Molecular Plant Breeding
Mojtaba Khayam Nekouei; Mohammad Reza Ghaffari; Mohsen Mardi; Zahra Ghorbanzadeh; Rasmieh Hamid; Mehrshad Zeinalabedini
Abstract
Today, using advanced technologies such as the global positioning system (GPS), agricultural drones, satellite mapping, remote sensors, and precision agriculture machinery provides farmers with a lot of big data during production. According to the reports, this can be considered a part of the digital ...
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Today, using advanced technologies such as the global positioning system (GPS), agricultural drones, satellite mapping, remote sensors, and precision agriculture machinery provides farmers with a lot of big data during production. According to the reports, this can be considered a part of the digital economy in precision agriculture and be economically exploited. The analysis of this data cannot be processed by traditional processing systems due to its complexity. Given the size and complexity of big data, artificial intelligence can transform this data into valuable information through machine learning algorithms. This technology is being used to performance prediction algorithms, reducing agricultural inputs such as fertilizers and poisons, monitoring the growing conditions, pest management, breeding, molecular studies and finally value chain management. Developing programs using artificial intelligence technology will soon be able to manage the time of agricultural products entering the market, in addition to determining the planting time in order to increase productivity. The production of bio fertilizer from agricultural waste can be another achievement of the development of algorithms based on artificial intelligence to reduce the negative environmental effects and increase the economic productivity of the remaining waste from agricultural products. This study discusses the important applications of artificial intelligence in agriculture and its impact on Precision agriculture.
Genetic Engineering and Gene Transformation
Mohamad Amin Neycee; Motahhareh Mohsenpour; Hassan Rahnama
Abstract
Safflower with low oleic acid content is one of the native plants of Iran. Generally,, high oleic acid oils have more oxidative stability than the oils with high linoleic acids . Genome editing technology enable us to obtain oilseeds with high oleic acid. In this research, two guide RNA sequences ...
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Safflower with low oleic acid content is one of the native plants of Iran. Generally,, high oleic acid oils have more oxidative stability than the oils with high linoleic acids . Genome editing technology enable us to obtain oilseeds with high oleic acid. In this research, two guide RNA sequences were designed to target of Fatty Acid Desaturase 2 (FAD2-1) gene, which were located within the coding region and at a distance of 640 base pairs from each other. The guide sequences along with the codon optimized Cas9 gene were cloned in the T-DNA region of the Agrobacterium construct and transferred to the safflower by the In-planta method. The resulting seeds were cultivated and the plants were screened to track changes in the fatty acid profile of the seeds. The results showed that the amount of oleic acid in the seeds of one of the lines reached 53.14% on average. This line had four amino acid changes (L66F, N204D, S236A and I238V) at the same time. This is while the amount of oleic acid in the control plant was measured as 11.62% on average. The results showed that in the segregating generation, the change in fatty acid profile occurred in the line with homozygous amino acid change, and the heterozygous plants have the same oil profile as the control plants. Also, the results of this research can indicate the possibility of increasing the amount of oleic acid in oilseeds by changing the FAD2 enzyme sequence and without gene knockout.
Genetic Engineering and Gene Transformation
Zahra Ghorbanzadeh; Mehrbano Kazemi Alamouti; Leila Pourhang; Seyyed Mohammad Mousavi Pakzad; Elahe Moatamed; Mona Mapar; Aliakbar Ebadi; Mohammad Reza Ghaffari; Ghasem Hosseini Salekdeh; Behzad Ghareyazie; Motahhareh Mohsenpour
Abstract
Improvement of the root architecture lead to higher grain yield and seed quality. This is achieved via improvement of the plant growth, better establishment in soil, higher absorption of water and nutrition resulting in the biosynthesis of the essential amino acids and hormones. It increases the efficiency ...
