ORIGINAL_ARTICLE
Expression pattern of the synthetic pathogen-inducible promoter (SynP-FF) in the transgenic canola in response to Sclerotinia sclerotiorum
Sclerotinia 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. Inthis study, the “gain of function approach” was adopted for the construction of a pathogen-inducible promoter.The synthetic promoter technique was used, which involved the insertion of the dimerized form of the cisactingelement (F) upstream of the minimal CaMV 35S promoter, which drives the expression of the β-glucronidase(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 promoteris responsive to methyl jasmonate and S. sclerotiorum treatments. The results of histochemical GUSexpression patterns showed strong reporter expression in leaf, flower and stem tissues of canola. Hence,the SynP-FF synthetic promoter, carrying fungal pathogen-inducible features, could be considered as avaluable tool for controlling the expression of transgenes to improve resistance against the same lifestylepathogens.
https://www.ijbiotech.com/article_7137_942d1fc0628c234f8cd5125c4dab5dfc.pdf
2011-01-01
1
10
cis-acting element
Synthetic promoter
Pathogen-inducible
Brassica napus
Fungal elicitor
Reporter gene
Sclerotinia sclerotiorum
Farhad
Shokouhifar
shokouhifar@um.ac.ir
1
National Institute of Genetic Engineering and Biotechnology, P.O. Box 14965/161, Tehran, I.R. Iran.
AUTHOR
Mohammad Reza
Zamani
2
National Institute of Genetic Engineering and Biotechnology, P.O. Box 14965/161, Tehran, I.R. Iran.
LEAD_AUTHOR
Mostafa
Motallebi
motalebi@nigeb.ac.ir
3
National Institute of Genetic Engineering and Biotechnology, P.O. Box 14965/161, Tehran, I.R. Iran.
AUTHOR
ORIGINAL_ARTICLE
Altitudinal genetic variations among the Fagus orientalis Lipsky populations in Iran
Nuclear simple sequence repeats (nSSRs), together with 16 different enzyme loci, were used to analyzegenetic diversity and differentiation among beech (Fagus orientalis Lipsky) populations along two altitudinalgradients in Hyrcanian forests of Iran. Both enzymes and nSSR analyses revealed a high level ofgenetic 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(AMOVA) indicated that there was a relatively low genetic differentiation among populations, and just 1%and 5% of the genetic variation occurred among populations by nSSR and enzyme data, respectively.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, thanbetween transects; a process that could question altitudinal adaptation.
https://www.ijbiotech.com/article_7139_d789cb94348de2f641d0827763e29672.pdf
2011-01-01
11
20
Fagus orientalis Lipsky
genetic structure
altitudinal gradient
Microsatellite
enzyme gene flow
Parvin
Salehi Shanjani
psalehi@rifr-ac.ir
1
Research Institute of Forests and Rangelands, P.O. BOX 13185-116, Tehran, I.R. Iran.
LEAD_AUTHOR
Giovanni
Giuseppe Vendramin
2
Institute of Plant Genetic, CNR, Via Madonna del Piano, I-50019 Sesto Fiorentino, Firenze, Italy.
AUTHOR
Mohsen
Calagari
3
Research Institute of Forests and Rangelands, P.O. BOX 13185-116, Tehran, I.R. Iran.
