations, usage value, and informant consensus factor. This study aimed to enrich the understanding of native knowledge and traditional uses of medicinal plants. By documenting this knowledge, the research aimed to contribute to the identification and potential applications of medicinal plant resources. This could significantly benefit the Chamestan community's healthcare and well-being.
In this study, medicinal plants collected belonged to 21 families, with the families Lamiaceae, Rosaceae, Apiaceae, Amaryllidaceae, and Brassicaceae being the most abundant with 28.6%, 19.19%, 19.19%, 14.3%, and 9.5% respectively. The most relative frequency of quotation (RFC) was observed for Thyme, Borage, Nettle, and Tarragon, in descending order. Regarding the International Classification of Primary Care (ICPC), the largest level of agreement among informants about gastrointestinal, neurological, and renal diseases. The aerial parts (leaves and flowers) were the most commonly used plant parts, constituting 30% and 13% of the remedies, respectively, while the least used parts were the roots and skin, accounting for 3%. According to the findings, the plants with the highest RFC were Thymus serpyllum and Urtica dioica. The highest consensus of knowledge was found among informants regarding gastrointestinal, neurological, and renal conditions, with IFC values of 0.6, 0.5, and 0.3 respectively. The most consumed species were Thymus serpyllum, Echium amoenum, Urtica dioica, and Mentha mozaffariani. The Lamiaceae family, represented by Thymus serpyllum and Mentha mozaffariani, exhibited positive effects on gastrointestinal issues. In addition to medicinal uses, the Chamestan people consume numerous plants as raw vegetables or in combination with rice, making their diet rich in valuable constituents. Other plants used include Mentha pulegium, Allium ampeloprasum, Ficaria fascicularis, Allium ursinum, Rumex elbursensis, and Amaranthus retroflexus.
Due to the rich diversity of medicinal plants and the close relationship of indigenous people with nature, extensive use of herbal remedies and meeting life's needs was expected in this region. However, the complete transfer of knowledge between generations has not occurred, possibly due to changes in people's lifestyles and urbanization. Considering the abundant flora and local indigenous knowledge, supportive measures are recommended for identifying, preserving, and increasing the number of valuable medicinal plant species. Collecting and documenting this region's knowledge, especially from elderly individuals, is crucial.

Savory (Satureja hortensis L.) is one of the most important medicinal plants that grows in different parts of the world, including Iran. Water deficit has adverse effects on physiological processes such as photosynthesis, nutrient absorption, development and division of cells, and accumulation and transfer of nutrients, thus reducing the performance of plants. In contrast, biofertilizers play a prominent role in the growth and development of plants by improving the absorption of nutrients and sufficient water through changes in the physicochemical properties of the soil. This study aimed to investigate the effect of drought stress, growth-promoting bacteria (PGPRS) and vermicompost on the yield, amount of nutrients and essential components of the savory plant.
A factorial experiment based on a randomized complete block design with three replications was conducted at the Research Farm at the Mohaghegh Ardabili University, Iran, from 2016-2017. Treatments included three levels of drought stress (Full irrigation (S1), Water cut at 50% of flowering (S2), and water cut in early stages of flowering (S3), four vermicompost levels (including 0, 1, 1.5 and 2 t. ha-1) and included inoculation with microorganisms at three levels (non-inoculation, inoculation with Streptomyces and Pseudomonas fluorescens p15). Before planting, the seeds were inoculated with biological stimulants. Then, seeds were cultivated in rows at a distance of 25 cm between the rows, 20 cm on rows and at a depth of 0.5 cm. To apply the irrigation treatment, the field's soil was investigated in the horizon of the plant roots up to a depth of 20 cm by digging a profile. The weight method was used to control water. Field capacity (FC) and permanent wilting point (PWP) were measured using intact samples 24 hours after soil saturation for field capacity and placed in a suction device with 15 atmospheres of negative pressure for permanent wilting point. The corresponding values for the soil at a depth of 0 to 30 cm were determined according to the layering of the soil, and from their difference, the amount of usable soil water (AW) was determined. During the test, the usable water (depending on the plant's growth stage) was determined by the soil sampling cylinder and transferred to the oven in the form of moisture percentage by weight. The examined parameters included plant height, shoot dry weight, root dry weight, and concentration of phosphorus, potassium, and essential oil components.
Based on the obtained results, drought stress, vermicompost, and inoculation with PGPRS had a significant effect on the measured parameters. So that the maximum plant height (62.36 cm), shoot dry weight (28.72 grams) and root dry weight (6.35 grams), phosphorus (1.3 percent) and potassium absorption (3.3 percent) were obtained under non-stress conditions, application of vermicompost two tons per hectare of and inoculation with Streptomyces bacteriaand the lowest amount of plant height (22.33 cm), shoot dry weight (18.33 grams), dry weight roots (3.07 grams), phosphorus (0.85 percent) and potassium (1.2 percent)  were observed in the planting beds without vermicompost application and non-inoculation with biological stimulants and in severity drought stress. Also, the highest amount of carvacrol (62%) and gamma-terpinene (26.9%) was obtained in the treatment of severe drought stress and beds containing two tons per hectare of vermicompost in plants inoculated with Streptomyces.
Vermicompost with growth-promoting bacteria can be a sustainable agricultural fertilizer containing beneficial microorganisms that improve the growth performance and essential components of savory plants under drought stress and non-stress conditions.