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Improvement of the root architecture lead to higher grain yield and seed quality. This is achieved via improvement of the plant growth, better establishment in soil, higher absorption of water and nutrition resulting in the biosynthesis of the essential amino acids and hormones. It increases the efficiency of the nutrition usage and the stress tolerance. Drought conditions are a serious challenge in Iran; therefore, improving crop tolerance has a major importance. In this study, we investigate the presence of DRO1 gene, which is involved in the modification of the root growth angle, in rice cultivar Hashemi and compared to the Kinandang Patong cultivar. We further analyze the simultaneous presence of DRO1 and a second gene, OsCKX4, which is involved in the improvement of root structure. DRO1 and OsCKX4 are cloned together in a single construct under the control of the ubiquitin and the root specific promoters, respectively. The resulting construct, pUhrCkDro is transformed into the Agrobacterium tumefactions strain EHA105 and used for the gene transformation into Hashemi cultivar. Putative transgenic plants, survived on 50 mg. L−1 Hygromycin during tissue culture steps, are transplanted into the Yoshida solution and then into the pots until they set seeds. Construct specific and gene specific PCR analysis are used to confirm the transgenic plants. Transgenic plants show stronger root structure compared to the non-transgenic ones. Molecular analysis in the T1 and T2 generations leads to the homozygous events. The multi-genic construct used in this study, can be introduced into other crops for the aim of root structure improvement and drought tolerance. It is hoped that the production of transgenic rice with enhanced root structure results in improving drought tolerance, reducing water consumption and enhancing yield under drought stress conditions.
Genetic Engineering and Gene Transformation
Saeed Soheilivand; Amir Mousavi; Mohammadreza Safarnejad
Abstract
Sour lime (Citrus aurantifolia L.) is one of the most important woody plants is widely known for its recalcitrance to genetic transformation. We aimed herein to evaluate effective factors influencing the transformation efficiency and the reduction of chimeric transgenic shoots in sour lime. Epicotyl ...
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Sour lime (Citrus aurantifolia L.) is one of the most important woody plants is widely known for its recalcitrance to genetic transformation. We aimed herein to evaluate effective factors influencing the transformation efficiency and the reduction of chimeric transgenic shoots in sour lime. Epicotyl and internode explants were genetically transformed with different Agrobacterium tumefaciens strains e.g., LBA4404, GV3850, and GV3101, harboring the vectors pBI121 and pCAMBIA3301 containing β-glucuronidase (GUS) as a reporter gene. The effect of the following factors was evaluated: Agrobacterium concentration (OD600=0.3, 0.5 and 1), during inoculation (5 seconds, 10 minutes and 30 minutes), co-culture (2 and 3 days), and the selection regime (phosphinothricin at 1, 3, 5 and 10 mg/l and kanamycin at 25, 50, 75 and 100 mg/l). In following, transformation efficiency and the chimeric transgenic shoots rate were respectively confirmed by PCR and GUS assays. The results showed that Agrobacterium strain LBA4404, at the OD600 of 0.5, with 5 seconds (for epicotyl) and 10 minutes (for internode) inoculation at two-day co-culture period, were identified the most suitable treatments for both explants. The transformation frequencies ranged from 0.93% for internode on DKW medium containing 1.0 mg/l of phosphinothricin to 14.29% for epicotyl on DKW medium containing 50 mg/l of kanamycin. Inclusion of the high-level of selective treatments, improved the transformation rate through decreasing frequency escape and chimeric transgenic shoots. These findings provide novel insights into the appropriate procedure to constitute non-chimeric lime transgenic shoots.
Genetic Engineering and Gene Transformation
Elham Saboori-Robat; Mahmood Solouki; Ali Akbar Habashi; Motahhareh Moshenpour; Abbasali Emamjomeh
Abstract
Soybean is considered as one of the best source of protein for the nutrition of humans and mammals, and also is cultivated as an economic source of both vegetable oil and protein. Soybean like other Leguminosae, contains low levels of S-amino acids (methionine and cysteine). Using an appropriate selectable ...
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Soybean is considered as one of the best source of protein for the nutrition of humans and mammals, and also is cultivated as an economic source of both vegetable oil and protein. Soybean like other Leguminosae, contains low levels of S-amino acids (methionine and cysteine). Using an appropriate selectable marker can be effective in the regeneration of transgenic plants and increasing gene transfer rate. Glyphosate is a widely used non-selective herbicide with broad spectrum of weed control around the world. The aim of this study is constructing of two-genes construct consists of 11 kDa delta zein and EPSPS genes to improve the methionine content and induce resistance to glyphosate herbicide using Agrobacterium-mediated method in soybean. After experimental processes tissue culture, gene transformation and regeneration, plants produced by gene transformation showed glyphosate resistance at 3.5 mM concentration of glyphosate herbicide. Chlorophyll and shikimic acid content analysis also revealed that these two indexes in lines produce by gene transformation compared to wild type were significantly altered after glyphosate application. Complementary analyses are under progress.
