مجله پژوهش های سلولی مولکولی (زیست شناسی ایران)
سال سی و سوم شماره 2 (تابستان 1399)

Antibacterial activity of alcoholic extracts of Thymus spp. and Aloysia spp. against Pseudomonas syringae in laboratory condition Abstract: P. syringae pv. syringae (Pss), the causal agent of bacterial canker and blast of stone fruit trees, is one of the most important plant pathogen in the world. This research was conducted to survey antibacterial activity of plant extraction on P. syringae pv. syringae bacterial strains. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) activity for bacterial strains were determined. Inhibitory concentrations (MICs) of alcoholic extract of Thymus spp. and Aloysia spp. were 0/65 and 1/31 and Minimum Bactericidal Concentration (MBC) activity was 1/31 and 2/62 mg/mL respectively. Pseudomonas syringae was recognized susceptible to Tetracycline, Erythromycin, Gentamicin and nalidixic acid with halo diameter 25, 15, 25 and 27 and medium susceptibility to Polymyxins with 15 mm and finally 15 mm were observed for Copper oxychloride. According to the results, plant extraction showed significant effect on control of the above mentioned bacteria in laboratory condition and with further research, it can be used instead of chemical compound. Keywords: Aloysia, canker, Pseudomonas, Thymus

TGF β1 is a multifunctional protein that is involved in several biological processes. This protein is initially synthesized as 390 amino acids biologically inactive latent precursor that prior to binding to the TGF β receptors; undergo proteolytic cleavage, to separate mature form of TGF-β1 from its propeptide. The mature form of this protein comprise of the C-terminal 112 amino acids from its precursor. In this study, the potential of two agrobacterium GV3101 and GV3101 (pMP90RK) strains were first investigated for the transformation of Nicotiana benthamiana with pTRAkc-ERH expression vector by creating the pTRAkc/CJG138 (including GFP) as a control construct. The results were analyzed by western blot technique and confocal imaging. Then, in order to transient expression of TGF β1 from its precursor sequence in the tobacco plant, recombinant pTRAkc/ DCF6 and pTRAkc/ DCF7 constructs (containing endoplasmic reticulum retention signal (SEKEDL) and the lacking of this sequence, respectively) were designed and generated. Tobacco plants agro-infiltrated with these constructs alone or in combination with another recombinant construct; harbouring the coding sequence for Furin protease. Results showed that the both of these constructs were successfully expressed in tobacco plants. However, more efficient processing of TGF β1 precursor in infiltrated plants with pTRAkc/ DCF7 construct show that the accumulation of TGF β1 precursor in endoplasmic reticulum prevent to its processing. In addition, in this research was revealed that a Furin-like cleavage between the mature TGF β1 and its propeptide does not occur in N. benthamiana.

In the most organisms, signaling photoreceptors have been made to percept and interpret of environmental signals. They can absorb light through their chromophores, trigger different signaling pathways and result in appropriate responses in the cells. Optogenetics, a new field of science, try to use the photoreceptors as molecular instruments in order to exact control of the cellular events. Determination of the new molecular instruments with special characteristics is one of the ways to develop the optogenetics. For this goal, investigation on the predicted genes ortologous with determined photoreceptors is very useful. Two Cryptochrome/ Photolyase family genes were predicted based on the results of Volvox carteri (the model algae) genome sequencing. The goals of this investigation are as follow: the exact determination and isolation of coding sequences related with these two predicted genes, insertion into the expression vector (pGEX2TK) and determination of the subclude of their open reading frames in the Cryptochrome/Photolyase family. Volvox carteri has been grown up in optimum condition. Then this two coding sequences were amplified by using RT-PCR technique. These coding sequences were inserted in the pGEX2TK by genetic engineering techniques. Produced recombinant vectors were transferred to the Escherichia coli strain Top10 and sequenced after plasmid extraction. In current study, it was found that documented mRNA sequences for these two genes in NCBI are different with our mRNA sequences.

With fast advances in next generation sequencing technologies, they has become powerful and low-cost tools for transcriptome studies, Nowadays; de novo assembly of transcriptome data, has caused the formation of the new pathway in the study of non-model genome species. With the expansion of this technology and increasing the number of assembly softwares, choosing the best software for assembling transcriptome sequencing data has become a challenge for biologists. For the first time in this study, we used transcriptome sequencing data of Dracocephalum kotschyi in order to reach the appropriate parameters and superior software; so here we used Oases-velvet, SOAPdenovo-Trans, Trans-ABySS and Trinity softwares with two different values of K parameter; K=25 and K=32. The results of assembly by each software were compared to others in the term of several criteria. The result suggests the superiority of Trinity and Trans-ABySS softwares. Finally, the output of the best assembly was used to estimate abundance of various isoforms and Gene Ontology analysis as regards to the pharmaceutical properties of this plant and the high amount of secondary metabolites, the highest frequency of sections in the biological processes was related to the metabolic activity.

