احسان شکری

Salmonella is a common and persistent pathogen in soil environments, posing a significant threat to safe food production worldwide. Given the vital role of soil in agriculture, it is crucial to be cautious about the spread of Salmonella in soil and to employ effective methods for its detection and control. The increasing prevalence of Salmonella can be attributed to the rapid expansion of agriculture and industry, leading to the contamination of fertilizers and water sources with the bacterium. The survival of Salmonella in soil is influenced by various physical, chemical, and biological factors, leading to the continuous colonization of plant organs. Consequently, given the importance of healthy agricultural products, there is an increasing demand for new methods to investigate and identify bacteria in these foods. Several techniques exist for the identification of harmful bacteria in soil. However, using nanosensors as an advanced tool for bacterial detection is very promising, as it can effectively overcome the limitations of other methods. This review study examines the mechanisms of Salmonella contamination in soil and its interaction with plants, highlighting the importance of using biosensors for faster and more accurate detection of this bacterium in soil.

The use of activated carbon in company with metal nano catalysts for pesticides removal from the environment has been considered by researchers, recently. This study aimed to remove diazinon residues from water using Nano-MnO2/PAC composite mading from Pinus eldarica activated carbon containing manganese dioxide nanoparticles.

Initially, the powdered residues of pine cone were treated using phosphoric acid and then converted to activated carbon by chemically heat method under ordinary atmosphere. Then manganese dioxide nanoparticles were synthesized in its substrate. The chemical structure and carbon appearance of the resulting pine fruit and nanocomposite were described by SEM, TEM, XRD, and IR characterization methods. Adsorption tests are performed to evaluate the removal efficiency of diazinon from aqueous solution by applying operational variables including pH (2-10), temperature (16-42 0C), contact time (2-120 min), and at initial concentrations (0.05-100 mg/L) of diazinon was studied.

Microscopic images and spectroscopy showed that manganese dioxide nanoparticles with an approximate size of 37.5 nm were present in the nanocomposite. The results showed that small amounts of nanocomposite (3 mg/L) were able to remove 94.6% of the diazinon with an initial concentration of 40 mg/L. The best description of the adsorption process at optimal pH 4, with fit in the Langmuir isotherm model with a correlation coefficient of 0.985.

According to results, the presence of manganese dioxide nanoparticles improved the removal efficiency of diazinon by 13.7% compared to activated carbon of pine fruit.

C4 plants have very high photosynthetic efficiency under drought stress due to lower water usage and do not require the opening and closing of stomata. This advantage has been created by complementary mechanisms of primary carboxylation in leaves mesophilic cells. In species which are initial carboxylation of mesophilic CO2 during C4 photosynthesis and crassulacean acid metabolism done, the PEPc enzyme catalyses the irreversible reaction of phosphoenolpyruvate in the presence of bicarbonate ions to produce oxaloacetate. The aim of the present study was to partial isolation of pepc coding sequence from halophytic grass Aeluropus littoralis, studying codon bias pattern and also quantitative analysis of its expression in 0 (control) and 600 mM NaCl relative to beta actin gene expression as internal control using Real-time PCR. As a result, a 678 bp fragment of pepc gene coding sequence was isolated and registered in NCBI gene bank under KP122945 accession number. The isolated fragment showed highest homology (in protein level) with Eragrostis minor and Zoysia japonica species and its CAI index was estimated 0.82. Finally, the quantitative analysis of gene expression under 600 mM salt stress showed that the level of pepc gene transcription was increased about 3 times (compared to control) in response to stress.

In the present study in order to investigate the photosynthetic mechanism of this plant under salinity stress , and 600 mM NaCl were applied for 10 days then plant responses in physiological and molecular levels were assessed and compared to the control plants ( 0 mM NaCl). For this end, the coding sequence of this gene was isolated from Aeluropus littoralis for first time and the bioinformatic investigation were done. Results showed that the chlorophyll amount, the starch content and Sodium excretion of leaves in 600 mM NaCl were significantly (P

