Nova Biologica Reperta

The joint effect of hydro-alcoholic extract of nettle root and metformin on ovarian tissue of diabetic model of Wistar rat

Mahnaz Azarnia

Diabetes mellitus is known to be resistant to insulin, to dysfunction of beta cells and to increase liver glucose production. Poor glucose control during hyperglycemia causes damage to the tissues and creates dangerous consequences, such as infertility. Chronic hyperglycemia has harmful effects on the growth of follicles, which is essential for normal female sexual function. Metformin is the most widely prescribed drug in diabetes, but chemical drugs, despite their undeniable benefits, have destructive effects, so alternative strategies for current modern diabetes medications are essential. Herbal medicines are widely used by patients, based on non-medical recommendations and as blood-glucose-lowering agents, including the nettle. Nettle contributes to lower plasma glucose levels by secreting insulin and increasing the proliferation of beta cells in the pancreas. Therefore, the aim of this study was to determine the effect of nettle extract as a supplement of metformin on ovarian tissue of diabetic model. In this experimental study, 30 female Wistar rats were used. Animals were weighed and randomly divided into 5 groups (n=6). 1) control group 2) diabetic group who were diabetic with intravenous injection of alloxan (150 mg / kg) 3) diabetic group + Nettle root extract (150 mg / kg) 4) diabetic + metformin (150mg / kg) 5) Diabetic group + Metformin(150 mg / kg) + Nettle root extract(150 mg / kg). At the end of treatment, the effect of metformin and nettle root extract on ovarian tissue and biochemical factors such as blood glucose and sex hormones were compared and the data obtained were analyzed by SPSS. Hyperglycemia and body weight loss after metformin and nettle root increased for 4 weeks. Simultaneous administration of metformin and extracts of nettle root significantly increased the primordial, primary, secondary, and corpus luteum and reduced the atretic follicles and significantly increased FSH, LH and testosterone levels as compared with metformin alone. The results of this study showed that the root of nettle with its antioxidant compounds and other properties could be a complement to metformin with a corrective effect on hyperglycemia and the improvement of ovarian disorders.

Investigation apoptotic effects of silver nanoparticles coated with Achillea biebersteinii extract on A2780 ovarian cancer cells

J-avad Baharara

In recent years using of silver nanoparticles due to its unique properties was increased. Medical plant, the Achillea biebersteinii is rich in anti-cancers compounds. The aim of this study was to investigate the cytotoxicity effects of AgNPs synthesized using Achillea biebersteinii extract on human ovarian cancer A2780 cells. Cytotoxic effects of AgNPs with MTT test was performed at 48 hours in concentration of 2, 4, 6, 8, 16 and 32 µg/ml. To study the cell death induced by AgNPs DAPI, acridine orange (AO)/ Propidium iodide (PI) staining and Annexin v/ Propidium iodide assay and activation of caspase 3 and 9 were assessed. Results: The results showed that the synthesized AgNPs decreased cell viability dose dependently, calculated inhibitory concentration of 50 % (IC50) was 4 µg/ml. The results from AO / PI, DAPI staining, Annexin V / PI showed that in treated cells the percentage of apoptotic cells compared with control increased. In addition, the treated cells showed increased activation of caspase 3/9. Hence silver nanoparticles induced cell death through the caspase dependent pathway. The results suggest that AgNPs synthesized using Achillea biebersteinii extract exert their cytotoxic effect by inducing apoptosis.

Study of GH and Ghrelin genes expression during the larvae developmental period in Danio rerio

Hamed Paknejad

Zebra fish is an important species in genetics and considering the proximity of its genome to the human genome, investigating the expression of some of the growth and appetite genes during its larvae development is essential. Genes coding growth and appetite (GH and ghrelin) hormones are involved in the synthesis and release of growth hormone, which can be considered to be economic genes in pisciculture. Given the importance of these genes during the early larvae development stages, this study was performed to assess their activity. Samples were collected at 4, 7, 10, 15, 30 and 45 post-hatching days. Samples were immediately placed in liquid nitrogen (-196 degree centigrade) and then stored in a freezer at -80 degree centigrade until RNA extraction (using RNX-Plus kit). To analyze normal expression of target genes, reference gene β-actin was used by 2^-^∆∆Ct method. The expression of genes associated with the growth and appetite was significantly different at various stages of the development of zebra fish, as the gene expression of GH on day 4 and ghrelin gene on day 10 after hatching were significantly higher compared with other samples (P<0.05). Overall, the expression of GH and ghrelin genes at the early stages of growth is higher than other fish developmental stages due to their great importance in the survival of larvae at these stages of life.

