Background and objective: Shigellosis causes the most contagious diseases, acute diarrheal disease in the world. The icsA gene plays a key role in pathogenesis in bacterial. The aim of this study was cloning of icsA gene and developing a mutant constructpGEM:: DicsA (Catr) in order to induction recombination in a native shigella for generation a live attenuated vaccine candidate strain.Materials and methods: by use of biochemical tests, monoclonal Antibody and PCR the native shigella strain was examined. The entire icsA gene specific primers were designed, the DicsA gene was cloned in pGEM-7zf vector and the nucleotide sequence was determined. According to the data of sequencing, digestion mapping of pGEM DvirG was obtained, after digested of DicsA gene by EcoRV restriction enzyme a chloramphenicol antibiotic resistance (catr) gene Cloned in amidst of DicsA gene.Results: The native shigella strain by using biochemical tests was confirmed. Sequence of the icsA gene in native strain was sequentially identical with the strains submitted in the Gene-Bank database.Structure accuracy using specific primerspGEM:: DicsA (Catr) was confirmed.Conclusion: Using the technique of allelic exchange based on the incident of recombination in bacteria is one of the most effective methods to develop a disruption in the target genes. This mutant construct can be applied in development of a live attenuated Shigella flexneri2a vaccine candidate.

In order to evaluate the genetic diversity in 115 rice mutant genotypes of Tarom Jalodar, a randomized complete block design with three replications was conducted at Sari Agricultural Sciences and Natural Resources University. Eleven quantitative traits were evaluated. Analysis of variance showed that there is a significant difference between the genotypes for the morphological traits. The correlation coefficients between traits showed that flag leaf width, number of tillers, number of grains and 100-grain weight had a positive and significant correlation with grain yield. Based on cluster analysis, rice genotypes were divided into three groups. Stepwise regression analysis showed that panicle length, 100-grain weight and number of unfilled grain explain 58. 9 % of variation of yield Results of path analysis showed that the most direct and positive effects obtain from number of tillers (0. 59) and 100-grain weight (0. 37). Results of the main principle component analysis (PCA) revealed that the four main components justify 70. 15% of the total data variation. In general, the results showed that genotypes 109, 64 and 75, due to the more number of tillers, higher length of panicle, the more100-grain weight and less number of unfilled grain were chosen as desirable genotypes, they can be used in the following program.

Genetic diversity is the base of plant breeding. Hence, 33 M7 soybean mutant lines, which were evolved by γ ray from cultivar L17 irradiated with doses of 150, 200 and 250 Gray (absorbed dose), with L17 cultivar and two commercial cultivars (Clark and Williams) were evaluated in view of some morphological traits (number of leaves/plant, pods/plant, seeds/pod, seeds/plant, 100-seed weight, dry weight of aerial parts, dry weight of roots, plant yield, harvest index, nodules/root and dry weight of nodules) under completely randomized design with three replications in greenhouse of Agricultural, Medical and Industrial Research School, Nuclear Science and Technology Research Institute, Atomic Energy Organization of Iran, Karaj, Iran. All traits in the studied mutant lines, except number of seeds per pod, showed a significant difference at a=1% and a=5% in comparison with L17 and commercial cultivars. Mutant line number 13 (M13) was recognized as the top line in view of the studied traits. Seed yield per plant showed the highest correlation (0.886) with harvest index (P<0.01). Cluster analysis of the studied traits along with Ward method resulted in separation of the lines into four independent groups. It can be inferred from the results that irradiation did induce significant genetic variability with regard to majority of the studied traits, such as number of nodules per plant and harvest index.

In order to evaluate the physiology and biochemistry of sixth generation of rice mutant tolerant lines under salinity stress, an experiment was carried out as split plot arranged in a Completely Randomized Design with four replications. Main factor of experiment included three levels of salt stress (0, 45, 75 mmol/l) and sub factor included 5 local rice mutant lines contain Tarom Hashemi 1, Tarom Hashemi 2, Tarom Hashemi 3, Tarom Chaloosi and promising Nemat. The results of analysis of variance of interaction salt stress and rice mutant lines showed that affect of interaction on the studied traits was significant. In comparision with other lines, the results of experiment showed that Nemat under all levels of salt stress had the highest content of Proline that was accompanied with decrease of protein activity in this line. Catalase and Gayacol peroxidase activity under stress condition increased and and there was fewer amount of Malondialdehyde and ionic leakage. Considering the positive and significant correlation between dry weight of rice lines with Proline content (r = 0. 36 * ** ), Catalase (r = 0. 49 ), and Guaiacol peroxidase (r = 0. 51 ** ) and negative correlation with Protein content (r =-0. 29 * ) and Malondialdehyde (r =-0. 36 * ) it seemed to suggest that the Nemat mutant could be as the most tolerant line, since the resistance mechanism of this line involves osmotic regulation and increasing the amount of proline amino acids through protein degradation and catalase enzymatic system, so these traits can be considered as basic mechanisms for tolerance to salinity.

