فصلنامه ژنتیک نوین
سال هشتم شماره 3 (پیاپی 34، پاییز 1392)

Calcitonin gene-related peptide (CGRP) is a 37 amino acids neuropeptide in mammals. The pathological, CGRP is involved in regulating the release of inflammatory factors such as TNFα, IL-1β and norepinephrine - epinephrine. Since CGRP is involved in releasing of inflammatory factors and during illness, the concentration of that would change in the body, then it can be used as a diagnostic factor of disease. To investigate the expression of CGRP in the liver, at first liver sample was taken from the cows (2 cow up to 60 days and 8 slaughtered cows including bulls and cows in dry period), then the samples was added to nitrogen and immediately transferred to lab. In there RNA was extracted, and the quality of the extracted RNA was determined, then cDNA was synthesized and RT-PCR was performed. The results of this study showed that CGRP gene was expressed in the liver tissue of dairy cows up to 60 days of lactation. However, it seems this gene could not be capable to express in the liver tissues of bulls and cows in dry period. Also, in this study CGRP amino acid variety were performed by DNAMAN, MEGA and Multialine software according to its amino acids sequence in NCBI. The CGRP variety of amino acids in cattle had the most homology with sheep, mice, rats and dogs. The homology of CGRP in cattle and human is high (94 %), thus it probably can be used the human or rat kit to determine CGRP gene expression or concentration of CGRP in blood.

One of the limits for tobacco (Nicotiana tabacum L.) farming extension is short time for growth in many parts of the world. One solution to this issue is the use of precocious cultivars. The plan was performed to determine suitable molecular markers for detection of precocity gene on the basis Bulk Segregation Analysis. To achieve this goal, a precocious cultivar (Urumieh 2) and a serotinous varietie (Burley Ree 103) from the Burley type was crossed and the F2 generation was grown on the farm. One hundred plants were selected, contained precocious and serotinous. This plants was sampled and DNA was extracted. The annealing temperatures of 60 RAPD primers were determined and were tested on parents and precocious and serotinous Bulks. Two primers (OPC09 and OPB08) showed polymorphism and after the test on 96 samples from the F2 generation two markers were selected (OPB08- 1050 and OPC09-1900). That showed 19.7 cM and 15 cM distance from precocity locus respectively.

The Prunus genus belongs to the Rosaceae family comprises important species distributed in the northern hemisphere like Iran. In recently novel and industrial pomology, using interspesific hybrids as rootstocks in Prunus genus was attended for useful rootstock obtaining. Rootstocks characterization and selection possibility were prepared with molecular markers improvement. The aim of this study was evaluation of genetic diversity, structure and genetic relations of Prunus rootstocks using fluorescent-AFLP markers. Therefore we studied genetic diversity of 89 genotypes of the Prunus genus, including plum× almond, prune × almond, peach × almond, prune × apricot, plum × apricot hybrids, almond, apricot, peach, prune and plum genotype and other diverse interspecific hybrids. Eight primer combinations showed 728 polymorphic bands with clearly band pattern between 20 candidate primer combinations. Using gCLUTO v. 1.0, SplitsTree4, Structure and R 2.14.1 softwares most (peach × almond) hybrids segregated from the other hybrids that have plum or prune as a parent and parental genotypes separated in the different species. Also Licor system and labeled AFLP markers showed effective results in comparison with encient AFLP, method in this research. Therefore this study can identify promising hybrids based on the parents and can be used to make molecular certificate to register them.

This study investigated the effects of genetic damage, cell contamination and apoptosis, nanoparticle silver (nano-Ag), in vitro conditions by using fibroblast cell line rainbow trout gonad (RTG-2 cell line) as a model. Experiment was done with 9 treatments of different concentrations of silver nanoparticles (NanoCid) with average size of 10 nm, containing 0 (control group), 0.01, 0.05, 0.1, 0.5, 1, 5, 10 and 50 μg/ml. Cells exposed to the desired concentration of silver nanoparticles, and after 24 hours, were measured of toxicity with the cytotoxicity test and the effects of genetic damage tests micronucleus assay. The effect of silver nanoparticles on the growth and cell morphology also were evaluated at times 24, 48 and 72 hours after being exposed to silver nanoparticles. Results showed that silver nanoparticles have strong effects on genetic damage, cell contamination and Apoptosis on celles fish and this effects are concentration-dependent. The results of cytotoxicity test showed that the concentration of nanoparticles increases the cytotoxicity effects at concentrations of 50 μg / ml so that all cells were destroyed. Also results indicated an increase in the abundance of micronucleolus in all treatments in compared with control group, So that treatment 1 μg / ml with 232 % (P<0.05), treated 5 μg / ml with 251 % and treated 10 μg / ml with 261 % increase in frequency of micronucleolus showed significant differences in compared with control group. In general, it can be concluded that due to widespread production and consumption of products containing silver nanoparticles, the release of nanoparticles in the aqueous environment, is not far-fetched. Therefore, studying environmental impact of releasing nanoparticles and strategies to prevent or reduce these effects is required.

