National Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30439120110101Expression pattern of the synthetic pathogen-inducible promoter (SynP-FF) in the transgenic canola in response to Sclerotinia sclerotiorum1107137ENFarhad ShokouhifarNational Institute of Genetic Engineering and Biotechnology, P.O. Box 14965/161, Tehran, I.R. Iran. Research Center for Plant Sciences, Ferdowsi University of Mashhad, P.O. Box 91775-1653, Mashhad, I.R. Iran.Mohammad Reza ZamaniNational Institute of Genetic Engineering and Biotechnology, P.O. Box 14965/161, Tehran, I.R. Iran.Mostafa MotallebiNational Institute of Genetic Engineering and Biotechnology, P.O. Box 14965/161, Tehran, I.R. Iran.Journal Article20110101Sclerotinia sclerotiorum is a phytopathogenic fungus which causes serious yield losses in canola. A pathogen inducible-promoter can facilitate the production of Sclerotinia-resistant transgenic canola plants. In<br />this study, the “gain of function approach” was adopted for the construction of a pathogen-inducible promoter.<br />The synthetic promoter technique was used, which involved the insertion of the dimerized form of the cisacting<br />element (F) upstream of the minimal CaMV 35S promoter, which drives the expression of the β-glucronidase<br />(GUS) gene. The pGFF construct containing this synthetic promoter (SynP-FF) was used for stable transformation of the canola plant. Fluorometric GUS expression analysis indicated that the SynP-FF promoter<br />is responsive to methyl jasmonate and S. sclerotiorum treatments. The results of histochemical GUS<br />expression patterns showed strong reporter expression in leaf, flower and stem tissues of canola. Hence,<br />the SynP-FF synthetic promoter, carrying fungal pathogen-inducible features, could be considered as a<br />valuable tool for controlling the expression of transgenes to improve resistance against the same lifestyle<br />pathogens.<br /><br />https://www.ijbiotech.com/article_7137_942d1fc0628c234f8cd5125c4dab5dfc.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30439120110101Altitudinal genetic variations among the Fagus orientalis Lipsky populations in Iran11207139ENParvin Salehi ShanjaniResearch Institute of Forests and Rangelands, P.O. BOX 13185-116, Tehran, I.R. Iran.Giovanni Giuseppe VendraminInstitute of Plant Genetic, CNR, Via Madonna del Piano, I-50019 Sesto Fiorentino, Firenze, Italy.Mohsen CalagariResearch Institute of Forests and Rangelands, P.O. BOX 13185-116, Tehran, I.R. Iran.Journal Article20110101Nuclear simple sequence repeats (nSSRs), together with 16 different enzyme loci, were used to analyze<br />genetic diversity and differentiation among beech (Fagus orientalis Lipsky) populations along two altitudinal<br />gradients in Hyrcanian forests of Iran. Both enzymes and nSSR analyses revealed a high level of<br />genetic diversity in natural populations of F. orientalis. The genetic diversity, estimated by expected heterozygosity, was 0.19 (by enzymes) and 0.65 (by nSSRs). Genetic variation across both markers did not reveal genetic structuring along altitudinal transects. There was less genetic variation among altitudinal gradients within transects compared to transect sites. Differentiation assays and analysis of molecular variance<br />(AMOVA) indicated that there was a relatively low genetic differentiation among populations, and just 1%<br />and 5% of the genetic variation occurred among populations by nSSR and enzyme data, respectively.<br />Mantel tests showed that there was not a significant correlation between the genetic distances among populations and the distance of elevation. The results of the present study indicate that the relatively low genetic differentiation among F. orientalis populations at different elevations was not caused by ecological factors. These patterns suggest that higher rates of gene flow along altitudinal gradients within transects, than<br />between transects; a process that could question altitudinal adaptation.<br /><br />https://www.ijbiotech.com/article_7139_d789cb94348de2f641d0827763e29672.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30439120110101Identification and mapping of quantitative trait loci associated with salinity tolerance in rice (Oryza Sativa) using SSR markers21307141ENJafar AhmadiDepartment of Agricultural Biotechnology, International University of Imam Khomeini, P. O. Box 34149-16818, Qazvin, I.R. Iran.Mohammad-Hossein FotokianDepartment of Plant Breeding, University of Shahed, P.O. Box 3319118651, Tehran, I.R. Iran.Journal Article20110101Salinity stress is one of the most widespread soil problems next to drought, in rice growing areas. Reducing<br />Sodium (Na+), while maintaining Potassium (K+) uptake in rice are traits that would aid in salinity tolerance.