The effect of alginate extracted from brown algae Sargassum ilicifolium on the shelf life of rainbow trout roe trout roe

Moazameh Kordjazi

Nowadays, brown macroalgae are considered to be a source of various nutritional and pharmaceutical compounds. In addition, fish roe is very susceptible to spoilage due to its high amount of protein and unsaturated fatty acids. In order to postpone the deterioration of roe-related products, salt and preservatives are utilized. Natural preservatives are recommended because of the adverse effects of chemical additives. Thus, different concentrations of pure alginate (0.1%, 0.2%, 0.3%, 0.4%, 0.5%) were chose and then the 0.2% alginate was selected on the basis of the sensory evaluation of the results. Moreover, the treatments including control (salt-alginate-free), A₁ (containing 2.5% salt) A₂ (containing 0.2% alginate), A₃ (containing 2.5% salt and 0.2% alginate) were used to investigate their preservative effects on fish roe. The quality of rainbow trout roe was assessed within 0, 15, 30, 45 days at refrigerated storage. The treatment A₃ showed significant differences in microbial, chemical and sensory results compared with other treatments (p≤0.05). According to the results, salting 2.5% and alginate 0.2% method gave good flavor and increased shelf life to the roe.

The effects of the combination of bioplastic and its degrading bacteria (Genus Acidovorax) on the metabolic activity of anaerobic bacteria in Siberian sturgeon (Acipenser baerii) fingerlings hindgut by using CLPP

Ebrahim H. Najdegerami

Community Level Physiological Profiles (CLPP) is novel method to evaluate microbial activity and diversity in ecosystems. According to the previous findings, poly-β-hydroxybutyrate (PHB) as a bio-control product, increases bacterial diversity in some aquatic animals. In this study, the effects of four experimental diets (control, combination of two PHB degrading bacteria, 2% PHB, bacteria+ 2% PHB) on the anaerobic activity of cultivable bacteria in Siberian sturgeon fingerlings hindgut was investigated. Community level physiological profile (CLPP) was used to analyze anaerobic metabolism by using Biolog™ Ecoplate microplates. The results indicated that anaerobic metabolic potential of the bacterial community was different in the 2% PHB-treated fishes as compared with other groups and PHB improved anaerobic metabolism of bacteria in fingerlings. Also anaerobic metabolism of bacteria was calculated on the different carbon sources (amino acids, carboxylic acids, carbohydrates and polymers) in Ecoplate microplates and the results showed that fish treated with PHB had the highest metabolic activity in mentioned carbon sources. The results of this research revealed that replacing of the diets with 2% PHB increases anaerobic metabolism of culturable bacteria in Siberian sturgeon hindgut while adding PHB degrading bacteria did not change this parameter in fingerlings.

Callus induction in Portulaca oleracea L. by different hormone concentrations and explant types

Hossein Moradi

Purslane (Portulaca oleracea L.) contains valuable secondary metabolites such as Dopamin, Noradrenaline and Omega-3. This plant is used in various medicinal, food and hygienic industries as well as the treatment of different diseases such as diabetes, heart disease and pain relief. Callus induced from medicinal plants are used to increase the production of secondary metabolities in cell suspension culture and gene transfer. The purpose of this experiment was the study of different concentrations of BAP and 2,4-D of two explants from leaf and shoot tips to produce callus. Leaf and shoot tip explants were used in MS with different concentrations of BAP at three levels (0, 1 and 2 mg/L) with 2,4-D at three levels (0, 0.5 and 1.5 mg/L). Results showed that interactions between hormones and explants were significant in the percentage of callus induction, fresh weight and callus diameter at 1% level. The best result which was the leaf explant with 100% callus induction, 121 mg fresh weight and 5.106 mm callus diameter was obtained by the combination of BAP 2 mg/L and 2,4-D 0.5 mg/L. Shoot tip explants with 75% callus induction, 106 mg fresh weight and 3.03 mm diameter was obtained by the application of 1 mg/L BAP and 0.5 mg/L 2,4-D.

