Showing 221 results for Type of Study: Research
Ms Mohammadamin Almasi, , , ,
Volume 1, Issue 1 (4-2012)
Abstract
Potato leaf roll virus is an important virus that causes economic loss in the yield and quality of potato tubers. One of the primary methods of managing infection in potato crops is using certified virus-free tuber as ‘seed’ for planting. Early and efficient detection of virus is essential for production of PLRV infection free tubers. There are several techniques to detect the virus including serological test and molecular methods. LAMP is a new method to identify pathogens that is used in this study for detection of potato leaf roll virus. Potato plants with symptoms similar to PLRV were collected from Zanjan province and were subjected to a serological test. Total RNA was extracted and RT-LAMP reactions were carried out. Different methods were used to confirm performance of this reaction. Positive reaction was confirmed based on the resulting turbidity and loading product on agarose gel and using SYBR and ethidium bromide florescence dyes. The advantages of this new method include the speed (75 min), ease and safety of the method compared to other methods.
Saeid Navabpour, Hossein Saburi,
Volume 1, Issue 1 (4-2012)
Abstract
In order to search for signaling factors altering the expression of barley metallothionein promoter, the mutant screening technique was used. The promoter region of metallothionein in barley (cv. Hordea) was used to drive the gus gene and it was transferred to Arabidopsis thaliana. The M0 seeds was mutagenized using four doses (10, 20, 30, 40 Grays) of fast Neutron radiation. The chemical compound of 3-Amino-1,2,4-Triazole (3-AT) at 20 mM was chosen as the best activator using CRD statistical design. Inheritance of GUS activity was evaluated using histochemical GUS assay. 48 hours after spraying by 20 mM 3-AT the inhibition model of 3:1 GUS expression: non expression was tested. 20 potential mutant lines were selected by carrying out several screening steps. In each stage 30 seedlings of 1500 M2 lines were checked by GUS staining. The arrangement of promising lines included 3, 5, 9 and 3 which were referred to 10, 20, 30 and 40 Gray of fast Neutron treatments respectively. These lines were analyzed using fluorometric b-glucuronidase technique and were selected as potential mutants in genetic pathways of metallothionein promoter activity for further studies.
Naghmeh Abiri, Saeed Aminzadeh, Mohammad Reza Bihamta ,
Volume 1, Issue 1 (4-2012)
Abstract
Chitin, poly-β-1,4-N-acetyl-D-glucosamine (GlcNAc), is the second most abundant organic polymer in nature after cellulose and it is the main part of insects cuticle and crustaceans that includes in cell walls of most fungi and some algae and nematodes. One of the enzymes that are responsible for disintegration of chitin is Chitinase. Bacteria produce Chitinase for digesting chitin, principally as a source of carbon and energy. With the objective of achieving the maximum production of Chitinase in an Iranian thermophile strain of Cohnella sp. A01, five different media were examined. We partially purified this enzyme from a native strain. Bacteria were transferred to a pre-culture media after an initial culture. After 24 hours, the bacteria were transferred to five different media containing colloidal chitin as the main component. Maximum enzyme production reached in a treatment that contained (NH4)2SO4 0.05% and Agar 0.1%. We performed Chitinase assay using the DNS (dinitrosalicylic acid) and precipitated proteins by Ammonium sulfate. After precipitation of proteins with Ammonium sulfate, enzyme activity was increased. Completely Randomized Design was used and statistical analysis was performed using SAS software. Anti-fungal effect of Chitinase partially purified in this study was tested on five plant fungal pathogens and was confirmed in four cases.