Madder (Rubia tinctorum L.) is a perennial and herbaceous plant. Madder's root is known as its most important economic part, and it is rich in anthraquinone derivatives. Madder is a plant with medicinal, nutritional, color and health benefits. Recent pharmaceutical studies have shown that the anthraquinones of the madder root have selective inhibitory effects on cancer cells. In this research, according to the water and soil limitations of the tested area, the use of biodegradable calcium carbonate and phosphate fertilizers to produce madder was suggested. So, considering climate change and water shortage as severe risks to sustainable development, the cheapness of madder production and its multiple uses, the planting and production of this plant are mainly used in semi-arid areas. 
In field conditions in Ardakan, Yazd, this research was tested in factorial form in a randomized complete block design with two factors and four replications. Calcium carbonate agent was supplied at five levels of 0, 20, 50, 100 and 150 kg hˉ1 and phosphate from triple superphosphate fertilizer at four levels of 0, 50, 100 and 150 kg hˉ1. In each four-by-four-meter plot, four rows of seeds were planted, two side rows and two plants from the sides of the remaining lines were removed as marginal effects, and the rest of the plants were considered statistical populations. Irrigation water's electrical conductivity was measured during the growth period, and its average was 11.6 ds mˉ2. The plots were irrigated immediately after planting the seeds. Four days later, irrigation was done again. In the fall of the second and third years, soil was again given to the plants along with the desired treatments. Madder plants were harvested in the third winter year. Root fresh-weight biomass was weighed immediately after harvesting. After drying the roots at 104 °C, the dry weight of the roots was measured. A spectrophotometer also determined the content of anthraquinone and hydrogen peroxide in the roots.
The results of bivariate analysis of variance of the data showed that the simple and reciprocal effect of different calcium carbonate and phosphate treatments on functional indicators of biomass, root fresh weight, root dry weight, anthraquinone content and root hydrogen peroxide in this research was significant at P≥ 0.05. The comparison of averages showed that the highest values of biomass, root wet weight, root dry weight, anthraquinone and hydrogen peroxide were observed in the combined treatment of 20 kg h-1 of calcium carbonate and 100 kg h-1 of phosphate, compared to the control. The lowest value of performance indicators investigated in this research was observed in the combined treatment of 150 kg h-1 of calcium carbonate and 150 kg h-1 of phosphate, compared to the control. The simultaneous use of these two fertilizers had a complementary effect on each other. This was because the application of 20 and 50 kg h-1of calcium carbonate and 100kg h-1 of phosphate had an increasing impact on the anthraquinone and hydrogen peroxide content in madder roots. The comparison of the averages showed that in the combined treatment of 20 kg h-1of calcium carbonate and 100 kg h-1of phosphate, compared to the control, the highest wet weight of the root increased by 71%, the highest dry weight of the root increased by 91%, and the highest anthraquinone content increased by 142% was observed. The comparison of the averages showed that in the combined treatment of 50 kg h-1 of calcium carbonate and 100 kg h-1 of phosphate, compared to the control, the highest biomass index was observed, with an increase of 79%.
The results showed that the functional indices of root fresh weight, root dry weight, anthraquinone content and biomass of madder increased significantly under the combined treatment of 20 kg h-1 of calcium carbonate and 100 kg h-1 of phosphate. It is suggested that madder be cultivated using calcium carbonate and phosphate fertilizers to increase the anthraquinone compounds under the influence of biodegradable and inexpensive external environmental stimuli in regions similar to the climatic conditions studied in this research.

