Journal of Molecular and Cellular Researches
Year:2019 | Volume:31 | Issue:4|Papers Count:13

Cadmium sulfide nanoparticles (CdS NPs) have many commercial applications in making dye-sensitized solar cells, light emitting diodes (LED) and nanobiosensors due to their unique properties. In this work, screening of aquatic bacterial strains for their ability in the green extracellular synthesis of CdS NPs was studied. The prepared nanoparticles were examined using Optical emission and absorption spectroscopy analyses, Energy Dispersive X-Ray Spectroscopy (EDS) Scanning electron micrograph analysis and Fourier-transform infrared spectroscopy (FTIR). 32 cadmium-resistant aquatic strains were isolated by using enrichment. Among isolated strains, Pseudomonas pseudoalcaligenes is capable to synthesize CdS NPs. The extracellular nanoparticles produced by the strain Cd11 were investigated under optimal culture conditions. The results showed that culture supernatant of strain Cd11 treated with CdSO4 (3 mM) was able to catalyze the extracellular synthesis of CdS NPs in the range of 12-19 nm with an average size of 15. 5 nm after 10 h of incubation at pH 8. 0 and temperature 35 ° C. To evaluate the antibacterial activity of nanoparticles, agar well diffusion method was tested. Based on obtained results, CdS nanoparticles had an inhibitory effect against all tested microorganisms. Additionally, particles size distribution of CdS nanoparticles synthesized by culture supernatant of strain Cd11 had a good monodispersity and homogeneity for the specific applications.

HMGB4 protein which was identified in 2008 is a member of mammalian non-histone HMGB protein family. All HMGB proteins bind to DNA through two tandem DNAbinding motifs named HMG-box A and HMG-box B. HMGB proteins are known to improve the anticancer properties of platinum-based drugs such as cisplatin by high affinity binding to the platinum-DNA adducts and retarding the process of DNA excision repair. The results from previous studies have revealed in comparison with full-length HMGB1, the full-length HMGB4 binds with higher affinity to platinum-DNA lesions and inhibits the excision repair of the lesions much more efficient. It seems the hypersensitivity of testicular germ cell tumors to cisplatin is originated from strong expression of HMGB4 in this tissue. In the present work, the tertiary structure of human HMGB4 was determined using two methods based on homology modeling (MODELLER software and I-TASSER server). The results show that models created by both methods have good quality and stability. Although I-TASSER model in comparison to the modeller better explain the differences between HMGB1 and HMGB4 binding to the platinated DNA. The results obtained from the work demonstrate that HMGB4 binds to DNA by intercalating of Valine17 and Phenylalanine38 from box A and Leucine101 and Valine120 from box B between base pairs of DNA minor groove. The results also show the C-terminal part of HMGB4 possesses disordered tertiary structure. In general, the results will be helpful to understand the structure-function relationship of the protein and to design new anticancer drugs.

Hyaluronic acid (HA) is a major component of the extracellular epithelial, neural and connective tissue of vertebrates, as well as of extracellular microorganisms. This polysaccharide has specific applications in ophthalmology, orthopedic, cosmetics and tissue engineering due to features such as high viscosity and biocompatibility. Animal tissues and streptococci have a major role in producing hyaluronic acid, but because of problems such as protein contamination and the presence of endotoxin, attention has been drawn to the generally recognized as safe (GRAS) bacteria for producing hyaluronic acid. Lactobacilli are probiotic bacteria that have diverse uses. The production of biochemical macromolecules by this group of bacteria is a priority because they are GRAS. Hyaluronic acid production for the first time in these GRAS bacteria including Lactobacillus acidophilus PTCC1643, Lactobacillus rhamnosus PTCC1637, Lactobacillus casei PTCC1608 and Streptococcus thermophilus PTCC1738 were investigated. In the first step, Hyaluronic acid was isolated from the specific culture medium and partially purified by ethanol precipitation daily for 96 hours. Carbazole method was used to measure the production of hyaluronic acid. Based on results, Lactobacillus rhamnosus produced about 0. 4 g/l of hyaluronic acid at 96 hours. The increase of HA was also investigated using optimization culture medium by Taguchi design. In this method, optimal points of temperature, urea, lactose and trace elements were determined at 37 ° C, 5 g / l, 30 g / l and 1. 5 ml / liter, respectively, and production of hyaluronic acid was increased by 162 %.

