hasan marashi

Small non-coding RNAs known as microRNAs are essential for responding to biotic and abiotic stresses. A few microRNAs that exhibited varied expressions in the face of drought stress are miR393, miR159, miR169, and miR319. This study examined how these four microRNAs helped petunia deal with drought stress. A 50% drought stress was applied to the petunia plants in the 4 to 6-leave stage, and RNA was isolated from both stressed and control plants. The expression level of microRNAs was examined using the qRT-PCR method under two conditions: control and stress. The data obtained indicated that the expression level of miR319 remained unchanged, while miR393 showed a decrease in expression, and miR169 and miR159 showed increases of 4 and 6.5-fold, respectively.

Begonia is one of the most important ornamental plants around the world. Begonia species are grown for their attractive leaves and flowers in indoor and outdoor conditions. The tissue culture technique is an alternative method for the propagation of begonia species. It can overcome the difficulties of vegetative propagation and lead to high-quality and uniform planting material under disease-free conditions irrespective of the season and weather. In begonia tissue culture, different species showed diverse responses to the medium composition, especially plant growth regulators. Also, lack of information on large scale micropropagation of begonia in Iran highlights research on optimization of begonia species propagation on in vitro culture condition. Thus, in this study the effect of plant growth regulators on shoot regeneration and proliferation of three begonia species (B. elatior‌, B. soli-mutata, and B. tiger), the impact of Auxin and sucrose on rooting of B. elatior‌and diferent pot substrate on acclimation were survived.

In this experiment, petioles of three begonia species (B. soli-mutata, B. elatior, and B. tiger) were used as explant sources. So explants were washed under tab water for 30 mins and sterilized with 1.5% sodium hypochlorite for 15 mins. After sterilization, thin Cell Layers (TCL) with 2 mm thickness were prepared from petiols and cultured in MS medium with different concentrations (0.2, 1, 2 mg/L) of kinetin (kin) or thidiazuron (TDZ) in combination with NAA (0, 0.2 mg/L). Next, the effect of auxin type (1 mg/L of IAA, IBA, and NAA) and sucrose concentration (30 and 40 g/l) on rooting of B. elatior plantlets were determined on in vitro condition. For plantlet acclimation, sand, CocoPeat perlite mixture (1:1 ratio), and Peat moss as pot substrate were evaluated.

The results showed the shoot regeneration response of begonia species to the plant growth regulators of the medium, and there was a significant difference between them in all of the species adding the Auxin to the medium increased shoot regeneration. In B. elatior and B. tiger lacking of NAA to the medium caused no shoot regeneration. Maximum shoot regeneration (100%) was achieved in B. soli-mutata species in medium containing 1 or 2 mg/L of kin and 0.2 mg/l NAA, in B. elatior at 1 or 2 mg/L of TDZ in combination with 0.2 mg/l NAA and in B. tiger at 2 mg/L of both plant growth regulator (TDZ and Kin) and 0.2 mg/l NAA.

For rooting B. elatior, MS medium containing 1 mg/L IAA plus 30 g/L sucrose was the best and favorite acclimation was acheived in cocopeat perlite mixture and peat moss pot substrate.

Regarding high potential of green plants for development of recombinant vaccines, this research was conducted to evaluate expression of a novel recombinant vaccines against Foot and Mouth Disease (FMDV) in tobacco plant. For this purpose, a synthetic gene encoding 129-169 amino acids of foot and mouth disease virus capsid protein VP1 was transferred to tobacco plant via Agrobacterium-mediated genetic transformation. Considering codon usage optimized for tobacco, ribosome binding site and endoplasmic reticulum signal peptide were included in the synthetic gene to enhance expression level. Expression of the synthetic gene in tobacco seedlings was analyzed at transcription and translation levels and production of recombinant protein was quantified. Moreover, an in vivo immunization assay was carried out to verify immunogenicity of the expressed peptide in model animals. Results showed that expression of the recombinant protein in two lines of transgenic plants was a high as 0.65% and 0.72% of total soluble protein. The recombinant protein was able to induce immunogenic response when parenterally administered in rabbit.

Begonias are popular ornamental plants in the world so introducing of various forms of this plant is very important. Nowadays plant modification through biotechnology and genetic engineering for production of new genotypes with varied forms is highlighted in this genus. So optimization of regeneration in in vitro culture condition and gene transferring is in priority to provide a suitable condition for gene transferring in this plant specially those responsible in flower color.
 
In the first step, thin cell layer (TCL) explants from petioles of B. soli-mutata were assayed in MS medium containing Kin or TDZ (0.2, 1 and 2 mg/l) in combination with NAA (0 and 0.2 mg/l). For co-culturing of petiole TCL explants with Agrobacterium tumefacians strain LBA4404 containing GUS and NPTII genes MS medium supplemented with 1 mg/l Kin and 0.2 mg/l NAA was used. Transformed plantlets were confirmed by PCR and Gus assay test.
 