Saffron (Crocus sativus L.) is one of the most expensive spices in the world, and due to the high demand for it in the world markets, its cultivation is developing strongly. In addition to genetics, the region's climate and soil characteristics also affect the quantity and quality of the product. Therefore, identifying regions, whose climatic and soil conditions are close to the optimal conditions for saffron production will help us achieve the most favorable quantitative and qualitative performance.
In the present study, saffron was cultivated in six different regions of Kurdistan province (Sanandaj, Kamyaran, Ghorveh, Dehgolan, Divandarreh, and Marivan) as a randomized complete block design, and their functional and phytochemical characteristics were evaluated during three years (2020-2022). The saffron corms used in this research were prepared from Torbat-e Heydarieh city. In each region, land with an area of 25 m2 was selected. After preparation, three plots with dimensions 2x2 m with 1m distances were created. Each experimental plot consisted of 10 rows, and in each row, 20 holes were made with a depth of 15 cm and a distance of 10 cm from each other, and then one corm was planted in each hole. Irrigation was done twice a year (late September and late October). During the flowering season, saffron flowers were harvested, and their functional and phytochemical characteristics (number of flowers, dry flower yield, dry stigma yield, stigma length, crocin, and safranal content of the stigma) were measured. Total monthly precipitation and average monthly temperature for each region during the research period were obtained from nearest meteorological station. The physicochemical properties of soil samples from the studied regions were evaluated in the laboratory. Finally, the mean comparison of data (using Duncan’s multiple range test) and correlations between climate and soil data with functional and phytochemical traits (using the Pearson method) were obtained by SPSS software.
The results indicated that the maximum stigma yield in all three years belonged to saffron cultivated in Kamyaran (0.13±0.009, 1.27±0.085 and 1.54±0.052 g/m2 in 2020, 2021, and 2022, respectively). In all three years, the highest crocin content was obtained from the stigmas harvested from Divandarreh farm (53.01±15.77, 92.54±26.6 and 143.07±5.12 mg/g dry stigma in 2020, 2021 and 2022, respectively). The highest mean of safranal content in the year 2020 (4.09±0.38 mg/g dry stigma), 2021 (2.48±0.44 mg/g dry stigma), and 2022 (5.05±0.14 mg/g dry stigma), belonged to the saffron of Kamyaran (no significant difference with other regions except Marivan), Marivan (no significant difference with Dehgolan and Divandarreh) and Divandarreh (no significant difference with other regions except Kamyaran), respectively. Dry flower yield and stigma yield showed a significant negative correlation with clay (r= -0.329* and r= -0.356**, respectively), organic matter (r= -0.422** and r= -0.428**, respectively) and nitrogen (r= -0.437** and r= -0.444**, respectively), and a significant positive correlation with potassium (r= 0.544** and r= 0.561**, respectively) and phosphorus (r= 0.410** and r= 0.435**, respectively). The crocin content of stigma had a significant positive correlation with clay (r= 0.557**) and pH (r= 0.438**) and a significant negative correlation with sand (r= -0.461**). Safranal had no significant correlation with soil characteristics and altitude. Also, significant correlations were obtained between the measured traits and average monthly temperature and total monthly precipitation.
Generally, regions of Kurdistan province with less rainfall in spring, summer and early autumn; more rainfall in late autumn and during winter; higher average monthly temperature; with medium-textured soils containing more phosphorus and potassium, lower nitrogen and organic matter, and slightly alkaline pH, were identified as more suitable regions for saffron production. Based on all measured quantitative and qualitative traits, among the studied regions, Kamyaran had the most suitable conditions for saffron production.

