مجله بیوتکنولوژی کشاورزی
سال سیزدهم شماره 2 (پیاپی 43، تابستان 1400)

The use of herbal medicines is even more common in East. Besides, more than half of all spinal cord injuries happen in individuals aged 15 to 30, most of whom are males (80 percent). Following spinal cord injuries, male fertility especially sperm parameters such as sperm movement decreases. Male fertility depends on different parameters, such as sperm count, motility, viability and epididymal sperm reserves as well as expression levels of cation channel sperm-associated protein 1, 2 (CATSPER1, 2). Additionally, Blepharis.persica seed extract traditionally used to promote male infertility. The purpose of the present study was to measure the effect of Blepharis.persica aqueous extract on some semen parameters and gene expression in male rats after SCI.  

This experimental study was performed on 30 male Wistar rats which divided into 5 groups: one control, and four experimental groups. SCI rats orally treated with plant extract (100, 200 and 300mg Kg-1) daily for 15 days. Body weights, morphological features of testes (diameters of the seminiferous tubules), sperm parameters (motility, viability and epididymal sperm reserves) and CATSPER1, 2 gene expressions were assayed at 30th and 60th days and compared with control and SCI groups. The data were analyzed using SPSS software by one‐way ANOVA test and through Tukey's test (p < 0.05) was considered statistically significant.  

The results of the present study demonstrated that the sperm viability, diameters of the seminiferous tubules increased significantly (4.89%) at 300 mg Kg-1 of Blepharis.persica aqueous extract. Expression levels of CATSPER1, 2 genes were significantly increased in experimental groups (p < 0.05).  

These results indicated that Blepharis.persica seed extract can improve the recovery of male fertility. This study could provide insights into consumption of plant extract in treatment of male infertility with SCI.

Melon charcoal rot is one of the most important diseases that significantly reduces the yield of melon (Cucumis melo L.). The main objectives of this study were: isolation of actinomycetes from the rhizospheric soil of different melon cultivation regions located in the Kerman and Sistan and Baluchestan provinces of Iran; investigating the antagonistic effects of actinomycetes against melon black stem or charcoal rot disease; evaluation of siderophore production by isolates in vitro; identification of potential isolate by PCR, and investigating their biocontrol efficacy against Macrophomina phaseolina in melon under greenhouse condition.

Eighty actinomycete isolates were isolated from the soil of selected different melon cultivation areas and their antifungal activity against Macrophomina phaseolina was investigated. Potential isolates were evaluated for biological activities. Moreover, the efficacy of selected actinobacteria in order to biocontrol charcoal rot disease was investigated under greenhouse condition.

Three actinomycete isolates (R1.6, R5.52 and R5.56) were revealed the highest inhibition zone size against Macrophomina phaseolina and selected for further investigations. All three isolates were able to colonize melon roots, produce extracellular enzymes and control disease in the greenhouse. The actinomycete isolate R5.56 was identified by sequence analysis of small ribosomal RNA subunit (16S rRNA) and based on the results this isolate had the highest similarity (98%) with Streptomyces species.

Biological control of plant pathogens, unlike the application of chemical pesticides, does not work quickly, but in successful cases, has more long-lasting effects compare to chemical pesticides. Biological control should be mentioned as a key component of the integrated pests and plant diseases management system approaches to minimize the environmental side effects and risks as the consequences of over usage of chemicals. This study is a prelude to further studies such as the use of these antagonists against Macrophomina phaseolina in the field, which must be done for at least three years.

The present study was performed to investigate the effect of salicylic acid at different times on the expression of three genes involved in the morphine production end pathway (T6ODM, COR and CODM) in Opium Poppy capsules. It should be noted that these genes play an important role in the production of morphine as a medicinal alkaloid.

Seeds of Papaver Somniferum were grown in pots under greenhouse conditions. When the plants reached the flowering stage, two to three days after the petals fell; a superficial scratch was created in the plant capsule and then treated with salicylic acid at a concentration of 50 μM. Capsule sampling was performed at three times zero (a few minutes after spraying salicylic acid solution on the capsule), three and 12 hours after salicylic acid treatment, compared to the control plant. RNA was extracted from the capsule samples, followed by cDNA, and finally the results obtained by qRT-PCR were analyzed using GenEX software.