Aspergillusniger is one of the microorganisms that produce the pectinase enzymes group of the at high levels. In this fungus, PgaA genes code Glycoside hydrolase family 28 that secrete polygalacturonase enzymes. Green algae (Chlorophyceae) and blue-green algae (Myxophyceae) having pectin, cellulose and hemicellulose compounds such as xylan, arabinoxylan in their structure could act as a precursor to the production of cellulase, xylanase and pectinase enzymes in fungi. In this work, the effect of green algae extract ; Spirogyra sp. at 500 and 1000 mg/mL concentration on the rate of pectinase enzyme secretion and the relative expression of PgaA gene at 4 and 8 day after adding algae extract to culture medium was evaluated using qRT-PCR analysis. . The highest level of enzyme secretion was observed at 8th days after algae extract adding to culture medium at 1000 mg/mL concentration( 4.28 U/ml). Gene expression analysis of PgaA gene showed that the highest expression level was relted to 4th days time interval after adding alge extract at 1000 mg.mL (10/142). According to positive effect of spirogyra algae compounds on expression level of PgaA gene responsible to pectinase production and increase of enzyme secretion, it is concluded that microalgaes could be used as important sources of enzyme production from favorable fungi and could be useful in biotechnological industry

T-cell acute lymphoblastic leukemia is a lymphoid malignancy affected by oncogenic transformation of immature T-cell progenitors. Recently, it has been reported that microRNAs play a role in various leukemias including T-ALL. The aberrant expression of these microRNAs in proliferation, invasion, and apoptosis has been reported through targeting signaling pathways. miR-34a is a tumor suppressor in many cancers, including T-ALL.

In this experimental study, Jurkat T-cell acute lymphoblastic leukemia (T-ALL) cells were maintained in RPMI 1640. miR-34a mimic was transfected using jetPEI in vitro DNA transfection reagent. RNA extraction and cDNA synthesis were performed after 24 h. Expression of miR-34a and BACH1, PTEN and SIRT1 genes were examined using qRT-PCR.

qRT-PCR analyses showed that in Jurkat cells after transfection with miR-34a mimic the expression of BACH1 and SIRT1 genes were reduced and the expression of PTEN was increased.

Low expression level of miR-34a in associated with metastasis and tumor invasion in different cancers. In current study, our results showed that the expression levels of BACH1 and SIRT1 genes which are oncogenes reduced and the expression levels of PTEN which is tumor suppressor increased after the transfection with miR-34a. 

Non-enzymatic glycation of proteins and the formation of its advanced glycation end products (AGEs) are one of the important factors involved in pathogenesis of diabetes chronic complications. Since these events damage the proper function of proteins, antioxidant compounds with plant origins can be used as a therapy to prevent such chronic effects. In this research, the effect of G. tournefortii L. hydroalcoholic extract of Lorestan province on the inhibition of the non-enzymatic glycation process of bovine serum albumin (BSA) in diabetes models was investigated. For this purpose, at first the phenol and flavonoid contents of the plant were determined. To investigate the antioxidant activity of this plant, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging test was used. Then, for the AGEs formation, BSA was glycated in vitro by using the glucose and the effect of different concentrations of G. tournefortii hydroalcoholic extract on the AGEs formation was evaluated by using various assays. The results showed that the hydroalcoholic extract in a dose-dependent (25- 400 µg/ml) manner has DPPH radical scavenging ability as well as antiglycation activity. In addition, the hydroalcoholic extract inhibited benzoate hydroxylation during glucose autoxidation. Concerning the involvement of oxidative reactions in AGEs formation under hyperglycemia condition and also regarding the high phenolic and flavonoid contents (antioxidant compounds) of the plant, it can be concluded that the G. tournefortii hydroalcoholic extract probably through its potent antioxidant activity inhibits the protein glycation process.

L-asparaginase is an enzyme which hydrolyze the amino acid asparagine into aspartic acid and ammonia. L-asparaginase used for leukemia and lymphoma treatment is produced mainly from bacteria such as Escherichia coli and Erwinia chrysanthemi and search continues to find new sources for this enzyme. In this study, isolation of bacteria producing L-asparaginase from wastewater of potato processing was performed.The wastewater containing all the soluble compounds in potato juice, such as significant amounts of asparagine amino acids. Bacterial isolation and screening were performed on nitrogen agar and M9 media. Two isolates, called PW4 and PW5, were able to produce ammonium in the M9 medium. The results of molecular identification and biochemical tests showed that these isolates belong to the genus Escherichia and Enterococcus, respectively. Quantitative and qualitative measurements of the enzyme production of these isolates were carried out in different conditions. The highest enzyme activity of 10 IUml-1 was seen in PW4 isolate growth in the in a medium containing glutamine. The maximum amount of enzyme activity of PW5 isolate, 2.33IUml-1 was observed as a result of bacterial culture in a culture medium containing asparagine. The enzymes produced by both isolates did not show glutaminase activity. The maximum activity of the enzyme produced by PW4 isolate was evaluated at 37 ° C and enzyme activity decreased at lower and higher temperatures. These results suggest that this enzyme has a potential to be further studies for use in drug design .