Maintaining medicinal plants' quality and effective compounds is a fundamental challenge. Salvia L. has antioxidant, anti-inflammatory, and antimicrobial properties and is a rich source of biological compounds that destroy free radicals. Salvia virgate Jacq. has several biological activities and is used to treat wounds and various skin diseases and prevent blood cancer. Cadmium is one of the most toxic heavy metals due to its long biological half-life, significant mobility in soil, and capacity to be absorbed by plants. Cadmium's negative effects on plant growth, and physiological and biochemical activities reduce plant quality and performance. Sodium nitroprusside is involved in biological processes and responds to various stresses by releasing nitric oxide (NO).
To investigate the effect of different levels of sodium nitroprusside on improving the response of salvia virgata to cadmium stress under in vitro conditions, an experiment was conducted in the Research Institute of Plant Sciences of the Ferdowsi University of Mashhad with a factorial design based on a completely randomized design with four replications. The studied factors included different levels of cadmium (0 (control), 25, 50, and 100 µM) and sodium nitroprusside (0 (control), 15, and 30 µM). Murashig and Skoog culture media were used for seed cultivation. In this experiment, different amounts of cadmium nitrate were added to the culture medium before adjusting the pH. Sodium nitroprusside was added to the medium after autoclaving and a sub-laminar filter due to its sensitivity to high temperatures. At first, the seeds were sterilized with 70% alcohol and 2% sodium hypochlorite; then, they were planted in a culture medium. After three months of growth, the seedlings were removed from the culture medium. Growth traits include the fresh and dry weight of the aerial part, root, and whole seedling, and physiological and biochemical traits including photosynthetic pigments (measurement of chlorophyll a, chlorophyll b, carotenoid, total chlorophyll, and total photosynthetic pigment), flavonoid, total phenol, carbohydrate, proline, protein, and antioxidant activity were measured. The data obtained from this research were analyzed by Minitab software. The mean comparisons were done by the Bonferroni test at the five percent probability level.
According to the results of the analysis of variance of the data, the interaction effect of cadmium and nitroprusside on seedling fresh weight, chlorophyll a, chlorophyll b, carotenoid, total photosynthetic pigments, total phenol, carbohydrate and proline at the probability level of one percent and seedling dry weight, total chlorophyll, flavonoid, and protein were significant at the five percent probability level. The interaction effect of treatments on antioxidant activity was not significant, but their simple effects were significant. The results of the mean comparison of the data indicated that cadmium stress decreased the fresh and dry weight of seedlings, chlorophyll a, chlorophyll b, total chlorophyll, carotenoids, and total photosynthetic pigments in S. virgata, and the application of sodium nitroprusside led to the improvement of these traits; so that the application of 30 μM sodium nitroprusside under cadmium stress with a 50 μM concentration showed the highest fresh and dry weight of seedlings and the proline content, and the application of 15 μM sodium nitroprusside under cadmium stress with a 25 μM concentration showed the highest amount of photosynthetic pigments. Also, this research indicated that cadmium stress increased antioxidant activity, total flavonoid, total phenol, soluble carbohydrate, proline, and protein in the S. virgata plant. However, sodium nitroprusside application improved these negative effects of cadmium stress. So, the application of 30 μM sodium nitroprusside led to an increase in total flavonoid, total phenol, soluble carbohydrate, and protein under cadmium stress conditions at 25 μM concentration.
The results showed that cadmium stress reduces plant growth and physiological and biochemical functions, but sodium nitroprusside application could improve cadmium's negative effects. These findings show that sodium nitroprusside can be used as an effective instrument in cadmium stress management in medicinal plants and help to improve the growth and quality of these plants. This research, in addition to enhancing our knowledge about the mechanisms of plants' response to cadmium stress, helps to analyze the effect of sodium nitroprusside in exposure to environmental life pollutants.

Punica granatum L. var. pleniflora is an ornamental/medicinal shrub native to Iran. Gulnar extract, like morphine, reduces pain in patients. It is also used against various diseases such as prostate cancer and breast cancer due to triterpenoids. In traditional medicine, it has been applied to cure diarrhea, bleeding, mucous secretions, and vaginal secretions. In external use, it has been used as a gargle to cure tonsillitis. Atherosclerosis is one of the most dangerous factors in cardiovascular disease. Due to antioxidant compounds, Gulnar reduces artery-clogging by almost 70%. Therefore, it is critical to enhance Persian Gulnar growth and flowering, a natural source of antioxidants.

