مجله تحقیقات گیاهان دارویی و معطر ایران
سال بیست و ششم شماره 4 (پیاپی 50، زمستان 1389)

Application of Biofertilizers in a sustainable agriculture system improves sustainability of yield especially in medicinal plants production. Anise (Pimpinella anisum L.) is a medicinal plant containing essential oils especially in its seeds. In this reserach, the effects of vermicompost and phosphate biofertilizer on yield and yield components in anise including plant height, number of umbels per plant, 1000 grain-weight, biological yield and seed yield were studied. The experiment was carried out at Homand research station - Iran in 2009. The factors were vermicompost (0, 5, 10 ton/ha) and phosphate biofertilizer (non-inoculated, inoculated seed and inoculated seed + spraying on soil in stem elongation). The experiment design was factorial experiment based on randomized complete blocks design with nine treatments and three replications. Mean comparison was carried out using Duncan multiple range test (P<0.05). Results showed that the highest plant height, umbel no./plant, biological yield, and seed yield were obtained from consumption of 10 ton/ha vermicompost but 1000 grain- weight were not affected by vermicompost significantly. Phosphate biofertilizer also showed significant effects on umbel no./plant, biological yield and seed yield (except plant height and 1000 grain-weight). The maximum umbel no./plant, biological yield and seed yield were obtained from two times consumption of phosphate biofertilizer. There were positive and synergistic interactions between factors like interactions between factors on biological yield. According to the results of this study, application of 10 ton/ha vermicompost and two times consumption of phosphate biofertilizer were determined as the most suitable treatments.

In this research, proline, soluble sugars, catalase and glutathione peroxidase levels which change in response to water deficiency and the effect of ascorbic acid as protecting factor in Nigella sativa L. were surveyed. Plants were cultured in greenhouse and irrigated based on the following treatments: 1) Control with irrigation equal to field capacity, 2) two third of field capacity, 3) one third of field capacity, 4) two third of field capacity along with 10 mM ascorbic acid, 5) two third of field capacity along with 1 mM ascorbic acid, 6) one third of field capacity along with 10 mM ascorbic acid, 7) one third of field capacity along with 1 mM ascorbic acid. Proline and soluble sugars levels in shoots and roots were significantly different for all treatments except for the treatment with 2/3 field capacity along with 10 mM ascorbic acid. Enzymes levels in 1/3 and 2/3 treatments were significantly different in both shoots and roots. One mM ascorbic acid along with different field capacity showed no reducing effect. However, 10 mM ascorbic acid especially in 2/3 field capacity could reduce the effect of water deficit through reduction in level of metabolites produced in response to low irrigation.

Drying is one of the important pre-processing methods of medicinal plants after harvest that has a major role in quantity and quality of their active substances. To determine the effect of different drying methods on drying time, essential oil content and composition of Savory (Satureja hortensis L.), the experiment was carried out during 2008. The experimental design was randomized complete block design with three replications and treatments were: two temperatures: 50°C and 70°C, six microwave powers: 100, 180, 300, 450, 600 and 900 w and drying in shade and under sun light. The drying process was continued until the mass of the sample reduced to a moisture content of about 0.10 on a dry weight basis or 10% on a wet weight basis. Results indicated that different treatments of drying method had significant effects on drying time and essential oil content. The maximum essential oil content (3%) obtained from drying at 70°C and drying in shade while minimum content (0.9%) obtained from drying under sun light. One hundred and 300 w microwave powers had an average content of essential oil (2.3%). Carvacrol content percentage in treatments of microwave drying was more than other methods as maximum percentage (63.9 %) was obtained from 300 w microwave drying with a little difference with 180 w microwave drying treatment. Drying by low microwave powers is recommended for Satureja hortensis L. due to the reduction of drying time and considerable maintenance of essential oil content and composition. Maximum γ-terpinene content (28.2%) obtained from drying at 70°C with a low difference with 50°C, 100 and 300 w microwave.

Because of the economic and medicinal importance of Rosa damascena Mill. the propagation of this plant was considered. In order to protect the selected genotypes, this experiment was performed by simple layering method during autumn and winter seasons 2007 on new growth twigs of Rosa damascena mature shrubs. The branches were laid in sand bed, soaked for 24 h by different concentrations of IBA and NAA hormones. The results showed that the rate of rooting was high in sand bedding material in autumn and winter. The highest rooting percentage was %100 with 50 mgl-1 concentration of IBA in autumn and winter seasons and the lowest of rooting percentage was %53.33 with 50 mgl-1 concentration of NAA in autumn season. The rooting branches were cut from the parent plants after 3 months and planted in pots containing soil: sand: peatmoss complex (1:1:1) in greenhouse conditions, survival percentage was 90%.

