a. ismaili

The chlorophyll biosynthesis pathway is a main target for genetic modification to change plant photosynthesis and growth rate to support a greater demand for food in the growing world population. In this study, the effect of overexpression of GSA gene one of gene involved in biosynthesis pathway of chlorophyll on physiological condition of tobacco plant was investigated. 5-Aminolevulinate (ALA) is product of GSA gene. ALA is a precursor for all tetrapyrrole, these components have impotent roles in living cell, as pigments, light receptor (Phytochrome), prosthetic group of many different proteins (like cytochromes, hemoglobin, myoglobin, and leghemoglobin) and enzymes (for example Catalase, Ascorbate, Peroxidase and etc.). Nowadays, ALA has received wide attention for its widespread usage in agriculture, forestry and medication. ALA at low concentrations increases photosynthesis, growth, development, yield and productivity, also promoted fruit color appearance and quality and taste of products in treated plants under both normal and stressful conditions. ALA also improves antioxidant features, absorption of nutrient, water use efficiency and osmotic balance in plants.

 In this research, according to bioinformatics studies, MsGSA gene cDNA of Alfalfa (Medicago sativa L. cv. Isfahani) was selected to transfer to Xanthi tobacco (Nicotiana tabacum) plant, the binary expression vector pBI121 which has Kanamycin antibiotic resistance gene for selection in bacteria and plants, cutting sites for SacI and BamHI enzyme, CaMV35S promoter (cauliflower mosaic virus promoter), nos transcription termination sequences and ß-glucuronidase (GUS) reporter gene was used. After constructing the gene construct pBI121-GSA and confirming the transfer of the construct using PCR cloning methods, enzymatic digestion and sequencing were performed, then the corresponding construct was transferred to Agrobacterium tumefaciens strain LB4404 using Agrobacterium with the gene construct. The corresponding gene was transferred to the tobacco plant genome and the transgenic plants were selected on the medium containing kanamycin and the presence of the gene was confirmed by performing PCR in the regenerated plants. The rooted transgenic sprouts were transferred to the soil. The level of GSA gene expression in the resulting transgenic plants was evaluated by real-time PCR, and their growth rate and biochemical content were also evaluated.

 It was observed that the growth of transgenic plants increased significantly depending on the level of GSA gene expression, and the content of ALA (aminolevulinic acid) and chlorophyll a, b and total chlorophyll, which are the products of the corresponding gene expression. The results also showed the content of anthocyanin, flavonoids and phenol of plants have significantly increased in proportion to the increase in GSA gene expression compared to wild type tobacco plants.

 The growth rate as well as the content of chlorophyll a, b, total chlorophyll, ALA, anthocyanin, flavonoids and phenol of transgenic GSA plants is proportional to the increase in the expression of the GSA gene. The results from this study indicate that an increase in transgenic growth rate as well as an increase in the secondary metabolites content in transgenic plants were influenced by GSA transferred gene and an increase in the content of ALA. These results imply that transgenic tobacco plants expressing MsGSA gene had higher resistance potential to stresses than the wild type tobacco plants.

miRNA genes consider as an important class of gene expression regulators that have diverse responses to environmental stresses and play a key role in regulating gene expression during post-transcriptional regulation. Hypericum perforatum, is well-known medicinal plant for its application in the treatment of depression due to the effects of its bioactive compounds named hypericin and hyperforin. The present study was performed to identify the conserved miRNAs and their target genes in the transcriptome of H. perforatum. First, RNA-seq data were obtained from the European Nucleotide Archive (ENA) of EMBL-EBI database and then the transcriptome (RNA) sequences were assembled. The non-coding transcripts were identified and considered as miRNA precursors candidate sequences. Finally, six miRNAs named Hp-miR395, Hp-miR845d, Hp-miR414, Hp-miR159, Hp-miR159e, and Hp-miR156c were identified from the candidate sequences using of C-mii software after applying strict filters.  Investigation of the protein-protein interaction network determined the relationships between the target genes and target proteins especially in transcription factors. In the next step, to confirm the target genes for the identified miRNAs, the foliar application of Methyl Jasmonate (MJ) with concentrations of 0 (control) and 200 μM was performed on H. perforatum plants and the expression pattern of two target genes was investigated. In the analysis of qRT-PCR expression changes at 12, 24, 48 and 72 hours after applying MJ, the relative expression level of Hyp-1 (DN121523_c1_g4_i2) and HD-Zip (DN121003_c0_g1_i1) transcripts increased after 72 hours of application of MJ. In general, regarding to the regulatory role of the identified miRNAs in the present study, these genes could be used to better and more accurately identify the biosynthetic pathway of hypercin and hyperforin.