Cellulases are important industrial enzymes for the most bioconversion processes. In this study, Bacillus aerius isolated and identification as the best thermophilic cellulose degrading bacterium from Gorooh hot spring and was identified as Bacillus aerius AV10 based on 16S rRNA sequence homology and microbial-biochemical tests. Cultivation conditions of cellulase production by Bacillus aerius AV10 in solid-state fermentation (SSF) were investigated. The moisture content, yeast extract, MgSO4 and initial culture pH were identified by Plackett-Burman design (PBD) as the significant factors for endoglucanase, exoglucanase and filter paper (FPA) activities. Response surface Methodology and central composite design by 5-level and 4-factor were used for the optimization of enzyme. The optimal ranges of moisture content, yeast extract, MgSO4 and initial culture pH were 61. 4%, 1. 65%, 0. 29% and 6. 2, respectively. Under the optimized conditions, maximum CMCase production was 561. 12 U/ml. The optimal pH and temperature of the enzyme for wheat straw hydrolysis were determined to be 7. 0 and 60 ° C.

Reducing fossil fuels, increasing fuel prices and carbon dioxide emissions, and concern about climate change are encouraging factors for biofuels production. Microbial biofuels are liquid and gaseous fuels which produced by microorganisms. Bioethanol is a suitable alternative biofuel for petroleum based fuels. For the industrial and high-scale production of bioethanol, the first step is isolation of suitable yeast strain with high tolerance to ethanol and high-level ethanol production. Hence, the aim of this study was Isolation of industrial strain Saccharomyces cerevisiae with high tolerance to ethanol from Iran's alcohol manufactures. Sampling from several Iran's alcohol manufactures was carried out for the isolation of Saccharomyces cerevisiae with the highest ethanol production and tolerance. Selected yeast strain was examined by morphological, biochemical tests such as sugars fermentation, absorption of nitrogen compounds and carbon compounds, and also molecular test. The selected yeast strain produced 8% ethanol and tolerated 12% ethanol. The morphological, biochemical and molecular tests confirmed that the yeast strain is Saccharomyces cerevisiae, and this strain was named Saccharomyces cerevisiae Sahand 101. The isolated industrial yeast strain with the suitable characteristics in terms of ethanol production and tolerance to ethanol can be used for further studies and increase ethanol production by optimization methods at the level of flask and bioreactor.

Mentha Spicata is an Iranian native plant with therapeutic effects. In this study, the effects of M. Spicata essential oils were investigated through considering oxidative injury parameters as well as expression of COX-2 inflammatory gene in lung tissue of experimental cecal ligation and puncture (CLP) rat model. Rats were divided into 5 groups: negative control (lapratomy), CLP, treatment groups with M. Spicata E. O (50 and 100 mg/kg b. w) and indomethacin. Then, 24h after CLP induction, LP, GSH, GST, FRAP, MPO, PGE2 and COX-2 expression were measured in plasma and lung tissues. The data indicated that the sepsis induction reduced GSH, FRAP and increased LP, MPO, PGE2 and COX-2 levels but didn`t affected GST activity. Treatments of rats with E. Os as well as indomethacin were been effective in increasing the GSH, FRAP and decreasing the LP, MPO, PGE2 and COX-2 levels. Histopathological examinations indicated that sepsis caused lung tissue damage which was decreased by E. O treatments. In conclusion, sepsis causes oxidative lung tissue damage and use of M. Spicata can prevent injuries through modulation the oxidative stress/antioxidant parameters.