Genetic Engineering and Gene Transformation
Katayoun Zamani
Volume 8, Issue 22 , September 2018, , Pages 65-79
Abstract
Transient expression is widely used as an alternative method for stable transformation of plants, especially cereals, trees and recalcitrants. In this method, multiple copies of transgene are introduced in to the plant cell and then are transcribed and translated. As introduced gene is not integrated ...
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Transient expression is widely used as an alternative method for stable transformation of plants, especially cereals, trees and recalcitrants. In this method, multiple copies of transgene are introduced in to the plant cell and then are transcribed and translated. As introduced gene is not integrated in the genome, its expression is not affected by epigenetic factors and the location of insertion. Transient expression is a convenient method for functional analysis of genes including overexpression, silencing, gene expression networks, protein localization, promoter analysis, identification of biosynthesis pathways and so on. Additionally, this method is applied in biotechnological industries like production of recombinant proteins. Transient transformation is performed by different methods, however, many of them have overlap. So, a combination of these methods can be used for transient expression. In this review, various transient transformation techniques, their advantages and disadvantages, latest progress regarding transient expression in model and crop plants and potential and limitations regarding the application of transient expression technique in science and industry will be discussed.
Genetic Engineering and Gene Transformation
Masoumeh Fallah Ziarani; Masoud Tohidfar
Volume 8, Issue 21 , June 2018, , Pages 71-79
Abstract
Today, Change the color of the flower due to its commercial importance is one of the goals of the researchers. Creation of the variation in flower color of ornamental plants can be highly profitable for the country and facilitate the way for export to other parts of the world. In the past, efforts have ...
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Today, Change the color of the flower due to its commercial importance is one of the goals of the researchers. Creation of the variation in flower color of ornamental plants can be highly profitable for the country and facilitate the way for export to other parts of the world. In the past, efforts have been do in the traditional way and genetic engineering for change the color of the flower, but with slowdown. With the discovery of Crispr's system, the ability to make targeted changes at the genome level took less time. In order to target change by Crispr, the target gene and area must be identified, that is done by bioinformatic tools. The desired change through the design of gRNA is done. In following, the normal protein and mutated protein were checked for the function. Today, donor DNA is used to enhance the performance of the Crispr system. In this system, by homologous recombination can be enhanced Crisper's performance by replaces a healthy gene or knocked out and prevented the creation of non-specific changes in the genome.
Genetic Engineering and Gene Transformation
Amin Abedi; Mohammad Mehdi Sohani; Reza Shirzadian
Volume 5, Issue 12 , February 2016, , Pages 67-78
Abstract
The monoterpenoids comprise a family of structurally and pharmaceutically diverse alkaloids. Strictosidine synthase is a key enzyme in monoterpenoid indole alkaloid biosynthesis pathway. In spite of the apparent lack of complex alkaloids in Arabidopsis, a gene family called Strictosidine synthase like ...
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The monoterpenoids comprise a family of structurally and pharmaceutically diverse alkaloids. Strictosidine synthase is a key enzyme in monoterpenoid indole alkaloid biosynthesis pathway. In spite of the apparent lack of complex alkaloids in Arabidopsis, a gene family called Strictosidine synthase like (SSL) has been found in the genome. SSL6, a member of SSL family, has been induced significantly by various stresses and signaling molecules. An overexpressed mutant is a powerful tool for functional characterization of an unknown gene. In view of that, SSL6 overexpressed mutant have been generated in order to study the possible role of the gene in Arabidopsis defense against biotic and abiotic stresses. The open reading frame from SSL6 was amplified and cloned into intermediate pJET vector before subcloning into pPZPY122 plant vector. Plant transformation was made by floral dip method using Agrobacterium tumefaciens strain GV3101 (PMP90). The putative transgenic plants were isolated on selective MS medium containing gentamycin. Transgene integration was further analyzed by PCR using SSL6 and gentamycin resistance gene specific primers. The transcription level of SSL6 in the T2 plants was measured using q-PCR and indicated an overexpression in transgenic compared to wild type Col-0 plants. Segregation ratio of plants in T2 and T3 generation on selection medium proved that some of the genotypes contain single T-DNA insert. The SSL6 Expression level in response to salt stress was measured in Col-0 and indicated an up-regulation of the gene 3, 6 and 12 hrs after treatment.
