نشریه مهندسی ژنتیک و ایمنی زیستی
سال دهم شماره 1 (پیاپی 19، بهار و تابستان 1400)

Introducing an effective antibacterial agent is very important in controlling pathogenic and antibiotic-resistant bacterial strains. Recent advances in nanotechnology, especially the ability to fabricate biological nanoparticles, have led to the development of a wide range of antimicrobial agents. Therefore, the aim of this study is to investigate the antibacterial properties of ZnO nanoparticles (NPs) in inhibiting resistant strains of Acinetobacter baumannii. Ten samples of Acinetobacter were collected from clinical centers of Khorramabad city in 2020 and were identified based on biochemical tests. After treatment of MDR strains with sub-minimum inhibitory concentration (MIC) of ZnO NPs, the expression level of efflux pump genes (AdeA, AdeC) was evaluated using real-time PCR. After applying the effect of ZnO NPs on AdeA gene, 80% of the samples showed reduced expression. 20% of the sample also had incremental expression. Also in AdeC gene, 40% of the samples showed reduced expression after the effect of ZnO NPs. 60% of the samples also had incremental expression. According to the results, the optimal inhibitory effect of biosynthesized ZnO NPs on the expression of AdeA and AdC genes, which play an important role in drug resistance of the Acinetobacter species, was evident. On the other hand, due to the increased expression in 60% of the samples for the AdeC gene, the possibility of mutations in the regulatory genes was raised. It seems that ZnO NPs can be used as a pharmaceutical candidate in the pharmaceutical industry in the future.

Chrysanthemums (Chrysanthemum morifolium) are an important ornamental plants in Iran and the world with a high commercial value. Among different doses of gamma irradiation, the irradiation at 40 Gy by a Cobalt-60 source to four-week-old seedlings of a purple commercial cultivar of chrysanthemum, i.e. ‘Rambla’ produced three mutants with dark purple, pale purple, and Indian red colors. To explore the effect of variations in the expression levels of the structural genes involved in the anthocyanin biosynthesis pathway on the color of the mutants, the RNA of the mutants and control plants was extracted from the petals of the M1V3 generation and studied by the RT-qPCR method. The significant changes in the relative expression levels of F3'H, DFR, ANS, and 3GT in the mutants versus the control showed the role of these genes in the petal color change. The total anthocyanin content had significant correlation with the relative expression levels of DFR, ANS, and 3GT. Stepwise regression for the total anthocyanin content as the dependent variable and the relative expression level of the structural genes involved in the anthocyanin biosynthesis pathway as independent variables showed that 3GT had the strongest impact on dependent variable. F3'H relative expression caused significant changes in the mutants versus the control and it was introduced as an appropriate key gene for genetic manipulations to create color diversity in chrysanthemums.

ABC transporters (ATP-binding cassettes) are among the largest gene families found in the three domains of bacteria, eubacteria, and eukaryotes. Most known ABC proteins are ATP-dependent, and membrane transporter and transport a wide range of molecules. These carriers play an important role in various biological processes. Since no comprehensive study has been performed on the identifying of ABC transporters in the potato plant, in this study, by searching the NCBI database and screening the identified genes, 228 ABC transporter genes were finally identified. Analysis of gene ontology (GO) showed that about 53% of ABC transporters in biological processes are responsible for transporting materials. In the analysis of molecular function and cell content, it was found that 32% are ATP-binding and 43% of them are inner membrane actuators, respectively. Also, the Cis elements in the promoter of ABC transporter genes was investigated, which finally identified 13 elements in drought stress and three elements in salinity stress. Finally, the expression pattern of eight selected ABC transporter genes under drought and salinity stresses was investigated using qPCR method. The results showed the effect of the studied genes under stress. StABC-050, StABC-084 and StABC-124 genes in drought stress and StABC-084 and StABC-001 genes had high expression in salinity stress. Then, to ensure the application of the applied stresses, the amount of proline and RWC and Na+/K+ were measured. According to the results, with increasing drought stress, the amount of proline in different developmental stages increased and RWC decreased, so that this decrease showed a significant difference at the level of 1%. The aim of this study was to identify and evaluate the expression pattern of selected ABC transporter genes under drought and salinity stresses in potato.

