Journal of Molecular and Cellular Researches
Year:2016 | Volume:29 | Issue:4 |Papers Count:10

Despite of the increasing functional and molecular studies in the sunflower family (Asteraceae), the study of generative biology that consists of flowering developmental stages in necessary. In this research, Embryological characteristics including of ovule and pollen grains development of Echinops ilicifolius from Asteraceae family were studied. The flowers and buds in different developmental stages were removed, fixed in FAA70, stored in 70% ethanol, embedded in paraffin and sectioned with a thickness of 5-7 μm by microtome. Staining was carried out with Eosine and Hematoxylin. Then slides were studied using light microscope and were photographed. Results indicated that anthers were Tetra-Sporangiate and tapetum was of the periplasmodial type. Pollen tetrads were tetrahedral and tetragonal. In anther tissue interface particles were crystalline needles. Pollen grains were 2-celled at the time of Diffusion. Ovule was anatropous, unitegmic and tenuinucellate. Endothelium was uniseriate and connected with embryonic sac. Embryo sac development was of the Polygonum type Antipodal cells increased up to 6-8.

Apple (Malus orientalis.) distributed throughout the Hyrcanian forest from lowland regions to steep and mountainous areas. For evaluation genetic diversity, leaf material were collected from 104 individual of were 14 population. DNA was extracted and DNA polymorphism were considered using four primers (GA)8YC, (AC)8YA,(AG)8AT and (AC)8G with ISSR-PCR method. The results showed that for four primers was detected 385 allele with 0.049 heterozygosiyt.The mean heterozygosity was 0.049 and varied from 0.031 in “Afratakhte” population to 0.059 in “kodir” and “Lamzer” populations. The maximum Nei genetic distance was belong to “Siahbill” and “Masal” populations and the minimum was related to “Yoush”, “Afratakhte” and “Dinekooh”. the AMOVA result indicated that the intra and inter population diversity were 94% and 6%, respectively that indicate significant within population diversity of this species.Despite the different habitat conditions and long geographical distance among populations, The low genetic differentiation (excluding Siahbill population) and similar heterozygosity within populations suggest high gene flow among populations of Malus orientalis in north of Iran.

Plant tissue and cell culture is being used as a stable method to investigate plant secondary metabolites widely. Because of the important position of Pennyroyal (Mentha pulegium) in food industries, sanitary and pharmaceutical fields, cell culture is done to investigate effectiveness of different concentrations of yeast extract and salicylic acid on the cell survival and the amount of β-Caryophyllene and Iso-Pulegone secondary metabolites. MS medium with 1 mg/l 2, 4-D without agar was used with different concentrations of yeast extract and salicylic acid to investigate amount of secondary metabolites. after ten days, the amount of these two secondary metabolites were investigated by GC-MS and the cell survival was investigated by tetrazolium test. In the cell culture, percentage of cell surviving was decrease by increasing density of two elicitors and amount of b-Caryophyllene secondary metabolite significantly increased compared to the natural plants (maximum amount of this metabolite was obtained in the cell culture containing yeast extract 80 mg/l). Given that increasing the amount of secondary metabolites is the main purpose of the plant cell culture, the use of high concentrations of elicitor to increase b-Caryophyllene is justified despite the decrease in cell viability. Percentage of Iso-Pulegone secondary metabolite was in the natural plant more than the cell culture and also in the cell suspension culture, amount of this metabolite was decrease by increasing density of yeast extract and salicylic acid elicitors, thus cell culture containing these two elicitors is not recommended to increase amount of this secondary metabolite.

