Nova Biologica Reperta

Evaluation of genes expression changes in the mitochondrial pathway of apoptosis and cell cycle in AsPC-1 cells treated with Fluorouracil

Javad Baharara

Pancreatic cancer is one of the most deadly and aggressive cancers; Fluorouracil induces apoptosis and cell cycle arrest in cancer cells. In the present study; the effect of Fluorouracil on different stages of the cell cycle and the expression of genes involved in the internal pathway of apoptosis in the AsPC-1 cell line (human pancreatic cancer) were investigated. In order to do so, MTT assay was used to evaluate the cytotoxic effect of Fluorouracil on AsPC-1 cell proliferation; The type of induced cell death and cell cycle changes were investigated by flow cytometry; changes in the expression level of genes (BAX, Bcl-2, APAF-1, Caspase-3, Caspase-9, p53, p21) were examined by Real-time PCR. Quantitative data were analyzed at the significant level of (p<0.05). The MTT assay results showed that Fluorouracil decreased AsPC-1 cell proliferation in a concentration-dependent manner. The results of flow cytometry analysis showed that increased percentage of apoptotic cells in the treated cells; Fluorouracil induces S phase cell cycle arrest in AsPC-1 cells and reduced distribution in the G1 phase. The Real-time PCR results in treated cells showed an increase in the expression of genes in the mitochondrial apoptotic pathway as well as genes effective in regulating the cell cycle. Fluorouracil reduces cell proliferation and induces apoptosis by increasing the expression of genes involved in the Intrinsic apoptotic pathway in AsPC-1 cells; Fluorouracil also caused cell cycle arrest in these cells by regulating the (p53, p21) genes.

An investigation on the mollusc fauna in west and south of Tehran Province, Iran

Elham Ahmadi

Molluscs are important pests of natural and agricultural ecosystems, by feeding on different parts of leaves, stems, flowers, buds, fruits and roots of different plant hosts, reducing the marketability of annual agricultural products and damage plant species in different habitats in the country. Moreover, they are intermediate hosts of human and cattle parasites, and therefore are of medicinal and veterinary importance. In this study, snails and slugs were sampled from nine designated stations located in the south and west regions of Tehran province, considering their natural condition, vegetation, location of gardens and fields, soil or subsoil types, from the beneath of rocks and trunks, water bodies and other areas related to the habitat of snails and slugs. Samples were made using sampling nets or gathered directly by hand from plants, shrubs and river banks. Overally, 17 species belonging to 12 genera from nine families were collected and identified. Identification was done according to morphometric characters including shape, size of shell, dextral and sinistral position of the shell, operculum, aperture, radula, mucus colour, respiratory pore, keel, renal ridge and reproduction system. The highest and lowest mollusc species diversity among the stations studied were turned to be 16 and 6 species belong to station 9 (west of Tehran province) and station 2 (south of Tehran province), respectively. The most commonly diversed family was Hellicidae with 5 species, i.e., Monacha cantiana, Euomphalia pisiformis, Helicella krynickii, Helicella derbentina, Caucasotachea atrolabiata. Also, Lymnaea truncatula is identified as the most frequent species, which has medical importance. Monacha cantiana was identified in the present study and reported for the first time from Iran. All the samples and their respective spatial and temporal information were made available in the museum of the Agricultural Zoology Research Department in Iranian Research Institute of Plant Protection.

Investigating the bioremediation of heavy metals by Pleurotus ostratus

Mehdi Rahimi

Heavy metal contamination (HMs) of water and soil is the most serious problem caused by industrial and mining processes and other human activities. Mycoremediation is a biotechnological method that employs fungi to remove toxic contaminants from the environment in an efficient and cost-effective manner. Pleurotus species are considered to be the most popular and widely cultivated species worldwide, and this may be due to their low production cost and high yield. It has been indicated that Pleurotus species may improve plant growth in metal-contaminated soils through enhancing nutrition uptake or by alleviating toxicity of the metals. In this experiment, the fungus was grown in vitro in liquid and solid media for 3 weeks on five different concentrations (0, 15, 30, 45, 60 ppm) of five heavy metals (Cd, Zn, Ni, Pb, Mn) as sulphate and the effect of these metal on radial growth, biomass production and metal content of fungal biomass were determined. Based on the results, this fungus showed a great variety of tolerance against the metals, as that growth parameters were increased in the case of two metals contamination (Zn and Mn) and inhibition of growth was observed even at the low concentrations of nickel, cadmium and lead (15 mg/liter). The amount of metal accumulation in the fungal mycelium also increased with the increase of the metal concentration in the culture medium. In this research, for the first time, the growth parameters and the amount of accumulation of heavy metals in axenic conditions have been investigated and described.