The embryology of Ailanthus altissima (Simaroubaceae): an invasive species from Iran

Fateme Mousavi

Simaroubaceae is a monophyletic family in the order Sapindales with 22 genera and 109 species. Ailanthus altissima (Mill) Swingle, also known as the tree of heaven, is a highly invasive species, which systematically belongs to Simaroubaceae. Reproductive characteristics are essential for the deployment and maintenance of invasive species in new environments and examination of the reproductive system of these species is interesting. To date, no embryological investigations had been directed to A. altissima and our knowledge on the embryology of this species was very limited. A study was carried out on the embryology of A. altissima and the comparison of anther development in male and female flowers in order to gain insights into defining embryological features in this species. According to the results of histological analysis, male flowers have rudimentary gynoecia and anthers are tetrasporangiate. Cytokinesis in meiosis is simultaneous resulting in tetrahedral tetrads. Anther wall development follows the dicotyledonous type and the tapetum is secretory. Female flowers have staminodes and microspore mother cells are degraded too early before entering the course of meiosis. Ovules are anatropous, bitegmic, and crassinucellate. The development of the embryo sac follows the polygonum type. Embryos follow the Onagrad type. The endosperm development is nuclear type. A well-developed hypostase is present and a nucellar cap is formed. These observations will advance our understanding of embryology and embryo anatomy in A. altissima as an invasive species.

Investigating the effect of antioxidant compounds and various concentrations of BAP and NAA on the improvement of in vitro stem and root formation of Nepeta binaloudensis Jamzad

Ali Ganjeali

Nepeta binaloudensis Jamzad is a medicinal plant endemic to Iran. It is an endangered plant due to habitat destruction and intensive harvest. We investigated the effect of antioxidants and different concentrations of BAP and NAA on in vitro stem and root formation of N. binaloudensis. Stem explants were cultured in ½ MS medium supplemented with BAP (0.5 mg/L) and different concentrations of ascorbic acid and reduced glutathione. The effect of different concentrations of BAP on the regeneration of this plant was then evaluated. Moreover, root formation of regenerated stems was investigated in the ½ MS medium supplemented with different concentrations of NAA. The results showed that the combination of antioxidants in ½ MS medium supplemented with BAP (0.5 mg/L) had a significant effect on regeneration in vitro culture. The reduced-glutathione (2 μM/ L) in comparison with other antioxidant treatments increased the stem regeneration in explants. The levels of BAP hormone (1 and 1.5 mg/L) had a significant (p-value<0.05) effect on the stem regeneration rate and the number of produced branches. The NAA (2 mg/L) increased root formation and root height average. We recommend the use of these treatments for in vitro propagation of this endangered plant.

The effects of Ascophyllum nodosum extract on the stimulation of germination indices in chickpea (Cicer arietinum) under drought stress

Saeed Reza Hosseinzadeh

In this study, the effects of seaweed extract (Ascophyllum nodosum) on seed germination stimulation and growth indices in chickpea were studied. A factorial experiment was performed on the basis of a completely randomized design with 3 replications. Treatments consisted of seaweed extract at four levels (0, 1.5, 2.5 and 3.5%) and drought stress at four levels (0, -0.3, -0.6 and -0.9 MP). Drought stress was induced by PEG 6000. Results showed that drought stress at -0.6 and -0.9 MP reduced the germination traits significantly compared with the controls. Seaweed extract at 2.5 and 3.5% resulted in a significant increase in the germination percentage (+18%, +24%), germination rate (+15%, +20%), seedling length (+13.3%, +15.7%), radicle area (+10.4%, +12.8%), radicle diameter (+14.2%, +25%), plumule length (+14%, +16.6%) and plumule dry weight (+10.3%, +12.6%). Under non-stress and -0.3 MP, seaweed extract levels at 2.5 and 3.5% could significantly increase the germination vigor, seed vigor index, consumed endosperm, radicle length and radicle dry weight. The application of seaweed extract at 3.5% improved the germination vigor, seed vigor index and radicle dry weight under -0.6 MP conditions. It also improved the consumed endosperm, length and radicle dry weight in samples affected by -0.9 MP conditions. According to the results, the use of seaweed extract (Ascophyllum nodosum) in the seed germination of beans is recommended for the reduction of the negative effects of drought stress.