The Clustered Regularly Interspaced Short Palindromic Repeats associated Cas9/gRNA system (CRISPR/Cas9) is a novel and an efficient targeted genome-editing technique are used to achieve new traits and crops with high quality and quantity yields. Recently, CRISPR/Cas9 mechanism has been used extensively to genome editing, gene knockouts, reducing off-target effects and modulation of gene transcription with high specificity in several animal and plants through mediating double strand breaks (DSBs) in genomic DNA and making nucleotide changes in target gene. In this review, we have summarized the broad applicability of Cas9 nuclease-mediated targeted genome editing of plants for development of genetically edited crops, climate resilient and bio-energy agriculture and to develop non-GM plants along with regulating the uncertainty and the social acceptance of this new plant breeding technique. The development of GE crops, which is similar to conventional breeding crops show the high stability and efficient properties compared to genetically modified (GM) crops in different climates, social acceptance and environmental and human health concerns.

Endophytic bacteria live inside tissues of their plant host without causing visible symptoms, and in more cases are reported as plant growth promoting bacteria. This research was conducted to determine plant growth promoting affects of endophytic bacteria associated with Soybean (cultivar L17). In order to isolate endophytic bacteria from various parts of soybean (cultivar L17), samples were collected from the research farm of Agriculture faculty in this research endophytic bacteria were isolated from leaf, stem, seed and root of. After genomic DNA extraction, 16S rDNA gene was amplified using PCR. Then, the PCR product was sequenced by BLAST. Strains were surveyed for IAA and GA production ability was carried out using a randomized complete design in three replications. Analysisofvariance and mean comparison was performed using Duncan Test with SAS software. The isolated bacteria were able to produce IAA in various amount 20/03-45/33 without tryptophan and in the presence of it, 26/76-51/17 microgram/milt. GA producing abilities were also 12/97-17/15 microgram/milt for these isolates. Based on the 16S rDNA sequence studies, this bacterium belonged to Pseudomonas fluorescensand indicated 99% similarity to type strain. This study is the first report of isolation ofPseudomonas fluorescensfrom Soybean (L17 cultivar). The endophytic bacteria isolated in this study can be used to promote plant growth.

The development of nanotechnology is making the intrest of researchers towards synthsis of nanoparticles for the bio application. In the present study, copper nanoparticles (CuNPs) have been synthesized by the reduction of copper sulfate using aqueous extract of Postia puberula Flora. The green synthesized CuNPs were characterized using UV-Vis absorption spectroscopy, Scanning Electron Microscopy (SEM) and Enerrgy Dispersive X-ray analysis(EDX) The antimicrobial activity of CuNPs was determined by disc diffusion and broth microdilution methods against some selected species of bacteria. After addition of flower extract, the change in colour of Copper Sulfate solution was noted from light blue to sea green. The UV-Vis absorption spectra confirmed the formation of the CuNPs with the characteristic peak 575 nm. XRD and SEM analysis of copper nanoparticles indicated that they exist in amorphous in nature and with size ranged from14 to 39 nm. Fruit extract acts as both reducing and capping agent. The synthesized Cu nanoparticles exhibited activity against of both gram positive and gram negative ba cteria.

Reducing fossil fuels, increasing fuel prices and carbon dioxide emissions, and concern about climate change are encouraging factors for biofuels production. Microbial biofuels are liquid and gaseous fuels which produced by microorganisms. Bioethanol is a suitable alternative biofuel for petroleum based fuels. For the industrial and high-scale production of bioethanol, the first step is isolation of suitable yeast strain with high tolerance to ethanol and high-level ethanol production. Hence, the aim of this study was Isolation of industrial strain Saccharomyces cerevisiae with high tolerance to ethanol from Iran's alcohol manufactures. Sampling from several Iran's alcohol manufactures was carried out for the isolation of Saccharomyces cerevisiae with the highest ethanol production and tolerance. Selected yeast strain was examined by morphological, biochemical tests such as sugars fermentation, absorption of nitrogen compounds and carbon compounds, and also molecular test. The selected yeast strain produced 8% ethanol and tolerated 12% ethanol. The morphological, biochemical and molecular tests confirmed that the yeast strain is Saccharomyces cerevisiae, and this strain was named Saccharomyces cerevisiae Sahand 101. The isolated industrial yeast strain with the suitable characteristics in terms of ethanol production and tolerance to ethanol can be used for further studies and increase ethanol production by optimization methods at the level of flask and bioreactor.