Plants normally change their hormone levels in response to biotic and abiotic stresses. Since RON1 affects either signaling or biosynthesis of hormones abscisic acid (ABA), Indol Acetic Acid (IAA) and Ethylene (C2H4), therefore, The effect of change in RON1 activity on several morphological characteristics including vein pattern, lateral root formation, hairy root, stem, root and apical has been investigated. Hereby, morphological and molecular characteristics of ron1-1 mutant was evaluated and compared with the Ler-0 wild type plants in response to different concentrations of hormone IAA, ABA and ACC in vitro conditions. Based on the obtained results, there is a significant difference between primary root length, lateral root number, leaf number and rate of germination in mutant seedlings ron1-1 when compared with the wild type seedlings in response to Different concentrations of ABA, ACC, and IAA. Moreover, Investigation of cotyledon and leaf veins in ron1-1 mutant showed that secondary veins give open networks in ron1-1, while genotype Ler-0 has joined the network of veins. Overall, the results here indicate that a considerable increase in sensitivity of ron1-1 and old101 mutant plants to hormonal -induced stress has been observed in mutant seedlings comparison with the Ler-0 plants. RON1 enzyme activity has gone in ron1-1 plants, which causes PAP accumulation and possibly IP3 in mutant, changing the morphological characteristics associated with the mutant seedlings.

Citrullus colocynthis is a very drought tolerant cucurbit species which widely distributed in the deserts of Mediterranean region and Middle East. One of the possible mechanisms involved in the tolerance response of this plant could be overexpression of Rboh gene which its protein product are involved in the production of ROS signal molecules. In this research, Rboh gene expression during drought stress, were studied in C. colocynthis and watermelon. Seeds after sterilization and germination in germinator transfer to pots containing sand and 5 leave plants treated by PEG 8000 15%. At times zero, 3, 9, 18 hours after treatment relative water content of leaves and Quantitative gene expression of Rboh by Real Time PCR method was studied. The results showed a significant decrease in relative water content in both species but in C. colocynthis this decrease was less remarkably. After PEG treatment the expression of Rboh increased in two species but expression level in C. colocynthis was more than watermelon. Expression of Rboh which play an important role in response to stress stimulus, was rapidly and strongly induced by PEG treatment in C. colocynthis rather than C. lanatus. The result can be a confirmation that Rboh Play a critical role in C. colocynthis tolerant against the drought stress.

Reactive oxygen species, which lead to oxidative stress and related diseases via both intra- and extracellular pathways, could be identified and cleaning through some of the bacterial enzymes. Bearing in mind, the assessment of the structure, function and hosting of the antioxidative enzymes while lead to disclose species of bacteria with probiotics capability, provides approaches to designing genetics constructs. In this regard, the nucleotide and protein sequences of the katA, katE, katE*, sodA, sodA*, gshR, gshR1, gshR4, trxB1 and trxR genes were retrieved from databases. Then the structural, functional,topological and physicochemical properties of the protein sequences of related enzymes were investigated. Moreover, their hosting in bacterial microorganisms were explored. The results of this study whilst disclosed the physicochemical properties of these enzymes reveal that KATE, KATA, KATE* and SODA are secretory capacity. Structural monitoring of these enzymes introduced collaborative and pragmatic domains with the ability to remove reactive oxygen species, hydrogen peroxide decomposition and regulation of redox reactions as well as immunomodulatory effects and ammonia removal in some of them. In this regard, examination the binding affinity of these enzymes to oxidant agents revealed high binding affinity of them, in particular KATA, to O2-. Additionally, checking the host of these enzymes revealed the presence of homologous sequences especially sequences like to TRXB1 and TRXR in different species of Weissella, Pediococcus, Leuconostoc, Tetragenococcus, Peptoniphilus and Listeria. Meanwhile, similarity search lead to detection seven encoding sequences with antioxidative capacity in the genomic context of the Pediococcus acidilactici, Lactobacillus pentosus and Lactobacillus plantarum.