Based on the regeneration results, Kin was better than TDZ, also adding NAA to the medium has positive effect on both of cytokinins. In transformation process, out of 2000 explants which infected with Agrobacterium, after 4 months of culture only 17 plantlets were adopted. Of these 17 plantlets, only 7 plantlets were confirmed by Gus assay test and PCR as transgenic plants.
 
The result showed in this species transformation percentage is low and optimization of transformation condition is important.

Quality of bread baking is affected by gluten genes and balance between their expressions. Hence, it is necessary for a comprehensive research to study and compare all gluten genes and their regulating elements simultaneously.
The aim of this study was to evaluate the molecular mechanism of bread quality at the level of coding genes and regulating elements via comparative transcriptome analysis of two extreme wheat cultivars.
RNAs were extracted from the grain of two wheat cultivars with high (Pishtaz) and low (Navid) bread making qualities, collected during endosperm development at five stages. mRNAs were sequenced and gluten transcripts were assessed to find differentially expressed genes. Then, transcription factors interacting with gluten genes were detected and evaluated for expression.
Results showed that Ɣ-gliadin and LMW-GS genes had a higher expression in Pishtaz and Navid, respectively. Most identified transcription factors were active at the early stage of growth and it seemed that NAC and ERF transcription factors had significant roles in regulating genes with different expressions. There was no significant difference in the expression level of NACs between two cultivars. It is proposed that the ERF transcription factor which classified as BREB2C transcription factor could control the expression of LMW-GS genes in two cultivars and functionally act as a repressor for their target genes.
The priority of Pishtaz wheat cultivar in bread quality originated from high expression levels of Ɣ-gliadin gene and ERF transcription factor.

In consideration for the increasing widespread use of genetically modified (GM) crops, one of the important issues for assessment is the effect of GM crops on soil microbial communities In this study, T2 chitinase-transgenic cotton (line #57) and its non-transgenic line were investigated for bacterial and fungal dynamics during its development stages. The assessments were performed by viable plate count and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) assays. Viable plate count analysis showed an increase in community structures and the number of culturable bacteria in rhizosphere of both transgenic and non-transgenic cultivars as compared to bulk soil. PCR-DGGE confirmed results of viable plate count assays of the changes in bacterial and fungal communities for all cotton development stages in rhizosphere and bulk zones. No significant differences in number of functional bacteria were observed between rhizosphere soil of chitinase transgenic and non-chitinase transgenic cotton at one particular stage. The results indicated that T2 chitinase-transgenic cotton (line #57) might have no adverse effects on community structures and total number of culturable bacteria and fungi in the rhizosphere.

Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture medium of known composition and widely used to produce clones of a plant in a method known as micropropagation. Plant research often involves growing new plants in a controlled environment. These may be plants that we have genetically altered in some way or may be plants of which we need many copies all exactly alike. These things can be accomplished through tissue culture of small tissue pieces from the plant of interest. These small pieces may come from a single mother plant or they may be the result of genetic transformation of single plant cells which are then encouraged to grow and to ultimately develop into a whole plant. Tissue culture techniques are often used for commercial production of plants as well as for plant research. Tobacco (Nicotiana tabacum L.) is one of the most important model plants used in the physiologic, genetic and tissue culture studies. The manipulation of tobacco genetic structure requires an efficient technique of gene transferring and regeneration. Whereas, the tobacco plant is a very effective bioreactor in the production of recombinant proteins, in this research we optimized the best tissue culture system and also, genetic transformation process of this plant.
Our plant tissue culture protocols, Include helpful information for Murashige and Skoog media, plant growth regulators, plant growth hormones, plant transformation systems, and other products for plant tissue culture. For this purpose, different concentrations of sucrose and 4 combinations of growth regulators (BAP and NAA) on callus induction, direct shoot regeneration and rooting were examined in a factorial experiment based on completely randomized design with 3 replications. The sensitivity of tobacco explants to kanamycin was examined through the cultivation of them on the selective medium with different concentrations of antibiotic. For genetic transformation, agrobacterium tumifacious (GV3101) harboring plasmid pBI121 was used and the transgenic plants were confirmed by PCR analysis.
The results of variance analysis and the means comparison showed that the best medium for callus induction was M1 (0.1 mg/l NAA and 1 mg/l BAP) with 15 g/l sucrose in the leaf explants, while the most direct shoot regeneration rate was obtained on the M1 medium with 30 g/l sucrose concentration. High-frequency of rooting was also influenced by 0/1 mg/l NAA and 60 g/l sucrose. So, supplementing the medium with NAA and BAP at different concentrations facilitated induction of multiple shoots from explants. NAA was proved to be the best and the number of shoots increased with increase in the concentration up to (0.1 mg/l), and exceeding this concentration resulted in decline in percent response as well as number of shoots was recorded shoot regeneration. The concentration of BAP was further increased a linear increase in the number of shoots was observed up to an optimal level (1 mg/l). Beyond the optimal concentration (1 mg/l), a decrease in the response as well as number of shoots was recorded due to profuse basal callusing. The effect of cytokinins on multiple shoot regeneration, higher concentrations of NAA found to be inhibitory for shoot regeneration because of huge callusing which hampered the growth and development of new shoots. Also different concentrations of sucrose have a different effect on the shoots and callus. The concentration of sucrose had significant effect on direct shoot regeneration. The main effect of sucrose concentration, concentration of 30 grams per liter, compared with a concentration of 15 grams per liter had the highest direct shoot regeneration. Concentration of 50 mg/l kanamycin could completely prevent the regeneration of untransformed explants so was used in the selective culture medium. Subsequently, the presence of nptII gene (798 bp) in the transgenic plants was confirmed and the transformation efficiency obtained by using the agrobacterium-mediated transformation was more than 95%.
Conclusions In present research, an efficient in vitro regeneration protocol has been developed for tobacco, where different factors including the age of the explant and plant growth regulators were optimized for maximum propagation of tobacco. The results showed that regeneration and transformation method described here is highly efficient and fast for the introduction of any foreign gene directly in tobacco plant.

