Background: Infectious diseases are an important cause of mortality in the world. Because of resistance of some human pathogenic bacteria to antibiotics, use of natural and herbal medicine is increasing to control these factors. The aim of this study is to investigate the antibacterial effects of RAMALINA HYRCANA Sipman lichens against many Gram-positive and Gram-negative bacteria.Materials and Methods: The lichen ethanol extract was prepared by using a rotary machine. Nine standard strains of Gram- positive and Gram -negative bacteria were cultured in nutrient broth. The minimum inhibitory concentration (MIC) was determined by the micro dilution method.Results: The results of this study showed that the lichen R. HYRCANA extract in different concentrations inhibited both Gram-positive and Gram-negative bacteria. The results showed that the lichen extract of all concentrations inhibited of Staphylococcus aureus. After S. aureus bacteria inhibition, the minimum inhibitory concentrations are for Hafnia and Acinetobacter (12.5 ppm).Conclusion: According to the results and other reports in this area, lichen R. HYRCANA extract could be introduced as a biological control agent. However, for the use of different lichen extracts for clinical application, the chemical analysis of extracts and clinical research is necessary.

Background: Buxus HYRCANA is species of Buxus genus. Buxus specien are known an The are of sterois alkaloids with biologic activity. They are widely Ysed in Traditicnal Medicine to Treat rhematism gout, skin diseases and Malaria nesistant to common therapies . therapies. The present study was carries out to extract steroid alkaloids from Buxus HYRCANA gathered in uoshahr, Iran, and assessed its phytochemical chara cteristics.: Materials and Methods: For This exploratory study, The plant was dried regardless of exte heat or light. Then was powdered and ulraclecl using metharel. Turllur experiments enealed thal the produst contained alkaloids, and flavonoids. To txtract alkaloids , The product was exposed to Hcl s so thal ets PH reached 2. then chlorofcrn was osed to divide The chlovofornic phase contained non - alkaloids, wheras the olter phase contained and ets PH was reached Q By means of arncniurn , Finally chloroform wan wed for extraction During The Column chranatography of the chloro formic phase, an unpurified material was esolated that finally purified by thin loyer chromatography Results: The purified material had The following characteristics: UV absorption, Melting point of 170-173° C . molecular formula of And molecular mass of 385 Meanuhile, The presence of ketonic, alcoholic and amin groups as cyclopropanic cyclen wene all Cenfirmed.

Karyological studies of seven populations of Buxus HYRCANA Pojark. from different habitats are presented. We found one usual basic chromosome number in this species x=14. All populations were diploid (2n = 2x = 28). The results of kryotypic analysis made it possible to categorize different populations of a species and make a logical comparison among them.

In this research, the application of Artificial Neural Network or MLP method in the process Assignment of relevé-groups/ plant communities allocation was evaluated using Buxus HYRCANA forests database. For this purpose, firstly, the ecological and sociological groups of B. HYRCANA were determined using TWINSPAN and Braun-Blanquet method, respectively. The results of both numerical and expert based classification dendrogram of the B. HYRCANA communities, which included seven levels of classification as primary groups/plant communities, were introduced to MLP. Then, with assignments in three sets of training (70%), test (15%) and validation (15%), the MLP classification was performed on each level of the two dendrograms. The results showed that by increasing the level of classification, the degree of adaptation of the MLP result with primary ecological and sociological groups of TWINSPAN (99% to 60%) and Braun-Blanquet (98% to 68%) decreased from the cutoff level of 1 to 7. Results of sensitivity and kappa cross tab coefficients, except in 7 cut level, imply that the quality of MLP groups based on TWINSPAN primary ecological group is better than the primary Braun-Blanquet groups. The MLP results in Buxus HYRCANA plant communities classification were consistent with the results of TWINSPAN (90%) and Braun-Blanquet (89%) ecological/syntaxa groups at the fifth cut level of both dendrograms concluding the reliability of MLP application for classification of plant communities. So, our result confirms that MLP can be introduced as a suitable method for the assignment of releves to plant communities.