In order to evaluate sثرثقشم ecological factors and morphological traits and also to determine ploidy levels and chemical composition of essential oil of Thymus pubescens Boiss. & Kotschy ex Celak two natural habitats, Bostan-Abad and Yam habitats, were chosen in East Azerbaijan province. To study and evaluate quantitative morphological characters, five complete plants and flowering branches were collected from each locality at flowering stage for essential oil extraction. Seeds also were collected in seed production season to study seed characters and chromosome counting. Ecological data of both localities also were recorded. Vegetative and reproductive traits of each population were studied and essential oil of each locality was analyzed by combination of GC-FID and GC-MS. At least 10 mitotic cells of each population were studied to determine chromosome number and ploidy levels. Results revealed that specimens from “Yam” had the longest flowering stems, the largest leaves and flowers, and also the highest number of flowers per inflorescence. Chromosome count results showed that specimens from Bostan-Abad were hexaploid (2n = 6x = 90) and those of Yam were tetraploid (2n = 4x = 60). The total yield of oils based on dry weight was 1. 3% and 2. 2% (w/w) from Bostan-Abad and Yam, respectively. Thymol was the main valuable phenolic compound similar to most of the species of genus Thymus (Bostan-Abad: 57. 6% and Yam: 44. 9%).

Oliveria genus belongs to Umbeliferae family and only has one species, Oliveria decumbens Vent endemic to Iran. On the other hand, essential oils are of secondary materials in plants widely used in the food, hygienic and pharmaceutical industry, and also as antimicrobial compounds. Oliveria decumbens was collected from northwest mountains of Kuh-dasht in Lorestan province. Afterward, plant materials were dried in shade and hydro distillated using a Clevenger apparatus. The oil was analyzed by capillary GC and GC/MS. The anatomical studies were carried out using distaining method with Bismark brown and methyl green. GC and GC/MS analysis of the essential oil resulted in identification of 16 compounds representing 96.4% of the total oil. Anatomical studies showed that the surface of leaves was covered by non-glandular hairs while in cross section of the stems, the secretory canals were observed in cortex region.

Castor bean (Ricinus communis L.) is one of the most important medicinal plants used in pharmaceutical, cosmetics and hygienic industries in many developed countries. Oil obtained from plant's seeds is of the most valuable aperients in medicine. The purpose of this research was determining the effects of climatic factors (seven different locations) on oil content and fatty acids composition of castor bean plant and increasing quantity and quality of oil. The results showed that climatic factors had significant effects on oil content and fatty acids composition and there were significant differences among the different climates (P<0.01). The highest oil content was reported from West of Tehran (52%) and Marand (51%) regions. Based on the results of fatty acid analysis by Gas Chromatography (GC), eight major fatty acids were observed in castor oil. Ricinoleic acid (18:1 Δ9c-12OH) was known as the major oil fatty acid in Castor bean. The highest Ricinoleic acid was reported from Urmia region (88.94 percent) and the lowest Ricinoleic acid was recorded from Shabestar region (85.72 percent). Other fatty acids identified from various regions including linoleic acid (2.25 to 4.73 percent), palmitic acid (0.9 to 2.13 percent), oleic acid (3.05 to 4.22 percent), stearic acid (0.55 to 1.54 percent), linolenic acid (1.35 to 2.88 percent), dihydroxystearic acid (0.51 to 0.85 percent) and eicosanoic acid (0.86 percent).