The aim of this manuscript is investigation, extraction and expression pattern evaluation of lifeguard genes in fresh water polyp Hydra vulgaris. For this purpose, about 100 freshwater polyps were cultured and total DNA extraction was performed. Specific primers were designed for all genes and PCR reaction applied to amplify Lifeguard genes. In order to investigate the gene expression pattern, In situ hybridization experiment used. The experiments revealed that there are three lifeguard genes in hydra, which are called Lifeguard-4 and invertebrates’ Lifeguard-1. In situ hybridization shows that Lifeguard-4 expressed in endodermic layer of polyps in all budding stages and also in different stages of oogenesis. In Conclusion, Phylogenetic tree indicates that lifeguard genes are well conserved and present at the base of Euometazoa like freshwater Hydra polyp. Since apoptosis signaling pathway is present during Hydra gametogenesis (oogenesis and spermatogenesis), expressions of Lifeguard-4 as an apoptosis regulatory gene, in places were apoptosis occur is logistic and confirms the regulatory function of these genes.

Androgenesis is an effective method of doubled haploid plant regeneration in breeding programs. For this purpose, anther and microspore cultures of two medicinal plants, Satureja khuzistanica Jamzad and S. rechingeri Jamzad, were studied. As the first step, uninuclear developmental stages of microspores were determined in relation to flower size, using acetocarmen staining. In anther culture, the effects of different temperatures, hormonal treatments and carbohydrate sources (maltose, sucrose) on androgenesis were studied in N6 induction medium. The ploidy level of calluses was determined using flow cytometry. Isolated microspores were also subjected to three induction media (NLN-13, FHG, A2-60), different temperatures and hormonal pretreatments. The highest percentage of callus induction was obtained in hormonal treatment of 0. 5 mg/l of BA, 2, 4-D, and Kin with temperature treatment of 30 º C for 3 days. Based on flow cytometry results, ploidy level of almost all calli was similar to that of donor plants and only some calli of S. rechingeri in the media containing 0. 5 mg/l of BA, 2, 4-D, and Kin were haploid. In the regeneration medium (MS), only roots but no shoots were observed. In microspore culture of both species, the highest numbers of multi-cellular structures were observed in NLN-13 and FHG induction medium with temperature treatment of 30 º C for 8 days. Heart-shaped embryo like structures were only observed in NLN-13 induction medium. The finding of this research will be a good base for haploid plant regeneration of S. khuzistanica and S. rechingeri.

Powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt) is one of the most important diseases of wheat that make yield damage annually. In recent years, the use of induction elicitors has been widely welcomed. Therefore, in this research in order to examine the ability of chitosan as a biotic elicitor, Flat were selected as susceptible cultivar to powdery mildew, and the leaves were sprayed with chitosan in the two-leaf stage. Then treated plant together control plants were inoculated with Bgt pathogen. All stages of the were conducted in 2016 in the genetic laboratory in Gonbad-e Kavouse University. Finally, The expression rate of MLO, B-I1 and NPR1 genes was examined in using Real Time PCR technique at 4 time courses and in 3 independent replicates. Results showed that in both chitosan treated plants and the controls, rate of gene expression were increased after infection for all genes, So that, in the first hours after inoculation, treated plants showed NPR1 expression more rapidly than control. Maximum expression level of this gene was observed at 24 hours after infection. But, in treated plants, the expression rate of BI-1 and MLO were less than the control and they showed minimum expression at 24 hours after infection. These results indicated these genes by decreasing their expression rate in order to increase the level of active oxygen species causes induction of cell death in the plant to inhibition of fungal growth and development in the early stages after infection. The gene expression rate in plants treated with 250 mg/L was more indicative than those treated with chitosan 500 mg/L. Overall; results showed that chitosan could induce systemic resistance in susceptible wheat against powdery mildew in low concentration.