Respiratory diseases and pneumonia are among the most common diseases affecting more than 50% of the world's population. Various drugs have been developed to prevent and treat these diseases. rT1 peptide is a plant peptide isolated from the medicinal plant of Hibiscus sabdariffa. This 27-amino-acid peptide with inhibitory properties of human neutrophil elastase has been shown to have drug potential in the prevention and treatment of inflammatory lung diseases. Due to the high economic value of the compounds with neutrophil elastase inhibitory properties, in this study, for the first time, recombinant expression vector pBI121-rT1 was constructed with the aim of expressing rT1 peptide in plants. Because of the lack of rT1 gene sequence in databases, first, its appropriate gene sequence was designed and synthesized based on the Nicotiana tubaccum codon usage, the vectors used in this research, the structure and stability of its transcript and characteristics of the produced protein. For the designed sequence, the Codon Adaptation Index (CAI) and the GC frequency were 0.92 and 42.32%, respectively. In comparison to the main peptide, the stability of the designed peptide in human cells increased 25 times and its instability index calculated to be 20.90. Following amplification of the synthesized sequence by PCR reaction, it was cloned into pMCS5 vector. In order to express in plant-based systems, sub-cloning was performed in the expression vector pBI121 under the CaMV 35S promoter.

Salinity is one of the most determinative abiotic stresses in wheat cultivation. In order to overcome this concern, understanding how genes respond and act under stress conditions can provide useful information for plant breeding to withstand environmental stresses. The aim of this study was to identify genes with significant differences between two normal and stress conditions in bread wheat (Triticum aestivum). Gene expression patterns of pyrroline-5-carboxylate synthetase, serine-threonine protein kinase and Aldehyde dehydrogenase family 3 member I1, chloroplastic-like showed that: under salt stress, the genes expression levels have increased while the expression of Proline dehydrogenase 2, mitochondrial like, Putative calcium binding protein CML29 and GDSL_esterase / lipase_At2g40250-like genes decreased compared to control. Proline content and expression of key genes involved in proline biosynthesis under stress conditions showed significant changes in comparison with control conditions. Salinity stress induced P5CS expression and increased proline levels, which could indicate the crucial role of pyrroline-5-carboxylate synthetase as valuable gene transfer from stress tolerant to sensitive plant to increase plant tolerance under salinity.

Despite the negative effects of salinity on plants, due to the limited area under cultivation, planting a variety of plants in saline lands is inevitable. Among them, okra is one of the plants that few studies have been done on its response to different levels of salt. Therefore, in order to study the effect of salinity levels on Okra (Abelmoschus esculentus Moench), an experiment was designed based on a randomized complete block design with five treatments including concentration of 0, 50, 100, 150 and 200 mM sodium chloride in three replications at the greenhouse of the faculty of agriculture, Shahid Madani University of Azarbaijan. The results showed that by increasing salinity levels, potassium to sodium ratio, chlorophyll a, relative water content, plant dry weight, number of fruits per plant, fruit weight per plant, number of seeds per plant, seeds weight per plant and 1000 grains weight have decreased but amount of sodium and proline have increased. During the experiment, the plants under 200 mM treatment have died shortly after salinization, but plants under other treatments survived until the end of the experiment and produced the yield. Maintaining high potassium content, low reduction in chlorophyll content and no noticeable change in relative water content were factors that led to continuance of plant growth, dry matter and fruit production at different salt concentrations. Although in terms of number of fruits per plant, there was no significant difference between salinity levels and the control, but a significant decrease in fruit weight per plant was recorded at high salt concentrations, especially under 150 mM, which can be an important factor affecting economic yield of okra in saline environments with EC higher than 10 ds/m.

