1 2383-1367 Lorestan University 23 Plant improvement Identification of Genomic Oregions Controlling Iron Concentration and Content in Shoot of Barley in A × B Doubled Hoploid Mapping Population Gheitaran Poorsahrigh Shiva b Abolghasem Mohammadi Seyed c Sadeghzadeh Behzad d b Former M.Sc. Student, Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Tabriz University, Tabriz, Iran c Professor, Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Tabriz University, Tabriz, Iran d - Assistant Professor, Dry land Agricultural Research Institute, Maragheh, Iran 1 5 2014 1 1 1 12 13 07 2014 13 07 2014 Iron is one of the essential micronutrient, which has an important role in nitrogen fixation and activity of some enzymes such as catalase, peroxidase and cytochrome oxidase. To map QTLs related to accumulation of iron in shoot of barley at five leaves and maturity stages, 148 doubled haploid lines derived from a cross between Clipper and Sahara3771 varieties were evaluated under greenhouse condition and single plant iron concentration and content were measured. For QTL analysis by linkage map including, 26 retrotransposone markers IRAP and REMAP, 246 SSR and EST-SSR, 238 RFLP and one morphological markers was used. Analysis of variance revealed significant difference between lines for all the studies traits and presence of trangreesive segregation for all the traits and indicated presence of desirable parental allele combinations in the progenies. In total, 511 markers in 7 linkage covered 1099.09 cM of barley genome with an average distance of 2.37 cM between two adjacent markers. For single plant iron concentration, eight and four, iron content in single plant, six and three QTLs were identified at vegetative and maturity stages, respectively. Negative additive effects of the most QTLs indicate the role of Sahara3771 alleles in increased iron accumulation in offspring. One common genomics regions was detected for QTLs of single plant iron concentration and content at maturity which could be due to linkage between the QTLs or the pleiotropic effect of a single QTL.
24 Plant improvement Genetic Diversity Structure of Aegilops cylindrica Accessions Revealed by Genomic ISSR Markers Mohammadi Samira e Ashraf Mehrabi Ali f Arminian Ali g Fazeli Arash h e Former M.Sc. Student, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Ilam University, Ilam f Assistant Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Ilam University, Ilam g Assistant Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Ilam University, Ilam h Assistant Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Ilam University, Ilam 1 5 2014 1 1 13 26 13 07 2014 13 07 2014 Genetic diversity among 35 accessions of Ae. cylindrica using 17 ISSR primers were investigated. Totally, 190 alleles were amplified and 188 alleles (98.95%) o were polymorphic. Number of Amplified alleles ranged from 6 to 20 with average 11.18 alleles for each primer. Polymorphic information content (PIC) varied from 0.10 (primer UBC841) to 0.35 (primer UBC836). Marker index criterion ranged from 0.6 (primer UBC841) to 6 (primer 15). Cluster and Principal Coordinate Analysis could not completely separate accessions and showed no association between molecular diversity and geographic diversity of genotypes, indicating that there is high genetic diversity among accessions. West and south-west genotypes showed more diversity than genotypes from north and north-west of the country. Therefore, the center of diversity and origin of Aegilops cylindrica might be the western and south-western regions of country and this species might transferred from these regions to the northern parts of the country. Results of this study showed that ISSR markers are useful tools for management of genetic resources of wheat and their wild relative species. 26 Plant improvement Cloning, Transformation and Stable Expression of a Fusion of Human Interferon Gamma and bar Genes in Tobacco Plant (Nicotiana tobaccum cv. xanthi) Zakerghoran Bahareh i Rajabi Memari Hamid j Nabati Ahmadi Daryoosh k Siahmard Marzieh l i M.Sc. Student, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Shahid Chamran University, Ahwaz j Assistant Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Shahid Chamran University, Ahwaz k Assistant Professor, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Shahid Chamran University, Ahwaz l M.Sc. Student, Department of Agronomy and Plant Breeding, Ramin Agriculture and Natural Resources University, Ahwaz 1 5 