Journal of Molecular and Cellular Researches
Year:2017 | Volume:30 | Issue:4 |Papers Count:10

Phytic acid is the main phosphorus storage form that is not digestible for monogastric animals such as poultry, fish and even humans. Phytase breaks down phytic acids that are found in vegetable products and increases the availability of phosphorus. The aim of this study was to produce phytase from rice bran extract as a rich and inexpensive source of phytic acid using Bacillus spp. isolated from sediments of the Caspian Sea.Using thermal treatment of the sediment samples, phytase producing Bacillus isolates were screened on PSM agar. In addition, after a quantitative assay of phytase production using the ammonium molybdate method, phytase production of the isolates was investigated in the MGE broth containing rice bran extract. Screening results of 40 isolates demonstrated that the four isolates MGH2, MGH3, MGH4 and MGH7 showed high levels of phytase production with a clear halo zone forming around the colony.Morphological and physiological characteristics indicated that all of the selected isolates belong to Bacillus. Maintaining a clear halo zone, after staining with ammonium molybdate, confirmed the production of phytase in all selected isolates. In addition, the results of the quantitative assay of phytase production in the isolates showed that maximum and minimum phytase production relate to MGH4 with 10.58 U ml-1 and MGH3 with 9.34 U ml-1, respectively. When the production of phytase in MGE broth and PSM broth were compared with one another, a 55-60 percent increase in phytase production was examined in the MGE broth. The results of present study indicated that rice bran extract can be considered as a suitable and affordable substrate with high levels of phytase production.

In order to investigate of cis -regulatory elements' role in the promoter region in the expression pattern of genes Catalase 2 (CAT2) and Thioredoxin H5 (TRX5) in Arabidopsis in response to different biotic and abiotic stresses, the promoter sequences of the genes, cis -regulatory elements in them, data relating to the expression of these genes in the different stresses were received using Phytozome, Plant CARE and the Bio-Array Resource for Plant Biology, respectively. Significant differences in the number of repetitions of eachcis -regulatory element among promoters were determined by a variety of statistical methods available at bioinformatics IDEG6 tool. The results showed that eight cis-regulatory elements, including ARE, CAAT-box, ABRE, Unnamed-4, Box-W1, TGA-element, W box and TATA-box had significant and different occurrences between two promoters. It can be suggested that geneTRX5 unlike geneCAT2 is able to respond to biotic stress due to the presence of a greater number of specific regulatory elements in response to pathogen (Box-W1 and W box) and auxin (TGA-element). Also, a part of the expression pattern of these genes in response to abiotic stresses can be justified due to the presence of different number of repetitions of specific regulatory element in response to auxin (TGA-element) on theTRX5 gene promoter and anaerobic regulatory element (ARE) and abscisic acid regulatory element (ABRE) on theCAT2 gene promoter. Based on these results, TRX5 plays a role in response to biotic and abiotic stresses. Identification of these genes with functional response to biotic and abiotic stresses can be considered a huge progress in order to select candidate genes for genetic manipulation to enhance plants stress tolerance.

Shoot apical meristem (SAM) and root apical meristem (RAM) are regions in the plant which contain stem cells. SAM provides cells for development of all above-ground structures such as leaves, flowers, branches and inter nodes and RAM is responsible for formation of plant underground organs. Furthermore, root is major component of plant nutrient and use of water in the plant. Therefore, the growth and development of the plant depends on the function of meristematic regions in shoot and root apex. In this study, the transcriptome of the shoot and root tip of soybean were investigated to identify genes that are highly expressed in this regions and likely to have major role in these areas. Hence, RNA-seq data were analyzed using the CLC Genomics workbench.Results revealed 1956 and 1725 genes were specifically expressed in the shoot and root tip, respectively. Moreover in the shoot tip in comparison with the root tip, 3750 genes of differentially expressed genes had a fold change of two or greater and 3710 of down regulated genes in the shoot tip had a fold change of two or greater. Functional analysis of differentially expressed genes revealed that these genes are involved in catalytic, transcription, signaling and enzyme regulator activities.