Aeluropus littoralis is one the most successful monocot grasses in terms of high salinity tolerance, that contains forage quality and has valuable physiological features. The goal of this study is to introduce and provide quick access to DNA-based molecular markers for valuable halophyte, Aeluropus littoralis to assessment of genetic diversity. For this, a total number of 110 EST-SSR markers derived from wheat, rice and barley were amplified using A.littoralis genomic DNA to identify markers were being transferable. Accordingly, the total transferability of gramineous EST-SSR markers in A.littoralis was calculated about 44 %. This result showed that gramineous SSRs pool can be used easily for development of molecular markers in the valuable related species by spending less time and cost.Then, 12 pairs of transferable markers were used to evaluate the effectiveness of identifying markers in diversity assessment and genetic clustering of A. littoralis plants. Overall, 74 alleles of which over 90% were polymorphic were detected with an average of 6.1 alleles per marker. The polymorphism information content (PIC) values ranged from 0.26 to 0.38 with an average of 0.33. Cluster analysis using Unweighted Pair Group Method with Arithmetic Means (UPGMA) and Dice´s coefficient similarity grouped the Aeluropus ecotypes into seven groups. In this regard, ecotypes were rather separated in terms of geographical distances and the polymorphism among individuals and differences between groups was highly consistent with visual observations and variation of samples.

EST sequences recorded in the Gene bank database are an important source for microsatellite discovery in coding genome. These sequences give us the opportunity to discover new genes, as well as a resource for development of markers. Unlike genomic microsatellite markers (noncoding), EST-based microsatelite markers (EST-SSR) are effectively extracted from free publicly EST libraries with less time and expense and show the good performance. In this research using the halophyte grass, Aeluropus littoralis EST library and insilico analysis microsatelite motifs were identified and based on that 16 polymorphic markers (ALES), was developed for genomic studies. The results showed that in this plant (6.7%), such as rice (4.7%), there is a high rate of microsatellite in the coding regions. Also, tg, (gcg- gaa) and ttga motifs constitute the major variety of SSR motifs in a set of Aeluropus EST records. The polymorphism information content (PIC) values ranged from 0.27 to 0.37 with an average of 0.32 that higher values goes to ALES number 3,1,11,13 and 15. An average similarity coefficient (Jaccard) between samples was reached to 55%. These results suggest a great genetic variability in genomic coding regions of A. littoralis plant. Finally, cluster analysis grouped the A. littoralis ecotypes into six groups.

Codon bias refers to the differences in the frequency of occurrence of synonymous codons in coding DNA. Pattern of codon and optimum codon utilization is significantly different between the lives. This difference is due to the long term function of natural selection and evolution process. Genetics drift, mutation and regulation of gene expression are the main reasons for codon bias. In this study, the codon bias analysis was done on photosynthesis and respiratory related genes of phosphoenolpyruvate carboxylase (PEPC), NADP-malic enzyme (NADP-ME), pyruvate orthophosphate dikinase (PPDK), glycerate kinase (GK) (nuclear genes), rubisco, NADH-dehydrogenase subunit F and cytochrome-C (chloroplast genes) from Aeluropus littoralis plant. Nuclear gene sequences were obtained after partial isolation and for chloroplast genes obtained from nucleotide database. Calculation of codon adaptation index (CAI) showed that studied genes with direct or indirect association with photosynthesis, had high level of gene expression and had also a tendency to optimum codon utilization. The results also showed the difference in codon bias between genes encoded in nucleus and chloroplast for some amino acids.

H+-ATPase proton pump is one of the major proteins found in plasma membrane of plants which plays an important role in molecular physiology of stress response. Aeluropus littoralis grass due to resistance to some environmental stresses like salinity and heavy metals is a good model for studying the structure and function of the plasma H+ATPase pump. In this study as a starting point، the complete coding sequence of the gene were identified and cloned from Aeluropus littoralis halophyte. Preliminary data showed that nucleotide sequence of A. littoralis plasma membrane proton pump contains a 2856 bp full-length ORF region that shared a significant homology with Sorghum bicolor، Oryza sativa and Zea mays. Also the predicted peptide of this gene consists of 165 amino acid residues with a molecular weight of 104567 Da which is in terms of domains type and structure، consistent with most plant and fungal plasma membrane H + ATPase protein type III (type III) from a large family pump ATPase p-type. Comparative alignment of amino acid sequence of A. littoralis plasma membrane H+ATPase with some species indicated that nucleotide substitutions did not targeted critical and functional positions of protein and these regions remained conserved. At last the sequence was submitted to GenBank with accession number of GI: 346230720.