Seyedeh Maedeh Feizbakhsh, Masuod Tohidfar, Seyed Hasan Marashi, Nasrin Moshtaghi, Motahare Mohsenpoor, Mohsen Mardi,
Volume 1, Issue 1 (4-2012)
Abstract
Mexican lime is one of the most important and economic fruit crops in the southern regions of Iran. This crop is susceptible to Citrus Tristeza Virus (CTV) and Witches Broom Disease of Lime (WBDL). Therefore, optimization of regeneration and transformation system for this plant is necessary for its improvement. In this project, the effect of different factors such as internode and epicotyl explants; two different regeneration media containing different concentrations of BAP and NAA, different Agrobacterium strains: “LBA4404” and “EHA105”, each harboring a binary vector pBI were studied in a factorial experiment. The results showed that the etiolated epicotyl was the best explants for regeneration and callus production. There was no significant difference between two regeneration media. Furthermore, EHA105 was the best Agrobacterium strain for this purpose. Polymerase Chain Reaction (PCR) using gus-specific primers has been carried out on DNA extracted from all regenerated plants. Twenty-one shoots were carrying this gene but only 8 shoots out of these 21 showed the expression of this gene. Furthermore, the lack of Agrobacterium-related infections has been confirmed using virG-specific markers.
Saeideh Keyarsalan, Seyed Elyas Mortazavi, Behzad Ghareyazie, Sakineh Mehranie,
Volume 1, Issue 1 (4-2012)
Abstract
In order to produce a marker-free transgenic rice with improved tolerance to salinity and drought stresses, expression vectors pABRII-Chl and pABRII-Cyt containing "choline oxidase" gene (with or without leader sequence respectively) were constructed from pChl and pCyt and pTRA132 for co-transformation. The pChl and pCyt vectors were digested with HindIII-BamH and BamHIEcoRI enzymes. Then the resulting sequences were ligated and inserted into expression vector pTRA132, in which the HindIII-EcoRI fragment (hph gene) had been deleted. The constructs pABRII-Chl or pABRII-Cyt and pTRA132 (containing hph gene) were introduced into embryogenic calli derived from the mature seeds of a rice cv. Hashemi by biolistic transformation method. Then putative transformants were screened after 3 rounds of selection on N6 medium containing increasing concentrations of Hygromycin B from 60 to 80 mg/L. Finally, Hygromycin resistant calli were regenerated on MS medium supplemented with 50 mg/L Hygromycin B. Putative transgenic rice plants were analyzed by polymerase chain reaction PCR. Then, four of the transgenic plants were analyzed using Southern blotting. Each transgenic plant received one copy number of both choline oxidase and hph genes. Expression of the transgene was confirmed by reverse transcription PCR. The high frequency of transformation rate in this study showed that co-transformation method is a reliable method for stable transformation with the goal to make markerfree transgenic plants in subsequent steps.
Bentol Hoda Modirroosta, Masoud Tohidfar, Jalal Saba, Hossein Hadavand,
Volume 1, Issue 1 (4-2012)
Abstract
One of the main goals of modern biotechnology is the production of higher yielding quality varieties to help attaining food security. In spite of numerous benefits, genetically modified crop plants have raised concerns for some of the consumers. Although some of these concerns have no scientific basis, safety evaluations of transgenic plants could reduce them. One of the studies to this end is the metabolic analysis of transgenic plants. Anions and cations (Na, K, Mg, acetat, chloride, nitrate, phosphate, sulfate, succinate and oxalate) of transgenic plants and their non transgenic counterparts were measured using Ion chromatography. After planting, sampling and extraction, Anions and cations of transgenic cotton (chitinase and Bt cotton) were measured. Significant differences in the amount of oxalate, sodium and ammonium in cry1Ab expressing cotton lines and oxalate and sodium in chitinase over expressing lines were observed.
Samira Kahak , Ali Mohammad Shakib, Jalal Saba, Siamak Asadi, Mana Ahmadraji,
Volume 1, Issue 2 (10-2012)
Abstract
Oilseed rape is one of the important oil plants that its cultivation has been expanded in Iran in recent years. Seed shattering before and during harvesting is a problem of cultivation of this plant can causes up to 50% yield decrease depending on the environmental conditions. Production of seed shatter tolerant plants can increase the yield. Different genetic and environmental factors can affect this trait. The FUL gene is one of the genetic factors in that its over expression could prevent silique opening and seed shattering. Rapeseed transgenic plants of three cultivars containing the BnFUL gene were produced and molecular studies were carried out on T2 generations transgenic plants using PCR, Southern blotting and real time PCR. PCR showed that the BnFUL transgene has been inserted in the transgenic plants in addition to the internal gene. The copy number of transgene in transgenic and control plants were analyzed by Southern blotting which confirmed presence of at least one intact copy of the transgene in transgenic plants. Real time PCR showed that BnFUL gene expression in transgenic plants in comparison with control plants has been increased. Differences between the expression of transgenic plants in different lines could be because of position effect and copy number.
