مجله فنآوری زیستی در کشاورزی
سال بیستم شماره 1 (بهار و تابستان 1400)

Black cumin is belong to Ranunculaceae family that it has important secondary metabolites especially thymoquinone that used in the pharmaceutical industry. The purpose of this study was optimization of conditions for callus induction and cell suspension culture with use salicylic acid and methyl jasmonate elicitors to enhance some important secondary metabolites, especially thymoquinone. Callus induction experiment was carried out in factorial arrangement within CRD template with three replications. Three factors included explants, BAP and IAA. Cell suspension experiments were carried out in completely randomized design (CRD) with two replications with used of stem callus of the best plant growth regulator. In the first experiment of cell suspension culture, salicylic acid in four concentrations (0, 125, 250 and 500μM) and in the second experiment, methyl jasmonate in five concentrations (0, 50, 100, 200 and 300μM) were used and the results of the cell suspension for secondary metabolites were analyzed by GC-MASS. The results of callus induction experiments showed that the best explants for callus induction (100%), was stem in combination with 1/5 mg/l BAP and 2mg/l IAA. The results of cell suspension experiments showed that in both of the cell suspension experiments, different concentrations of elicitors have different effects on the increasing or decreasing in some secondary metabolites. The results indicated that thymoquinone content was maximally increased by 33% times, in comparison to control, when salicylic acid (500μM) were added and maximally increased by 43% times, in comparison to control, when methyl jasmonate (300μM) were added as abiotic elicitors.

Endophytes are microorganisms that spend at least one stage of their life cycle into plants without causing any symptoms. This study was conducted to identify and survey diversify of endophytic fungi in Capparis spinosa. Sampling was performed from 10 areas. Four healthy plants were selected from each region based on a systematic random sampling design and Samples were taken from different plant tissues (including leaves, stems, roots and fruits). After surface disinfection, the samples were placed on medium and the obtained fungal isolates were purified. Isolates were identified by sequencing ITS1-5.8S-ITS2. Totally, 72 isolates were obtained, which include 6 orders and 10 genera. Among isolates, 95.83% belonged to Ascomycota and they were divided into six orders, namely; Eurotiales, Sordariales, Pleosporales, Diaporthales, Hypocreales and Mucorales. In this study, Alternaria sp. and Aspergillus tubingensis had the highest abundance and about half of the endophytic fungi (49.25) were isolated from the two mountainous regions of Khodafarin (East Azerbaijan) and Ab-Bar (Zanjan) which it seems to be the presence of endophytic fungi in plants in colder and mountainous areas is more than others. Also, the total percentage of colonization was 11.25% and the highest percentage of colonization was seen in root tissue (50.00%) and the lowest in fruit tissue (4.17%), which can increase the probability of the effect of coexistence of soil fungi in the vicinity of the root. Among the isolated species, Chaetomium globosum, Diaporthe foeniculina, Mucor circinelloides and Stemphyliu vesicarium are reported for the first time from Iran for Caper, and fungal species identified for the first time in the world are reported as Caper endophytic fungi.

Tomato bacterial canker disease is one of the most important diseases with severe economic damage in the world and Iran. In order to better understand the molecular basis of the resistance mechanisms to this disease and consequently resistance genetic improvement, RNA sequencing data obtained from resistant wild tomato (Solanum arcanum) and susceptible commercial tomato (Solanum lycopersicum) leave samples at 0, 8, and 24 hours after inoculation with the bacterial canker were analyzed via various bioinformatics tools. The results led to the identification of 4306 and 1177 genes with significant differential expression in resistant wild tomato at 8 and 24 hours after inoculation, respectively. Gene enrichment analysis indicated the presence of biological processes, including “stimulus response” and “stress response” at 8 hours after inoculation, as well as biological processes such as ‘response to wound”, “regulation of the jasmonic acid mediated signaling pathway” and “regulation of the defense response” at 24 hours after inoculation in the wild tomato. In addition, 45 genes encoding the transcription factor showed more than threefold expression change, 24 h after inoculation in the wild tomato genotype. Among them, 9 transcription factors in the wild genotype had a significant increase in expression, while their expression changes were not significant in susceptible tomato cultivar. Also, the analysis of promoter regions of selected transcription factors led to the discovery of a number of regulatory elements related to hormonal signaling pathways in biotic stresses and defense responses. Due to the difference in expression patterns between resistant and susceptible genotypes, as well as the existence of biotic stress-related elements, these transcription factors are proposed as potential candidates in the defense response to bacterial canker.

