Molecular Genetics and Genetic Engineering
Zahra Fathi; Katayoun Zamani; Solmaz Khosravi; Mohammad Malboobi
Abstract
Breeding crops with a higher ability in using soil minerals is one of the biotechnology researchers’ goals. Genetic engineering methods provide considerable advances in crop breeding by transferring and creating desired traits for further production under normal or stress conditions. In these procedures, ...
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Breeding crops with a higher ability in using soil minerals is one of the biotechnology researchers’ goals. Genetic engineering methods provide considerable advances in crop breeding by transferring and creating desired traits for further production under normal or stress conditions. In these procedures, Design of efficient gene constructs is of particular importance and requires promoters with proper function to specifically express the gene of interest in the target tissue and at the appropriate time to develop desired traits such as tolerance to biotic and abiotic stresses or other aims. Specific expression of phosphate-transporter genes in the roots and their induced levels in phosphate deficiency shows the potential of this gene-family promoters utilization in transgenic plants, particularly for the use in phosphate absorption from soil. Bioinformatics analysis showed that the 1826-bp promoter fragment of AtPHT1;1 gene carries several motifs leading to root-specific expression in Arabidopsis thaliana. The expression of a secretory acid phosphatase gene, AtPAP17, as a reporter gene in rapeseed transgenic plants indicated that the AtPHT1;1 promoter retains its root-specific criteria in rapeseed such that it could be used as a regulatory region for the specific expression of desired genes in transgenic rapeseed plant roots.
Molecular Genetics and Genetic Engineering
Marouf Khalili; Mohammad Hasso Mohammad; Hamze Hamze
Abstract
To study the effect of different levels of melatonin on the biochemical properties and the amount of expression of genes related to the activity of antioxidant enzymes in bread wheat, a split plot experiment was conducted based on a randomized complete Triticum aestivum block design. Irrigation levels ...
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To study the effect of different levels of melatonin on the biochemical properties and the amount of expression of genes related to the activity of antioxidant enzymes in bread wheat, a split plot experiment was conducted based on a randomized complete Triticum aestivum block design. Irrigation levels (normal (FC = 80%)), mild stress (FC = 60%), and severe stress (FC = 40%)) were allocated to the main plots and melatonin foliar spray (zero, 50, 100, 150, and 200 μM) were assigned to subplots. The results showed that the flavonoid content and ascorbate peroxidase enzyme activity increased with the intensification of dehydration stress. The level of 100 µM melatonin had the highest flavonoid content, ascorbate peroxidase enzyme activity. The mean comparison interaction treatments showed that the highest content of proline, phenol, superoxide dismutase and catalase was assigned to the 100 µM melatonin foliar spray under conditions of extreme stress of dehydration. Also, the lowest amount of malondialdehyde was observed for the 50 μM melatonin foliar treatment under normal irrigation conditions. The highest content of chlorophyll a, chlorophyll b, carotenoid and grain yield were recorded in the treatment of foliar spray of 100 μM melatonin and normal irrigation conditions. In this investigation, the maximum expression of superoxide dismutase, ascorbate peroxidase, polyphenol oxidase and catalase genes were determined in the foliar spray of 100 and 150 μM melatonin levels under conditions of extreme stress. . The foliar spray of melatonin, especially at the level of 100 μM, could moderate the effect dehydration stress on grain yield by improving biochemical and antioxidant properties.
Molecular Genetics and Genetic Engineering
Sepideh Charkhandaz; Karim Sorkheh
Abstract
The yield and acceptable quality of the oil of canola make this plant as one of the most important plants for providing edible oil. Due to the hot and dry climate in Iran and water necessary for seed germination, discovery of the solution to improve germination and seedling establishment is required. ...