Effect of drought stress on MYB gene expression and osmotic regulator levels of five durum wheat genotypes (Triticum turgidum L.)

Plant growth is greatly influenced by environmental stresses including water deficit, salinity and extreme temperatures. Therefore, the identification of genes, especially regulatory ones whose expression enables plants to adapt to or to tolerate these abiotic stresses, is very essential. MYB proteins, a superfamily of transcription factors, play regulatory roles in developmental processes and in defense responses in plants. Members are characterized by a structurally conserved DNA-binding domain, i.e., the MYB domain. Thus, a factorial experiment in a randomized complete block design with three replications was carried out to investigate the effect of different levels of drought stress on the relative expression of TaMYB73 transcription factor gene using Real Time PCR method at Biotechnology Research Institute of Zabol University. The experimental treatments included durum wheat genotypes (Shabrang, Behrang, Karkheh, Aria and Dena) and drought levels (5, 10, 15, 20 and 25% of field capacity). Seedlings of genotypes
were cultured in pots and drought stress was exerted after 45 days (four to five leaves). Data analysis was performed using Ratio =2 ^-ΔΔCT and SAS software version 9.1. The results of the 2-way ANOVA for the effect of genotype, drought stress and cross-effects of drought stress on the relative expression of TaMYB73 gene and the amount of osmotic regulators (Proline and Carbohydrate) at different stress levels (20, 15, and 5% of field capacity) were significant in comparison with the normal condition (25% of field capacity) at 1% probability level. With the increase of drought stress level from 5 to 20% of crop capacity, relative to the normal level (25% crop capacity), the relative expression of TaMYB73 gene and osmotic regulators of proline and carbohydrates increased in Beharang, Karkheh and Dena genotypes. Therefore, Karkheh and Dena genotypes showed greater resistance to drought stress among the 5 genotypes studied.

The effect of water deficit stress on chlorophyll fluorescence, photosynthetic pigments, trigoneline and grain yield in fenugreek in response to zeolite and nitrogen

Seyed Ali Mohammad Modarres-Sanavi

In order to determine the effects of water deficit stress, nitrogen fertilization and zeolite on chlorophyll fluorescence, pigments, trigonelline content and seed yield in, a split factorial experiment was laid out in a randomized complete block design with three replications. Five irrigation regimes were randomly applied to the main plots. Subplots included six treatments and consisted of a factorial combination of three nitrogen fertilization (untreated plots, vermicompost at a rate of 2.7 ton ha^-1 and nitrogen chemical fertilizer at a rate of 11 kg.ha^-1) and two zeolite rates (0 and 9 ton.ha^-1). The results demonstrated that (F_m), (F_v), (F_v/F_m), chl a, b, total chl content and also seed yield were significantly reduced by water deficit stress, wheras minimum fluorescence (F₀), carotenoid and trigonelline concentrations were increased. In addition, the highest F_v/F_m, chl a+b content and the lowest F₀ and carotenoids were observed when irrigation was done after unloading 40% of ASW. In most treatments vermicompost increased the yield (by 25.51% and 98.32% in 2014 and 2015, respectively) and grain trigonelline concentration (7.46% in 2014) in Fenugreek. Mild water stress with vermicompost treatment is recommended for the production of trigoneline from Fenugreek seeds.