<br />Therefore, the identification of quantitative trait loci (QTLs) associated with those for Na+ and K+<br />uptake, will enable breeders to use marker-assisted selection to transfer QTLs into elite lines in rice<br />improvement programs. In view of this, 62 advanced backcross-inbred lines (BILs), at the BC2F5 generation,<br />derived from the cross of Tarome-Molaei (salt tolerant) and Tiqing (Salt sensitive), were used to identify<br />the QTLs involved in salinity stress tolerance, using SSR markers. Advanced backcross inbred lines along<br />with their parents were evaluated for six parameters viz. Sodium (Na+) and Potassium (K+) in roots and<br />shoots and the Na+/K+ ratio, using the modified Yoshida’s nutrient solution at an electrical conductivity<br />of 6 and 12 dS/m. A total of 114, out of 235 simple sequence repeats (SSRs) markers that showed polymorphism in the parents, were used to genotype the BILs. A linkage map was constructed with an average<br />interval of 15.3 centiMorgan (cM) between the markers, spanning 1747.3 cM across all 12 rice chromosomes.<br />Using the composite interval mapping (CIM) and a minimum logarithm of the odds (LOD) threshold<br />of 3.0, a total of 14 QTLs were detected as follows; on chromosome 1 (5 QTLs), 3 (1QTL), 4 (3 QTLs), 5 (2<br />QTLs), 6 (1 QTL), and 8 (2 QTLs) for all six traits except, Sodium (Na+) in the shoot. The phenotypic<br />variation explained by these QTLs ranged from 9 to 30% of the total variation. A QTL (QKr1.2) for K+ content<br />in the root was identified with the highest LOD score (7.8), on chromosome 1. This QTL explicated 30% of the total variation and was identified as a major QTL conferring salt tolerance in rice.<br /><br />https://www.ijbiotech.com/article_7141_30a733abac56a6562571265fd105157e.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30439120110101A study on evening-primrose (Oenothera biennis L.) callus regeneration and somatic embryogenesis31367143ENAzim GhasemnezhadDepartment of Horticultural Sciences, Gorgan University of Agricultural Sciences and Natural Resources, P.O.Box 49138-15739, Gorgan, I.R. Iran.Seyyed Javad MousavizadehDepartment of Horticultural Sciences, Gorgan University of Agricultural Sciences and Natural Resources, P.O.Box 49138-15739, Gorgan, I.R. Iran.0000-0002-2549-5906Kambiz MashayekhiDepartment of Horticultural Sciences, Gorgan University of Agricultural Sciences and Natural Resources, P.O.Box 49138-15739, Gorgan, I.R. Iran.Journal Article20110101Evening primrose (Oenothera biennis L.) is a wild flower with high and valuable oil content. High seed<br />shattering, indeterminate inflorescence and heterogeneous germination limit the commercial cultivation of<br />this plant. Besides agronomical research, breeding programs can also remedy the above mentioned problems.<br />Since traditional breeding methods take a long time, using techniques such as tissue culture and<br />somatic embryogenesis accelerate the breeding process. In the present study, callus formation was<br />accomplished in MS (Murashik and Skoog) medium, but no embryogenesis was observed in the presence<br />or absence of plant hormones like 2,4-D (2,4-dichlorophenoxy) acetic acid. In contrast, B5 (Gamborg) medium containing 2,4-D induced embryogenesis. Different parts of plants exhibited good callus<br />production potency and the hypocotyl was found as the best plant explants. In B5 medium, various concentrations of 2,4-D (0,2.26 μM, 4.52 μM, and 9.04 μM) were applied as a complete randomized design<br />experiment with 4 replications. For embryogenesis, mhypocotyls (1cm long) were cultured in B5 liquid medium<br />at the induction phase. After 3 weeks, induced organs were sub-cultured to realization phase and the<br />number of embryos (different stages of embryogenesis) was counted 4 weeks later. The results indicated<br />that variation in hormone concentrations caused significant differences with respect to somatic embryogenesis.<br />Embryo development were not observed in hormone-free media. The highest numbers of globular,<br />heart, torpedo, cotyledonary, and total embryo was recorded at 9.04 μM of 2, 4-D. Histological studies of<br />embryos after the realization phase revealed largenuclei and abundance of starch grains indicating the<br />presence of embryonic cells in evening primrose hypocotyls.https://www.ijbiotech.com/article_7143_5cbbe1c9791885bc85974a2e83ab3999.