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The yield and acceptable quality of the oil of canola make this plant as one of the most important plants for providing edible oil. Due to the hot and dry climate in Iran and water necessary for seed germination, discovery of the solution to improve germination and seedling establishment is required. Seed priming and treatment of seedlings with the extract of the seaweed Ascophyllum nodosum is one of the solutions for uniform germination and establishment of seedlings. The currently study have been explore the effect of application of Ascophyllum nodosum seaweed extract on seed germination and changes in the relative expression pattern of some important key genes related to gibberellic acid and brassinosteroids biosynthesis pathway using qRT-PCR. For this purpose, germination indexes were measured by the application of treatment 1% v/v treatment of Ascophyllum nodosum extract in four levels of treatment (0, 10, 15 and 30 days) until flowering stage. The leaf of sampling in each treatment was performed in three biological replications. The results of this study showed the effectiveness of seaweed extract treatment on increasing germination and seed vigor percentage, so that 100% germination of treated seeds obtained on the 3-day. The relative expression of algae extract showed an increase in the expression of EXP, ATI, ENTKO, AEC and BINSP genes, which are all important and as the key genes in the germination process compared to the control treatment. The use of seaweed extract treatment showed the highest expression in ATI (4.39) and ENTKO (5.70) genes in 30-day treatment, respectively. The lowest expressions for GIB and EXP genes in all three treatments (10, 15 and 30 days) were obtained. Also, there was the highest positive correlation between EEN and GIB (1), ENTKO and BINSP (0.98), AEC and BINSP (0.96) genes at the 5% significantly level. The correlation of these genes due to the use of algae extract ultimately led to a positive effect on the germination process of rapeseed seeds. Also, the results of heat map showed the genes of EXP and AEC (-0.57), GIB and ATI (-0.50) genes and XEN and ATI (-0.49) have a negative correlation at an error level of 5% and the increase of one led to the decrease of the other. This indicates that’s the activity of the final product of each of these genes is able to affect the germination process and promoting plant growth and development.
Molecular Genetics and Genetic Engineering
Parisa Daryani; Fatemeh Farzaneh Piralger; Nasser Zare; Zahra-Sadat Shobbar; Rasool Asghari Zakaria
Abstract
WRKY gene family encodes a large group of transcription factors regulating biotic and abiotic stress-responsive genes. In order to identify the WRKY gene family members in rice (Oryza sativa ssp. japonica), multiple searches were done in the related databases. Rice WRKY-conserved sequences were used ...
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WRKY gene family encodes a large group of transcription factors regulating biotic and abiotic stress-responsive genes. In order to identify the WRKY gene family members in rice (Oryza sativa ssp. japonica), multiple searches were done in the related databases. Rice WRKY-conserved sequences were used as the templates for tBLASTN searches in datasets for finding new members. An HMM profile of WRKY domain was also used to find WRKY gene family. Multiple sequence alignment was done using clustalW software, and phylogenetic trees were drawn using MEGA10 software based on a neighbour-joining method with a 1000 repeats bootstrap index. According to the results, 165 members of the WRKY gene family were found in rice, of which 63 were new members. Sequences were divided into three main groups based on the number of WRKY domains and the structure of zinc-finger motifs. Conclusively, there were 21 proteins with two WRKY conserved domains in group I, 53 proteins with one WRKY conserved domain and Cx7Cx23HxC zinc-finger motif in group III and 82 proteins with one WRKY conserved domain and Cx4-5Cx22-23HxH zinc-finger motif in group II. The chromosomal location of OsWRKYs was detected on the rice genome. The different groups were distributed on various chromosomes. The greatest number of OsWRKY genes (32 members) were located on chromosome 1. Following complementary research and identification of promising candidate genes involved in tolerance to each stress, they can be used to increase tolerance to the desired stresses and provide food security using genetic engineering or molecular breeding approaches.
Molecular Genetics and Genetic Engineering
Zahra Fathi; Katayoun Zamani; Mohammad Malboobi
Abstract
Phosphite is a reduced form of phosphate, wherein an oxygen replaces a hydrogen atom, and this substitution has a significant effect on its performance in living organisms. Phosphite is readily transfered into plant cells through phosphate transporters. However, plants do not have the ability to use ...
