مجله تحقیقات گیاهان دارویی و معطر ایران
سال سی و پنجم شماره 4 (پیاپی 96، مهر و آبان 1398)

Diabetes mellitus is associated with physical, physiological, and pathological changes in the liver tissue. The aim of this study was to investigate the protective effects of hydroalcoholic extract of Crataegus monogyna Jacq. on histopathology and liver function in streptozotocin (STZ)-induced diabetic rats. In this experimental study, Wistar male rats were randomly divided into six groups of eight including control, diabetic, diabetic treated with 100, 200, and 400 (mg/kg) doses of C. monogyna extract and positive control (diabetic treated with captopril). Diabetes was induced by intra-peritoneal injection of a single-dose of STZ (60 mg/kg). After observation of the diabetes symptoms (blood glucose level above 250 mg/dL and polydipsia), treatments were done using extract and drug. After three weeks of treatment with extract and drug intra-peritoneally, the serum level of liver enzymes and albumin was determined and liver tissue samples were prepared for histopathologic examination. In diabetic rats, there was a significant decrease in serum albumin and a significant increase in serum glutamic-pyruvic transaminase (GPT) and gamma glutamyltransferase (GGT) in comparison with healthy rats (P<0.05). Treatment of diabetic rats with different doses of C. monogyna extract increased albumin and decreased GGT in serum significantly (P<0.05). In diabetic rats, hepatocytes necrosis, complete destruction of the liver lobules and Remak bundles and hyperemia were observed, and the treatment with C. monogyna extract prevented the pathologic changes. The results of this study showed that C. monogyna extract can prevent liver tissue degradation and its dysfunction in diabetic rats.

Domestication, cultivation and crop production of Bunium persicum (Boiss.) B. Fedttsch. are important to preserve genetic resources of the species. In this regard, in order to identify the proper medium and nutrient solution for the seedling establishment of B. persicum, a factorial experiment in a completely randomized design was carried out under greenhouse conditions with three replications. Experimental factors consisted of culture medium at three levels of cocopeat, sand and soil, and Hoagland solution in five concentrations of 0, 25, 50, 75, and 100%. The results showed that the highest seedling emergence percentage (37.7) was observed in cocopeat without Hoagland solution. The highest seedling vigor index (773) and also green area of seedling (2.14 cm2/plant) and the lowest seedling vigor index (205) were observed in cocopeat and soil media (both with 75% Hoagland solution), respectively. Maximum tuberization percentage (100) was recorded in the sand with 50% Hoagland solution. The highest tuber weight (76.0 mg) and volume (79.2 mm3) were obtained in the sand with 75% Hoagland solution without any significant difference with the results of sand with 50% Hoagland solution treatment. Generally, the results of this study indicated that using a medium with suitable porosity and water holding capacity and also irrigation with at least 50% Hoagland solution can improve seedling emergence percentage, seedling establishment, and production of tuber with appropriate size in B. persicum.

The study of the plant compounds efficacy for pest control is one of the expanding researches in most agricultural systems. In this research, the efficacy of aqueous and ethanolic extracts of four plants including dwarf elder (Sambucus ebulus L.), common nettle (Urtica dioica L.), blackberry (Rubus fructicosus Boiss.) and eagle fern (Pteridium aquilinum (L.) Kuhn) was assessed against the adult female of Tetranychus urticae, one of the most important pests of crops and greenhouse plants. The acaricidal effect of the extracts and their long-lasting effectiveness were studied as well as their oviposition deterrent effect. Two experiments included the long-lasting effectiveness (with the concentrations of 0.5, 1 and 1.5 mg/cm2) and oviposition deterrent effect (with the concentrations of 0.4, 0.7 and 1 mg/cm2). The study of extracts was conducted in a completely randomized design. The aqueous extract of blackberry did not have any acaricidal effect on the pest, and this effect varied from 9.2 to 49.8 % for other aqueous extracts. The LC50 values for ethanolic extracts of dwarf elder, common nettle, blackberry, and eagle fern against the adult females of the pest were obtained to be 0.54, 0.82, 0.76 and 0.72 mg/cm2, respectively. The study of long-lasting acaricide effectiveness of the ethanolic extracts showed that the common nettle extract had high long-lasting effectiveness of 48 hours, after which the extracts of blackberry, dwarf elder, and eagle fern were respectively placed. The blackberry ethanolic extract had the highest oviposition inhibitory effect in comparison with other extracts (94% at 1 mg/cm2). The results demonstrated the appropriate efficacy of the ethanolic extracts to control T. urticae pest in comparison with the aqueous ones, and accordingly, evaluating their control capacity under field conditions will provide some practical valuable data. Amongst the ethanolic extracts, dwarf elder displayed the most acaricidal effect.