Aeluropus littoralis، a monocot halophyte، plays an effective role in the conservation of water and soil resources and the production of forage for livestock in saline lands. In the current study، physiological characteristics of Aeluropus littoralis was investigated in response to different salinity levels. After disinfection، the seeds of Aeluropus littoralis were cultured in acid-washed sand in a growth chamber under controlled condition، temperature of 25/16 °C and photoperiod of 14/10 h for day/night. After 45 days of start culture، salinity treatments (0،100،200،300،400mM NaCl) were applied. The experimental design was completely randomized design with five treatments and three replications. After 14 days of last salinity treatment، plant materials were harvested. Results showed that the highest amount of shoot dry weight and ash content was observed in 200 mM NaCl. According to the results، potassium content of the shoot unlike sodium decreased with increasing salinity، while secreted sodium and potassium from plant aerial parts increased. Root carbohydrate content and the ratio of chlorophyll a to b increased in response to salinity. The concentration of starch and total soluble sugars of shoot demonstrated no significant relationship with the ratio of chlorophyll a/b at the highest salinity level. Our results suggested that the management of detrimental ions along with the regulation of plant assimilations effectively involved in the performance of Aeluropus littoralis to avoid the inhibitory effect of saline condition.

Thin layer chromatography (TLC) method was used to determine the diversity of gamalinolenic (GLA) fatty acid in sixteen ecotypes of Iranian ox tongue flowers. In assessment firstly oil content of root, leaf and seed organs was extracted by soxhelt system and hexane solvent. Isolation. And identification of GLA were conducted via TLC on stationary phase of silicagel 60f 254 and the movable phase of (hexane, diethil ether – glacial acetic acid) by using phosphomolibdic acid reagent. The data obtained from TLC spots areas has been analysed by spss15.0 and clusters of GLA diversity traced. It was based on Squared Euclidean distance and Agglomerative Hierarchical algorithm. Within group average linkage was used for tracing dendrogram. Refrence line at eleventh value of genetic distance, clustered mentioned ecotypes in three main gorupes. The result of this project not only can be used in plant breeding programs for fatty acid improvment of Iranian oxtongue flower but olso can be used as a biochemical data in chimotaxonomy field for discrimination of intra and inter specific of Echium genius.

Agrobacterium rhizogenes induced hairy roots gain of the potential of production and accumulation of plant secondary metabolites. In recent decades, many researchers have focused on the biosynthesis of valuable secondary metabolites in hairy roots, because genetically modified roots produce such compounds with more genetic and biosynthetic stability than cell suspension cultures. Chicory (Cichorium intybus L.) is a biennial or perennial medicinal plant from Asteraceae. The plant contains many important metabolites including inulin, scoline, coumarin, vitamins, flavonoids and aromatics. In this study, with the aim of optimizing hairy root culture establishment, cotyledons of Chicory plants inoculated with different strains of Agrobacterium rhizogenes were cultured on different plant tissue culture media. Numbers of hairy roots produced in different experiments were recorded and molecular confirmation of hairy root clones was performed through specific amplification of rolB and rolC genes in PCR reactions. Based on the efficiency of hairy root induction in different experiments, the best combination of bacterial strain-culture medium for establishing hairy root cultures of Chicory were determined. Agrobacterium rhizogenes strains A4, A13 and 15834 induced hairy roots in Chicory and MS medium was considered as the best culture medium for developing and establishment of hairy root cultures.