To investigate the effect of the amino acids tryptophan, phenylalanine, and methionine on the morphophysiological and phytochemical characteristics of the Persian Gulanar, an experiment was conducted in the research greenhouse of Zanjan University in a completely randomized design including ten treatments in three replications and 3 pots for each experimental unit in the research greenhouse, Faculty of Agriculture of Zanjan University. Different levels of treatment included methionine 0.5, 1, 2 mM, phenylalanine 0.5, 1, 2 mM, tryptophan 0.5, 1, 2 mM. The extraction conditions were optimized for total phenol, total flavonoid, antioxidant activity, and antimicrobial activity tests. To extract and optimize the extraction using a dry flower sample, an ultrasonic device was used and an experiment was designed to determine the appropriate solvent, temperature, and duration of extraction, as well as to determine the optimal ratio of plant dry matter to solvent, and a comparison was made between two water solvents and alcohol at temperatures of 30-50-70 degrees Celsius and different amounts of plants 0.2, 0.35, and 0.5 grams per ten milliliters of solvent and durations of 5, 15, 25 minutes. Based on these cases, the initial tests by recommended software. To investigate the antimicrobial activity of the flower extract and Persian Gulnar's red color, a good penetration method was used. Plates containing a solid culture medium of Mueller Hinton agar were used in this method.

The treatments significantly affected the dry weight of flowers and flavonoids at a 5% probability level. In addition, they affected the number of flowers, anthocyanin content, antioxidant activity (FRAP), and chlorophyll b at a 1% probability level. The flower dry weight increased by 97.09% in the presence of 2 mM tryptophan compared to the control. The most anthocyanin was observed with an increase of 56.05 and 50% in the concentration of 0.5 mM phenylalanine compared to the control. The highest amount of antioxidant activity (FRAP) was obtained with a 110% increase compared to the control at a concentration of 2 mM tryptophan. The highest flower number was at 1 mM phenylalanine and 2 mM methionine. The most positive correlation (*r=0.68) was between flower dry weight and branch growth, and the most negative correlation (*r=0.80) was between chlorophyll b and anthocyanin. There was a significant positive correlation between chlorophyll a and total flavonoids, chlorophyll b, and branch growth. Also, there was a significant positive relationship between the number of flowers with anthocyanin and total phenol. The best extraction point was related to the temperature of 70 ºC, 70% ethanol solvent, 15 minutes, and the ratio of plant dry matter to solvent was 0.2 grams of dry weight in 10 ml of solvent. The red flowers showed a higher inhibitory effect against Escherichia coli, and the variegated flowers had a higher inhibitory power against Staphylococcus aureus.

According to the results, in addition to confirming Gulnar's antibacterial activity and determining the optimal extraction point, it was found that the levels of 0.5 mM amino acids in most of the traits, including branch growth, flower dry weight, total phenol, and anthocyanin brought the best results from the treatment. It also justifies them in the field. Since amino acid fertilizers are applied in small amounts, they are economically justified.

In recent years, Urtica dioica L. has attracted the attention of researchers due to its significant effect on many diseases, especially diabetes. However, more research needs to be done in the field of tissue culture and the effect of elicitors on antioxidant properties and the production of its secondary metabolites. This research aimed to investigate the best hormonal combination and light conditions effective on nettle callogenesis and the effect of methyl jasmonate elicitor on its biochemical properties.