The results of this study showed that with increasing the time after treatment of the capsules with salicylic acid, the expression level of all three genes decreased compared to the control plant. The expression of T6ODM gene at zero time increased about 1.84 times the expression of gene compared to the control plant, but the expression of this gene at 3 and 12 hours decreased by 46% and 21%, respectively, compared to the gene expression in the control plant. The results also showed that the expression of COR gene increased by about 1.26 times in zero hours compared to the control plant and decreased by 15% in three hours and by almost 30% in 12 hours compared to the control plant. . The expression level of CODM gene at zero hours was almost equal to that of the control plant and at three and 12 hours their expression level was almost zero. Also, the graphs related to T6ODM, CODM and COR genes showed that there is a significant difference in terms of gene expression between different times in each gene.

It is generally inferred that salicylic acid treatment reduces the expression of the three end genes involved in the morphine production pathway by increasing the time from zero hours to 12 hours.

Among Iran's climate-well-adapted oily plants, safflower (Carthamus tinctorius L.) is considered as a top-ranked plant since it is well-adapted to areas with water shortage. Safflower is known as a model crop with a variety of fatty acids with a relatively high tolerance to salinity and drought, and also due to its high nutritional value that is related to the composition of 90 percent unsaturated fatty acids in its oil content. It has always been considered as a desirable and valuable oilseed plant and can play an important role in expanding the area under planting of oilseeds in the country. Although, Iran with its diverse climate, is one of the richest regions in the world in terms of safflower genetic resources, safflower planting is not very common in our country. One of the main reasons is probably the lack of promotion and low grain yield of its cultivars. In this study, For the first time the floral dipping was performed in Agrobacterium suspension in order that the glufosinate gene would be transferred to the safflower plant. Actually, the access to appropriate engineered cultivars will play a critical role in the development of safflower planting.  

Nine safflower lines were selected from a world-wide collection genotypes based on the different agronomic and phenotypic traits. They were submerged in Agrobacterium suspensions with optical density (OD600 nm) of 1 and in the presence of two different amounts of silwet-77.  

The genomic analysis of T1 generations using glufosinate resistance gene primers indicated the success of glufosinate transfer in safflower plant through flower immersion.  

Among 9 studied genotypes and 17 transformed plants using the floral dipping method, 3 transgenic plants were obtained with sequencing confirmation: one seedling belongs to Mahali Yazd genotype, one seedling belongs to the Mahali Kerman and one seedling belongs to the Paeize 12 genotype. All transgenic plants were inoculated with 40μl concentration of silwet-77.

Biotechnology is one of the most important technologies in particular novelty ones that has many applications in entrepreneurship and knowledge-based businesses. Yet, because of not equipping biotechnology graduates with entrepreneurial skills being one of the important reasons, the process of transferring knowledge from universities to industries unfortunately is too weak. Therefore, the main purpose of this study is identification and strategic planning of the entrepreneurship skills, which are essential for biotechnology graduates to promote their abilities and then entering to the labor market.  

The data collection method has been documentary and survey and the tool used in the survey method was a questionnaire, as 22 questionnaires were analyzed which completed by biotechnology experts in Kerman province. Analytical Hierarchy process (AHP) was used to prioritize the main dimensions and indicators of each dimension. Then, to comprehensively review and strategically plan for developing the entrepreneurial skills of biotechnology students, the SWOT technique utilized.  

Based on experts’ opinions, the results showed that technical skills had been the most prominent and entrepreneurial, managerial and personal maturity skills were placed in the next priorities, respectively. Investigating the result of the internal and external factors matrix demonstrated that contingent and competitive strategies had been the appropriate ones for biotechnology students to develop their entrepreneurial capabilities.  

Considering that higher education has an important role in equipping graduates with different dimensions of entrepreneurial skills, universities can play a vital role in developing the entrepreneurial skills of graduates by identifying internal and external factors affecting entrepreneurship. As a result, it is necessary that policy makers and planners pay special attention to the importance of curriculum improvement, use of applied content and personal skills development; additionally, they must create a suitable environment for students in order to increase entrepreneurial opportunities in society and industry.