Banafsheh Fattah, Mohammad Mehdi Sohani, Abdollah Hatamzadeh, Alireza Afsharifar, Behrooz Goleyn, Mohammad Hossein Rezadoost, Mohammad Reza Mirzaei, Amir Hossein Zamani,
Volume 1, Issue 2 (10-2012)
Abstract
In this study transgenic plants of sour orange (C. aurantium) that is an important citrus rootstock were produced by Agrobacterium-mediated transformation. Epicotyl and hypocotyl segments-derived explants were co-cultured with Agrobacterium strain EHA105 carrying pFGC5941 plasmid containsing CTV coat protein (p25) gene. One of the main objects of present research was to improve the direct in vitro organogenesis efficiency in C. aurantium. Therefore different combination of BAP (0, 1, 2 mg/L) and NAA (0, 0.25, 0.5 mg/L) were used in selective medium to culture transformed explants. The highest regeneration (57%) was obtained from explant treated with 2 mg/L BAP and 0.25 mg/L NAA. Effects of wounding and vacuum infiltration on transformation efficiency were evaluated either. The best transformation efficiency (11.25%) was obtained from explants that were vacuum infiltrated during transformation and subsequently were cultured in medium containing 2 mg/L BAP and 0.25 mg/L NAA. PCR analysis using two different genes were performed to confirm transformation. Micro grafting of transformed shoots were carried out on non-transgenic, in-vitro grown seedlings.
Athar Yaghtin, Mokhtar Jalali Javaran, Ghasem Karimzadeh,
Volume 1, Issue 2 (10-2012)
Abstract
The use of plants as a source of medicine is traced back to long time ago. Modern biotechnology
proviedes the possibility of production of valuable protein such as pharmaceutical protein in
plant. In this study human Interferon gamma-oleosine genes under the control of Napin promoter
were transferred to Carthamus tinctorius L. (safflower) by Agrobacterium tumefaciens strain LBA4404.
Cotyledonal explants from safflower plant (Carthamus tinctorius L.) Padideh cultivar were used for
transformation. The transformed plants were screened on MS medium containing 0.09 mg L-1 NAA, and 1
mg L-1 TDZ containing 40 mg L-1 kanamycin. Presence of transgenes was confirmed using polymerase
chain reaction (PCR). Characterization of the transgenic plants is going on.
Leila Sarmadi , Abbas Alemzadeh , Behzad Ghareyazie ,
Volume 1, Issue 2 (10-2012)
Abstract
This study was carried out to detect the presence of genetically modified maize in imported into Iran using molecular approaches. Five samples of imported maize from Argentina in the second half of 2010 were obtained from Bandar Imam Khomeini custom. Using specific primers for CaMV 35S promoter and nos terminator, PCR was performed. In this study Invertase gene of maize was used as internal control. The results showed that maize samples imported from Argentina were genetically modified and they have regulatory regions of CaMV 35S and nos in their genome. The shipment was not labled and there was no indication in the accompanying documents that the
shipment “may contain living modified organisms”.