For this purpose, using stem explants, the effect of different levels of auxin hormones, naphthalene acetic acid (NAA) (1.5 and 3 mg/l) and 2,4-dichlorophenoxy acetic acid (2,4 D) (0.5 and 1.5 mg/l) and cytokinin hormone, benzylaminopurine (BAP) (0.5 and 1.5 mg/l) were evaluated in two light conditions (light and dark) by measuring fresh weight, dry weight and percentage of callogenesis. The nettle plant's callus growth curve was obtained using liquid culture. After starting the suspension culture, methyl jasmonate treatment was applied with three levels of 1, 0.1 and 0.01 mM, and sampling was done at 24, 48 and 96 hours. The enzyme activity of phenylalanine ammonialyase and polyphenol oxidase and total protein content by extracting in 50 mM Tris-HCl buffer with 8.8 acidity and total phenol, total flavonoid content by extracting in 80% methanol was measured using spectrophotometric and standard methods for control samples and those under different treatments of methyl jasmonate. Also, three metabolites of quercetin, kaempferol and rutin were measured using an HPLC device with a UV detector and 18C column stationary phase. Data from the callogenesis experiment were analyzed in a two-factor experiment in a completely randomized design. The data from the biochemical and HPLC experiments were analyzed in a split-plot in time design, and the means were compared with Duncan's test.

According to the results, the best hormonal combination to produce fresh and dry weight in light conditions includes 1.5 mg/l of NAA and 1.5 mg/l of BAP and in dark conditions, it includes 1.5 mg/l naphthalene acetic acid and 0.5 mg/l of benzyl aminopurine. Also, according to the results, naphthalene acetic acid (62.5%) performed better than 2,4-D (43.5%) in the nettle callogenesis percentage. Lighting was determined as the best light condition for callogenesis in nettle, so the amount of fresh and dry weight and percent of callogenesis in light compared to darkness increased by 4.79, 4.91 and 1.92 times, respectively. There was an increase in polyphenol oxidase enzyme activity, total phenol and total protein in some samples treated with methyl jasmonate compared to the control, and no significant increase was observed in the activity of phenylalanine ammonialyase enzyme and total flavonoid content. According to the HPLC results, the highest amount of rutin metabolite was obtained in 96 hours after the addition of the elicitor and at the concentration of 0.01 mM methyl jasmonate.

The naphthalene acetic acid hormone was more effective than the 2,4-D hormone and light conditions than darkness in nettle callogenesis. Methyl jasmonate elicitor increased the antioxidant property of nettle (48 hours after treatment) by increasing the polyphenol oxidase enzyme activity. However, compared to the control, it had no significant effect on the phenylalanine ammonialyase enzyme activity and total flavonoid. It is suggested that other concentrations of methyl jasmonate be tested to reach optimal conditions for increasing the production of secondary metabolites in nettle in vitro culture conditions.

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.

In different ecosystems, factors such as altitude, rainfall, temperature, lighting, humidity, and soil nutrients are essential variables that affect the metabolism and accumulation of secondary metabolites. Therefore, it is vital to know the factors affecting the active substances of medicinal plants. Therefore, researchers try to increase the production of effective substances by different methods. We can mention the research conducted in the field of the effect of ecological factors on the essential oil compounds of different populations of Nepeta crispa, Salvia sharifii, Satureja rechingeri and Stachys pilifera. This study investigated the relationship between climatic, topographical, and edaphic parameters as necessary factors that influence the morphological traits and essential oil compounds of the medicinal endemic species Dracocephalum ghahremanii Jamzad.

Three habitats of D. ghahremanii were selected in Semnan province with different environmental conditions, and 15 plots of 4 m2 were established by systematic random method. In each plot, the plant height, canopy diameter, cover, biomass and leaf area of the studied species were measured. Flowering branches were collected from natural habitats with three replications, and essential oils were extracted using a hydrodistillation method using a Clevenger apparatus. GC-FID and GC/MS identified the essential oil compounds. Five soil samples were randomly taken from inside the plots in each habitat, and some soil physicochemical parameters, including pH, EC, OM, N, P, K, lime and soil texture, were measured. Analysis of variance and comparison of data mean was done using SPSS software. The most important environmental factors affecting morphological traits and essential oil compounds were determined using the correlation analysis method in SPSS software and the principal component analysis (PCA) method in Minitab software. The conservation status of this species was determined using the IUCN method and based on the criteria of EOO and AOO using GeoCAT software, as well as data related to population size and habitat quality.