Genetic Engineering and Gene Transformation
Atena Mozafari; Rouhollah Kazemi; Jafar Amani; Mahyat Jafari; Ali Hatef Salmanian
Volume 5, Issue 11 , December 2015, , Pages 1-13
Abstract
Diarrheal diseases have been considered as one of the most major world health problems in all age categories. Among all the pathogens caused to diarrhea, entrohemorrhagic Escherichia coli (EHEC) and Vibrio cholerae are the most important agents. E. coli O157:H7 as the most important serotype of EHEC ...
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Diarrheal diseases have been considered as one of the most major world health problems in all age categories. Among all the pathogens caused to diarrhea, entrohemorrhagic Escherichia coli (EHEC) and Vibrio cholerae are the most important agents. E. coli O157:H7 as the most important serotype of EHEC bacteria caused diarrhea by producing STX2 toxin. In vibrio cholerae the cholera toxin (CTX) also is the main virulence factor that leads to diarrhea. Both of these toxins are belong to AB5 family and due to non-toxicity and natural immunogenic characteristic of B subunit, it could be introduced as an appropriate candidate for immunogenicity against these toxins. Plant seeds are an effective biological bioreactor for production of recombinant immunogenic antigens and also foreign proteins can be expressed in plants with the native structure. In this research, the production of recombinant protein composed of binding subunits of STX2 and cholera toxin was evaluated in canola seed as an oral vaccine candidate. The SC construct composed of (stx2B and ctxB) was subcloned to plant vector under the control of canola seed specific promoter (fae) and transformed to Agrobacterium tumefaciens. Then the recombinant vector was transformed to plant host via Agrobacterium mediated transformation. Transgenic plants was evaluated and the amount of chimeric protein expressed in transgenic canola seed was determined by semi quantitative ELISA and subsequently the amount of chimeric proteins was estimated 40 milligram per 1 gram seed. The results showed that the canola seed can efficiently produce recombinant protein.
Genetic Engineering and Gene Transformation
Kasra Esfahani; Ali Hatef Salmanian
Volume 4, Issue 7 , December 2015, , Pages 59-69
Abstract
Binary vectors such as pBI121 are widely used for introducing genes of interest into plants via Agrobacteriun-mediated transformation method. These vectors usually have low number of unique recognition sites of restriction enzymes in their multiple cloning sites. They also lack elements such as sequences ...
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Binary vectors such as pBI121 are widely used for introducing genes of interest into plants via Agrobacteriun-mediated transformation method. These vectors usually have low number of unique recognition sites of restriction enzymes in their multiple cloning sites. They also lack elements such as sequences necessary for protein purification. In this article, a new plant expression vectorwith a developed multiple cloning site including recognition sites of 13 restriction enzymes, a sequence for coding 8xHis-Tag to purify recombinant proteins and the sequence of M13 reverse sequencing primer, is introduced. Construction of new vector was confirmed by PCR, digestion pattern analysis, and sequencing. The T-DNA regions of the new vector and pBI121 as a control sample were transformed to Nicotiana tabbacum L. cv. Samsun by using Agrobacterium tumefaciensstrain LBA4404. Integration of T-DNA of the vector to the genome of plantlets, which were regenerated on selective culture, was analyzed by PCR. GUS assay also confirmed expression of the transgene in transgenic seedlings. These evidences indicated that the new plant expression vector, pBI105, is efficient for plant transformation.
Biotic and Abiotic stress
Marjan Bahrabadi; Farhad Shokouhifar; Mohammad Ali Ebrahimi
Volume 4, Issue 6 , October 2014, , Pages 11-20
Abstract
Synthetic pathogen-inducible promoters are suitable alternatives for native promoters in plant genetic manipulation to the purpose of resistant crop production. An ideal pathogen-inducible promoter would only be activated in response to target pathogens. Furthermore, it should express the transgene locally ...