Autophagy is one of the defensive response pathways under stress and non-stress conditions in different organisms. In autophagy, damaged compounds and organelles are sealed into double-membrane vesicles as an autophagosome. Autophagosome has been directed to the vacuole to be degraded by hydrolytic enzymes into initial substances. ATG8 proteins are the hallmark of this pathway. These proteins are involved in autophagosome formation and target cargo location inside the autophagosome. In selective autophagy, cargo receptors play an important role in trapping the cargo inside the autophagosome. There is the ATG8 binding motif called AIM or LIR in Cargo receptors. This motif makes able the cargo receptors interact with LDS, LIR docking site in ATG8. Any modification in the AIM motif or LDS could abolish the interaction between ATG8 and cargo receptors. In this study, ATG8B mutant in LDS has been generated in Marchantia polymorpha for future studies to identify the possible cargo receptors. In this order, the protein sequence of ATG8B has been identified in the marchntia.info database. Aligning ATG8A from Arabidopsis with ATG8B from Marchantia leads to identifying the residues involving in ATG8B folding and LDS formation. Two residues, Lysin 51 and Arginine 70, have been replaced with Alanine residue by site-directed mutagenesis. The GreenGate has been used for fusing the GFP to ATG8B mutant fragments. To consider the same condition with wild-type ATG8B lines presented in the lab, by using the Gateway adapted primers, ATG8B-LDS mutant has been cloned in the expression vector, pMpGWB303. After transforming to Agrobacterium GWB303+pSOUP strain, it has been transformed to the plants. Analyzing the transformed plants in western blot experiments has shown that the only form of modification that can express and fold as a stable protein is Lysine to Alanine modification, however, in the proteins with the mutation of Arginine to Alanine, protein is not stable, and it cannot be expressed.

Soil salinity is one of the most crucial agricultural problems that in addition to having negative effects on crop production, causes hormonal imbalance and growth reduction in the plant. Plant growth-promoting rhizobacteria (PGPR) are a range of bacterial strains from different groups that live in plant roots and rhizospheres and can cause the plants to withstand stress by causing complex alterations in their growth and development. PGPR alters plant metabolism, signaling, and hormone homeostasis. The present study aimed to evaluate the effect of two auxin-producing Bacillus strains on vegetative traits and the relative expression of the ARF15 gene of two genotypes of chickpea (Cicer arietinum) (MCC77 and MCC68) under salinity stress of 50 and 100 mM sodium chloride in greenhouse conditions. The number of sub-branches of MCC68 has been significantly increased in the inoculation of strains at the concentrations of 0.5 and 1 in the presence of 50 mM sodium chloride. Moreover, the inoculation of Bacillus amyloliquefaciens (FZB42) and Bacillus. cereus at a concentration of 0.5 and 1 led to a significant increase in the fresh weight of MCC77 root. In addition, FZB42 at the concentration of 1 caused a significant increase (28.20%) in the height of MCC77 plants in the presence of 100 mM sodium chloride. In the inoculation of Bacillus sp. at the concentration of 0.5 and the presence of 50 mM sodium chloride, the relative expression of the ARF15 gene of MCC68 genotype showed a 9-fold increase. Both FZB42 and B. cereus strains affected the vegetative traits of MCC68 and MCC77 genotypes, which indicated the role of PGPR in increasing the supply of enzymatic and non-enzymatic metabolites, and the expression of plant genes in response to abiotic stresses.

Plants utilize different strategies to combat abiotic stress, depending on the species and growth stage. One of these strategies is to increase the remobilization of water-soluble carbohydrates (WSC) under stress, which can be important sources of carbon to fill the grain in response to drought stress. Drought is one of the most important abiotic stresses affecting yield of agricultural products. In addition, drought stress is one of the main limiting factors in plant growth, it can also prevent respiration, photosynthesis and opening and closing of plant stomata. As a result, it affects the growth and physiological metabolism of the plant. In response to drought stress, plants activate drought response mechanisms such as morphological and structural changes as well as the expression of responsive-drought genes, the synthesis of hormones and osmotic regulators to reduce drought stress. Drought initiates the senescence of cereal leaves, including changes in the expression of thousands of genes that ultimately affect grain protein content, grain yield, and nitrogen utilization efficiency. Also, under drought stress, soil nitrogen availability is reduced causing initiation and acceleration of the leaves senescence. Leaf senescence is strongly influenced by plant hormones and environmental factors including the availability of nitrogen. During maturity or drought stress, reduced nitrogen uptake can cause nitrogen to be redistributed from leaves and stems to seeds, eventually leading to leaf senescence. Under these conditions, genes involved in the fructan biosynthesis pathway and in chloroplast degradation and proteases show increased expression. For example, genes involved in protein degradation (proteases) and transcription factors (NAC, WRKY) are expressed in the process of cells senescence. In this paper, it was shown that the genes involved in the fructan biosynthesis pathway, chloroplast degradation, protein degradation (proteases), and transcription factors (NAC, WRKY) during the aging process show increased expression.