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Phosphite is a reduced form of phosphate, wherein an oxygen replaces a hydrogen atom, and this substitution has a significant effect on its performance in living organisms. Phosphite is readily transfered into plant cells through phosphate transporters. However, plants do not have the ability to use phosphite as a phosphorus resourc such that this property has limited the use of phosphite as fertilizer; however, phosphite has been used as a fungicide and biostimulant in agriculture. Some bacteria are able to oxidize phosphite into phosphate to cover for various cellular functions. In the last decade, the molecular mechanism of this biological oxidation has been elucidated to occure by the enzyme phosphite oxidoreductase or phosphite dehydrogenase. Phosphite is produced in large quantities in various chemical industries as a by-product or waste that is not recycled. The identification of the enzyme phosphite dehydrogenase, that catalyses the oxidation of phosphite to phosphate, has opened a new path for the recycle of this waste. Recently, there have been reports for the production of transgenic plants expressing ptxD gene. In practice, ptxD gene can be used as a marker in the selection of transgenic plants. By producing these transgenic plants, phosphite can be used as a herbicide and even as a phosphorus fertilizer.
Molecular Genetics and Genetic Engineering
Seyede mehri Javadi; Zahra-Sadat Shobbar; Asa Ebrahimi; Maryam Shahbazi
Abstract
Drought is the most important environmental stress that reduces crop yield. Therefore, research toward developing tolerant varieties is of great importance. In this study, microarray data analysis was used for identification of drought stress responsive genes and relevant hub genes in the reproductive ...
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Drought is the most important environmental stress that reduces crop yield. Therefore, research toward developing tolerant varieties is of great importance. In this study, microarray data analysis was used for identification of drought stress responsive genes and relevant hub genes in the reproductive stage of barley, and then their promoter analysis was performed. To achieve the goal, all the differentially expressed genes (DEGs) at drought conditions with fold changes ≥+2.5 and ≤-2.5 were identified between two microarray data-series in barley using FlexArray software. Bioinformatics analysis indicated that 559 genes were drought responsive at reproductive stage. The hub genes were distinguished using three Cyto-Hubba computational algorithms by Cytoscape software. Based on the hub analysis results, 10 unique (non-redundant) genes were identified as the most effective genes in response to drought stress. According to the gene ontology analysis of DEGs and hub genes, regulation of transcription were among the major groups indicating the importance of transcription factors (TFs) at drought tolerance mechanism. Amongst the hubs, several TFs such as HvCBF6, HvDRF1.3, LFL1, VP1, ABI5 and WRKY71 genes (belonged to AP2, WRKY and bZIP families) were observed. Promoter analysis was also revealed that some TF families including AP2, AT-hook family, bHLH, NAC, bZIP and MYB had binding site in 85% of promoters of the drought responsive genes and the hub genes in barley. Studying these transcription factors can help in better identification of drought tolerance mechanism in barley.
Molecular Genetics and Genetic Engineering
Ali Saeedpour; Soodabeh Jahanbakhsh; Tahmineh Lohrasebi; Kasra Esfahani; Ali Hatef Salmanian; Khadijeh Razavi
Abstract
Conventional pBI121-based binary vectors are widely used in transformation of higher plants mediated by Agrobacterium but they are useless in transformation of some monocots because of inefficiency of kanamycin in selection, while, hygromycin resistance gene is an important selectable marker that usually ...
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Conventional pBI121-based binary vectors are widely used in transformation of higher plants mediated by Agrobacterium but they are useless in transformation of some monocots because of inefficiency of kanamycin in selection, while, hygromycin resistance gene is an important selectable marker that usually used in transformation of several plants, especially monocots. The aim of this study was to improve the pBI121 vector for transformation of monocot plants. For this purpose, the hygromycin resistance gene with the 35S terminator were isolated from p6-ubi-rnai plasmid and cloned into pBlueScript intermediate vector via SmaІ and NotI restriction enzyme digestion. The CaMV 35 promoter was isolated from pBI121 vector by using SmaI and HindIII enzymes and cloned upstream of the gene. By using HindIII and Eco53KI enzymes, the complete hygromycin resistance gene cassette was replaced the kanamycin resistance gene cassette (which digested by HindIII and MssI) of pBI121 vector. Construction of this vector was confirmed by PCR method, digestion pattern analysis, and sequencing. Due to the popularity of pBI121-based vectors than other binary vectors and the researchers' familiarity with their manipulation, the vector which is introduced in this study could be used in gene transfer studies of monocot plants.