Box tree (Buxus HYRCANA Pojark. ), is an ornamental tree and shrub species that has application in various industries. Growth and development of box tree is very slow, its rooting is hard and is under danger of extinction. Thus, the purpose of this research was investigation of the effect of different concentrations of BAP and IBA (0, 0. 5, 1, 1. 5 and 2 mg l-1 form each two) on micropropagation of box tree. The experiment was carried out as factorial based on a randomized complete block design in four replications. Results of the present research showed that the largest number of shoots (6. 200/plantlet) was obtained in apical buds expalnts treated with 1 mg/L BAP along with 0. 5 mg/L IBA. Apical bud explants were produced the maximum node number (4. 100/plantlet) in medium containing 1 mg/L BAP and the largest number of leaf (with average of 6. 566/plantlet) along with 1. 5 mg/L IBA. Also, the largest number of root (6. 466/plantlet) was calculated in explants treated with 1 mg/L BAP plus 1. 5 mg/L IBA. Plantlets were transferred to pots containing peat and perlite with ratio of 1: 1 for acclimatization. The pots were kept in a greenhouse with temperature of 24-26º C and relative humidity of 70% and periodic irrigation. Around 90% of those were healthy. These acclimatized plantlets were similar to mother plants.

Lichens are small environments containing two fungus and algae symbiosis. They are one of the richest sources of natural ingredients with anti-tumor, antibiotic, and antioxidant properties, some of which are used as drugs and for the treatment of certain diseases. This research was aimed to identify and determine the amount of betolin and betulinic acid as well as evaluation of the antioxidant activity of lichen Betula pendula Roth. Roth. and its symbiotic fungus. It was found that the lichen isolated from the bark of birch contained bioactive compounds entitled Raminalina sinensis. The symbiotic fungus (Arthrinium arundinis) was detected by the molecular method. This fungus belongs to the ascomycetes, registered for the first time in the NCBI website with MG198621 code. In the extract of lichen and its symbiotic fungus, the amount of betulinic acid and betulinic acid was determined using HPLC. Betulinic acid and betulin, belonging to the terpenes, are known as a strong anticancer agent. The mentioned ingredients were significantly found in the lichen tissue (2. 17 and 0. 075 percent, respectively) and in its symbiotic fungus A. arundinis (1. 6 and 0. 025 percent, respectively). In this way, the lichen and its symbiotic fungus are introduced for the first time as new sources containing these two active ingredients. Measurement of antioxidant properties of lichen extract and its symbiotic fungus was examined by methanol and ethanol solvents. It was found that the lichen extract and the symbiotic fungus, besides having alternative metabolites, have antioxidant properties, which are able to remove toxic free radicals. These results could have valuable effects in medicine and industry.