An Agrobacterium-mediated transient gene expression assay was carried out in alfalfa (Medicago sativa) leaves for expression of a chimeric gene encoding a part of capsid protein of Foot and Mouth Disease virus called VP1. The plant leaves were transformed via agroinfiltration procedure. The presence of the foreign gene and its expression in transformed plants were evaluated by polymerase chain reaction (PCR), real time PCR, protein Dot blot and ELISA. Moreover, gene expression in the transformed leaves was quantified by ELISA method. The results obtained in this investigation indicated high level of gene expression in alfalfa leaves, showing that transient gene expression can be applied as an effective and time-saving procedure for the production of recombinant proteins. The procedures for transformation, detection of recombinant protein and its application for molecular experiments are described in the study.

Orobanche aegyptiaca, commonly known as Egyptian broomrape, is a holo-parasite that causes high economic damages on tomato production in Iran. Breeding for resistance is the most economic and feasible method to reduce broomrape infestation. 20 tomato cultivars and four wild tomato species were screened for O. aegyptiaca resistance. Tomato yield reduction, dry weight reduction of shoot and root, total number of attached broomrapes, dry weight of broomrapes and tolerance index were widely varied among tomato genotypes. The results showed that LA2530 (Ora mutant) was a very susceptible genotype while it was previously introduced as O. aegyptiaca resistant. According to the reports, the resistance phenotype is controlled by a dominant gene (Ora) and it seems that it is broken by the parasite because the overcoming of monogenic resistances by parasites is more likely than polygenic resistances. High levels of resistance were found in two wild species Solanum chilense TL00798 and S. pimpinellifolium L00134 respectively. The results suggested that wild relative species of tomato are promising diverse genetic resources for developing resistant crops to broomrape.

Seedless barberry (Berberis vulgaris L. var. asperma) is one of the few crops that is only cultivated in eastern parts of Iran. As a new crop there has not been any study to identify phylogenetic relationships of this plant with other related species existing in Iran. In this study, Amplification fragment length polymorphism (AFLP) markers based on four selected primer combinations (EcoRI/Tru1I) were used to evaluate genetic variation and phylogenetic relationship among wild and cultivated barberry populations belonging to north east and eastern Iran. Two other species of ornamental barberry and one species of Mahonia aquifolium were also taken in this study. An Unweighted pair group method with arithmetic averages (UPGMA) dendrogram based on genetic distances clearly clustered each species, confirming phylogenetic relationships at the molecular level. These results can clarify the ambiguity in the relationship between Mahonia and Berberis genera. The heterozygosity index, principle coordinates analysis (PCoA), Fst Index and analysis of molecular variance (AMOVA) revealed a significant difference among wild barberry populations. As expected, observed variation within the cultivated barberry population was very low and close to zero. Moreover, morphological markers were used to evaluate variation and phylogenetic relationships among Berberis populations compared to results from AFLP markers by means of the Mantel correspondence test. No significant value was found by the mantel test between AFLP data and morphological markers. The lack of correlation between AFLP and morphological markers suggests low efficiency of identification key of Flora Iranica for classification and phylogenetic consideration of the Berberis family. Further molecular and morphological investigations are necessary to improve understanding of the relationships within species and genera of the Berberis family.