In this research, the effect of different harvesting stages on essential oils content and composition of Mentha piperita L. and Salvia officinalis L. was studied. The rhizome of Mentha piperita was planted at 5 cm soil depth in the field of "Jangalkar Ferdos Company", 2008. The salvia seeds were received from Isfahan research center. Their seedlings were transplanted on the furrows of the main field. The experimental design was randomized complete blocks and the treatments included early, full and after flowering stages. The experiment included six treatments and three replications in a total of 18 experimental units. After drying the samples in room air condition and removing all woody parts, essential oils of both species were obtained by water-distillation method in three replications and then analyzed by GC & GC/MS. According to the results obtained from analysis of variance, the effect of different harvesting stages on oil yield of Mentha piperita (P<0.05) and Salvia officinalis (P<0.01) was significant. Comparison of treatment means showed that the highest percentage of essential oil (2/8%) was recorded in full flowering stage of M. piperita while S. officinalis had the highest percentage of essential oil in early flowering stage (3%), with a significant difference in comparison to the other stages. Analysis and Identification of essential oils components showed that the major constituents of essential oil in S officinalis were α-thujene (20/8%, 27/1%, 35/9%) camphor (29/2%, 14/6%, 17/2%) and β-thujene (15/1%, 14/6%, 4/1%) in early, full and after flowering stages respectively. The major constituents of essential oils in M.piperita at early, full and after flowering stages were menthol (27/7%, 26/9%, 27/0%), menthon (37/0%, 21/9%, 17/2%), and menthofuran (16/0%, 22/0%, 25/3%).

Plantago species generally are grassy plants that belong to Plantaginaceae, with large lamina on the ground surfaces. Plantago major L. grows widely in Asia, Europea, Africa, and North American and contains chemical compounds such as polysaccharides, lipids, coffeic acid derivatives, flavonoids, terpenoids, ascorbic acid, benzoic acid, ferrolic acid, galactoronic acid and so on. In this study, Plantago major L. plants were cultured in several pots irrigated with hougland solutions and after 20 days, during 3 leaves stage, irrigation was continued with hougland solutions containing different concentrations of CuSO4, 0 (control), 100, 300, 500, 700 and 100µM. Antioxidant enzymes activities (catalase, peroxidase, poly phenol oxidase, ascorbate peroxidase) and phenolics content in leaves and roots were measured. The aim of the study was the effect of copper concentrations on antioxidant enzymes and phenolic compounds content in plantago major L. The result shows that by increasing concentration of copper in the medium, activity of catalse, ascorbate peroxidase and peroxidase in the root and leaves decreased significantly (0.05) in comparison with control. Polyphenol oxidase enzyme activity in roots and leaves increased, that was only significant in roots. Phenolics content increase in roots that was not significant but the decrease in leaves was significant.

Essential oil from aerial parts of Salvia bazmanica Rech. f. & Esfand, an endemic species to Iran, was extracted using hydro-distillation method and analyzed by GC and GC/MS. Twenty-one compounds were identified in essential oil of this species through comparing retention indices and mass spectra of each compound with that of standard compounds. β-pinene (37.5%), β-phellandrene (15.3%), α-thujene (9.5%) and α-pinene (8.9%) were determined as major constituents in the essential oil. The oil was richer in monoterpenes than in sesquiterpenes.

Dragonhead (Dracocephalum moldavica L.) is an annual plant that belongs to the (Labiate or Lamiaceae) family. The essential oil of dragonhead has natural antibacterial and antimicrobial substances and having wide usage in industries like pharmaceutical, dietary and many others. In this research, effects of different harvest times on content and composition of essential oil were studied in Research field at Khoy Agricultural Research Center in West Azarbaijan province. Dragonhead seeds from both landrace and modern cultivars were planted simultaneously. The aerial parts of plant were harvested in six stages. After drying the plant materials in shade, their essential oils were obtained by hydro-distillation. The oils were analyzed by GC and GC-MS. According to the results, maximum and minimum essential oils percentage for both landrace and modern cultivars were respectively observed in full flowering (0.53) and yellow-maturity stage (0.07). In addition, there were some differences between oil compositions of these dragonhead accessions at different harvest times. The major oil components of dragonhead were the geranial, geraniol and geranyl acetate. The highest and the lowest sum total amount of geranial and geraniol were obtained at the early flowering (65.1%) and yellow-maturity (44.9%) for the landrace cultivar. The largest amount of geranyl acetate (50.7%) was gained for the modern cultivar at the early flowering stage while the least one (28.1%) was recorded for landrace cultivar plants 26 days after flowering when 10 to 30 percent of the flowers remained. The results showed that different harvest times had significant effects on essential oil content and composition.