Extended Abstract Background: The plant biodiversity of any ecosystem is directly affected by its vegetative characteristics and diversity of plant species, which always guarantees the ecosystem’s stability against variable environmental and biological factors. Biodiversity indices are measured to compare the biodiversity of different masses and estimate the changes in biodiversity over time at the mass level. The Hyrcanian forests of northern Iran are a legacy left from the third geological period, and today a large part of them has been destroyed in the plains due to destructive human activities. These forests have a special importance for the protection and management in the south of the Caspian Sea. Hyrcanian boxwood (Buxus HYRCANA Pojark.), which belongs to the Buxaceae family, is the only species of boxwood in the Hyrcanian forests of Iran. It has unique values from various aspects, including economics, tourism, preservation of biodiversity, etc. This valuable species has been threatened by human and natural factors for a long time and its area has decreased in the country. The present research investigates the floristic-physiognomic status of boxwood habitats in Cheshme Bulbul, Sangdeh, Si-Sangan, and Shafarood regions. Methods: In this research, the important habitats of Hyrkanian boxwood were sampled from the protected area of Cheshme Bulbul, Bandar Gaz, Golestan province, as the easternmost distribution area of this species in Hyrkanian forests to Shafarood forests in Gilan province. In total, 125 sample plots of 400 m2 (20 × 20 m) were planted in the four identified areas (35, 30, 40, and 20 sample plots in Bandar Gaz (Cheshme Bulbul), Frame (Sangdeh), Si Sangan, and Shafarood, respectively). To measure the herbaceous cover of the forest floor in each sample plot, five small plots of four m2 (2 × 2 m) were walked in the center and four corners of each sample plot to harvest their herbaceous cover. Biodiversity indices, including Shannon, Simpson, and Fisher diversity, Margalf and Menhinik richness, and Pilou, Simpson, and Shannon uniformity, were measured for each of the sample plots. The plants were identified very carefully using the Persian Flora of Iran, the Flora of Iranica, the Flora of Türkiye, and the Flora of Europe. The biological form of any plant was determined and recorded based on the Rankier method. The range of geographical distribution was determined according to the mentioned flora. Then, the belonging of each species to the existing phytochorions was determined and the geographical distribution diagram of the regional plants was drawn using floristic data in the division of the vegetative regions of the earth's surface.  Species diversity between habitats was compared with the one-way analysis of variance. Means were compared using the Tukey-HSD test in SPSS software. Graphs were drawn using Excel software (2013). Results: In total, 186 plant species belonging to 68 plant genera were identified in the studied regions. The hemi-cryptophytes (65 species, 34.94%), phanerophytes (40 species, 21.5%), geophytes (37 species, 19.90%), trophytes (36 species, 19.35%), and cryptophytes (5 species, 69.2%) dominated the vegetation composition of Hyrkanian boxwood habitats. Kamephytes were the rarest biological form in the region with a percentage (3 species). The results of the geographical distribution of plant species in the region showed that the entire flora was mainly of European Siberian (55 species) and multi-regional (38 species) elements, which accounted for more than 50% of the species (93 species). This was followed by Europe-Siberian/Turanian Iran/Mediterranean vegetation areas (25 species, 13.44%), Europe-Siberian/Turanian Iran (25 species, 13.44%), Turanian Iran (16 species, 60.8%), Europe-Siberian/Mediterranean (13 species, 6.70%), and cosmopolitan (8 species, 4.3%) species. The lowest presence in the region was observed for the vegetation areas of Mediterranean/Turanian Iran (5 species, 2.70%), considering that the northern forests are geographically located in the Auxin-Hyrcania state of the Pontic sub-region, which is a large vegetation area belonging to Europe-Siberia. The presence of species belonging to this geographical spectrum in the flora of these regions is not far from expected. Moreover, the presence of species in other geographical areas is a result of the natural patterns of the accidental presence of plant elements from other vegetation areas of the world. Conclusion: The results of this research show higher indices of diversity, uniformity, and species richness in Shamshad Frame and Shafarood habitats, which have a higher average height above sea level than Cheshme Bulbul and Sisangan habitats. The review of the sources shows that physiographic factors play an important role in the indicators of species richness and diversity. Changes in altitude above sea level often lead to changes in environmental conditions, such as temperature, precipitation, and soil characteristics. This diversity in habitat conditions increases its heterogeneity and allows a wider range of species to coexist. As a result, more habitat heterogeneity leads to higher species diversity, which is also reflected in the biodiversity index.

Precise evaluation of extent and distribution of genetic diversity in rare and endangered species is of particular importance for conserving and utilizing genetic resources. To study inter- and intra-population genetic diversity of three natural populations of boxus (Buxus HYRCANA Pojark.) in north of Iran (Bandar Gaz, Nowshahr and Gorgan) ISSR markers were used. Twenty samples per population were randomly selected, their DNA was amplified in PCR reaction using 10 ISSR primers and electrophoretic patterns were obtained. Then, population genetic parameters were calculated based on respective numerical scores. Results showed that there was a desirable genetic variation (34.1%) in the studied populations. However, contribution of intra-population genetic variation in total genetic variation was estimated over than that of inter-population genetic variation (73% vs. 27%). On the basis of Nei’s similarity, a relatively high genetic similarity (74 to 88%) was observed among the populations. The lowest genetic distance (13.2%) was observed between Bandar Gaz’s and Gorgan’s populations and the highest amount (30.1%) was observed between Gorgan’s and Nowshar’s populations. Based on cluster analysis using UPGMA method, populations of Bandr Gaz and Gorgan were grouped together, indicating their higher genetic relatedness and this grouping coincided to geographic distribution. Investigation of Hardy-Winberg genetic equilibrium showed that Bandar Gaz’s population had the highest genetic equilibrium at different loci (~78.9% of loci), while two other populations had a mild equilibrium (58% and 63% of loci in Nowshahr and Gorgan populations, respectively). Gene disequilibrium in the latter populations is probably due to harder endangering forests and losing the hereditary resources of boxus species in the regions. Altogether, on the basis of the results of this research it can be concluded that ISSR markers are efficient and reliable markers for studying genetic diversity and population genetics of Buxus HYRCANA species in forests of north of Iran.