Abbas Salahi Ardakani, Mohsen Morovvati, Mehrnaz Entesari,
Volume 1, Issue 2 (10-2012)
Abstract
Insistence on opposing transgenic plants and genetic engenearing by some individuals and/or organizations, hasresulted high levels of application of dangerous chemical pesticides in Iran. Samples of cucumber and tomato fruits were collected from various fields of Kohgiloyeh and Boyerahmad province and the residue of Endosulfan and Diazinon were determined in them. The highest amount of residue was of Diazinon in cucumber. The average residues of Diazinon in cucumber were 0.462, 0.669, and 0.205 mg/kg in Gachsaran, Boyerahmad and Kohgiloyeh respectively. However the internationally accepted maximum residue level of Diazinon in cucumber is 0.1 mg/kg. The residues of Diazinon in tomato were 0.504 and 0.534 mg/kg in Gachsaran and Dena respectively which are higher than the international levels. According to this research, the average residue of Diazinon in cucumber in the whole province was 0.355 to 3.5 times more than the maximum residue level. The residue of Endosulfan in tomato fields of Boyerahmad regions like Tangari, Keveshk, Tang Tamoradi and Sepidar. It was also higher than the international limits in Khairabad region at Gachsaran country. The residue of Endosulfan on cucumber was higher than the international levels in Tangari and Dornkore regions of Boyerahmad country and also Dehre, Dehkhalife and Shain brakan regions of Gachsaran, Delirech region of Dena country and in Zarghamabad of Kohgiloye country. The results showed not only the presence of high levels of Diazinon and Endosulfan residues in tomato and cucumber, but also showed that the period between pesticide use and marketing was also very short. It also shows that number of spraying, recommended dose and the interval between the spraying, were not taken into consideration. Continued restrictions on the application of transgenic plants is therefore considered as acceptance of the continued application of Endosulfan and Diazinon and their presence in food basket in Iran.
Somayeh Rahimnahal, Jamal Fayazi, Khalil Mirzadeh, Mohammad Taghi Beigi Nassiri, Hedayat Allah Roshanfekr,
Volume 1, Issue 2 (10-2012)
Abstract
MHC locus codes antigens and leukocyte surface proteins which have roles on immune reactions and identification of foreign proteins. In cattle this locus is known as Bulla and is composed of three classes of genes: CLASS І, CLASS ІІ and CLASS ІІІ and is located on short arm chromosome 23. Each of these classes of genes has complex and various genes and each gene may have dozens of alleles. The objective of this study was to study the variation of exon 2 of locus BoLA-DRB3 in buffalo population in Khuzestan province. In this study, the Heminested-PCR method was used to amplifing this exon. In order to determain the level of polymorphism, blood sample were collected from 80 buffalos in Shadegan, Ahwaz, Dashte azadegan, Dezfoul and Shoshtar cities. DNA extraction and exon 2 of the MHC gene was amplified by specific set of primers for this gene to produce a 284 bp fragment. The amplified fragments were digested with HaeIII and RsaІ restriction endonuclease. Digested products were separated and were stained by vertical electrophoresis on 8% Polyacrylamide gel. After digestion with HaeIII and RsaІ nine and ten alleles (restriction digestion pattern) were obtained at this locus respectively. Alleles a and b with 34.37 and 23.75 percent were the most frequent alleles when digested with HaeIII and alleles a and b with 20 and 30 percent frequency were the frequent alleles when digested with RsaІ. We identified 17 genotypes using each of the restriction enzymes in this population.
Masoud Tohid Far, Pezhman Azadi,
Volume 2, Issue 2 (10-2013)
Abstract
classic plant breeding has increased the beauty and utility of ornamental plants, but through the application of biotechnology completely new traits could be developed in plants. Creation of blue color petal in carnation and rose are examples demonstrating the optional of biotechnology in creating novel traits out of the scope of the conventional hybridization. Application of genetic engineering in ornamental plants is not limited to changes in color. Resistance to pests and diseases, enhancement of flowering, extended cut flowers life and modification of flower structure are examples of other important traits that could be modified using modern biotechnology. It is predicted that genetic engineering will revolutionize ornamental plants industry in the future. With the introduction of any new technologies, concerns are raised about their safe use and deployment. Concerns have also been raised about the possible negative impacts of commercialization of genetically modified (GM) ornamental plants on the ecosystem. The concerns have several different origins and providing responses to these concerns and not addressing them is the main objective of this paper. This article considers GM ornamental plants in the context of current ecological risk assessment principles, research results achieved, and current regulatory frameworks.This article can be very useful for biotechnologists, biosafety specialists, and authorities concerned.