Betulin and betulinic acid are from the most important anticancer and anti-HIV metabolites, and the birch species (Betula spp.) bark is considered as the primary source of these metabolites. Due to the extinction of these tree species in Iran, it is necessary to replace the metabolites extraction from the birch bark with modern methods such as cell and tissue culture to produce the metabolites. The aim of this study was to determine the effect of explant type on the amount of betulin and betulinic acid produced in calli of two birch species B. pendula and B. litwinowii under in vitro conditions compared to the amount of metabolites extracted from the tree bark. Bark and leaf explants of two mentioned species were cultured in WPM medium containing 1 mg/L BAP and 0.1 mg/L 2,4-D for callogenesis. The amount of betulin and betulinic acid in three-month calli was measured using the HPLC technique and compared with the amount of these metabolites in one-centimeter stem bark samples taken from nature. The bark explant was more successful in callogenesis, and calli derived from this explant had more active ingredients. The amount of betulin and betulinic acid from the extract of bark sample taken from nature was respectively obtained to be 5.23 and 2.91 percent for B. pendula, and 5.65 and 2.52 percent for B. litwinowii. Moreover, calli derived from the bark explant of B. pendula and B. litwinowii contained 0.023 and 0.016 percent of betulin and 0.053 and 0.057 percent of betulinic acid, respectively. Generally, the results indicated that the bark explant was more capable of callogenesis and secondary metabolite induction than the leaf explant in both birch species under in vitro conditions.

Tomato early blight disease, caused by Alternaia solani fungus, is one of the most important diseases of tomato in the world, and also in Iran. In order to investigate the antifungal activity of some medicinal plants to control this fungus, separate experiments were conducted in a completely randomized design under in vitro and greenhouse conditions. In the first experiment, the effect of concentrations of 200, 400 and 600 ppm of hydro-alcoholic extract (70% methanol and 30% water) of Thymbra spicata L., Thymus eriocalyx (Ronniger) Jalas, Satureja khuzistanica Jamzad and Oliveria decumbents Vent. with the negative (distilled water) and positive (Daconil, 3 g l-1) controls on the growth of A. solani fungus was studied under in vitro conditions. The results of the first experiment indicated complete inhibiting of fungal colony growth in all treatments. In the second experiment, 72 hours after contamination of tomato plants with A. solani fungus in an isolated greenhouse, contaminated plants were sprayed with the above-mentioned treatments and monitored for the disease symptoms 14 days later. In the third experiment, the protective effect of the above-mentioned treatments was studied. The results of the second and third experiments showed that with an increase in the extract concentration, the inhibitory and protective effects of the treatments increased. Although all treatments reduced the disease severity compared to the control, the concentrations of 600 ppm of T. spicata, 600 ppm of T. eriocalyx and 400 ppm of T. spicata, respectively had the most effective inhibitory and protective effects on the growth of fungus studied. Therefore, the use of the above treatments is recommended as a practical method for biological control of A. solani.

Drought stress, as one of the most important abiotic stresses, plays an important role in decreasing the yield of medicinal plants. In order to investigate the effect of nitric oxide on morphophysiological and biochemical characteristics of basil (Ocimum basilicum L.), the native population "Shahre-ray", a factorial experiment was conducted in a completely randomized design under drought stress conditions with three replications in the research greenhouse of Mohaghegh Ardabili University in 2017. Experimental factors included drought stress at three levels of complete irrigation (drip irrigation at three-day intervals throughout the growing season) and complete stop of irrigation at early flowering stage (60 days after planting at 10-12 leaf stage) and 50% flowering (75 days after planting at 14-15 leaf stage) and nitric oxide spray with sodium nitroprusside at four concentration levels of zero (leaf spray with distilled water), 0.5, 1, and 2 mM. The foliar application was carried out from the eight-leaf stage of the plant, with intervals of two weeks to four times. Results showed that stopping irrigation at both mentioned growth stages significantly reduced plant height, plant dry weight, chlorophylls a and b, total chlorophyll, carotenoid, and relative water content, and significantly increased electrolyte leakage rate, leaf proline content, and essential oil percentage. Foliar application of 2 mM nitric oxide significantly increased plant growth under irrigation interruption, especially at 50% flowering. Increasing the concentration of nitric oxide produced the highest essential oil percentage (1.4%) at the early flowering stage under irrigation interruption conditions and significantly increased the content of essential oil components including methyl chavicol, linalool, geranial, geraniol, α-pinene, β-pinene, and myrcene. In general, the results showed that foliar application of nitric oxide at 50% flowering stage under irrigation interruption improved the growth of basil and increased the percentage and components of essential oil via osmotic adjustment, preventing chlorophyll degradation and maintenance of leaf water content.