2014 1 1 27 36 20 07 2014 20 07 2014 The production of human gamma interferon in eukaryote expression systems refers as a therapeutic recombinant protein which has significant impact in medical studies. Unique com position of gamma interferon makes such protein as a suitable tool against cancer. It is documented that phosphinothricin (PPT) classified as non-selective herbicide group of bialaphos acts an inhibitor for glutamine synthetase. The bar gene is encoding the phosphinothricin-N-acetyltransferase (PAT) enzyme. This enzyme capable of boosting resistance against PPT herbicide, thus it can be selected as a selective marker within plant population. Then various colony PCR techniques, enzymatic digestion and sequencing were used to confirm the accuracy of fusion of IFNγ-bar gens within expression transporter. Using freezing and thawing method to transfer the pCAMBIA1305.1- IFNγ-bar construction into strain of LBA4404 of agrobacterium, then disc leaves was used to integrate into the genomic of tobacco plant. The transgenic plants were selected under selector condition which possess 30 mg/l of hygromycin. After the developed roots were transferred into soil, and PCR technique was used to confirm the presence of IFNγ-bar in the genomic of plants. Dot blot analysis was applied to detect IFNγ-bar protein in transgenic of to tobacco plants. 27 Plant improvement Investigation of Esterase and Peroxidase Isozymes in Improved and Iranian Landraces of Alfalfa and Their Relationships with Agronomic and Morphological Traits Ahmadi Maryam m Valizadeh Mustafa n Tourchi Mahmoud o Moghaddam Vahed Mohammad p Mohammadzadeh Jalaly Hossein m Former M.Sc. Student, Department of Plant Breeding and Biotechnology, Agriculture Faculty, University of Tabriz, Tabriz n Professor, Department of Plant Breeding and Biotechnology, Agriculture Faculty, University of Tabriz, Tabriz o Professor, Department of Plant Breeding and Biotechnology, Agriculture Faculty, University of Tabriz, Tabriz p Professor, Department of Plant Breeding and Biotechnology, Agriculture Faculty, University of Tabriz, Tabriz Professor, Department of Plant Breeding and Biotechnology, Agriculture Faculty, University of Tabriz, Tabriz 1 5 2014 1 1 37 50 20 07 2014 20 07 2014 For evaluation of genetic diversity among improved alfalfa varieties and Iranian landraces, 12 populations including five improved varieties (Kaysari, Kadi, Ranger, Mesmir, Seariver) and seven landraces (Gharayonje, Amozeynadin, Rahnani, Tazekand, Shazand, Hamedani, Yazdi) were evaluated using agronomic traits and enzyme markers. Thirty-five individuals of each variety were grown and analyzed in separate pots in a unbalanced completely randomized design (CRD). Analysis of variance for agronomic traits showed significant differences for most of the traits among improved and landrace varieties. For esterase and peroxidase enzymes based on presence or absence of enzyme bands (1, 0) eleven polymprphic isozyme bands were detected. For improved and landrace varieties Shanon index mean was 0.48 ± 0.246 and 0.519 ± 0.193, respectively, furthermore Nei genetic diversity index mean for improved and landraces was 0.327 ± 0.181 and 0.352 ± 0.148 respectively, suggesting no difference between improved and landrace varieties was found. Analysis of relation between isozyme markers and agronomic traits showed that there are significant differences between the presence of POX-4 and wet and dry yield in improved varieties. 28 Plant improvement Molecular Analysis of Genetic Diversity and Relationships of Barley Landraces Based on Microsatellite Markers Shuorvazdi Ali Mohammadi Seyed Abuolghasem Norozi Majid Sadeghzadeh Behzad Former M.Sc. Student, Department of Plant Breeding and Biotechnology, University of Tabriz, Tabriz, Iran Professor, Department of Plant Breeding and Biotechnology, University of Tabriz, Tabriz, Iran Assistant Professor, Department of Plant Breeding and Biotechnology, University of Tabriz, Tabriz, Iran Assistant Professor, Dryland Agriculture Research Institute of Iran, Maragheh, Iran 1 5 2014 1 1 51 64 20 07 2014 20 07 2014 Due to their adaptation to different environment conditions, landraces are valuable genetic resurces for incresing diversity of breeding germplasms and are potential resources for biotic and abiotic stress resistant genes. In the present study, genetic diversity and relationships of 119 barely landraces from different countries along with 25 commerical varieties and breeding lines were assessed, using 