The COVID-19 outbreak has turned into a serious pandemic emergency, and presently is a global concern for humans worldwide. Lack of timely management and control of this disease could lead to a human catastrophe. Accordingly, various therapeutic interventions are being conducted for the COVID-19 remedy worldwide. The use of herbal medicine is one of the proposed remedy options in China and many other countries. The high homology of this viral genome with SARS (80%), and the presence of conclusive evidence of herbal medicine applications in the treatment and prevention of SARS are promising. This article focuses on the use of herbal medicine with potential application for COVID-19 remedy, and assessments of the inhibitory effects of these compounds on vital coronavirus proteins with molecular docking simulations. We also provide an opportunity to reveal the application of artificial enzymes for COVID-19 treatment. The use of artificial enzymes for the COVID-19 remedy is proposed for the first time in this study. Our molecular docking simulation results demonstrated that Glycyrrhetinic acid has an impressive inhibitory effect on all studied coronavirus proteins. Furthermore, carbonyl groups were identified as the key functional groups in the ligand-protein interactions. These can be subjected to the artificial enzyme designing for the COVID-19 remedy.

The National Biosafety Law was approved by the Islamic Consultative Assembly (the Parliament) and entered into force in 2009. According to the law, the National Biosafety Committee was established and tasked with the coordination of policies and harmonization of national competent authorities identified in the Law. However, other organizations with no legal involvement according to the Biosafety Law have played a more significant role; mostly in the prevention of the implementation of the Law. Therefore, in practice, the players and organizational chart were different from what has been approved by the parliament. We attempt to create a real map of the institutions playing role in the implementation of the National Biosafety Law. Besides, we evaluate the performance of National competent authorities listed in the Law. This research has been conducted based on the framework of political evaluation and qualitative research method. Data were collected through semi-structured interviews with stakeholders and authorities as well as the scientific Society members in the field including University Professors. Complementary data was collected through a structured literature review and content analysis of all related laws and regulations. The collected data was analyzed using Atlas TI software. The findings show that among the competent authorities, it was only the “Ministry of Health and Medical Education” (MHME) that was able to fulfill its duties to the “acceptable” level. MHME performance in implementing the law can be characterized by two main features: 1) dissociation from ideological evaluation and adhering to the text of the law and scientific evaluation, and 2) not accepting illegal pressures exerted by pressure groups including a couple of the security organizations and related media. But the performance of the Ministry of Agriculture and the Environmental Protection Organization has been criticized in particular by the scientific society members. The Ministry of Agriculture and the Environmental Protection Organization has reportedly failed to carry out their legal duties, mainly due to the ideological stand of the authorities, their position, non-compliance with the law, and obedience to the illegal demands of an unauthorized organization and media under their control.

Using genetic engineering for gene transformation through crossable species is known as Cisgenesis and Intragenesis. These two technologies have been developed to respond to the considerations expressed in relation to Transgenesis. These technologies have technical limitations compared with Transgenesis. In these technologies, it is not possible to transfer a gene to the recipient organism from species that their crosses are rare in nature. Cisgenesis and intragenesis are different in some aspects such as the removal of introns and the selection of promoters and terminators from other genes. These restrictions have been met in comparison to transgenesis, in the hope of exempting from biosafety regulations. The issue of reduced biosafety evaluations of these technologies has been debated in various countries. Some countries accepting the reduced biosafety evaluations and others rejecting it for some reasons. Considering that definition of Living Modified Organisms (LMOs) produced in Cisgenesis and Intragenesis such as transgenesis is in full compliance with the definition of LMOs in Cartagena Biosafety Protocol, products of these technologies would not be exempted from Biosafety regulations in international level. In addition, several studies have shown that gene transfer from distant species is possible in nature, and also some other concerns remained unanswered, exempting these products from biosafety regulations is not advisable.