In this study in order to achieving high frequency regeneration of sunflower effect of genotype (Record, Progress, Gabor, AF81-196 A line and CMS19 A line), Proximal and Distal section of the cotyledon explant and 10 plant growth regulator combination have been investigated. Seeds without pericarps were surface sterilized and germinated on half strength MS basal medium. Cotyledons from 2-day old seedling were cut from the middle and three explants of each section were transferred to regeneration medium. Regeneration medium consisted of full strength MS medium supplemented with two levels of BAP (1 and 2 mg/l) in combination with three levels of NAA and IAA (0, 0/5 and 1 mg/l). Experiment was conducted in a factorial completely randomized design with three replications. Statistical analysis showed that there were significant differences among the genotypes, explant and plant growth regulator treatments on both organogenesis parameters including regeneration rate and the average number of shoots per explant. Our results showed that the shoot regeneration in sunflower is genotype-dependent, and proximal section of cotyledon explant was greater potential for shooting. CMS19 and Progress were the best responding genotypes with a regeneration rate of 51.21% and 41.21%, respectively. The highest rate of shoot regeneration was observed with 2 mg/l BAP in combination of l mg/l IAA.

Septoria tritici blotch (STB) caused by Septoria tritici is one of the most important wheat diseases in Ilam Province. In order to determine genetic diversity of pathogen, 44 samples were collected from wheat fields of different regions in Ilam province. Molecular test was carried out with a set of five pairs of SSR primers after purification and identification of isolates. The SSR primers amplified a total 22 alleles among isolates. The average of allele number was 4.4 per each primer. Genetic diversity of the populations ranged from 0.138 to 0.166 with an average of 0.154. Cluster analysis based on UPGMA method and Dice coefficient, divided the isolates into 18 groups at 0.54% similarity level. Result of AMOVA showed that 98% of genetic diversity was within populations and only 2% was between populations from different geographical regions. Therefore there is the high genetic similarity between isolates from different regions. High genetic similarity can be attributed to emigration of gene or genotype as a result of various factors.

Root rot is the most important disease of sugar beet caused by Rhizoctonia solani fungus which lives in soil. Many soil bacteria such as Pseudomonas spp. by production of materials such as siderophores could control soil borne pathogens. So in this study, after identifying Pseudomonas fluorescents in the rhizosphere of Sabzevar sugar beet fields and the assessment of the bacteria to inhibit R. solani growth, detection of siderophore biosynthetic gene was performed. For this purpose, plant samples along with its rhizosphere soil were collected. To obtain specific Pseudomonas, Pseudomonas agar F medium was used. After 2 subcultures on the same medium, three Pseudomonas fluorescent strains based on microbial tests were identified. To evaluate the growth inhibition of R. solani by Pseudomonas fluorescent, dual culture tests were done. Based on inhibition areola, it was shown that C7 and C3 isolates have severe and moderate deterrence respectively but C5 strain has no deterrent effect. PCR results also showed that only C7 isolate contained siderophore biosynthetic genes.

Cancer has become an epidemic disease. Scientists are trying multiple methods of chemotherapy, radiotherapy and surgery to improve patient’s health. However, the use of these methods of treatment are associated with severe side effects, which make the treatment process very painful. Therefore, researches are looking for new treatment options with fewer side effects using natural compounds such as peptides. In this study, we used different concentrations of GL-9 peptide to evaluate its cytotoxic effect on human lung adenocarcinoma cells (A549) and normal human and bovine lymphocytes and erythrocytes cells was investigated. 12, 25 and 50 mg/ml of GL-9 peptide were used to examine its cytotoxic effect on human lung adenocarcinoma cells (A549) after 24 h and 48 h via MTT assay. Furthermore, hemolysis and lympho-toxic assay were used to analyze GL-9 cytotoxic influence on human normal cells. Our results showed the cytotoxic effect of GL-9 peptide on A549 cell line within 48 h. Moreover, our results showed no hemolytic and cytotoxic effect on human and bovine red and white blood cells. GL-9 peptide had cytotoxic effect on A549 cell line after 48h with no toxic activity on human and bovine peripheral blood mononuclear cells (PBMC), therefore through further analysis and examination this peptide could be a replace for common cancer therapy drugs in the near future.