 Comparing the average morphological traits of D. ghahremanii showed that the highest and lowest values of morphological traits are related to the Tash region with an altitude of 3043 meters and Tang Kavard with an altitude of 2032 meters, respectively. Variance analysis of morphological traits also showed a significant difference between the plants of the three regions in terms of leaf area at the level of 1% and in terms of plant height, canopy diameter, canopy cover and biomass at the level of 5%. The analysis of D. ghahremanii essential oil showed that its chemical composition differs in three regions. In Tang Kavard region, (E)-β-Farnesene, trans-Cadina-1(6),4-diene and (E)-Caryophyllene, in Dibaj region,(E)-β-Farnesene, trans-Cadina-1(6),4-diene and Caryophyllene oxide, and Tash region, Caryophyllene oxide, α-Vetivone and Carvone hydrate were identified as dominant compounds. Analysis of the variance of common essential oil compounds of this species also showed a significant difference between the averages of these compounds at 0.1%. Physicochemical parameters of D. ghahremanii habitats had significant differences. Variance analysis of the values of these parameters showed that there is a significant difference between the three regions in terms of OM, N, P, K, lime, sand and silt at the level of 0.1%, pH at the level of 1% and EC and clay at the level of 5%. Correlation between the characteristics of this species with some environmental factors showed that altitude, annual precipitation, annual temperature, minimum temperature of the coldest month, maximum temperature of the hottest month, lime, OM, N, P, sand and silt have the most significant correlation with morphological characteristics and essential oil compounds. Considering that the area occupied by this species in the studied populations (AOO) is 0.750 km2 and its presence area in the province (EOO) is 691.046 km2, its conservation status in Iran was determined as Critically Endangered (CR).

As a general conclusion, the increase in altitude has increased the functional characteristics (morphology and phytochemistry) of D. ghahremanii. Despite the relative similarity of the dominant compounds in the essential oil of this species, the composition of the essential oil (percentage of major and minor components) was different in the three investigated regions. Among the environmental parameters, altitude, annual precipitation, OM, N, P, K and silt were the most critical factors affecting the morphological traits and the percentage of oxygenated terpenes, which can indicate the importance of high altitude habitats in terms of essential oil content and compounds with higher antioxidant properties for this plant. Based on these findings, the maximum temperature of the hottest month, the minimum temperature of the coldest month, and the absolute minimum temperature of lime and sand were the most important parameters affecting the percentage of hydrocarbon terpenes. These results can be significant for extracting D. ghahremanii essential oil from natural habitats or commercially cultivating this plant to extract the secondary compounds in its essential oil.

Considering the high damage caused by storage pests and the adverse effects of chemical pesticides, using plant compounds is considered one of the best options for controlling storage pests. Essential oils are an effective plant compound with a high fumigant effect. In this study, Oviposition deterrence and ovicidal activities of three native species of savory were investigated to select the savory essential oil with the highest insecticidal properties.
First, three native species of savory (Satureja sahendica, Satureja khuzistanica, and Satureja macrantha) were collected, and essential oils were extracted after drying. All three essential oils were injected into a gas chromatograph and gas chromatograph coupled with a mass spectrometer to determine chemical compounds. Experiments were performed in a completely randomized design and were evaluated under laboratory conditions (27±1°C, 65±10 R.H. and 16:8 L:D).To determine the oviposition deterrence of the oils, one pair of one-day-old adult moths was exposed to different concentrations of the oils for four days, and the number of deposited eggs was counted. Also, one-day-old eggs were exposed to three oil concentrations at 3, 12, and 25 μl essential oil/l air. The percentage of egg hatching was recorded after 96 hours. The experiment was performed in 5 replications.
The results showed that carvacrol content, as the principal active ingredient of S. khuzistanica, was 91.5%. Thymol (48.59%), p-cymene (59.5%) and γ-terpinene (18.57%) had the highest amount in the essential oil of S. sahendica, respectively. The dominant components of S. macranthaessential oil were p-cymene (59.5%), γ-terpinene and thymol (13.5%). All three essential oils had the Oviposition deterrent activity, and the rate of Oviposition deterrence increases with increasing concentration of essential oils. Different concentrations of three savory essential oils showed that at a concentration of one microliter with S. khuzistanica essential oil, more than 50% oviposition deterrent activity was observed. In comparison, oviposition deterrent activity was observed for two other savory essential oils at a concentration of 3 microliters. Among the investigated essential oils, S. khuzistanica had the highest effect of oviposition deterrent activity on Mediterranean flour moths. Also, there is a significant difference in the ovicidal activity of S. khuzistanica compared to the others, which can be attributed to its high carvacrol content. In contrast to the low insecticidal effect of the essential oil of S. macrantha, it has had a very suitable effect of oviposition deterrent activity.
All three types of savory essential oils had sound oviposition deterrence effects and ovicidal activity, but S. khuzistanicaessential oil in the lowest concentration had better oviposition deterrence effects and ovicidal activity than S. sahendica and S. macrantha, that it is also more acceptable from an economic point of view. Based on the results of this research, the essential oils of native Iranian savory spices can control storage pests; only high volatility and insolubility in water are the main limitations in the widespread use of essential oils. However, the nanoencapsuled essential oil, through the controlled release of active ingredients, overcomes the restrictions of plant essential oil usage in storage. Therefore, more studies are needed to use essential oils in pest control using this method