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30439120110101Biological removal of phosphate from synthetic wastewater using bacterial consortium37497145ENUsharani KrishnaswamyDivision of Environmental Management and Biotechnology, DRDO-BU Center for Life Sciences, Bharathiar University, Coimbatore -641 046, TN, India. Department of Environmental Sciences, Division of Environmental Engineering and Technology Lab, Bharathiar university, Coimbatore-641 046, TN, India. Department of Environmental Sciences, Division of Environmental Microbiology, Bharathiar university,Coimbatore-641 046, TN,
India.Muthukumar MuthuchamyDepartment of Environmental Sciences, Division of Environmental Engineering and Technology Lab, Bharathiar University, Coimbatore-641 046, TN, India.Lakshmanaperumalsamy PerumalsamyDepartment of Environmental Sciences, Division of Environmental Microbiology, Bharathiar University,Coimbatore-641 046, TN, India.Journal Article20110101The biological phosphorus removal is a microbial process widely used for removing phosphorus from<br />wastewater to avoid eutrophication of water bodies. The study was aimed to screen the efficient phosphate<br />reducing isolates and used to remove phosphate from synthetic wastewater using batch scale process. The<br />three most efficient phosphate reducers were isolated and screened from eutrophic lake water and forest soil<br />samples. The total heterotrophic bacterial analysis of the samples showed the presence of about 38 phosphate reducers based on the minimum inhibitory concentration (MIC) test. Among them, Bacillus sp RS-1,<br />Pseudomonas sp. YLW-7 and Enterobacter sp KLW-2 were found to be efficient in phosphate reduction.<br />Among the individual strains, Pseudomonas sp YLW-7 was noticed to be 68% removal in MSM with glucose<br />at neutral pH. The consortium with combination of Bacillus sp. RS-1, Pseudomonas sp. YLW-7 and<br />Enterobacter sp KLW-2 was effectively removed the phosphate in the synthetic medium when compared to<br />individual strains. The phosphate removal was observed to be maximum of 92.5% in mineral salts<br />medium (MSM) at pH 7and 5, and 63.4% in synthetic phosphate solution at neutral pH with lactose as a carbon source by the consortium after 72 h. Thus the microorganisms may use the contaminants as nutrients<br />and as energy sources or it may be utilized by cometabolism. Therefore, these bacterial isolates might<br />be used in the remediation of phosphate contaminated environments.<br /><br />https://www.ijbiotech.com/article_7145_9295b3aa49b0644eac6a52fb2ccde3f8.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30439120110101Genetic population structure of Hawksbill turtle (Eretmochelys imbricta) using microsatellite analysis56627147ENHossein ZolgharneinDepartment of Marine Biology, Faculty of Marine Science, Khorramshahr University of Marine Science and Technology, P.O. Box 669, Khorramshahr, Khuzestan, I.R. Iran.Mohammad Ali Salari-AliabadiDepartment of Marine Biology, Faculty of Marine Science, Khorramshahr University of Marine Science and Technology, P.O. Box 669, Khorramshahr, Khuzestan, I.R. Iran.Ali Mohammad ForougmandDepartment of Genetic, Faculty of Science, Shahid Chamran University of Ahvaz, P.O. Box 135, Ahvaz, Khuzestan, I.R. Iran.Somayeh RoshaniDepartment of Marine Biology, Faculty of Marine Science, Khorramshahr University of Marine Science and Technology, P.O. Box 669, Khorramshahr, Khuzestan, I.R. Iran.Journal Article20110101Information on the genetic structure of marine species is essential for stock improvement programs. In order<br />to analyses the genetic diversity of the Hawksbill turtle (Eretmochelys imbricte) by the microsatellite genetic<br />method, 64 samples were caught from the beaches located in Kish and Qeshm islands. Polymerase chain<br />reactions (PCR) of genomic DNA extracted from the samples were carried out using 5 pairs of microsatellite<br />primers. The results of this study indicated that all 5 pairs of primers were polymorphic. Average numbers<br />of real allele and effective allele were 4.90 and 2.99, respectively. Average rate of observed heterozygosity<br />was 0.570 and that for expected heterozygosity was 0.616. Study of the Hardy-Weinberg equilibrium was<br />shown the entire locus had not equilibrium except Cm3 and Ei8 locus in Kish area. Fst (0.166) and Rst (0.634)<br />calculated by the Analysis of Molecular Variance (AMOVA) test illustrated that there are separate populations<br />of Hawksbill turtle in this part of the Persian Gulf (Kish and Qeshm islands). It seems that Kish’s<br />turtles live under better conditions in contrast to their Qeshm counterparts. Diminution of genetic variation<br />within examined population decreases its adaptation to environmental alterations. We identified two different<br />E. imbricte populations from north of the Persian Gulf.