Sholeh Dah Pahlavan, Maryam Musavivand, Maryam Hashemi, Jalil Khara,
Volume 2, Issue 2 (10-2013)
Abstract
the present investigation was conducted to characterize enzymatic and molecular aspects of a beta 1 and 4 endoglucanase gene in Bacillus subtilis strain B5d.The gene was amplified with specific primers and its sequence was deposited in the NCBI (accession number ofKF670724). Sequence and phylogenetic analyses were performed using the Vector NTI and Mega 4 software. The PCR product comprised of 794 bp, encoding a peptid of 264 amino acids in length. Amino acid sequence analysis showed that the gene encoding beta 1 and 4 endoglucanase in B. subtilis B5d had multiple domains including cellulose binding domain belongs to family 3 and catalytic domain belong to glycoside hydrolase family 5.The enzyme activity was assessed in temperature range of 30-70 °C and pH 4.6. The results showed that the maximum catalytic activity of the enzyme occurred at 55 °C and was estimated at 461.83 U/L. The stability of the enzyme was measured at different pH and temperatures values and the results showed that the enzyme was stable at pH 5-10 and temperature values up 60 °C.
Diyana Heidari, Mohammad Hasan Shahhoseini, Zivar Salehi,
Volume 2, Issue 2 (10-2013)
Abstract
in the past two decades, biotechnology in genetic engineering has led the production of genetically modified organisms (GMOs) all around the world. The statistics of GMO cultivation has lightened the significant importance of GMOs worldwide, and shows that they are widely consumed by public. Maize is one of the most important transgenic crops which has widely cultivated and consumed in many countries. The aim of this study is the detection of genetically modified maize in Iran markets. First, three samples of transgenic maize were prepared from Shiraz’s Custom and their DNA was extracted using Cetyl Tri-methyl Ammonium Bromide (CTAB) method. The control PCR test of native maize was performed on Invertase (IVR) gene which is present in all corn types. For detection of genetically modified maize, the PCR test based on P35S promoter which present in most transformation vectors was optimized using specific primers (P35S1 and P35S2). Then, DNA of 37 maize samples collected from seven major cities of Iran and six maize samples from Jihad Agriculture Organization were extracted. PCR tests for the IVR gene and P35S were performed. Respectively, the 226 bp and the 195 bp amplicons were amplified. PCR products were cloned by T/A cloning in JM107 using pTZ57R plasmid for sequencing and preparing the positive controls. 46 samples collected from the market, Customs and Jihad Agriculture Organization were shown to be 100% positive for the native IVR gene and 56.5% (26 samples) positive for the exogenous P35S. This study showed that a high percentage of transgenic maize is present in Iran’s consumption market. However, no action has been taken toward labeling and the consumer awareness of these food products.
Mina Asgari, Mohsen Morowati, Sarab Eimani,
Volume 2, Issue 2 (10-2013)
Abstract
some of the top decision makers in the ministry of Jihad-e-Agriculture, ministry of Health and the Department of the environment emphasize on a ban on genetically modified products and are in favor of continuous use of synthetic pesticides while over 170 million hectare of the transgenic products are cultivated all around the world. To determine the diazinon residue level in sweet cherry, 40 samples were collected from Tehran Central Market. These products were produced in five main cherry producing cities such as Lavasan, Shahriar, Qazvin, Mashhad and Urumia. Samples were prepared and extracted by the standard method of QuEChERS. The GC/ MS analysis was used to determine the levels of Diazinon residue in the samples. The amount of pesticides measured was compared with the national and Codex MRLs. 10% of the samples contained Diazinon residue levels higher than the MRL value (0.05 mg/ kg) including 4 samples from Mashhad (0.3 mg/kg) and Lavasan (0.29 mg/kg). The remaining samples (90%) had no detectable residue levels of this insecticide. The results obtained in this study can be used for the future planning of pesticides application in the high risk areas of the country. Therefore, the continuous blockade of taking advantage of alternative products and methodologies which could reduce the pesticide usage like production of transgenic plants may result in the production of toxic food baskets for the consumers which may result in the spread of incurable diseases in the society.