Molecular Genetics and Genetic Engineering
Samaneh Bagheri; Barat-Ali Fakheri; Ali Mohammad Ahadi; Abbasali Emamjomeh
Abstract
Wheat streak mosaic virus (WSMV) is one of the most important wheat infectious which its prevalence is increasing in Iran. NIa protein as a key protein in WSMV, plays important roles in viral replication and proteolytic digestion of viral polyprotein. Considering the critical role of NIa protein in viral ...
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Wheat streak mosaic virus (WSMV) is one of the most important wheat infectious which its prevalence is increasing in Iran. NIa protein as a key protein in WSMV, plays important roles in viral replication and proteolytic digestion of viral polyprotein. Considering the critical role of NIa protein in viral infection, isolation, sequencing and determination of secondary and tertiary structure of this protein were the objectives of the present study. Also, the amino acids present in the NIa protein active site, which can be used as the targets for design and developement of new antiviral agents, were investigated. In this study, the gene encoding NIa protein was isolated from WSMV and its sequencing was done followed by cloning in pEST prokaryotic vector. The resulted nucleotide sequence was deposited in NCBI database with accession number MK335432. Investigation of physical and chemical properties showed that NIa protein is including 426 amino acids, 48.8 kD molecular weight and 8.81 PI. Prediction of secondary structure of NIa protein revealed that 53.29% of its structure composed of irregular loops, which was justified by the structural dirsorder of this protein. The results of the active site investigation of NIa protein based on homologous sequences alignment and three-dimensional structure of the protein showed a highly conserved site which was included of histidine, aspartic acid and cysteine amino acids.
Molecular Genetics and Genetic Engineering
Niloufar Peykari; Katayoun Zamani
Abstract
Producing transgenic plants need new promoters. Due to the advent of next generation sequencing technologies and the production of massive genomic data for various crop plant species paralleled by the development of bioinformatics tools, there is an opportunity to identify, isolate and characterize ...
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Producing transgenic plants need new promoters. Due to the advent of next generation sequencing technologies and the production of massive genomic data for various crop plant species paralleled by the development of bioinformatics tools, there is an opportunity to identify, isolate and characterize new promoters. Because of public concern about using viral promoters, there is a need for cloning and using plant promoters in transgenic food crops. Native plant constitutive promoters may be composed of non-specific elements that are simply more efficient at protein recruitment for transcription. Promoters are classified as inducible, constitutive and tissue specific according to the nature of gene expression they regulate. Housekeeping genes are the best and most important sources for isolating the constitutive promoters. In this study, -Glucoronidase reporter gene expression mediated by a polyubiquitin promoter (CaUBQ10) from chickpea (Cicer ariethinum) was analyzed in tobacco tissues. The functionality of the CaUBQ10 was confirmed in leaves, stems, and roots of stably transformed tobacco plants and suggested that the CaUBQ10 is a constitutive promoter and may provide a valuable choice for high-level expression of target genes during the life cycle of a plant.
Molecular Genetics and Genetic Engineering
Hadi Karimbeigi; Farhad Nazarian Firouzabadi; Saber Golkari; Ahmad Ismaili; Masoud Alirezaei
Abstract
Histamine, an important biogenic amine, is produced by many organisms and play a diverse role in living organisms. Many micro-organisms synthesis and stored histamine in food, especially in canned food products, posing health threat to human. Diamine oxidase or histaminase catalysis histamine in healthy ...
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Histamine, an important biogenic amine, is produced by many organisms and play a diverse role in living organisms. Many micro-organisms synthesis and stored histamine in food, especially in canned food products, posing health threat to human. Diamine oxidase or histaminase catalysis histamine in healthy individuals and reduces its harmful side effects. In this study, a full-length cDNA encoding an amine oxidase was isolated from a chickpea (Cicer arietinum) landrace (grit), deposited in Genbank database (KU058599) and cloned in a binary expression vector. The cloned cDNA had an ORF with 2013 bp length encoding a protein with 670 amino acids and a molecular mass of 75.7 KDa. Multiple sequence alignments analysis showed that the active site and important amino acids are highly conserved in E.coli, pea and grit histaminases. Bioinformatics analysis revealed differences in 4 amino acids between grit histaminase and that of GenBank deposited Cicer arietinum amine oxidase (CAA08855). Phylogenetic analysis grouped grit histaminase (KU058599) with plant, especially legume histaminases with 99% bootstrapping.