Objective DREB gene family is one of the effective transcription factors in plant tolerance to environmental stresses, including salinity stress. The aim of this study is to identify and evaluate the expression of DREB2 gene in different salinity concentrations in local wheat of Kalak Afghan. Materials and methods Salinity stress treatment was applied to wheat using NaCl salt. Experimental treatments included different salinity levels (0, 100, 150, 200, 250 and 300 mM). Then RNA extraction and cDNA synthesis were performed and the specific fragment of DREB2 gene was amplified with specific primers. The obtained transcript was then sent for sequencing and the sequence was analyzed to identify the DREB2 gene. Results Partial sequence of DREB2 gene of local wheat of Kalak Afghani was registered in NCBI database with access number KR106189. The results of sequencing showed that the obtained sequence is more than 95% similar to the sequence of this gene in Triticum dicoccoides and Agropyrum elongatum. Examination of the second structure of DREB2 protein sequence through PSIpred and SOPMA programs showed that this sequence has 46 alpha helices (32.86%), 11 beta helices (7.86%), 9 long strands (6.43%) and 74 is a random helix (52.86%). The results of evaluation of DREB2 gene expression showed that the concentration of 100 mM salinity treatment, has a significant effect on increasing gene expression. In general, the highest effect on DREB2 gene expression was 100 mM salinity treatment and the lowest value was 300 mM. Conclusions In general, the DREB2 gene increases plant tolerance compared to other stress-induced genes, which makes them a desirable target for genetic engineering and crop performance. It seems that the reason that the expression of DREB2 gene does not increase at high concentrations is that the salt load is greater than the cells' ability to transmit it. Under these conditions, salt is transferred to the cytoplasm and inhibits enzymatic activities. Also, a common consequence of the accumulation of high concentrations of salt is the accumulation of large amounts of Reactive oxygen species (ROS), which can cause protein oxidation, damage to cellular DNA, lipid peroxidation, and interaction with other vital cell components. Leads to cell toxicity.

Fritillaria imperialis from the Liliaceae family is an ornamental species in danger of extinction. Therefore, it is nessessary to conserve the plant genetic pool. Two proper methods to access this goal are micropropagation and germplasm conservation in in vitro freezing conditions. The presence of suitable pre-treatments is necessary for cryopreservation. The most important and the most application of pre-treatment is encapsulation-dehydration. The purpose of current research was in vitro proliferation of F. imperialis using plant growth regulators and its cryopreservation using encapsulation-dehydration pre-treatment. 

Bulb scales as explants and MS medium as basic culture medium were used. For micropropagation, kinetin (Kin) and α_naphthaleneacetic acid in concentrations of 0 (as control), 0.5, 1 and 2 mg l–1 were applied. For cryopreservation, explants were pretreated with encapsulation-dehydration followed by conservation in liquid nitrogen. Encapsulation was done using sodium alginate. Dehydration was carried out in air and using high level of sucrose.

The results of micropropagation showed that the maximum number of leaf (3.5) and root number (5.7) was obtained in explants treated with 0.5 mg l–1 Kin along with 1 mg l–1 NAA. The results of cryopreservation showed that encapsulation-dehydration in air as a pre-treatment had effective role on the survival and regeneration of explants (70.5%) after conservation in liquid nitrogen. Least explant survival was obtained in control (without pre-treatment). In vitro regenerated plantlets were cultivated in plastic pots containing peat moss and perlite (in ration of 1:1) and acclimatized with environmental conditions. The survival rate was 90% and the growth pattern of regenerated plants was similar to that of mother plants.

Present research proposes the use of 0.5 mg l–1 Kin together with 1 mg l–1 NAA for micropropagation and encapsulation-dehydration for germplam cryopreservation of F. imperialis. Micropropagation by direct and indirect organogenesis and embryogenesis is a proper approach for proliferation of ornamentals in danger of extiction. The success of these methods depends on type and concentrations of auxins and cytokinins applied in culture medium, singular or in combination. In order to protect rare and endangered ornamental species, modern biotechnological tools need to be utilized.

Acinetobacter baumannii is an important bacterium in causing nosocomial infections that has multidrug resistance. Several factors are involved in the resistance of these bacteria to drugs, the most important of which are the proteins OXA-24 beta-lactamase, OXA-23 beta-lactamase and OUTER MEMBRANE PROTEIN A (OMPA). In this regard, the aim of this study was to investigate the inhibitory effect of plant compounds in Salsify, Hyssop and Mangrove on these Acinetobacterproteins in silico.  