AUTHOR
ORIGINAL_ARTICLE
Identification and mapping of quantitative trait loci associated with salinity tolerance in rice (Oryza Sativa) using SSR markers
Salinity stress is one of the most widespread soil problems next to drought, in rice growing areas. ReducingSodium (Na+), while maintaining Potassium (K+) uptake in rice are traits that would aid in salinity tolerance.Therefore, the identification of quantitative trait loci (QTLs) associated with those for Na+ and K+uptake, will enable breeders to use marker-assisted selection to transfer QTLs into elite lines in riceimprovement programs. In view of this, 62 advanced backcross-inbred lines (BILs), at the BC2F5 generation,derived from the cross of Tarome-Molaei (salt tolerant) and Tiqing (Salt sensitive), were used to identifythe QTLs involved in salinity stress tolerance, using SSR markers. Advanced backcross inbred lines alongwith their parents were evaluated for six parameters viz. Sodium (Na+) and Potassium (K+) in roots andshoots and the Na+/K+ ratio, using the modified Yoshida’s nutrient solution at an electrical conductivityof 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 averageinterval of 15.3 centiMorgan (cM) between the markers, spanning 1747.3 cM across all 12 rice chromosomes.Using the composite interval mapping (CIM) and a minimum logarithm of the odds (LOD) thresholdof 3.0, a total of 14 QTLs were detected as follows; on chromosome 1 (5 QTLs), 3 (1QTL), 4 (3 QTLs), 5 (2QTLs), 6 (1 QTL), and 8 (2 QTLs) for all six traits except, Sodium (Na+) in the shoot. The phenotypicvariation explained by these QTLs ranged from 9 to 30% of the total variation. A QTL (QKr1.2) for K+ contentin 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.
https://www.ijbiotech.com/article_7141_30a733abac56a6562571265fd105157e.pdf
2011-01-01
21
30
QTL
Rice
Salinity
SSR
Jafar
Ahmadi
njahmadi910@yahoo.com
1
Department of Agricultural Biotechnology, International University of Imam Khomeini, P. O. Box 34149-16818, Qazvin, I.R. Iran.
LEAD_AUTHOR
Mohammad-Hossein
Fotokian
2
Department of Plant Breeding, University of Shahed, P.O. Box 3319118651, Tehran, I.R. Iran.
AUTHOR
ORIGINAL_ARTICLE
A study on evening-primrose (Oenothera biennis L.) callus regeneration and somatic embryogenesis
Evening primrose (Oenothera biennis L.) is a wild flower with high and valuable oil content. High seedshattering, indeterminate inflorescence and heterogeneous germination limit the commercial cultivation ofthis plant. Besides agronomical research, breeding programs can also remedy the above mentioned problems.Since traditional breeding methods take a long time, using techniques such as tissue culture andsomatic embryogenesis accelerate the breeding process. In the present study, callus formation wasaccomplished in MS (Murashik and Skoog) medium, but no embryogenesis was observed in the presenceor 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 callusproduction 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 designexperiment with 4 replications. For embryogenesis, mhypocotyls (1cm long) were cultured in B5 liquid mediumat the induction phase. After 3 weeks, induced organs were sub-cultured to realization phase and thenumber of embryos (different stages of embryogenesis) was counted 4 weeks later. The results indicatedthat variation in hormone concentrations caused significant differences with respect to somatic embryogenesis.Embryo development were not observed in hormone-free media. The highest numbers of globular,heart, torpedo, cotyledonary, and total embryo was recorded at 9.04 μM of 2, 4-D. Histological studies ofembryos after the realization phase revealed largenuclei and abundance of starch grains indicating thepresence of embryonic cells in evening primrose hypocotyls.
https://www.ijbiotech.com/article_7143_5cbbe1c9791885bc85974a2e83ab3999.pdf
2011-01-01
31
36
Evening primrose
Gamma linolenic acid
Medicinal plant
Tissue culture
Somatic Embryogenesis
Azim
Ghasemnezhad
1
Department of Horticultural Sciences, Gorgan University of Agricultural Sciences and Natural Resources, P.O.Box 49138-15739, Gorgan, I.R. Iran.
LEAD_AUTHOR
Seyyed Javad
Mousavizadeh
mousavizadeh@gau.ac.ir
2
Department of Horticultural Sciences, Gorgan University of Agricultural Sciences and Natural Resources, P.O.Box 49138-15739, Gorgan, I.R. Iran.
AUTHOR
Kambiz
Mashayekhi
kambizm@yahoo.com
3
Department of Horticultural Sciences, Gorgan University of Agricultural Sciences and Natural Resources, P.O.Box 49138-15739, Gorgan, I.R. Iran.