Persian black cumin is one of the most important endemic plants to Iran. Its seed is used in the daily diet of Iranians and also has medicinal properties and aromatic compounds. Unfortunately, there is little information about its genome and molecular markers. In this study, the transcriptome of Iranian black cumin was sequenced in the granulation stage and after extracting the microsatellites (SSR markers) related to this growth stage, their interpretation was done. After RNA extraction, the inflorescences and stems were sequenced. Sequencing depth and length were 6 GB and 150 bp pair-end, respectively. Sequencing data were analyzed using different softwares. De-novo assembly of Persian black cumin was performed by Trinity software. Microsatellite markers were extracted using MISA software. Finally, the functional interpretation of the microsatellite-containing sequences was performed by the WEGO database. The GC content of different samples was 47 to 50%. 8389 microsatellite markers were obtained from 45146 unigenes. At least 11% of unigenes contained microsatellites. A/T and AG/CT nucleotides had the highest percentage of microsatellite markers. Among the three nucleotide repeats, ATC/ATG and AAG/CTT had the highest frequency of microsatellites. The results of functional interpretation showed that unigenes containing microsatellite markers cover a wide range of biological activities of Persian black cumin and the genes involved in seed development and growth have a high level of expression. This study is the first report of microsatellite markers of Persian black cumin. The results of the present study can be useful in finding different markers for breeding programs and developing cultivars with different goals.

Allium includes more than 920 species worldwide. Iran is the main habitat of many wild Allium species. In this study, onion tissue of two species A. stipitatum and A. scabriscapum from two subspecies Melanocrommyum and Reticulatobulbosa were used to evaluate and identify the expression pattern of storage proteins obtained by the SDS-PAGE method. Two-dimensional gel electrophoresis and MALDI TOF/TOF MS mass spectrometry were used to identify protein spots and to observe the expression changes of the two species. Analysis of gels by two-dimensional electrophoresis revealed 138 protein spots. The results showed a slight similarity between the gels and a different expression pattern between the samples, based on which a small number of similar spots were obtained in both gels. The results showed that there were significant differences in the expression profile of onion tissue proteins of these samples. Using the T-test, 9 protein spots with the most significant changes were identified at the 5% probability level, and among them, 3 different spots were identified using Mass spectrometry was selected for identification. The results of mass spectrometry experiments and matching of the obtained data with NCBI and Uniprot databases and bioinformatics tools showed that the tested spots were consistent with Maturasek proteins, Agmatine coumaroyltransferase-1, and a protein with the unknown function called Hypothetical Protein. The findings of this study showed that since the samples belonging to A.stipitatum and A.scabriscapum are of the same genus, but the expression pattern of their stored proteins in onion tissue is very different from each other.

MicroRNAs (miRNAs) are one of the small and non-coding regulatory molecules, which regulate gene expression by transcriptional cleavage or translational suppression. miRNAs are involved in regulating a wide range of metabolic and physiological processes in plants.The mint family plants, especially Satureja khuzistanica, are well known herbs for its flavor, fragrance and medicinal properties. To date, no miRNAs have been identified in Satureja species. In present study, a computational approach based on homology search was used to identify miRNAs and their targets of Satureja khuzistanica. Non-coding unigenes were identified and considered for candidates of miRNAs precursor. After evaluating the general parameters such as GC percentage, minimum folding free energy (MFE), minimum free energy index (MFEI) and secondary structure, 58 miRNAs were identified, which among them, by applying plant specific parameters and filtring the miRNAs from several transcripts, overall ten miRNAs were identified. Then, 930 target transcripts were predicted using psRNATarget website and the annotation of them was performed by using BLASTx tools of NCBI Blast+ software (v2.6.0). Examination of target genes showed that auxin-responsive genes, GRAS, AGO2 and LAC family genes are the main targets of the identified miRNAs in S. khuzistanica. Pathway enrichment analysis of the target genes revealed that the secondary metabolic pathway is significantly among the targets of the identified miRNAs. This is the first study describing miRNAs and their role in the regulation of target genes in S. khuzistanica.