Fennel (Foeniculum vulgare Mill.) is one of the well-known medicinal and aromatic plants. To determine variability between 16 Iranian Fennel ecotypes based on molecular markers, an experiment was carried out based on completely randomized design with three replications at Payame Noor University of Kermanshah, Iran, in 2014. Ecotypes were evaluated based on 15 ISSR primer markers. All of the ISSR primers showed 77 visible bands. The ISSR13 and ISSR10 primers had the most number of bands with 10 and 9 bands respectively. The mean polymorphic information content (PIC=0.36), marker index (MI= 1.83), Effective multiplex ratio (EMR= 4.98) and polymorphism percentage (91%) indices were calculated for all primers. Total genetic similarity based on these primers was 0.567 percent. Cluster analysis classified all ecotypes to three groups. This clustering was confirmed by principal coordinate (PCo) and analysis of molecular variance (AMOVA). Genotypes were evaluated based on 15 ISSR primer markers. All of the ISSR primers showed 77 visible bands. The ISSR13 and ISSR10 primers had the most number of bands with 10 and 9 bands respectively. The mean polymorphic information content (PIC=0.36), marker index (MI= 1.83), Effective multiplex ratio (EMR= 4.98) and polymorphism percentage (91%) indices were calculated for all primers. Total genetic similarity based on these primers was 0.567 percent.

Colorectal Cancer is one of the most common malignancies and is one of the mojor causes of mortality. Todayes, researchers pay attention to use of herbal extracts, including phycocyanin and Citrus colocynthis extract for treatment of colon cancer. In this study the synergic effect of these compoundson colon cancer cells growth and expression of caspase-8 and Bcl2 genes was assessed. After treatment of human colon cancer cells (HT-29) in differnet times (24, 48 & 72h) and with different concentrations of aqueous and phycocyanin watermelon extract and also combination of aqueous extract and phycocyanin, percentage of live cells was calculated using MTT method. Then expression of caspase-8 and Bcl2 genes were measured by real-time PCR. The highest inhibition of growth of cancer cells was observed after treatment with high concentration of both compounds. In addition, significant differences were observed between the different treatment times with low concentration (1 and 2 g/ml). The aqueous extractand phycocyanin reduced the expression of Bcl2 gene by about 10.27 and 5.22, respectively. However, the combination of extract and phycocyanin was associated with a decrease of 7.31% in expression of this gene. The extract increased the expression of Caspase-8 gene by 2.3 fold. Citrus colocynthis extract induces apoptosis by strongly increasing the expression of caspase-8 gene. Additionally, the phycocyanin and the combination of extract and phycocyanin strongly decreased the expression of Bcl2 gene. Phycocyanin and extract and Phycocyanin combination had stronger anti-cancer effects than extract alone.

Rice is one of the most important crops in the world. Also, It is the main source of food and carbohydrates in the world. In this research, the relationship between the characteristics of 90 rice genotypes (Traditional and improved) with 11 SSR markers evaluated. This research was conducted as a lattice design with three replications in 2015-2016 at Gonbad Kavous University under normal and drought conditions. Multiple regression analysis between measured traits and SSR markers showed a significant relationship between data. The results of cluster analysis for agricultural traits in normal and drought stress genotypes were divided into four and three groups. The results of multiple regression analysis showed that the number of secondary branches and fertility, the most percentage of variation of explanation of yield variation, under normal and drought conditions, respectively. The results of this research can be used for breeding programs and selection of suitable and tolerant cultivar.

Seed of 22 populations of Tanacetum (12 species and 7 subspecies) and seed of Pisum sativum, Petunia hybrid and Triticum aestivum as internal standards were selected and were caltivated under the same greenhouse conditions. Genome size (C-value, mass of DNA per haploid nucleus) was estimated by flow cytometry. Data from the study were analyzed by SPSS 16 software. The result revealed genome size is positively correlated with pollen morphometric, shape and colour of capitule, type of inflorecences and corology of species, but is negatively correlated with size of seed and environmental factors such as altitude and humidity of the habitat. The variation in the 2C value (mass of DNA per diploid nucleus) was high, with a range from 3.84 pg in Tanacetum parthenium (Tehran) to 24.12 pg in Tanacetum polycephalum subsp. farsicum. As wellas a 6.28-fold variation in 2C value and 2.73 fold variation in C value were found. Key words: Altitude , Genome size, Pollen grains, Morphology, Flow cytometry.