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Synthetic pathogen-inducible promoters are suitable alternatives for native promoters in plant genetic manipulation to the purpose of resistant crop production. An ideal pathogen-inducible promoter would only be activated in response to target pathogens. Furthermore, it should express the transgene locally and temporarily. The absence of these characteristics in native promoters have drawn the attentions toward design and construction of synthetic promoters. Some components like cis-regulatory elements are used in construction of synthetic promoters and this provides high flexibility in determining the expression quantity and the inducibility type. One of the most common methods for a synthetic inducible promoter analysis is using Agrobacterium-mediated transient expression system. With this method, Functional analysis of the promoter can be performed in a short time by application of biotic and abiotic treatments and assaying the transgene expression. In this study, the SP-DD synthetic pathogen inducible promoter (containing of two copies of Box D cis-element derived from parsley PR2 promoter) fused with an intron-containing ß-glucuronidase reporter gene was transferred to tobacco leaves (Nicotiana benthamiana) by agroinjection. The promoter function was evaluated in response to salicylic acid treatment and environmental stresses like heat, cold and UV radiation. The results showed that the SP-DD synthetic promoter induced the ß-glucuronidase gene expression in response to salicylic acid and the expression amount increased over time from 2 hours to 24 hours post applicaion. Besides, the promoter showed slight sensitivity in response to heat and cold stresses but the ultraviolet radiation stress had no effect on the promoter induction.
Plant Disease and Biotechnology
Mahsa Banaee; Forogh Sanjarian; Gholam Reza Bakhshi Khaniki
Volume 3, Issue 4 , September 2013, , Pages 25-32
Abstract
Acetyl transferases are enzymes responsible for enzymatic transfer of an acetyl group to suitable receptor molecule by using acetyl CoA as donor. Acetyltransferase reaction is involved in biosynthesis pathway of some important secondary metabolites such as antibiotics, as well as in their detoxification. ...
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Acetyl transferases are enzymes responsible for enzymatic transfer of an acetyl group to suitable receptor molecule by using acetyl CoA as donor. Acetyltransferase reaction is involved in biosynthesis pathway of some important secondary metabolites such as antibiotics, as well as in their detoxification. Trichothecenes are significant secondary metabolites produced by the plant fungal phatogens Fusarium ssp. Such as F. sporotrichioides and F. graminearums. These fungi possess specific genes in their genomes encodeinge acetyl transferase enzymes are affecting trichothecene. In this study, the gene encoding acetyltransferase from the fungus F. sporotrichioides, TRI 101, was cloned and transferred into tobacco plant as model plants and the effect of the enzyme on the detoxification of deoxynivalenol (DON), a well known trichthecene, was investigated. In addition it was shown that, in comparison whit the roots of wild type plants, transgenic roots grew normally in the deoxynivalenol-contained medium.
Genetic Engineering and Gene Transformation
ali reza abbasi; Nahid Raanaian; Fariba Aboei; U. SONNEWALD; L. M. VOLL
Volume 3, Issue 4 , September 2013, , Pages 43-52
Abstract
Under stress conditions Reactive Oxygens Species (ROS) are produced and accumulate in plant leaves as a results of the oxidation of important cellular components like proteins, chlorophylls and lipids. Vitamin E is the collective term for tocochromonals. This lipid-soluble substance ...
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Under stress conditions Reactive Oxygens Species (ROS) are produced and accumulate in plant leaves as a results of the oxidation of important cellular components like proteins, chlorophylls and lipids. Vitamin E is the collective term for tocochromonals. This lipid-soluble substance acompanied by carotenoids, glutathion and other antioxidants and has an important role in keeping the integrity of photosynthetic membranes. Tocopherols and tocoterienols are amphipathic molecules and are able to detoxify ROS and lipid peroxyl radicals in lipophilic environments. Hemogentisate phytyl transferase (HPT) is the key enzyme in the vitamin E biosynthesis pathway. To silence HPT and invesitgate its role in tocopherol production RNAi thechnique was used and the results obtained revealed that tocopherol content was decreased in derived lines.