AUTHOR
ORIGINAL_ARTICLE
Biological removal of phosphate from synthetic wastewater using bacterial consortium
The biological phosphorus removal is a microbial process widely used for removing phosphorus fromwastewater to avoid eutrophication of water bodies. The study was aimed to screen the efficient phosphatereducing isolates and used to remove phosphate from synthetic wastewater using batch scale process. Thethree most efficient phosphate reducers were isolated and screened from eutrophic lake water and forest soilsamples. 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,Pseudomonas sp. YLW-7 and Enterobacter sp KLW-2 were found to be efficient in phosphate reduction.Among the individual strains, Pseudomonas sp YLW-7 was noticed to be 68% removal in MSM with glucoseat neutral pH. The consortium with combination of Bacillus sp. RS-1, Pseudomonas sp. YLW-7 andEnterobacter sp KLW-2 was effectively removed the phosphate in the synthetic medium when compared toindividual strains. The phosphate removal was observed to be maximum of 92.5% in mineral saltsmedium (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 nutrientsand as energy sources or it may be utilized by cometabolism. Therefore, these bacterial isolates mightbe used in the remediation of phosphate contaminated environments.
https://www.ijbiotech.com/article_7145_9295b3aa49b0644eac6a52fb2ccde3f8.pdf
2011-01-01
37
49
Phosphate removal
Synthetic waste water
Consortium- Bacillus sp RS-1
Pseudomonas sp YLW-7
Enterobacter sp KLW-2
Usharani
Krishnaswamy
1
Division of Environmental Management and Biotechnology, DRDO-BU Center for Life Sciences, Bharathiar University, Coimbatore -641 046, TN, India.
LEAD_AUTHOR
Muthukumar
Muthuchamy
2
Department of Environmental Sciences, Division of Environmental Engineering and Technology Lab, Bharathiar University, Coimbatore-641 046, TN, India.
AUTHOR
Lakshmanaperumalsamy
Perumalsamy
3
Department of Environmental Sciences, Division of Environmental Microbiology, Bharathiar University,Coimbatore-641 046, TN, India.
AUTHOR
ORIGINAL_ARTICLE
Genetic population structure of Hawksbill turtle (Eretmochelys imbricta) using microsatellite analysis
Information on the genetic structure of marine species is essential for stock improvement programs. In orderto analyses the genetic diversity of the Hawksbill turtle (Eretmochelys imbricte) by the microsatellite geneticmethod, 64 samples were caught from the beaches located in Kish and Qeshm islands. Polymerase chainreactions (PCR) of genomic DNA extracted from the samples were carried out using 5 pairs of microsatelliteprimers. The results of this study indicated that all 5 pairs of primers were polymorphic. Average numbersof real allele and effective allele were 4.90 and 2.99, respectively. Average rate of observed heterozygositywas 0.570 and that for expected heterozygosity was 0.616. Study of the Hardy-Weinberg equilibrium wasshown the entire locus had not equilibrium except Cm3 and Ei8 locus in Kish area. Fst (0.166) and Rst (0.634)calculated by the Analysis of Molecular Variance (AMOVA) test illustrated that there are separate populationsof Hawksbill turtle in this part of the Persian Gulf (Kish and Qeshm islands). It seems that Kish’sturtles live under better conditions in contrast to their Qeshm counterparts. Diminution of genetic variationwithin examined population decreases its adaptation to environmental alterations. We identified two differentE. imbricte populations from north of the Persian Gulf.
https://www.ijbiotech.com/article_7147_03c2880821dc3ccc4a1e42cf01722e0d.pdf
2011-01-01
56
62
microsatellites
Genetic variation
Polymorphism
Eretmochelys imbricte, Persian Gulf
Hossein
Zolgharnein
zolgharnine@yahoo.com;zolgharnein@kmsu.ac.ir
1
Department of Marine Biology, Faculty of Marine Science, Khorramshahr University of Marine Science and Technology, P.O. Box 669, Khorramshahr, Khuzestan, I.R. Iran.