<br /><br />https://www.ijbiotech.com/article_7147_03c2880821dc3ccc4a1e42cf01722e0d.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30439120110101Characterization of Iranian isolates of canine parvovirus in fecal samples using polymerase chain reaction assay63687136ENHadi Askari FiroozjaiiDepartment of Clinical Sciences, School of Veterinary Medicine, Shiraz University, P.O. Box 71345-1731, Shiraz, I.R. Iran.Sardar Jafari ShoorijehDepartment of Clinical Sciences, School of Veterinary Medicine, Shiraz University, P.O. Box 71345-1731, Shiraz, I.R. Iran.Ali MohammadiDepartment of Pathobiology, School of Veterinary Medicine, Shiraz University, P.O. Box 71345-1731, Shiraz, I.R. Iran.Amin TamadonDepartment of Animal Health Management, School of Veterinary Medicine, Shiraz University, P.O. Box 71345-1731, Shiraz, Iran and Stem Cell and Transgenic Technology Research Center, Shiraz University of Medical Sciences, Shiraz, I.R. Iran.Journal Article20110101Despite the widespread prevalence of canine parvovirus disease (CPV) in Iranian dog population,<br />molecular diagnosis of CPV variants, and investigation of the trends of its genetic changes is a new effort. In<br />this study 50 samples from dogs suspicious of infection with clinical signs of diarrhea and vomiting, and 25<br />samples from dogs suspected of infection with general symptoms such as depression and anorexia were<br />collected from dogs presented to the veterinary clinic of Shiraz University, Shiraz, Iran. Viral DNA was<br />extracted from feces. Three specific pairs of primers, P2, Pab, and Pb, were used in a PCR assay for differential diagnosis of the virus type. Pab primer pairs detect the new type-strains, CPV-2a and 2b. The<br />primer pairs P2 and Pb detect CPV types 2 and 2b, respectively. Our results showed that 44 individuals<br />with clinical signs of diarrhea and vomiting were positive for CPV-2. 39 individuals (89%) were positive for<br />CPV-2b and 5 individuals (11%) for CPV-2a. Therefore, the CPV-2b was identified as the predominant<br />virus type. All dogs without symptoms of diarrhea and vomiting were CPV-negative. The relationship of<br />breed, age and sex with PCR results was not significant (P>0.05). For the first time in the country, the<br />causative agent of CPV-2 was identified, and presence of new antigenic variants, CPV-2a and CPV-2b was<br />confirmed.https://www.ijbiotech.com/article_7136_34e68f932e2917571bd1349129e0e54a.pdfNational Institute of Genetic Engineering and Biotechnology of IranIranian Journal of Biotechnology1728-30439120110101A simple and rapid leaf genomic DNA extraction method for polymerase chain reaction analysis69717144ENJafar AmaniDepartment of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O.
Box 14965/161, Tehran, I.R. Iran.Roohallah KazemiFaculty of Agriculture and plant breeding, University college of Agriculture and
natural resources, Tehran university, P.O. Box 3158711167, Karaj, Iran.Ali Reza AbbasiFaculty of Agriculture and plant breeding, University college of Agriculture and
natural resources, Tehran university, P.O. Box 3158711167, Karaj, Iran.Ali Hatef SalmanianDepartment of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O.
Box 14965/161, Tehran, I.R. Iran.Journal Article20110101In plants, secondary metabolites and polysaccharides interfere with genomic isolation procedures and downstream reactions such as restriction enzyme analysis and gene amplification. The removal of such contaminants needs complicated and time-consuming protocols. In this study, a simple, rapid and efficient method for leaf DNA extraction was optimized. This method use small amount of plant material to reduce inhibitory agents (alkaloids, phenolic). The procedure involves homogenization of the plant leaf in extraction buffer, incubation at 60ºC, extraction by chloroform: iso-amyl alcohol and finally DNA precipitation by cold isopropanol. The results showed that the extracted DNA could be used directly for PCR.https://www.ijbiotech.com/article_7144_37c05c8092c9229060b05f685c0cad0a.pdf