Evaluation of osmotic regulation, total phenol, protein and antioxidant enzymes in four fennel populations under saline treatments using vermicompost

Siavash Hosseini Sarghein

Fennel is a medicinal plant; all of its parts were being used by humans in different ways. This plant is relatively sensitive to salinity. A factorial experiment as a randomized complete block design with three replications at the greenhouse level was executed to investigate the effect of vermicompost on the increase of salinity tolerance in four fennel populations (Mashhad, Urmia, Shiraz, and Bushehr) in the vegetative stage of the plant, emphasizing the evaluation of some osmotic and antioxidant protection indicators. Experimental treatments were designed with four levels of salinity (0, 40, 80, and 120 mM of NaCl) and two levels of vermicompost (0 and 5% v/v). After harvesting, the roots were separated from the stem to analyze the biochemical variables. Salt stress caused a decrease in the total soluble sugar and starch content in the shoot and an increase of those in the root of the studied populations. In addition, under stress conditions, the proline content of shoot and root, total free amino acid, total phenol, and activity of guaiacol peroxidase and catalase were increased in fennel populations, while total soluble protein and anthocyanin content were decreased. Vermicompost treatment increased the content of soluble carbohydrates, soluble protein, free amino acids, proline, total phenol, and anthocyanin, and decreased the activity of guaiacol peroxidase in the shoot, as well as the starch content in the roots of fennel populations under non-stressed and stressed conditions. Despite observing the complexity in the changes of the analyzed indicators which were dependent on the type of population and the dose of stress, our results showed that the application of vermicompost with a concentration of 5% can improve the osmotic and antioxidant protection in the studied populations of fennel under salinity stress.

Evaluation of freezing tolerance in chickpea promising genotypes (Cicer arietinum) using physiological traits and molecular markers

Omid Sofalian

Chickpea is one of the most important plants of the legume family and is very important in the diet. In order to investigate the genetic diversity of chickpea, an experiment was conducted with 18 chickpea genotypes in the form of a completely to investigate the genetic diversity of chickpea, an experiment was conducted with 18 chickpea genotypes in the form of a completely randomized block design. After acclimatization of plants to cold, freezing treatment was applied at temperatures of -6, -8 and -10 and their 50% lethality temperature (LT₅₀) was determined by probit transformation. Before and after the habituation stage, a leaf sample was taken and the relative content of leaf water, photosynthetic pigments, proline, soluble sugar, protein percentage, catalase, peroxidase, polyphenol oxidase and greenness index were measured. Genotype number 5 with the lowest LT₅₀ (-8.86) and the highest survival percentage (80%) was the most resistant genotype and genotype 10 with the highest LT₅₀ (-3.57) and the lowest survival percentage along with genotype 15 were recognized as the most sensitive genotypes. In order to evaluate genetic diversity, DNA extraction was utilized and 21 different ISSR primers were used in the investigation. The results showed the presence of polymorphism among the cultivars studied. A total of 101 clear bands were produced, of which 94 were polymorphic bands. Polymorphic information content (PIC) was in the range of 0.332 (initiator 7) to 0.049 (initiator 16). The amount of gene diversity was between 0.126 and 0.977 changes. Cluster analysis of genotypes was done using Jaccard similarity coefficient and UPGMA method

Phylogenetic placement of Stachys sect. Satureoides (Lamiaceae; Lamioideae)

Mahya Khakpash

Stachys sect. Satureoides includes four species characterized by being annual herbs having oblanceolate leaves, urceolate calyx in fruit and dense hairs at mouth and are distributed from Eastern and Southern Turkey, Syria, Northern Iraq to Western Iran. The main aim of present study is to determine the phylogenetic placement of S. sect. Satureoides based on Stachys melampyroides, as the only representative of this section in Iran. The present molecular study was conducted based on nuclear ribosomal ITS marker, using Maximum Parsimony and Bayesian Inference analyses. In total, 86 accessions (including nine outgroups and 77 ingroups) were included. Our results demonstrated that sect. Satureoides was placed in Olisia clade which is one of the most heterogenous and taxonomically challenging clades of Stachys as a paraphyletic genus. Moreover, not only sect. Satureoides was not found to be phylogenetically close to sect. Neurocalyx, as had been suggested by previous morphological studies, but also it was placed as a sister group of sect Fragilicaulis which was characterized by chasmophilus species with fragile stems and yellow flowers. Our results revealed that sect. Neurocalyx could be the closest group to the Satureoides+Fragilicaulis clade. In addition, based on the nrITS phylogenetic tree, annual species of Stachys showed affinity to perennial Stachys species. Hence, it seems that “Annual habit” have been evolved among their perennial ancestors, independently.