In this study, first the structure of plant compounds and proteins were obtained from PubChem and PDB databases, respectively. Then the physicochemical properties and mutagenic potential of plant compounds were predicted by Swiss ADME server and Toxtree-v2.6.13 software, respectively. ViewerLite, Chimera 1.14, Discovery Studio, AutoDockTools-1.5.6 and AutoDock Vina environments were used for molecular docking and interaction between plant compounds and proteins. The results were analyzed using three programs including  AutoDockTools, Visualizer DS and Ligplot.  

According to mutagenicity potential and inhibition cytochromes results, compounds related to H. officinalis had higher mutagenic potential and cytotoxicity than the other two plants. In addition, the results showed that A. marina compounds have higher digestive absorption compared to other compounds. A comparative study of the interactions showed that the compounds of H. officinalis and A. marina have stronger interactions with the studied proteins. In addition, it was found that the strongest interactions occurred between plant compounds and 1BXW protein. On the other hand, T. graminifolius compounds were found to have weaker interactions with the evaluated proteins.  

Based on docking results, among all compounds evaluated in all three proteins 1BXW, 3G4P and 4 K0X, the best docking results were related to cis-pinocamphone and 3,5-difluorophenyl ester 2,6-difluorobenzoic acid, respectively, from Hyssop and mangrove. In fact, these compounds with the most negative binding energy levels (-9 Kcal / mol and -8.8, respectively) have a greater tendency to bind to the key amino acids of the active site of all three proteins. Therefore, these compounds can be considered as important candidates for laboratory and in vivo testing of A. baumannii activity.

Application of molecular biology techniques to the production of new vaccines against different strains of the Newcastle disease virus (NDV) has been the subject of recent research reports. Development of improved techniques for genome sequencing has led to the recognition of protective mechanisms and the identification of possible candidate antigens. The present research aimed to design a recombinant chimeric immunogen against binding agents in Newcastle virus in birds.  

Therefore, we conducted this investigation to make a recombinant vaccine in silico in order to control and eliminate this disease using anti-leptospirosis epitopes, F, and HN antigens. To bind these epitopes, KK, as AAY linkers, were selected to bind the structure. This structure contained 309 amino acids. The important biological factors of this recombinant vaccine, such as physicochemical properties, different structures, stability, intrinsic protein disorder, solubility, and allergenicity of this vaccine structure were evaluated using immune system analysis.  

Various analyses confirmed the stability of this structure, and the epitopes predicted in the chimeric vaccine showed a high potential for inducing the immune response of B cell and T cell. Thus, immune system analysis revealed that the modeled multi-epitope vaccine could properly stimulate T and B cell immune responses and could potentially be utilized for prophylactic or control applications.  

The results of this study could be employed to control and eliminate leptospirosis in the future after confirming its effectiveness by experimental immunological assays

Evaluation and conservation of native chickensas valuable genomic resources is essential.This is the first study for discovering variants in Lari chicken by whole genome sequencing data. The study of genetic diversity of Lari chicken at genomic level can provide useful information for its preservation and breeding. In this study, genomic diversity of five Lari individuals was investigated using whole genome sequencing technique.

Blood samples were taken from five Lari chickens from cites of Shiraz and Zabol, Iran. Whole genome sequencing (paired end sequencing) was done by Illumina Company) Hiseq 2500). Data quality was determined by FastQC program. Whole genome sequencing datawere aligned with chicken genome reference(Gallus_gallus-5.0/galGal5) using MEM algorithm applied in burrows wheeler aligner program (BWA). Processing of bam files was done in several steps. PCR duplicates were removed using Picard program. The Percentage of alignment with the reference genome and coverage or depth were calculated using the flagstat and depth commands in samtools software. Single nucleotide polymorphisms (SNPs) and small insertions and deletions (INDELs) were identified by the genomic analysis toolkit (GATK)program. Annotation of SNPs and Indels was done using SnpEff program. Genetic diversityof five chicken genomes was calculated with VCFtools.

The mean percentage mapping of short sequences with the reference genome was 99.85% and the mean coverage depth was 7.65 X. In this study, 9.8 million SNPs and 10 million Indels were identified with the most counts of them in the intron and intergenic regions. The mean ofobserved and expected heterozygosity percentages for SNPs in five chicken genomes were 0.30 and 0.35, respectively.

Results from annotation showed that percentage of silentSNPs (74.38%) is higher than that nonsynomous SNPs (missense and nonsense, 25.62%) in Lari chicken genome. The lower observed genetic diversity than the expected genetic diversity, can be due to the forces such as inbreeding in the population of Lari chicken