Background: Shigellosis disease is the major causes of morbidity in children with diarrhea in Iran.ThevirG (icsA) gene plays a key role in pathogenesis and ability of invasion in shigella. The aim of this study was cloning, sequencingvirG gene and developing a mutant construct pGEMD virG in order to induction recombination in a native shigella for generation a live attenuated vaccine candidate strain.Materials and methods: Initially, by use of biochemical tests, the native shigella strain was detected.ThevirG gene was cloned in pGEM-7zf vector and the nucleotide sequence was determined.According to the data of sequencing, digestion mapping of pGEMvirG vactor was obtained and a part ofvirG gene by using enzymatic digestion was removed. Finally, pGEM D virG construct was transformed toE. coli by utilization of chemical transformation method.Results: The native shigella strain by using biochemical tests was confirmed. The result of sequencing virG gene (native strain) was submitted in NCBI Genebank database. The pGEMD virG construct contains a mutant construct ofvirG gene which 1751 bp was deleted through enzymatic digestion reaction and transformed inE. coli.Conclusion: Using the technique of allelic exchange based on the incident of recombination in bacteria is one of the most effective methods to develop a disruption in the target genes. This mutant construct can be applied in development of a live attenuatedShigella dysenteriae vaccine candidate.

Objective(s): Adeno-associated virus type 2 (AAV2) vectors are widely used for both experimental and clinical gene therapy. A recent research has shown that the performance of these vectors can be greatly improved by substitution of specific surface-exposed tyrosine residues with phenylalanines. In this study, a fast and simple method is presented to generate AAV2 vector helper plasmids encoding capsid proteins with single, double or triple Y®F mutations.Materials and Methods: A one-step, high-fidelity polymerase chain reaction (PCR) cloning procedure involving the use of two partially overlapping primers to amplify a circular DNA template was applied to produce AAV2 cap genes encoding VP1 mutants with Y→F substitutions in residues 444, 500 or 730. The resulting constructs were used to make the different double and triple mutant by another round of PCR (Y444500F mutant), subcloning (Y444730F and Y500730F mutants) or a combination of both techniques (Y444500730F mutant).Results: Nucleotide sequence analysis revealed successful introduction of the desired mutations in the AAV2 cap gene and showed the absence of any unintended mutations in the DNA fragments used to assemble the final set of AAV2 vector helper plasmids. The correctness of these plasmids was further confirmed by restriction mapping.Conclusion: PCR-based, single-step site-directed mutagenesis of circular DNA templates is a highly efficient and cost-effective method to generate AAV2 vector helper plasmids encoding mutant Cap proteins for the production of vector particles with increased gene transfer efficiency.