Genetic diversity of Iranian wolf populations and domestic/free-ranging dogs were examined using microsatellite markers. The Iranian wolf and domestic/free-ranging dog were sampled in 2010 and 28 wolves and 28 dogs were genotyped with 15 microsatellite loci. Parameters of variability such as allelic richness (Rs), expected heterozigosity (HE) and observed heterozigosity (HO) showed that genetic diversity in Iranian wolves is comparable in magnitude to that of other wild populations of wolf. The average of HE and Rs were 0.85, 11 and 0.83, 9.35 in wolves and dogs respectively. We identified 159 and 141 alleles in autosomal microsatellites in wolf and dog samples respectively. High genetic diversity may result from immigration of outside individuals and habitat diversity in Iran. The average inbreeding coefficient was - 0.148 and -0.122 in wolf and dog samples. The low and negative FIS values revealed no inbreeding problem in Iranian wolf and dog populations. We used Nei and Weir-cockerham F-statistics for investigating level of differentiation between wolves and dogs. Average values of FST, RST and Fit were 0.05, 0.36 and -0.084 reflecting relatively high genetic differentiation between two species. The neighbor joining tree between wolf and dog individuals based on ADS genetic distance showed wolves and dogs were split into two distinct clusters. Even though our sample did not cover the entire range of the wolf in Iran, our results emphasize the high genetic diversity of this species. Also, the results confirm that gene flow between wolves and domestic dogs is low and they show discrete genetic structure, although the two species are in contact around rural areas.

Transient expression of genes using Agrobacterium (Agroinfiltration) is a powerful tool for the analysis of foreign gene expression and function in different gene expression systems in plant before stable plant transformation. However, the production of stable transgenic plants is a lengthy process, in transient assays, it is possible to measure gene expression within a very short time. With simple infiltration of Agrobacterium cells carrying appropriate gene constructs into bean intact leaves, transient expression can be performed within 3 days without using expensive instruments or complicated procedures. In this study, We analysed different regulatory gene expression factors affecting transient expression following agroinfiltration in bean by injection suspensions of the Agrobacterium carrying the plasmid constructs of gene interest. After three days the amount of soluble protein in the leaves was measured and compared. This study proposed to evaluate the best systems expression in transgenic bean plants, using agroinfiltration technique. The results demonstrate that between the three systems expression the highest level of protein is producted by the system that makes the transfer and accumulation of proteins in the endoplasmic reticulumin this study. The efficiency of agroinfiltration was 24-50%.

Using molecular technology has resulted in discovering mutations with major effect on reproduction efficiency in sheep and goat. Most of these mutations have been found in BMP15, GDF9 and Booroola genes, for example, in GDF9, FecGH and FecTT have this effect. Due to the importance of prolificacy in goat, existence of mentioned and new mutations in GDF9 were investigated in both Beetal and Tali goats. A total of 50 and 60 blood samples were collected from Beetal and Tali goats, respectively. After DNA extraction, PCR reactions started for five PCR products (from individuals with different kidding records) from both Beetal and Tali goats were sequenced. Results from sequencing and phylogenic tree were analyzed by Vector NTI and MEGA4 software’s. 3-dimensional structures of GDF9 protein was evaluated by professional web site such as ps2. In Tali goats no nucleotide change has been found, but in Beetal goats with high kidding records a heterozygote mutation (C to T), substitution of an alanine with a valine was found Individuals with high kidding records were heterozygote but the others were wild type. Analyzing of special amino acid residues frequency in second structure shows that alanine has an intrinsic tendency to take part in α helix but valine has an intrinsic tendency to take part in β sheet. So, the mutation found in this research causes a change in third structure and probably function of the protein. In prediction of 3-dimension structure of protein, a difference was found between mutant and non-mutant sequence. Analyzing of special amino acid residues frequency in second structure shows that alanine has an intrinsic tendency to take part in α helix but valine has an intrinsic tendency to take part in β sheet that causes a change in third structure and probably function of the protein. Investigation of phylogenetic tree between some different types of animals showed that BMP15 protein sequence of goat is similar to sheep.

Septoria tritici Blotch (STB) was enumerated as the most important disease in which is responsible for abundant losses in wheat product. In this study, three spring wheat varieties and a resistant line of International Center Agriculture Research in Dry Area (ICARDA) were evaluated in a genetic design of the generation mean analysis, in order to study the existence of resistance to STB disease and genetic analysis of resistance at seedling stage in greenhouse. Therefore Six basis generations included parental varieties, F1, F2 and Backcrosses (BC1 & BC2) were planted in Completely Randomized Design (CRD) at the greenhouse. Result of generation mean analysis in resistant line×tajan cross indicated manifest roles of additative effects in controlling the necrose and necrose Area Under Disease Progress Curve (nAUDPC) traits. Also in this cross For picnid and picnid Area Under Disease Progress Curve (pAUDPC) traits the roles of dominance effects were more pronounced. In the crosses of the resistant line×Moghan and Morvarid×Moghan roles of dominance effects is much more than additive effect and noallelic interaction dominance×dominance was contributed significantly in all traits. High estimate of dominance and epistatic effects emphasized on hybrid seed production as trait breeding strategy, also indicated that selection should be delayed after several generations of selection such as single seed descent until a high level of gene fixation is attained. effective factors controlling resistance to STB disease was varied between 1 and 3 in various cross.