Artemisia genus belongs to Astraceae family with 34 species in Iran which two species, A. melanolepis Boiss. and A. kermanensis Podl, are endemic. In Iran and China this plant is highly noteworthy in traditional medicine. The purpose of this study is to determine the chemical composition in essential oil of Artemisia turcomanica Gand. Leaves after different hours of distillation and also identify essential oil composition of Artemisia turcomanica Gand. stems after four hours distillation. The essential oils of Artemisia turcomanica Gand. leaves were extracted after one, two and three hours of distillation by Clevenger apparatus and were analyzed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). Generally, thirty-nine volatile components were identified on the basis of mass spectra characteristics, retention indices and 13CNMR spectroscopy. Thirty compounds were identified after one hour distillation from which linalool (21%), 1,8-cineole (19%), camphor (17%) and cis-chrysanthenyl acetate (16%) were the major constituents. The results of GC-MS analysis of the essential oil after two hour distillation revealed the presence of twenty-five compounds from which the major constituents were cis-chrysanthenyl acetate (24%), bornyl acetate (15%), linalool (13%) and camphor (9.3%). After three hours, the analysis of the oil showed thirty compounds. The major constituents were cis-chrysanthenyl acetate (12%), selin-11-en-4-α-ol (8.7%), bornyl acetate (8%), cis-jasmone (4%) and camphor (3.6%). Eighteen compounds were identified in the volatiles from the stems of A. turcomanica which were rich in cis-chrysanthenyl acetate (29%), bornyl acetate (18%), camphor (9%) and 1,8-cineole (6.9%). The amount of oxygenated monoterpenes was the highest after one hour distillation (86.7%) which gradually decreased in the other two samples (72.7% and 38.2% respectively). In contrast, the content of sesquiterpenes and oxygenated sesquiterpenes increased in the last hour of distillation.

Ferula gummosa Boiss. is an endangered plant species of the family Apiaceae and endemic to Iran whose mass production is exposed to serious problems due to being monocarpic and prolonged seed dormancy. The first step to improve this precious medicinal, industrial and economic plant is ability in production of numerous sterile plantlets in order to prepare explants of appropriate vigour. In this survey, in vitro culture of galbanum was performed in MS 1/4 medium by a factorial trial in the form of completely random design including A factor in 2 levels (embryo culture horizontally; and vertically so that half part of the embryo is inside the medium and cotyledons stand upward) and B in 4 levels (containing 0, 0.5, 1, 1.5 g/L char-coal) in 3 repeats. Several growth traits were measured during 5 weeks. Seed culture was also studied in 4 medium including distilled water, solid MS 1/4, soil and moist filter paper. Embryo culture ways were resulted in vigorous and numerous plantlets in shorter time, in comparison with seed germination ways. Finally the best option was vertically culture of embryo in MS 1/4 medium containing 0.5 g/L char-coal. Obviously this method will provide the plant with a very similar condition to growth in natural environment.

The current study was performed to evaluate the effect of planting date and locality of seed on seed yield of Pimpinella affinis Ledeb. The experiment design was factorial done using a randomized complete blocks design with three replications. Treatments included locations from where seeds were collected (Khojir, Noshahr and Chalous Road areas) and planting dates (November 6th, December 6th and April 9th in 2006 and 2007). The experiment was performed in Alborz farm research complex located in Karaj. The results of variance analysis of the experiment in 2006 showed that the effect of planting date on seed yield was not significant while the effect of locality and interaction effect between planting date and locality was statistically significant at 1% significance level. Means comparison of treatments showed no significant differences between the yield seed of planting dates of November 6th and December 6th, while December cultivation's yield seed was higher (64.467 Kg/ha). The effect of locality on yield seed was significant as yield seed of Khojir locality (175.1 Kg/ha) was more than that of Chalous and Noshahr localities. Interaction effects between locality and planting date on yield seed were statistically significant at 5% significance level. Yield seed of December cultivation from Khojir locality (191.4 Kg/ha) was the highest amount. The results of variation analysis in 2007 showed that the effects of planting date, locality and their interaction on seed yield were significant at 1% significance level. Means comparison of treatments showed no significant differences between the yield seed of planting dates of November 6th and December 6th, but they had significant difference with April cultivation, while November cultivation's yield seed was higher (218.9 Kg/ha). The effect of locality on yield seed was significant as yield seed of Khojir locality (281.9 Kg/ha) was more than that of Noshahr locality. Interaction effects between locality and planting date on yield seed were significant as yield seed of November and December cultivations from Khojir locality (404.6 and 397.9 Kg/ha) was the highest amount. According to the results of cultivation for two years it can be concluded that autumn cultivation had more yield seed and biological function than that of spring cultivation due to having long growth period. In addition, seeds collected from Khojir locality had the highest yield seed and biological function among three studied localities.