Molecular Plant Breeding
Sara Kabirnataj; Elnaz Ghotbi Ravandi; Behzad Shahin Kaleybar
Volume 3, Issue 4 , September 2013, , Pages 69-76
Abstract
Using Agrobacterium rhizogenes to induce hairy root cultures is a useful method to increase the production of secondary metabolites especially medicinal compounds in vitro in various plant species. Hairy root cultures are fast growing and highly branching and due to the higher amount of metabolites synthesized ...
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Using Agrobacterium rhizogenes to induce hairy root cultures is a useful method to increase the production of secondary metabolites especially medicinal compounds in vitro in various plant species. Hairy root cultures are fast growing and highly branching and due to the higher amount of metabolites synthesized per unit of biomass, possess the same or greater biosynthetic potential for secondary metabolite production compared to the normal roots. In this research the amounts of total phenolics and chlorogenic acid were determined in hairy root clones induced in cotyledonary leaves of Cichorium intybus by A4, A13 and 15834 strains of A. rhizogenes, as well as in the control (untransformed) roots. The results obtained indicated that the absence of chlorogenic acid in all the studied clones led to a significant increase in total phenolics in hairy root clones induced by A4 and A13 but a significant decrease in the 15834-induced hairy root. This study revealed the role of bacterial strains in biosynthesis of phenolic compounds, and therefore, selection and application of the best strain of A. rhizogenes could be regarded as an important strategy for increasing phenolic compounds production in hairy root cultures of C. intybus.
Genetic Engineering and Gene Transformation
Nahid Ahmadi; Hasan Rahnama
Volume 3, Issue 4 , September 2013, , Pages 77-85
Abstract
Transient expression of foreign genes in plant tissues is a valuable tool for plant biotechnology and to shorten the time for gene functional analysis. Transient expression is a fast, flexible, simple and easy method that could be used in fully differentiated plant tissues. 2A11 promoter is a tomato ...
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Transient expression of foreign genes in plant tissues is a valuable tool for plant biotechnology and to shorten the time for gene functional analysis. Transient expression is a fast, flexible, simple and easy method that could be used in fully differentiated plant tissues. 2A11 promoter is a tomato fruit specific promoter whose expression occurs in fruit significantly. Cloning of fruit-specific promoter (2A11) is the main purpose of this study. 2A11 promoters were amplified from genomic DNA of tomato by PCR using specific primers. The promoter fragments were cloned into cloning vector PTZ57R/T. Recombinant plasmids were transferred into E. coli XLI-blue strain. Fragments of the interest were digested using restriction enzymes BamH1 and HindIII and were then purified and substituted in CaMV35S promoter in binary pBI121. Binary pBI121 was selected for cloning of 2A11 promoters as it is an ideal vector for agroinfiltration due to the presence of the CaMV35S promoter and the GUS reporter gene within its T-DNA region. Recombinant plasmids were transformed into Agrobacterium tumefaciens LBA4404 strain with freeze and thawing method. The expression of GUS gene was analyzed in tomato with agroinfiltration method. The results showed that, 2A11 promoter was found as efficient as CaMV35S promoter in expression of GUS gene specifically in tomato fruit. Then, we can use this promoter for tissue specific expression of recombinant proteins in tomato fruits.
Molecular Plant Breeding
Nahid Raanaian; Ali reza Abbasi; Hassan Zeynali-Khanghah
Volume 3, Issue 4 , September 2013, , Pages 149-156
Abstract
Exposing plants to both biotic and abiotic stresses could lead to increased reactive oxygen species and in turn could induce oxidative stress. In order to increase scavenging capacity of oxidative agents, the different plant antioxidant activities also are arising. Among those, vitamin E includes a group ...