The use of enzymes in organic solvents is the important for industrial and pharmaceutical industries. Reduced activity of enzymes in the presence of organic solvent could be due to the rigidity of the protein structure, denaturation and inhibition.Usually, the stability of enzymes in organic solvents decreases. To fix the problem, protein stabilization methods such as additives, protein engineering and chemical changes used. In this study, endoglucanase enzyme was produced, then the activity and stability of the enzyme in the different percentages of organic solvents, n-Propanol, Dimethylformamide (DMF) and dimethyl sulfoxide (DMSO) were studied. Finally, sucrose as additive was used for the stabilization of enzyme. The results showed that the solvent DMSO in concentrations of %5 and %10 increased the enzyme activity. The enzyme activity was reduced at higher concentrations of this solvent. Other solvents reduced the activity of enzyme from the low concentrations, among which DMF had the most effect on the reduction of enzyme activity. Stability of the enzyme by sucrose in the presence of the solvent DMSO was more in comparison with other solvents. Added sucrose had no effect on the stability of the enzyme in the presence of solvents npropanol and DMF.

Genetic variations were study for 12 accessions of Lolium perenne using molecular and biochemical markers. Soluble proteins were extracted for each population by Laemmli′s method and electrophoresis was done using polyacrilamid 12% gel. The numbers of 12 bands were observed for genotypes, which the numbers of 6 bands were polymorph.The highest number of band belonging to Tyrone with 12 bands and the genotype of Green Gold had the lowest number of band (6 bands). The results of cluster analysis and principal coordinate analysis were fall the genotypes in three groups. Genetic variation for accessions were surveyed using the number of 12 ISSR primers, that the number of 10 primers can be scored. The ISSR primers can be produced the number of 62 bands, which the polymorphism was showed for the number of 47 bands. The average of bands was 6.2 for each primer. The primer of IS9 showed the highest number of band (11 bands) and IS15 showed the lowest number of band (3 bands). A desirable polymorphism between genotype was observed based on ISSR markers, which the primers of IS9, IS10, IS13 and IS16 were determined for genetic variation study in Lolium perenneas desirable primers. Genetic variation based on ISSR marker had a average conformity with the results of protein surveys.

Hepatitis B virus (HBV) is of common viruses that transmitted by blood and blood products. The aim of this study is to design a polymerase chain reaction-based test to detect HBV in clinical samples. In this study, one blood sample with known load of HBV is prepared to assess the ability of designed primer pair. The specificity of the primer pair was somewhat confirmed by testing against three common virus types, in addition to bioinformatics analysis. The test was also used to screen applicants of blood transfusion organization of Kashan and its results compared with results of a current method used by this organization. The results of this study showed that the designed primer pair specifically amplifies the HBV genome and its sensitivity is 100 particle per ml of blood, which is more sensitive than the results of the ELISA method currently used in blood transfusion organization. In clinical samples, however, the results of the polymerase chain reaction (PCR) test were completely in accordant with the results obtained by blood transfusion organization after confirmatory tests. The designed test has no false positive results in contrast to ELISA, which, in addition to preventing the exclusion of healthy volunteer blood donors, could also have a significant role in reducing costs of the organization. So, it seems that it can replace the current screening method of this virus, that firstly the cost of doing it is much lower and secondly a part of bona fide person are not prevented from donating blood.

In this study, interaction between human serum albumin and CuO nanoparticle (NP) at different concentrations was studied using fluorescence method under the simulative physiological condition. The results showed the intrinsic fluorescence reduced in the presence of the NP as the manner increase NP concentration in media. According to the results of quenching rate constant (Kq), it deduced that the interaction take place through static mechanism. Thermodynamic parameters including enthalpy and entropy were calculated as -18.06 (kJ mol-1) and 0.0195 (kJ mol-1 K) respectively, which shows the importance of electrostatic interaction in HAS and NP interaction. Affinity of this particle to HAS deduced as the negative sign of free energy (DGo) that shows the interaction is exergonic process. Evaluation of ANS fluorescence intensity indicated that hydrophobicity of the surface protein increased in the presence of the NP. Based on the results it established, the protein structure changed in the presence of NP which may be affected its function.