A linear mixed model method for mapping quantitative trait loci (QTL) in inbred crosses was developed which treats the QTL (marker) effects as random. The method has two steps. First, significance of the variance associated with markers (genome variance) is tested using the Residual maximum likelihood ratio test (REMLRT). A significant genome variance component indicates the presence of at least one QTL in the genome. In the second step, the best linear unbiased predictions (BLUPs) of the markers are used to locate the QTL position (marker selection). The selected marker; having the highest BLUP's rank; is then fitted as fixed covariate and the above two steps are repeated until that there is no significant genome variance. The power of the suggested method was studied using simulated data. The results showed that the method is robust in detecting QTL in medium and relatively large populations derived from crosses between inbred lines. Although the power of the method is low in detecting QTL in small populations (as other available methods), the main advantages of the method are the ability to incorporate all of the markers in the analysis simultaneously and to overcome the multiple testing problem.

The production of pharmaceutical compounds having role in treating diseases like AIDS is necessary. In this regard, griffithsin (GRFT) (an algae lectin) can inhibit HIV entry to the cell at picomolar concentration. To production of recombinant GRFT protein, alfalfa, soybean and lettuce were used as hosts for transient expression. Effect of KDEL, signal peptide of carrot extensin and SP subunit of ZERA signal peptide that target protein to endoplasmic reticulum, apoplastic space and space around the cell wall, respectively was studied. 72 hours after agroinfiltration, qRT- PCR analysis showed high potential of lettuce to express of GRFT at transcript level while alfalfa had lowest transcription level. ELISA result using polyclonal anti-mouse anti-GRFT antibody confirmed that targeting of GRFT to apoplast space of soybean led to highest amount of GRFT. Lettuce despite of high level of gene expression, showed low level of GRFT accumulation which may be due to posttranscriptional and post translational modifications. Our results suggest using of soybean leaf tissue for fast and reliable production of pharmaceutical using transient expression.

to investigate the phylogenetic relationship of Schizothorax zarudnyi, Schizocypris altidorsalis, Barbus sharpeyi B. subquinciatus, B.xanthopterus and B.esocinus, the native barbinae species South of Iran, 10, 10, 5, 3 and 4 samples were collected from Chahnemeh’s reservoirs and Karoon River and transferred to laboratory. Genomic DNA was extracted from fin tissue by phenolchlorophorm method. PCR was performed using pair specific primers to amplification 980 bp mitochondrial cytochrome b (Cyt b) region. After checking the PCR products by electrophoresis, then they purified using purification kit and directly sequenced. Results showed that Schizothorax zarudnyi and Schizocypris altidorsalis form a monophyletic group that is sister group with Barbus sharpeyi than other species. Based on comparison Cyt b sequences of all barbus species suggested that Iranian barbinae were paraphylitic than other species. In fact, this position reflected that variation and diversion in Iranian barbinae take place in long time and independently than other barbinae in the world. The reported results could be of interest for management and conservation programs of this valuable native species.

yield and most morphological traits are controlled by the numerous of genes. In order to QTL mapping for yield and yield components traits, 169 barley recombinant inbred lines (F6 generation) and their parents “Igre” and “Arigashar”, were planted in a simple lattice design with two replications in Ghezlagh (research farm of Abouraihan college, Tehran University in 2007. The traits evaluated including: number of speckle, 1000-seed weight and yield. Analysis of variation showed that there was sufficient variation among the entries. Among traits in this study, the 1000 grain weight with the highest heritability is controlled mainly by additive effects. AFLP and SSR markers were used for preparing linkage map and 4 chromosomes of barley were mapped. Two QTLs identified for each trait on the number of 2 and 5 linkage maps. in and 5 linkage groups. Allelic effects (additive) justified the positive and negative correlation among traits. Pleiotropy of QTLs or linkage between them might lead to high correlation.

Artemisinin is a sesquiterpene lacton produced by glandular secretary trichome of Artemisia annua L. plant and is used against some disease including malaria and cancer. In this study effects of exogenous applications of MeJA and SA on known trichome gene expressions in Artemisia annua were investigated with Real time PCR assays. For more information about trichome gene responses to these elicitors, microarray data of Arabidopsis trichome genes in response to them in EBI database were studied. It was observed that MeJA affect trichomes by positive effect on expression of genes involved in the formation, growth, size and orientation of trichome, and SA has negative influence on them. Furthermore, Artemisia TTG1 gene was introduced as an appropriate candidate for further studies for use in trichome engineering in order to increase in artemisinin production.