Ecological-anatomical comparative adaptability of two gypsophylic Astragalus species of gypsum soils

Fatemeh Rabizadeh

Astragalus semnanensis Bornm. & Rech. belongs to sect. Semnanensis, of thorny astragali and A. fridae, of non-thorny astragali, belongs to sect. Incani are the only endemic gypsophyllic species in western Semnan. The morphological and systematical studies available on these two species are scarce. Nor have the anatomical, distribution and adaptive capabilities in relation to soil and elevation of these species been investigated yet. Therefore, this study is aimed to comparatively investigate these features. The effect of soil and elevation on the anatomy, morphology and micro-morphology of the leaves of both species, as well as associated plant species, were analyzed using RDA and CCA analytical methods, Canoco 4.5 software. Comparison of RDA analysis of both species indicated that gypsum had the highest effect on the abundance of both species, while elevation had a strongly positive correlation with A. fridae and a negative correlation with A. semnanensis. Both species adsorb gypsum on their epidermis and part of the gypsum absorbed is accumulated in leaf parenchyma cells as calcium sulfate crystals. Adaptability of leaf surface is in the form of long and dense hairs which, in A. semnanensis, are longer and basifixed and, in A. fridae, thicker and medifixed. Increase in soil Ec and Na, K and Mg content and content of CaCO₃reduce both species abundances. Species such as Acantholimon cymosum and Prunus lycioides were found to be associated with A. semnanensis and species such as Moltkia gypsaceae and Euphorbia gypsicola with A. fridae.

The study of antimicrobial effects of ethanolic, methanolic and aqueous extracts of six species of Fessia (Asparagaceae) on some pathogenic bacteria and Candida albicans

Zahra Salari

In this research, the antimicrobial effects of ethanolic, methanolic and aqueous extracts obtained from aerial parts, leaves and bulbs of six species of Fessia were investigated on the growth of some of micro-organisms, including Staphyloccocus aureus, Bacillus anthracis, Kelebsiella pneumonia and Candida albicans. Plant samples were collected from natural habitats and were then identified. The evaluation diameter of inhibitory zone, well and MIC (minimum inhibitory concentration) methods were used for the assessment. Aqueous and methanolic extracts obtained from the leaves and bulbs did not affect any of the studied micro-organisms. The results showed that ethanolic extract of the species Fessia khorassanica, had stronger inhibitory effect on the Candida albicans in comparison with the control group.

The production of Yarrowia lipolytica lipase powder by improved spray-drying method

Farshad Darvishi

Lipase is used in the production of foods, flavor enhancers, detergents, cosmetics and pharmaceuticals. A common impediment to the production of commercial enzymes is their low-stability aqueous solutions. In this study, the downstream process was investigated to obtain a stable spray-dried lipase powder of Yarrowia lipolytica. The enzyme solution samples were supplemented with different concentrations of Arabic gum and milk powder to spray-drying. Samples were dried in a pilot spray dryer at inlet and outlet temperatures of 175 and 85 °C, respectively, at a flow rate of 15 liters per hour. The best lipase powder obtained from spray-drying with 3% of Arabic gum and 9% of milk powder formulation as compared with other formulations. Results showed that spray-dried lipase powders of Y. lipolytica had a good yield suitable for various applications.

The identification of protein changes in Celeribacter persicus SBU1 after degrading phenanthrene

Mohsen Shahriari Moghadam

Organisms in different environmental conditions express different genes, which result in different protein expressions. These changes result from the adaptation of the organism to environmental conditions such as the presence of toxic substances. This study aimed to investigate the changes in protein expression in Celeribacter persicus SBU1 isolated from Nayband Bay mangrove forests, cultured in the medium containing phenanthrene as the sole source of carbon and energy. For this purpose, C. persicus SBU1 was cultured on mineral salt medium containing phenanthrene and sodium acetate as treatment and control, respectively. After the extraction of total protein, changes in protein expression were evaluated by SDS-PAGE. Proteins were identified by MALDI-TOF-TOF MS. After evaluating changes in protein content, two bands which showed greater variation in comparison with the control treatment (increased protein expression) were detected. The identified proteins included one ligand-gated channel protein and one unknown protein. In general, the results of this study showed significant changes in the protein content of C. persicus SBU1 after using phenanthrene. The up-regulation of ligand-gated channel protein signified the role of this protein in phenanthrene molecules transport in and out of the cells.