Feresht Ghadamgahi, Saeed Zerehdaran, Mokhtar Ali Abbasi, Mojtaba Ahani Azari, Mona Salehi Nasab,
Volume 2, Issue 2 (10-2013)
Abstract
Sara Maleki, Gasem Ali Garoosi, Rahim Haddad, Esmail Nezami Alanog,
Volume 2, Issue 2 (10-2013)
Abstract
pPistacia vera cv. Qazvini and UCB-1 rootstocks are two worldwide well-known pistachio rootstocks. They are valuable because of their tolerance to salinity and drought (Qazvini), and the resistance to cold and to Verticillium diseases (UCB-1). To improve vegetative and growth quality of pistachio and also to enhance the rooting efficiency, the effect of Scopoletin, iron source (Fe-EDDHA and Fe-EDTA( on growth characteristics as well as peroxidase activity was studied. In addition, the effect of the different concentrations of Ca2+ and Mg2+ cations, the temperatures and EDTA concentrations were investigated at various levels. The results indicated that Scopoletin increased the shooting percentage and enhanced the growth quality. It also affected the peroxidase activity (α=0.05) significantly in comparison with the control. Results also revealed that applying Fe-EDDHA instead of Fe-EDTA in the culture media increased the rooting efficiency up to two fold in comparison with the control. There was also a direct relationship between the rooting rate and the enzyme activity, suggesting peroxidase as a predictive marker in the rooting phase of Pistacia vera. The kinetic properties of enzyme revealed the positive role of cations, 30°C (as an optimum temperature) and a negative role of EDTA on the enzyme activity.
Somayeh Montazerinezhad, Mahmoud Solouki, Barat Ali Fakheri,
Volume 2, Issue 2 (10-2013)
Abstract
salinity is one of the undesirable environmental factors which affects both growth and crop production. The stresses cause a wide range of reactions in plants including the change in gene expression and cellular metabolism and the change in growth rate and production. To study the activity of the antioxidant enzyme Ascorbate Peroxidase (APX) and the expression level of its encoding gene in three Sistan melon Landraces (Cucumis malo L.), a factorial experiment was conducted in completely randomized design with three replications under control and salinity conditions (250mM and 350mM of NaCl). The relative expression of APX was compared by Real time PCR and enzyme activity was assayed in three melon landraces of Ghandak, Sefidak and Sefidak khatdar. According to the results, at 250 mM salinity level, the CmAPX gene expression showed an increase in comparison to control in all the three landraces and this increase in expression was significant at 5% in Ghandak and Sefidak landraces. At 350 mM salinity level, CmAPX gene expression was decreased in all the three studied landraces. At this level of salinity, the expression of CmAPX gene in the Khatdar Sefidak landrace significantly decreased (at 5% level) in comparison to control. More increase in antioxidant activity of APX with stable expression of CmAPX gene in Ghandak landrace was detected that shows this landrace could tolerate against salt stress better than the others. Decreased expression of this gene in Sefidak khatdar landrace with reduced enzymatic activity showed sensitivity of this landrace to salinity. We suggest that other genes related to antioxidant enzymes as well as their relationship with each others be examined in Sistan melon landraces.
Neda Mirakhorli, Afsane Heydari Reyhani, Behrooz Shiran, Sadollah Hooshmand,
Volume 3, Issue 1 (4-2014)
Abstract
since permanent transfer of genes and transgenic plant production, particularly in perennial plants such as almonds are time and cost consuming, optimization of various factors affecting the transgenic plants is critical. Transient genetic expression using Agrobacterium (Agroinfiltration) in plant is a useful tool to study gene function indifferent gene expression systems. Since optimization of factors that could affect the transgenic methods is necessary for gene transfer programs, Because This study was aimed at developing an efficient system for transient expression via Agrobacterium in almond leaves for rapid prediction of systems performance. Here the lichenasereporter gene was usedto assessthe parametersinfluencing thelevel oftransient expression. The main advantagesof using thisgene includeeasy operation, high sensitivity and biosafety. Three different constructs carrying lichenase gene and differentregulatoryelementswereinfiltrated intoleaves ofmamaei and shahrood 12cultivars,by two Agrobacteriumstrain, LBA4404 andGV3501,using syringewithoutneedle.After threedays,the amount of active proteinwas measured. Statistical analysis showedthat the highestamount ofprotein was obtainedin theleavesof mamaei cultivar transformedby theexpression systemcausing accumulation ofproteins in theendoplasmic reticulum by LBA4404 strain. So forrapidproduction ofrecombinant protein in Almond, theagroinfiltration method by using the related expression system, LBA4404 strain and mamaei cultivar, isrecommended.
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