Nowadays, due to the revelation of the harmful effects of chemical drugs for most users, the desire to use medicinal plants has increased significantly. Basil (Ocimum basilicum L.) is one of the most important medicinal plants of great interest in various pharmaceutical industries due to its valuable and vital secondary metabolites, such as essential oil. However, salinity is considered a major constraint for its productivity. Soil and irrigation water salinity is a major constraint for crop production. In salinity-rich soils, the osmotic pressure increases, and the plant must burn more vital energy to absorb water. Increasing the salinity tolerance of plants for sustainable food production, especially in arid and semi-arid regions, can reduce the need for irrigation and, thus, production costs. Since using calcium nanoparticles increases nutrient utilization efficiency, reducing soil toxicity, minimizing the adverse effects of over-fertilization and reducing the frequency of fertilizer application, the present study investigates the effect of calcium nanoparticles on physiologic and biochemical properties of basil medicinal plant under salinity stress.  The plants were treated with calcium nanoparticles in five levels by spraying with concentrations of 0, 0.25, 0.5, 1 and 1.5 g/l and salt concentrations of 0.75 and 150 mM along with Hoagland solution in a completely randomized design in a greenhouse environment with three replications. Some growth characteristics such as height, number of leaves, leaf area, fresh and dry weight, amount of chlorophyll a, chlorophyll b, total chlorophyll and carotenoid content of the plant were measured. Some biochemical characteristics such as flavonoid, soluble sugar, anthocyanin content and antioxidant enzymes such as catalase and peroxidase were also measured. The least significant difference (LST) test was used to compare the means in SAS software. The results of ANOVA showed a significant effect of calcium nanoparticles on most morphological traits. Also, different salinity levels alone significantly affected the studied traits (p<0.01). Following the reduction of the osmotic potential of the soil solution due to salinity stress, water absorption decreases and as a result of stomata closing, the photosynthesis and respiration decrease. The mean comparison of the simple effect of calcium nanoparticles on chlorophyll pigments showed that with increasing the nanoparticle level, chlorophyll content increased, and as a result, photosynthesis improved. The simple effect of different salinity levels on some traits (phenol, soluble sugar, peroxidase enzyme and anthocyanin) showed that increasing the salinity level caused an increase in these traits. So, at the concentration of 150 mM salinity, phenol content with 8.02 mg of gallic acid per g of dry weight showed the highest amount compared to the control sample of 10.5 mg. Also, the soluble sugar content was the highest at this salinity concentration, with 139.9 mg/g dry weight, compared to the control, with 110.9 mg/g dry weight. The interaction effect of calcium nanoparticles and salinity on basil's morphological characteristics and carotenoid content was insignificant. However, it was significant in the chlorophyll content, the amount of flavonoids and catalase (p<0.01). In fact, with nano fertilizers, the plant can absorb more nutrients, and while reducing the leaching of elements, the yield of the product increases. The results of this study suggest that the physiological and biochemical characteristics of Basil were affected by salinity stress, and the interaction of nano calcium with salinity stress reduced the effects of salinity stress. Therefore, spraying the nano-fertilizer in an appropriate concentration is suggested to improve the yield of basil in salinity stress conditions.