Aim and Due to evolving in stress habitats، Halophyte plants are the best options to describe the fundamentals of molecular physiology of salt tolerance and breeding of crops. Therefore، because of genetic relationship of Aeluropus littoralis with rice، wheat and barley، the present study was performed to investigate leaves proteome pattern changes in response to different salt treatment using proteomics methods To examine the effect of salinity on the Aeluropus proteome pattern، salt treatment in 3 levels (100، 200 and 300 mM NaCl) was applied. The mature leaves located in the middle part of plant were harvested for protein extraction after 21 days of salt exposure. Proteins were extracted according to TRIZOL approach and were separated by 2-DE using a nonlinear pH 4-7 IPG strips (24 cm) and 12. 5 % poly acryl amide gel. Statistical analysis revealed that among 550 repeatable detected spots، at least 95 protein spots showed a significant change during salinity levels. In addition، the most proteins with increased and decreased expression levels were obtained in comparison between control and 200 mM NaCl levels and between 200 & 300 mM NaCl treatments، respectively. Based on cluster analysis the whole responsive proteome was classified into 10 classes. Evaluation of these co-expression classes revealed that A. littoralis salinity responsive proteome follows four distinct expression patterns. These results showed that study of individual protein expression changes alone will not be fruitful. But also the identification of co-expression proteins collection and study of proteins gregarious changes pattern in response to different levels of stress was more importance and create better understanding.

Aim and Background. In plants, ions uptake management, translocation, exchange, compartmentalization and precise adjustment of ionic ratios were carried out through the activity of a series of carriers, ion channels and transporters. The vacuolar H+-ATPase (VHA), plays a crucial role in establishing and maintaining intracellular pH gradients across specialized organellar membranes, and influences the transport of cations into the vacuoles of plant cells. Activity of this pump provides the primary proton motive force for the execution and regulation of other ion transporters, semi-transporters and H+-dependent channels in the membrane vacuoles. Due to the apparent importance of this pump in improving salt tolerance, in this study, Aeluropus littoralis a monocot halophyte was used as a gene source. Materials and Methods. A cDNA clone with complete length of 498bp contains the necessary information to encode the c subunit of V-ATPase (AlVHA-c) isolated from the A. littoralis plant and was cloned in E. coli bacterium. Results. The nucleotide sequence of the gene (AlVHA-c) showed a significant homology with its counterpart in millet, rice, wheat and maize. In addition the various bioinformatics analysis was performed to clarify the evolutionary relationships and to investigate the possible amino acid and nucleotide sequences differences. Conclusion. Finally the complete coding sequence was submitted to GenBank with accession number of (JF504672).

Delta-6 desaturase (D6D), an enzyme of a lipid methabolic pathway, has been found as active form in some microorganisms and plants like different species of the genus Echium from family Boraginaceae. The enzyme converts essential fatty acids linoleic acid and α-linolenic acid into long-chain polyunsaturated ones, γ-linolenic acid and stearidonic acid, respectively. In this research, to isolate the d6d gene from Iranian borage (Echium amoenum), we followed total RNA isolation from immature seeds, total cDNA synthesis and PCR amplification using the gene specific primers designed according to the sequence of d6d in the other Echium species. Approximately a 1350 bp amplified fragment was inserted into the vector pBluscript SK(+) and cloned in Escherichia coli strain DH5α. Nucleic acid sequence analysis of the cloned fragment verified the successful isolation and cloning of the full length CDS of d6d gene from E. amoenum. The cloned CDS include 1347 nucleotides coding for 448 amino acids. The translated protein possessed high similarity of 94–96% to its counterparts previously reported in other Echium species. All the expected domains, i.e., cytochrome b5, fatty acid desaturase, HPGG motif and histidine rich conserved motifs were found in the sequence of translated protein. Furthermore, secondary structure, transmembrane domains and the hydropathy profile of the protein were highly similar to its counterparts in the other Echium species. The d6d gene is a promising gene to engineer the fatty acid metabolism in crop plants. Gene expression studies are necessary for further characterization of the cloned gene and its encoded enzyme. The sequence of the coding region of d6d gene from E. amoenum was submitted to GenBank under the accession number GU237486.

Pyrrolizidin alkaloeids (PÂs) are a main gorupes of plant toxin that 200 types of these materials had been identified at yet. Plants containing these alkaloeids are known to be significant causes of dead and desease in mammals such as humans. This research were conducted to assesment of PÂs quality and quantity in different organs of iranian Ëchium amoenum.