Obesity is currently a risk factor for many diseases and health problems that one of the main causes of its incidence are genetic factors. One of the genetic factors that recently considered for obesity is polymorphisms of GNB3. This study aimed to investigate the association between C825T polymorphism in GNB3 with obesity in the population of Ardabil. In this case-control study, 100 obese subjects (case group) and 100 healthy subjects (control group) were included basis of their body mass index.5 ml of blood was taken for molecular tests from subjects. DNA of all blood samples was extracted using standard phenol chloroform procedure and its amount was measured using spectrophotometer. Primers for copying GNB3 gene sequence were designed using GNB3 sequence and Primer3 software. The obtained PCR-RFLP products electrophoresed on 2% agarose gel and by the 100volt current for 30 min along with a normal or control DNA sample and then were visualized by Ethidium bromide. The TT genotype frequency in obese individuals was significantly greater than its frequency in healthy individuals (38% versus 13%, P=0.0001). Also, the T allele frequency in obese individuals was significantly greater than its frequency in healthy individuals (60% versus 39%, P=0.0029). The results of this study showed that C825T polymorphism of GNB3 gene is associated with obesity in Ardebil population and can represent one of the possible genetic causes of obesity incidence in this area.

Datura stramoniumas a well-known medicinal plant is rich in tropane alkaloids such as hyoscyamine and scopolamine and currently the vast majority of the studies have been focused on optimization of production conditions of these valuable materials in the world. In this research, the effect of sucrose (15, 30 and 45 g/L) as an abiotic elicitor was investigated on fresh weight (FW), hyoscyamine and scopolamine production, protein content, activity of catalase (CAT) and guaiacol peroxidase (GPX) ofD.stramonium. The amount of hyoscumine and scopolamine were measured by high-performance liquid chromatography (HPLC) and spectrophotometery method was applied for studying of total protrin content and activity of CAT and GPX. According to the results, the highest amount of fresh weight (0.82 g), production of hyoscyamine (5 mg/g FW), scopolamine (20 mg/g FW) and CAT activity (0.5 U/mg protein) were recorded on the root of 45 g/L-treated plantlets but the highest amount of protein (0.24 mg/g FW) was observed on shoot of 30 g/L ones. In addition, the maximum GPX activity was determined on root of plantlests treated with 30 g/L sucrose (1.52 U/mg protein). In conclude, the concentration of 45 g/L of sucrose can be a good elicitor to increase production of hyoscyamine and scopolamine with high medicinal value inD.stramonium which probably due to the fact that sucrose does not only act as a carbon and energy source but also induces signaling pathway of tropane alkaloids synthesis.

Lignans are a large group of important phenolic compounds and synthesized by many plants such as Linaceae family. In this research, the effects of different concentrations of mycelium extract and culture medium of Piriformospora indica on the levels of phenolic compounds in L. album hairy roots were investigated. The amount of lariciresinol, pinoresinol, podophyllotoxin and 6-methoxy podophyllotoxin were also analyzed by HPLC. To study the fungal elicitors’ mechanism of action, phenylalanine ammonia-lyase (PAL) enzyme activity was investigated. Results showed that maximum amounts of podophyllotoxin and 6-methoxy podophyllotoxin were at 1 % (v/v) of fungal medium, 248.9 mg/g and 59.59 mg/g DW, respectively and the highest amounts of lariciresinol and pinoresinol respectively were seen at 1 and 0.5 % (v/v) of mycelium extract, 144.68 and 72.68 mg/g DW. Maximum amounts of total phenol, flavonoid and lignin were analyzed at 5 % (v/v) and flavonol at 1 % (v/v) fungal culture treatments which were 3.3, 4.6, 1.6 and 2.1 -fold greater than those of control. The activity of PAL enzyme was induced by mycelium extract and fungal medium, reaching a peak at 1 % and 5 % (v/v), respectively. Consider to these results, fungal culture medium was more effective than mycelium extract for stimulation of phenolic compound production.