The vascular obstruction due to the growth of microbial agents, especially bacteria, is one of the most important reasons for reducing the quality and vase life of cut flowers in the postharvest stage. In order to investigate the effect of extract (1 and 2 %) and essential oil (EO) (25 and 50 ppm) of Thymbra spicata L. and Satureja bachtiarica L. on bacterial population and vase life of cut flower Gerbera jamesonii L. var. Pink Elegance, this experiment was designed as a factorial based on a completely randomized design with three replications. The results showed that the experimental treatments, by reducing the bacterial population at the end of flower branch, maintained the quality and increased the vase life of flowers (up to three days in treatment with 50 ppm of S. bachtiarica EO, compared with the control samples. Also, the antibacterial activity and vase life of cut flowers increased by increasing the concentration of EO and extract of both medicinal plants. After eight days of storage, the results showed that the maximum vase life (11 days), relative absorption of vase solution (8.95 ml/g FW per day), and the lowest bacterial population of the end of the flower branch were related to 50 ppm of S. bachtiarica EO treatment. However, in terms of relative fresh weight, total soluble solids, anthocyanin content, ion leakage, malondialdehyde content, and the activity of catalase, and phenylalanine ammonia lyase enzymes, there was no significant difference between two treatments of 50 ppm EO from medicinal plants T. spicata L. and S. bachtiarica L. Therefore, the use of 50 ppm EO of S. bachtiarica, and in the second rank of T. spicata, is recommended as a practical method in the process of postharvest and marketing of gerbera cut flowers.

Varroa destructor (VD) tick is one of the most important pests of honey bee colonies around the world, causing a lot of economic damage to the beekeeping industry in the world by feeding on bee larva and transferring some viruses. The use of chemical pesticides against this tick creates resistant populations and infects hive products. In this study, the level of repellent toxicity of different essential oil (EO) concentrations of Satureja khuzistanica Jamzad and S. spicigera (C. koch) Boiss., on the mortality of VD tick and honey bee (HB) was evaluated under laboratory conditions. Mature female ticks were treated with different EO concentrations including 0, 50, 75, 100, 150 and 200 ppm (at 32 °C, 50% relative humidity and darkness) for 5 and 10 hours, in four replicates, and after determining the equation of concentration vs. VD mortality, LC50 was calculated for concentrations of 20 to 1000 ppm. According to the results, the amount of LC50 at 5 and 10 hours after EO treatment was respectively determined as: for S. khuzistanica EO, 149.8 and 78.6 ppm for VD tick and 750.5 and 435.8 ppm for HB, and for S. Spicigera EO, 255.7 and 153.6 ppm for VD tick and 737.6 and 573 ppm for HB, indicating the more sensitivity of ticks in comparison with honey bees to applied EOs. The results showed that the EO of both species, especially S. khuzistanica, with the least mortality on HB, has a high potential for controlling VD tick. The main component(s) of essential oils was obtained as carvacrol (93%) in S. khuzistanica and carvacrol and thymol (43% and 29%, respectively) in S. spicigera based on qualitative analysis.

In this research, the effect of time (10, 15, 20, 25 and 30 min) and temperature (35, 45, 55, 65 and 75 oC) of ultrasonic waves, ethanol to water ratio (20, 35, 50, 65 and 80%) of extracting solvent and dry matter to solvent ratio (1:5, 1:10, 1:15, 1:20 and 1:25 mg/ml) was investigated on total phenol and flavonoids and antioxidant capacity (in two methods of DPPH and FRAP) of hydroalcoholic extract of celery (Apium graveolens L.) aerial parts by using the response surface method. Based on the results, ultrasound for 10 min at 55 °C, ethanol to water ratio of 80%, and dry matter to solvent ratio of 1:5 were considered as the best treatment. The highest amount of total phenol (471.1 mg GAE/g DW) and flavonoid (187.7 mg quercetin/gDW) content, and antioxidant activity of the extract was obtained under optimized conditions using DPPH (68.5%) and FRAP (4210.4 μM Fe+2/50 μl extract). In general, the high values of adjusted R2 and P-value in fitting models indicated that the drawn models of response surface were ideal. In other words, the high amounts of these parameters indicate the existence of a correlation between the observed values and the predicted ones.