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Exposing plants to both biotic and abiotic stresses could lead to increased reactive oxygen species and in turn could induce oxidative stress. In order to increase scavenging capacity of oxidative agents, the different plant antioxidant activities also are arising. Among those, vitamin E includes a group of Fat-soluble antioxidants, which their synthesis is limited to photosynthetic organisms including plants, alga and cyanobacteria. In this study, the coding region of Arabidopsis Tocopherol cyclase gene; which catalyzes 2,3-dimethyl-5-phytyl-1,4-benzoquinone (DMPBQ) into γ –tocopherol; was introduced into pBin vector containing the coding region of GFP protein. Then, the obtained construct was transformed into tobacco plant through agrobacterium-mediated method. Leaves of 2-3 weak old seedlings were selected as explant and direct regeneration was performed. In order to confirmation of transgenic plants, PCR by using specific primers was carried out. Due to the fact that the pBin vector contains the kanamycin resistant gene, seeds belonging to the T0 transgenic plants were planted on medium containing kanamycin and green seedling were selected as transgenic T1 plant. In order to evaluate the effects of transferred gens on physiological parameters, mature T1 transgenic plant as well as the wild type plants were subjected to drought stress and relative water content was measured.
Genetic Engineering and Gene Transformation
Hossein Maleki; Ali Hatef Salmanian; Jafar Amani; Alaodin Kordenaiej; Mahyat Jafari
Volume 3, Issue 5 , February 2013, , Pages 1-9
Abstract
E. coli O157:H7 is an important zoonotic pathogen that causing severe gastrointestinal disease such as hemorrhagic diarrhea and hemolytic uremic syndrome. The first step of bacterial pathogenicity is, attaching to the host cell. EspA is a protein molecule and forms as filamentous structure ...
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E. coli O157:H7 is an important zoonotic pathogen that causing severe gastrointestinal disease such as hemorrhagic diarrhea and hemolytic uremic syndrome. The first step of bacterial pathogenicity is, attaching to the host cell. EspA is a protein molecule and forms as filamentous structure that is transiently expressed on the outer membrane surface and interacts with the host cell during the early stage adhesion and forms bacterial biofilm, this filamentous structure make it a strong immunogenic. Tir is bacterial protein that translocated to host cell after expression in the bacteria by type III secretion system (T3SS) and integrated in to host cell membrane. Intimin can attach to the host cell by conducting to Tir. The purpose of this study is constructing bivalent gen which contains virulence factor mention before and transferring in to target plant in order to production edible vaccine against E. coli O157:H7.After bioinformatics investigation the two bivalent gen construction (espA-Tir,) were attached by peptide linker, and the gens were constructed. After codon optimization based on tobacco; construction were cloned in expression vector which contains CaMV35s promoter. After transformation and regeneration, the expressions of transgenes were showed in analysis.
Molecular Plant Breeding
M Ghaderi; A.R Abbasi; A.H Salmanian
Volume 2, Issue 2 , September 2012, , Pages 39-47
Abstract
Drought is by far the most important environmental stress in agriculture and many efforts have been made to improve crop productivity under water-limiting conditions.Minimizing the ‘yield gap’ and increasing yield stability under different stress conditions are of strategic importance in guaranteeing ...
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Drought is by far the most important environmental stress in agriculture and many efforts have been made to improve crop productivity under water-limiting conditions.Minimizing the ‘yield gap’ and increasing yield stability under different stress conditions are of strategic importance in guaranteeing food for the future. Expansin is a protein super family with 4 families in vascular plants. Expansions are cell wall proteins which mediate acidic cell wall loosening through breaking hydrogen bonds between cellulose and glycan matrix. In this study pBIEXPA1 construct was constructed. Arabidopsis plants were transformed by pBIEXPA1 which contained nptII gene and AtEXPA1 gene which was under the control of CaMV35s promoter. Transformation was done by floral dip which did not need any tissue culture and transformed plants were T1. Transformed seedlings were able to remain green on MS medium containing 50 mg/L kanamycin. Transgenic plants were confirmed by PCR. As expected two 753 and 1080bp band were seen in transgenic plants.
Genetic Engineering and Gene Transformation
S Nasr-Ramzi; M Sohani; H Hasani; J Asghari
Volume 2, Issue 2 , September 2012, , Pages 49-62
Abstract
Agrobacterium-mediated transformation technique is a powerful and essential tool for genetic transformation and transgenic rice plant production. In this study an in planta transformation method was used for rice plants transformation. Therefore, rice seeds were socked for two days and the mature rice ...