PÂs presence were considered in root، petal، leaves and seed by erlich reagent. Çolor apparence in extracts solation approves that PÂs exists in organs. Âlso، regarding senecionin in samples extract، the quantity of PÂs and these N-oxide were determined. This method is specific for alkaloids and other components with a non- basic unsaturated part (D- Pirolin ring).

Çollection of PÂs and N-oxide stimated in 500 mg of sample was 0. 031-0. 053 mg for root، 0. 369mg for leaves، 0. 026 mg for petals and not significantly detected in seeds. Çonclusion: Çompaired to senecionin LD50 (64. 12 ± 2. 24 mg/kg) and pyrroliziding alkaloeids lethal doses (2-27 mg/kg)، leaf samples of Ïranian Ëchium amoenum extracts are able to make toxical condition، unlike seed، petal and root samples PÂs which not able to be toxic، although using these material for long period، even at low level، can be dangerouse to body organs and cause hepatotoxical desease.

In this study, two important physical and chemical properties including percentage of total seed oil and fatty acid levels in two ecotypes of Iranian ox tongue flower (Echium amoenum Fisch & Mey.) collected from Kermanshah and Neka (Mazandaran province) were investigated. Oil extraction and identification of fatty acids were conducted by soxhelt system and Gas Chromatography (GC). All data were compared by statistical methods. Results showed that total oil percentage of (19.16 ± 0.31%) & (17.16 ± 0.26) were calculated for Kermanshah and Neka ecotypes, respectively. Also 8 types of fatty acids including palmitic, stearic, oleic, soxenic, linoleic, alpha-linolenic, gamma-linolenic and stearidonic acids were detected by GC. Soxenic acid and alpha-linolenic were respectively identified as the least (0.2±0.4%) and highest amount (46±48%) in each ecotype. The obtained data indicate that with regard to the nutritional values of essential fatty acids in seed oil, Iranian ox tongue flower has a good potential for producing nutritional supplements of essential fatty acids i-e., omega3 and omega6. According to the results of the study, obtained data from the current study could be applied as a reference in selection of suitable breeding methods to improve oil quality and quantity of Iranian ox tongue flower.

Random amplified polymorphic DNA (RAPD) markers was used to characterize genetic diversity among 16 ecotypes of Iranian Echium amoenum collected from western and northern regions of Iran. Total genomic DNA was extracted by STE method and polymerase chain reaction (PCR) was performed using twenty three RAPD primers. From 438 scoreable bands, three hundred and eighty five bands among different ecotypes were polymorphic (87/9%). RAPD products were scored for presence (1) or absence (0) of each amplicon. Binary method and subsequently genetic similarity was calculated by employing Dice index. Cluster analysis was carried out according to UPGMA algorithm. Cluster analysis, classified ecotypes into eight main groups. The highest similarity (0.68) was found between Gazvin and Lorestan ecotypes, whereas, the lowest was between Gannat Roudbar and Gorgan. The results showed there was a moderate level of genetic diversity among Echium amoenum ecotypes. Moreover, results indicated that RAPD technique is an efficient tool for assessing genetic diversity in Iranian Echium amoenum ecotypes.

Salinity resistant plants are important biotic components of arid and semi arid ecosystems.Aeluropus littoralis is recognized a valuable rangeland plant. Most species of genus Aeluropus are considered halophyte species. Several studies have been done to investigate effects of salinity on halophyte seed germination but there is limited literatures on effect of seed glume compounds on germination at different levels of salinity. Therefore, to understand the mechanism of germination in Aeluropus littoralis and also to find the effect of seed coat on germination rate at different concentrations of salinity, an experiment was done in complete randomized factorial design with three replications. For this purpose, six salinity levels (50, 100, 150, 200, 300 and 400 mM) and two seed coat conditions (with glume and without glume) were applied. The results showed thatincrease salinity levels leads to decrease in germination properties in both seed coatconditions.Seed without glumewas affected more than glume ones. The salinity-glume intraction was also significant at 5 % probability. The result of chemical analysis of methanol extract of glumes through GC-MS, revealed the existence of allogibberelic acid,a derivative of GA3 phytohormon in glumes. This compound was considered as an important factor for germination differences between glume-containing and without glume seeds under different salt treatments.