45 microsatellite primer pairs. In total, 225 alleles range from 2 to 14 and an average of 5 alleles per locus were amplified. Polymorphic information contenet (PIC) varied from 0.05 to 0.90 with a mean of 0.51. The minimum and maximum frequency of common allele belonged to EBMAC0788 (0.13) and GBM1411 (0.97) markers, respectively. Analysis of molecular variance (AMOVA) revealed a higher within group variation (94%) than between group. Maximum and minimum Shannon’s and Nei gene diversity indices were observed in Iranian and Egyptian landraces, respectively. Cluster analysis using Minimum Evolution algorithm and P-distance coefficient assigned the studied genotypes into three groups. This grouping was partly consistent with geographical origins of the genotypes. 29 Plant improvement Investigation of Thymus spp. Karyotypic Diversity in Different Regions of Iran Yousefi Valiollah Najaphy Abdollah Zebarjadi Alireza Safari Hooshmand Former M.Sc. of Plant Breeding, Razi University and Ph.D. Student, Department of Plant Production and Breeding, Faculty of Technology and Engineering, Imam Khomeini International University, Qazvin Assistant Professor, Department of Agronomy and Plant Breeding, Campus of Agriculture and Natural Resources, Razi University, Kermanshah Associate Professor, Department of Agronomy and Plant Breeding, Campus of Agriculture and Natural Resources, Razi University, Kermanshah Ph.D. Student, Department of Agronomy and Plant Breeding, Campus of Agriculture and Natural Resources, Razi University, Kermanshah 1 5 2014 1 1 65 76 20 07 2014 20 07 2014 Thymus, an aromatic medicinal plant, is a well-known, perennial and woody herb from Lamiaceae family. Thymus is taxonomically a very complex genus with a high frequency of hybridization and introgression among sympatric species, and some species of this herb are endemic to Iran. In the present study, in order to identification genetic variability in this medicinal plant seven Thymus spp. accessions collected from different regions of Iran along with London agricultural species were studied by karyotypic characteristics. The secondary basic numbers in all ecotypes was x = 15 that probably originate from a primary basic number x = 7. The ploidy levels of these ecotypes were diploid and tetraploid. The Thymus ecotypes under study occupied classes 1A and 1B of Stebbins’ karyotype classification, indicating the presence of a primitive symmetrical karyotype in these ecotypes. The mean chromosome length ranged from 1.03 to 1.53 µm. chromosome types were detected as metacentric “m”. Furthermore, the cluster analysis using chromosomal parameters and based on UPGMA assigned the ecotypes into four groups. 30 Plant improvement Expression of Recombinant Chimeric EspA-intimin Protein in Nicotiana tobaccum for Oral Vaccine Development Sahshorpour Mahdieh Amani Jafar Jafari Mahyat Salmanian Ali Hatef Former M.Sc. Student, Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran Assistant Professor, Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran Former M.Sc. Student, Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran Associate Professor, Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran 1 5 2014 1 1 77 94 20 07 2014 20 07 2014 One of the important pathogens which cause hemorrhagic colitis and hemolytic uremic syndrome in humans is enterohemorrhagic Escherichia coli (EHEC) O157:H7. Cattle are the most important reservoir of this bacterium. EspA and Intimin are two protein factors for bacteria colonization on intestinal epithelium and cause attaching/effacing lesion. The LEE pathogenicity islands code these proteins. EspA is part of type III secretion systems which delivers Tir to the host cell and integrate to membrane. Intimin encoded by eae gene and fused to Tir. In this research we supposed that chimeric recombinant form of EI gene containing EspA and Intimin were fused with a linker as an edible candidate vaccine would reduce colonization of E. coli O157:H7 in animal model. We constructed a synthetic gene EspA (E120) and intimin (Int282) fused by (EAAAK)4 sequence. The synthetic gene (EI) was codon optimized and subcloned into plant expression vector (PBI121) under CaMV35S promoter and then transferred to tobacco plant by agrobacterium mediated protocol. The presence of inserted gene in plant genome was documented by PCR and RT-PCR methods. The amount of EI protein in transgenic tobacco leaves were estimated 0.1% of the total soluble protein (TSP) by ELISA method.