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Agrobacterium-mediated transformation technique is a powerful and essential tool for genetic transformation and transgenic rice plant production. In this study an in planta transformation method was used for rice plants transformation. Therefore, rice seeds were socked for two days and the mature rice embryos was inoculated by means of an Agrobacterium coated needle. An experiment with factorial design including two strains of Agrobacterium tumefaciens (EHA105 and LBA4404) harboring pCAMBIAl105.1R, three levels of Acetosyringone (0, 100 and 200 Mµ), three cultivars of rice (Hashemi, Hasani and Gharib) and two treatments of vacuum and no vacuum operation was carried out in a Completely Randomized Design with three replications. Integration of the transgene into the genome of putative transgenic rice plants were confirmed using resistance of leaf tissues to Hygromycin, the histiochemical GUS assay and PCR with at least three different genes. Accordingly, EHA105 strain and Hashemi cultivar in the presence of 100µM acetosyringone using vacuum in a vir genes induction medium had a significant transformation efficiency (37.46%). The success of transformation was further confirmed by analysis of T1 generation with 21% transgenic.
Genetic Engineering and Gene Transformation
S Shahbazi; N Safaie; A Mousavi; F Sanjarian; A Alizadeh
Volume 2, Issue 2 , September 2012, , Pages 73-85
Abstract
Fusarium Head Blight (FHB), which is caused commonly by Fusarium graminearum, has ability to result significant reduce in yield and also cause indirect losses due to the accumulation of potent mycotoxins (trichothecenes) in harvested grain as secondary metabolites; which are hazardous for human and animal. ...
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Fusarium Head Blight (FHB), which is caused commonly by Fusarium graminearum, has ability to result significant reduce in yield and also cause indirect losses due to the accumulation of potent mycotoxins (trichothecenes) in harvested grain as secondary metabolites; which are hazardous for human and animal. Trichothecene mycotoxins (such as DON) are potent protein synthesis inhibitors for eukaryotic organisms. Trans expression of AYT1 gene from S. cerevisiae capable of trichothecene 3-O-acetylation and converts DON to a less toxic acetylated form. One of the detoxification and resistance to mycotoxins is acetylation. The main goal of this study was to evaluate transgenic tobacco model plants to deoxynivalenol (DON). In order to detect expression of the AYT1 transgene, we added cMyc tag via PCR-Tagging method and introduced it into the model tobacco plants through Agrobacterium-mediated transformation in an attempt to detoxify DON. After confirmation of integrations of AYT1-cMyc into the tobacco genome by molecular analyses, Immuno-blotting and serological protein studies and trichothecene acetyl transferase activity analyses confirmed the expression of AYT1 and tolerance to 10 ppm concentration of DON in transgenic lines was observed.
Genetic Engineering and Gene Transformation
Motahareh Mohsen Por; Masoud Tohid Far
Volume 1, Issue 1 , March 2012, , Pages 35-48
Abstract
A system was designed using E. coli heat shock promoter (groE) in plastid vector and a hybrid plant/bacteria sigma factor was constructed under control of a tissue specific promoter. This system was designed for overcome to deleterious effects on plant growth and fertility that may be caused by transgene ...
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A system was designed using E. coli heat shock promoter (groE) in plastid vector and a hybrid plant/bacteria sigma factor was constructed under control of a tissue specific promoter. This system was designed for overcome to deleterious effects on plant growth and fertility that may be caused by transgene overexpression. So that hybrid sigma factors contained N- terminal motives of tobacco sigma factors including chloroplast signal peptide and RNA polymerase interaction domains, composed by C-terminal motif of E. coli sigma32 that able to recognition and binding to groE promoter. Then this gene, HSig, was cloned in Agrobacterium vector after adding regulatory elements. The result vector was used for transformation of an Iranian variety of tobacco. Detection of transgenic plants was performed by PCR, southern blot and RT-PCR analysis. The Hsig gene expression and its targeting to plastid was confirmed after transformation of tobacco chloroplast using gene gun technique for targeting of green florescent protein (GFP) under control of groE promoter using pFNGi vector into transgenic HSig explants. We hope that the system that was designed and constructed in this study for GFP expression in chloroplast genome, be able to apply in molecular farming for expression of any other desired genes instead of GFP for specific gene expression in chloroplast.