AUTHOR
Mohammad Ali
Salari-Aliabadi
2
Department of Marine Biology, Faculty of Marine Science, Khorramshahr University of Marine Science and Technology, P.O. Box 669, Khorramshahr, Khuzestan, I.R. Iran.
LEAD_AUTHOR
Ali Mohammad
Forougmand
3
Department of Genetic, Faculty of Science, Shahid Chamran University of Ahvaz, P.O. Box 135, Ahvaz, Khuzestan, I.R. Iran.
AUTHOR
Somayeh
Roshani
4
Department of Marine Biology, Faculty of Marine Science, Khorramshahr University of Marine Science and Technology, P.O. Box 669, Khorramshahr, Khuzestan, I.R. Iran.
AUTHOR
ORIGINAL_ARTICLE
Characterization of Iranian isolates of canine parvovirus in fecal samples using polymerase chain reaction assay
Despite the widespread prevalence of canine parvovirus disease (CPV) in Iranian dog population,molecular diagnosis of CPV variants, and investigation of the trends of its genetic changes is a new effort. Inthis study 50 samples from dogs suspicious of infection with clinical signs of diarrhea and vomiting, and 25samples from dogs suspected of infection with general symptoms such as depression and anorexia werecollected from dogs presented to the veterinary clinic of Shiraz University, Shiraz, Iran. Viral DNA wasextracted 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. Theprimer pairs P2 and Pb detect CPV types 2 and 2b, respectively. Our results showed that 44 individualswith clinical signs of diarrhea and vomiting were positive for CPV-2. 39 individuals (89%) were positive forCPV-2b and 5 individuals (11%) for CPV-2a. Therefore, the CPV-2b was identified as the predominantvirus type. All dogs without symptoms of diarrhea and vomiting were CPV-negative. The relationship ofbreed, age and sex with PCR results was not significant (P>0.05). For the first time in the country, thecausative agent of CPV-2 was identified, and presence of new antigenic variants, CPV-2a and CPV-2b wasconfirmed.
https://www.ijbiotech.com/article_7136_34e68f932e2917571bd1349129e0e54a.pdf
2011-01-01
63
68
Polymerase Chain Reaction
Canine Parvovirus
Dog
Iran
Hadi
Askari Firoozjaii
1
Department of Clinical Sciences, School of Veterinary Medicine, Shiraz University, P.O. Box 71345-1731, Shiraz, I.R. Iran.
AUTHOR
Sardar
Jafari Shoorijeh
sjafari@shirazu.ac.ir
2
Department of Clinical Sciences, School of Veterinary Medicine, Shiraz University, P.O. Box 71345-1731, Shiraz, I.R. Iran.
LEAD_AUTHOR
Ali
Mohammadi
3
Department of Pathobiology, School of Veterinary Medicine, Shiraz University, P.O. Box 71345-1731, Shiraz, I.R. Iran.
AUTHOR
Amin
Tamadon
amintamaddon@yahoo.com
4
Department 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.
AUTHOR
ORIGINAL_ARTICLE
A simple and rapid leaf genomic DNA extraction method for polymerase chain reaction analysis
In 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
2011-01-01
69
71
DNA Extraction
Leaf tissue
Polysaccharide removal
Jafar
Amani
afar.amani@gmail.com
1
Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.
AUTHOR
Roohallah
Kazemi
2
Faculty of Agriculture and plant breeding, University college of Agriculture and natural resources, Tehran university, P.O. Box 3158711167, Karaj, Iran.
AUTHOR
Ali Reza
Abbasi
rezabbasi@ut.ac.ir
3
Faculty of Agriculture and plant breeding, University college of Agriculture and natural resources, Tehran university, P.O. Box 3158711167, Karaj, Iran.
AUTHOR
Ali Hatef
Salmanian
4
Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran.
LEAD_AUTHOR