In order to reduce the effects of salinity on germination and growth of Khuzestani savory by humic acid, two experiments were performed. The first experiment was done by priming the seeds with zero concentrations, 20, 40 and 60 mg / l of humic acid (H1, H2, H3 and H4, respectively) and germinating them in zero salinity (distilled water), 50, 25, 75 and 100 mM اNaCl (S1, S2, S3, S4 and S5, respectively) in petri dishes. In the second experiment in the greenhouse, humic acid at zero, 10, 20, 30 and 40 mg / kg soil (H1, H2, H3, H4 and H5, respectively) and salinity stress including zero, 50, 25, 75 and 100 mM sodium chloride (S1, S2, S3, S4 and S5, respectively). The results of the first experiment showed that germination percentage and rate were decreased by increasing in salinity, While, germination rate and percentage were improved by humic acid priming. Salinity also decreased the growth characteristics in pot experiment. While humic acid led to the improvement of these growth traits. With increasing salinity, electrolyte leakage increased, but it was reduced by humic acid. The positive results of using humic acid in reducing the harmful effects of salinity, in addition to laboratory (seed priming) were also proven in pots (soil application). Therefore, in general, according to the positive results of this study on improving the germination and growth indices of Khuzestani savory plant, the use of humic acid can be recommended to reduce the adverse effects of salinity.

High salt accumulation has severe adverse effects on soil characteristics. Humic acid can improve the soil structure, soil microbial communities, and absorption and maintenance of mineral nutrients. The aim of the present study was to determine the effects of humic acid on some physicochemical and biological soil properties in soils under salt stress.

The experiment was conducted as a factorial based on RCBD design with four replications. The first factor included humic acid in five levels (zero, 10, 20, 30, and 40mg kg-1 soil). The second factor was salinity stress in five levels (0, 25, 50, 75, and 100mM NaCl). The sampling was carried out in two stages, before and after harvest.

The results showed that S1H5 treatment had the lowest soil electrical conductivity (EC), soil reaction (pH), bulk density, and population of actinomycetes with average values of 0.26dS m-1, 6.21, 1.12g cm-3, and 516cell g-1 of soil and had the highest fungal and bacterial population with an average of 1525000 and 137500000cell g-1 of soil, respectively.

Salt stress has a significant effect on physicochemical and biological soil properties except for the population of actinomyces that their activity was better, at a high level of salinity stress, it had adverse effects on other properties. Although using humic acid improved soil properties. According to the results, using humic acid can be a good solution to reduce the adverse effects of salt stress.

Iran with rainfall of 240 mm per year belongs to arid and semi-arid regions of the world. Lentil (Lens culinaris M.) is one of the most important legumes around the world. In this study, levels of water stress (including irrigation up to 50% of field capacity and 25% of field capacity) for 7 days were applied on lentil seedlings in a completely randomized design with three replications in greenhouse. After treatment, shoots and roots were harvested and the expression of NAC genes was analyzed using qRT-PCR method. The expression analysis of NAC genes was performed in three biological and two technical replications. The means comparison by Duncan test showed a significant difference among all three gene expressions in the aboveground tissues. The GmNAC expression was significant in both treatments, but for MtNAC and MfNAC genes, significant differences were observed in 25% stress condition compared to control. Although MfNAC expression increased in both shoots and roots, the expression in shoot was much higher than root. In roots, significant differences were observed in 25% stress condition comparing to control. In shoots, expression of MtNAC gene was significantly increased in both stress treatments and in roots, only treatment 25% showed a significant difference with control. The increase in expression in the roots was less than shoot. In shoots, expression of GmNAC gene in the treatment of 50% was higher than treatment of 25% but the increase in expression in the root was same in both stress treatments. In general, MtNAC and MfNAC genes appear to be more important in the shoot. The current study is the first report of expression analysis of MfNAC, MtNAC and GmNAC genes in lentil under water stress conditions.