Extended Abstract Introduction and Objective: Salinity stress is one of the most important limitations of rice cultivation worldwide. On the other hand, the use of physical and chemical mutagens can be very important not only to the development and combination of new genes or alleles with agricultural importance but also to introduce genotypes more adaptable and stable to adverse weather and soil conditions. For this reason, with the development and progress of rice varieties that are tolerant and adapted to salinity, it is possible to increase the sustainable production of this stable food. Therefore, the present study was conducted with the aim of investigating the response of the tenth generation (M10) of rice mutants to salinity stress in the reproductive stage using agronomic and biochemical characteristics. Material and Methods: In this research, 13 mutant lines of Seng-e-Tarem, Hashemi and Khazar cultivars using gamma ray irradiation from Cobalt 60 which have been previously identified as tolerant lines in molecular studies were used along with the tolerant cultivars of Nonabukra and Deylamani and sensitive cultivars of IR29 and Sepidrood. The genotypes were exposed to three levels of salinity (0, 4 and 8 dS/m) from the source of sodium chloride. A factorial experiment based a randomized complete block design (RCBD) and a factorial split based a RCBD with three replicates were used to evaluate agronomic traits along with yield and biochemical traits, respectively. The measured parameters were tillers number (TN), the number of filled (NFG) and unfilled grains (NUG), 1000 grains weight (TGW), plant height (PH), Days to 50% of flowering (DTF) and the paddy yield (PY), and for the biochemical traits, the activity of catalase (CAT) and superoxide dismutase (SOD), protein (PN), proline (PR) and malondialdehyde (MDA) contents and electrolyte leakage (EL) percentage. Results: The results of the variance analysis showed that the effect of salinity and genotype and their interactions were significant for all the measured traits. A wide range of genetic diversity was observed among the genotypes for the agronomic and biochemical traits. Among the traits, PY was identified as the best and the most indicative trait to classify the tolerant genotype under salt stress conditions. Among the investigated mutants at different levels of salinity, the highest PY was recorded in MP6, MP10 and Deylamani genotypes, respectively. PY value in MP10 mutant at the salinity level of 8 dS/m was closed to Deylamani variety (tolerant check). With the increase of salinity from zero to 8 dS/m, the measured traits adversely affected. The cluster analysis diagram of studied genotypes based on agronomic traits and PY at a salinity level of 4 dS/m classified the genotypes into three separate groups. The genotypes MP2, MP3, MP4, MP9, and MP10 were grouped in the second group, in which Deylamani cultivar was also present as a tolerant check. The cluster analysis diagram of agronomic traits and PY at the salinity stress level of 8 dS/m divided the genotypes into four different groups. In the third group, two genotypes of MP9 and MP10 were included with Tarem Deylamani variety (tolerant check). The correlation coefficient between agronomic traits in 8 dS/m of salt conditions showed that PY had a positive and significant correlation with PH (r=0.51), the NFG (r=0.88) and TGW (r=0.63). Also, there is a negative correlation between DTF with PH (r=-0.70) and NFG (r=-0.62) and a positive and significant correlation with TN (r=0.60). Biplot analysis divided the studied lines into four groups based on agronomic traits at a salinity level of 4 dS/m. The first group includes six mutant lines as very sensitive group, the second group includes two mutant lines as sensitive group, the third group with three genotypes including lines MP9, MP10 and Tarem Deylamani variety (tolerant check) as tolerant group and the fourth group with six genotypes included lines MP11, MP12, MP13 and Nonabukra variety (international tolerant check) as well as Sepidrood and IR29 varieties as high salt tolerant group. Also, biplot analysis based on agronomic traits at a salinity level of 8 dS/m divided the studied lines into four groups. The first group consisted three mutant lines as very sensitive group, the second group included six lines as sensitive group, the third group involved five genotypes including MP11 and MP13 lines and Nonabukra varieties (tolerant check), Sepidrood and IR29 as the tolerant group and the fourth group with three genotypes included MP9 and MP10 lines and Tarem Deylamani variety (native tolerant check) as high salt tolerant group. For biochemical traits at 8 dS/m of salt stress, the highest amount of SOD was recorded in Deylamani cultivar (tolerant check), PN for MP2 mutant, PR for MP3 mutant, and the lowest amount of EL and MDA was recorded for MP2 and MP10 mutants, respectively. Conclusion: In general, PY of MP10 mutant line in salt stress conditions was closed to Deylamani as a native salt tolerant cultivar, therefore could be introduced as a superior line for further research in saline conditions.

Genetic diversity in strawberry is low due to its asexual propagation. One method of increasing genetic diversity, as an important tool in plant breeding, is the use of gamma rays to induce mutation. In order to determine the appropriate dosage of gamma irradiation for genetic diversity creation by mutation in strawberry, Kurdistan cultivar, and to study the influence of various doses of gamma ray (zero (control), 15, 30, 60 and 90 gray) on different traits of strawbery, this research was conducted in Faculty of Agriculture, Yasouj University, in 2012-2013. Results showed that irradiated plants had a lower rates in comparison to the control plants for most of the studied characteristics, except for number of leaves, fruit sugar and anthocyanin. Leaf area, fruit length, fruit diameter, fruit fresh and dried weight, and fruit volume, shape, and juice decreased with an increase in the gamma irradiation dosage. Moreover, multifoliate and dwarfism were increased in the irradiated plants. The appropriate dosage of gamma irradiation for mutation induction in strawberry, Kurdistan cultivar, for threshold 50% plants mortality (LD50) treated by gamma ray, was approximately 65-70 gray. Therefore, gamma ray intensity in the recommended limit could be used for the highest genetic diversity creation with the lowest plant mortality in strawberry breeding programs such as in fruit quality improvement.

Introduction & objective: The third common primary intracranial tumors are pituitary adenomas. Atypical pituitary tumors are characterized by MIB-1>3% & significant P53 nuclear staining. The aim of this study is to evaluate the relationship of the mutant P-53 & MIB-1 level with the outcome of these tumors.Materials & Methods: This Cohort study is conducted in both retrospective & prospective phases from April 2013 to October 2015, 83 patients were enrolled, and inclusion criteria were presence of radiological, clinical or endocrinologic evidence of functional or non-functional pituitary adenomas. Besides, all the patients underwent surgery, whether trans-sphenoidal or trans-cranial, for evacuation of adenomas. Follow-up radiologic and clinical evaluation were done 6 weeks, 6months, and one year after the surgery in all the cases.Results: The non-functional tumors were significantly more aggressive (82.2% versus 55.3% in functional adenomas) (P=0.008). No significant relationship was found between outcome MIB-1 & P53 levels (P=0.49, P=0.834 respectively). On the other hand, invasiveness was significantly related to outcome (39 patients (84.7%) with unaccepted outcome and 19 (51.3%) cases with accepted outcome were invasive, P=0.001).Conclusions: Among invasiveness & MIB-1 & P-53 level, the role reliable prognostic factors for anticipating the outcome seems to be invasiveness and the other factors can be just assumed as confounder factors.