The production of cellulase from Penicillium expansum MDFS2 on mixed substrates of barley straw and wheat straw (1:1) was investigated. The highest exoglucanase (6.72 U/g) and endoglucanase (40.75 U/g) activities observed on the 5th day of fermentation process. However, the highest β-glucosidase activity (15.47 U/g) was on the 6th day of fermentation process. The optimum temperature for exoglucanase and β-glucosidase activity recorded at 40°C and β-glucosidase retained more than 70% of its maximum enzymatic activity at 80°C. Also, the highest endoglucanase activity was recorded at 50°C. The highest activity for all the three major components of cellulase enzyme complex was observed at pH= 5.0. The enzyme activity reduced with increasing in temperature and incubation period. The results showed that β-glucosidase is more stable at 50°C in comparison of endoglucanase and exoglucanase and also 80% of β-glucosidase original activity was maintained 3-h after incubation period. According to the results of this study, Penicillium expansum MDFS2 could be suggested as a strain with good potential for the growth on the mixed substrates of barley straw and wheat straw for cellulase production.

The leaves of plants produce β-ocimen due to damage of herbivors. β-ocimen syntethase (OS) is the enzyme that produces it. There are no OS gene sequences in common bean databases. Nucleotide diversity of OS gene in resistant (Naz and KS41128) and susceptible (Akhtar and G11867) common bean genotypes was evaluated. Seeds were planted in pots and genomic DNA was extracted in 2-3 leaf stage and PCR was performed with a specific primer pair. PCR products were sequenced and compared with OS sequences in the NCBI database and their similarity was assessed. DNA sequencing showed that Naz and KS41128 were most similar. Phylogenetic analysis showed that the studied genotypes were in one group and NCBI samples were in the another group. Also, susceptible and resistant genotypes were separated from each other. The sequence of this gene in genotypes did not show single nucleotide polymorphism, which indicates the protection of this region of the gene in common bean.

Soil salinity is one of the most important factors in reducing the yield of durum wheat in Iran. high-throughput technologies advancement, while accelerating the production process of tolerant cultivars to ambience stresses, have made determining the function of newly discovered proteins a major challenge by generating large genomie informatics data. Therefore, the aim of this study was to predict the tertiary structure and function of unknown proteins responsive to salinity stress in durum wheat. The results of phyre software showed that the tertiary structure of the protein sequence has homology with A0A452XZB6 protein with 385 residues, 25 alpha helices and 23 beta sheets from the structural classification of Uniprot protein with E-value 1e-66 and 87 percent accuracy of assessment. The STRING site matched the protein function prediction with a localized Na+/H+ exchanger in the plasmamembrane. Prediction of transmembrane domains, identification of PTMs, and protein-specific motifs indicated that activation of this antiporter is probably accomplished by phosphorylation of serine threonine residues and plays an important role in salinity tolerance of durum wheat. This study provides a framework for future studies on the role of Na+/H+ plasma membrane exchangers and its relative contribution to the maintenance of cellular Na+ homeostasis in tolerance of durum wheat cultivars in saline soils.

Citrus Cachexia disease caused by Hop stunt viroid is one of the important economic diseases of citrus plants that can lead to non-compensable damage in citrus orchards. In order to detection and distribution of HSVD in orang orchards such as Valencia, Tamson, Washington and Blood variety oranges in the Mazandaran province sampling was done during 2019-2020 horticultural season. A 173 numbers was accidently harvested from sampling regions. RNA extraction was done from leaf tissue using Trizol reagents. Reverse Transcription Polymerase Chain Reaction (RT-PCR) method with specific primers was used to identification of viroid. The results of this research showed that viroid agents was practically identified in majority of sampling area. The average distribution of disease was determined as 45.06 percent that of all the maximum distribution (72.2%) was related to Saroo village. In the Bahanmir, Chaloos and Fereidonkenar regions, the viroid agent was not detected. Single Strand Conformation Polymorphism (SSCP) pattern showed the different isolates of viroid in sampling regions. Phyllogenetic analysis of obtained sequences showed the isolated are divided into 4 genetic groups that the first and second group are heterogenic and other two groups are monogeneic. Based on these results, it can be concluded that changes in the secondary structures of circular mRNA molecule of hop stunt viroid strain in the different host could lead to spread of viroid host range.