Two species of Nepeta assurgens Hausskn. & Bornm. and N. eremokosmos Rech., are endemic medicinal plants to Iran. This study was conducted to investigate the vegetative traits, aerial parts yield, the quantity and quality of essential oil, as well as evaluating the relationships between these traits in mentioned species under field conditions in Isfahan. The experiment was carried out in a randomized complete block design with three replications over 2017 and 2018. Results showed that the fresh and dry weight of aerial parts in N. assurgens (11353 and 2351 kg ha-1, respectively) was remarkably higher than that of N. eremokosmos (2638 and 527 kg ha-1, respectively). The average content and yield of essential oil in N. assurgens and N. eremokosmos was 1.60 and 0.64%, and 20.16 and 2.21 kg ha-1, respectively. Based on qualitative analysis of essential oil, the total amount of nepetalactone isomers (4aα, 7α, 7aα-nepetalactone, 4aα, 7α, 7aβ-nepetalactone, 4aα, 7β, 7aα-nepetalactone) in N. assurgens and N. eremokosmos was 78.6 and 62.8% on average, respectively. Correlation analysis showed that leaf dry weight, total fresh and dry weight, and large diameter of the plant had the highest correlation with essential oil yield. Stepwise regression analysis indicated that essential oil yield changes were mainly explained by leaf dry weight, essential oil content, total fresh weight, the number of stems and the amount of plant crown cover. In general, according to the results, the superiority of the N. assurgens species was significant and could be recommended as a valuable and high potential yielding plant for pharmaceutical purposes.

Licorice (Glycyrrhiza glabra L.) is one of the oldest and most important medicinal plants in Fabaceae, used for curing many diseases since 4000 years ago. This study was conducted to evaluate the genetic diversity of 22 different licorice populations based on morphological and yield traits at the research field of the Faculty of Agriculture, Zanjan University, during 2016 to 2018. Morphological and yield traits including plant height and width, leaf length and width, number, length and width of leaflets, number of lateral branches, main stem diameter, aerial parts fresh and dry weight, root fresh and dry weight, root to aerial parts ratio and aerial parts and root yields (per m2) were measured. Canonical discriminant (CDA) and cluster (CA) analyses were used to group the populations. In CDA, the first two canonical variables were significant. The first canonical variable included plant height and width, main stem diameter, leaf length and the number of leaflets, and the second one included aerial parts fresh and dry weight, root fresh and dry weight, root and aerial parts yields. The second canonical variable had the greatest role in population separation and grouping. Canonical variables divided populations into four main groups and confirmed CA clustering results. In general, the results indicated the good potential of canonical discriminant analysis in evaluating the genetic diversity and identifying the index traits in licorice.

Nowadays, RNA sequencing is an effective and fast choice to study the transcripts of non-model plant species used to identify gene networks and patterns of gene expression producing secondary metabolites in different plant organs. Terpenoids, flavonoids, and alkaloids are the main compounds in fruits and leaves of Citrullus colocynthis (L.) Schrad. In this study, the transcriptome of C. colocynthis fruit was performed using RNA-Seq technique, Iluumina HiSeq2500 platform. After quality control using FastQC and Trimmomatic software, 21,952,885 high-quality reads were produced and the de novo assembly with the Evidential-gene program resulted in the production of 55,311unigenes with an N50 equal to 927 base pairs. The sequence of assembled unigenes was loaded on the KAAS database. A total of 13,657 unigenes were annotated, matched into 134 biosynthetic pathways. The "terpenoid backbone" biosynthetic pathway with 93 unigenes was one of the most numerous identified pathways among 1,552 unigenes of secondary metabolites biosynthetic pathway. By examining the unigenes associated with the biosynthetic pathway of the terpenoid backbone, 29 gene identifiers (K number) of the pathway were detected, which contained all identified genes of the two main biosynthetic pathways, Mevalonic Acid (MVA) and Methylerythritol Phosphate (MEP) pathways, from the beginning of the path to the production of isoprenyl diphosphate (IPP). Identification of the genes in the biosynthetic pathway of the terpenoid backbone makes it possible to realize the commercial development of the medicinal plant products, providing the basis for further research on the identification of biosynthetic pathways of other specific metabolites, metabolite engineering, molecular breeding, and medical plants breeding.

In order to investigate the effects of growth regulators on yield and agronomical characteristics of saffron (Crocus sativus L.), an experiment was conducted in a randomized complete block design in the research greenhouse of Faculty of Agriculture, University of Zanjan. Experimental treatments included corms priming with gibberellic acid (GA) (250, 500 and 750μM), salicylic acid (SA) (700, 1400 and 2100μM), paclobutrazol (PBZ) (50, 100 and 150μM), chlormequat chloride (CCC) (500, 1000 and 1500 μM), hydropriming (HP) (distilled water), and control (treatment without any growth regulator). Among the treatments, the effect of GA was more pronounced than other growth regulators, so that the highest dry yield of flower and stigma was observed in GA treatment, especially at 500-μM level. CCC and PBZ caused the lowest flower and stigma yield and the highest amount of chlorophyll pigments compared to other treatments. In contrast, the use of GA and SA reduced the chlorophyll content. HP treatment did not show any significant difference with control. SA treatment increased safranal (perfume) and crocin (color) of the stigmas. The highest effect on picrocrocin (flavor) was obtained in CCC treatment.