Molecular Genetics and Genetic Engineering
Esmat Ashaar Ghadim; Shahrokh Gharanjik; Bahram Baghban Kohnerouz
Volume 6, Issue 15 , December 2016, , Pages 13-23
Abstract
Desaturase enzymes (EC: 1.14.19.3) catalyze dehydrogenation reactions and creates a double bond in the fatty acid chains. These enzymes classified into two groups, soluble and membrane each with different consensus motifs. Despite the Delta-6 desaturase is a key enzyme in the biosynthesis of unsaturated ...
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Desaturase enzymes (EC: 1.14.19.3) catalyze dehydrogenation reactions and creates a double bond in the fatty acid chains. These enzymes classified into two groups, soluble and membrane each with different consensus motifs. Despite the Delta-6 desaturase is a key enzyme in the biosynthesis of unsaturated fatty acids but due to lack of structural information of membrane-bound desaturase, it's three-dimensional and crystal structure has not been determined. Mortierella alpina is a rich source for production of unsaturated fatty acid arachidonic and has an active Delta-6 desaturases enzyme. The aim of this study was obtain encoding sequences of Delta-6 desaturase gene, and determination of functional domains using comparative analysis and molecular model and propose a membrane topology model for this enzyme. Thus, after extraction of total RNA and cDNA synthesis using gene specific primers, PCR product was cloned into pBlueScriptSK+ vector and then sequenced. Based on bioinformatics analysis by Swissmodel, Predict and Phobius servers presence of functional domains cytochrome b5 carrying motif (HPGG), three His-box motifs and transmembrane regions were identified. Then we proposed a membrane topology model for the Delta-6 desaturase enzyme.
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.
Molecular Genetics and Genetic Engineering
Masoud Tohidfar; Ebrahimi Ghorishi; Bartali Fakheri; Motahareh Mohsenpour
Volume 4, Issue 6 , October 2014, , Pages 47-59
Abstract
Abiotic stresses such as drought and salinity are the first factor of YIELD decrease in the world. In this study, betaine aldehyde dehydrogenase gene (badh) is used as both an abiotic stress marker gene and one of the abiotic stress tolerance candidate genes along with Flavodoxin (fld) in construction ...
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Abiotic stresses such as drought and salinity are the first factor of YIELD decrease in the world. In this study, betaine aldehyde dehydrogenase gene (badh) is used as both an abiotic stress marker gene and one of the abiotic stress tolerance candidate genes along with Flavodoxin (fld) in construction of chloroplast vector for rice. Thereby, adding the appropriate enzyme role to let gene cassette enter to the center. The first two parts of a specific target areaof Plastom rice (FR) the rice genome using PCR that they can be re-attached in cloning. Then, the gene cassetes were designed for fld and badh genes in regulatory regions of the chloroplast. So that the fld genes and with rbcl 5'UTR and badh genes with T7gene10 5'UTR were cloned the strong chloroplastsic promoter Prrn and the terminator rbcl 3'UTR. Finally, The complete two-gene cassettes fld/badh with the regulatory regions is separated from targeting chloroplast metabolism of rice in two sides cloned. The two specific chloroplast vector called pFrFB(-) and pFrFB(+) are potential to be attached to gene that are resistant to drought and salinity targeted with two different orientations relative to the inner regions of the chloroplast genome of rice plants and are able to be the goal of creating high resistance to salinity, drought and chill in transferring genes to use the chloroplast of rice plant via gene gun.
Biotic and Abiotic stress
Seyyedeh farzaneh Fatemi ardestani; Ghorbanali Nematzadeh; Hossein Askari; Hamidreza Hashemi
Volume 4, Issue 6 , October 2014, , Pages 73-83
Abstract
Soil salinity limits crop production by creating osmotic stress and disruption of ion homeostasis, leads to damage at the molecular level and finally cell death. In this study, gene expression analysis based on cDNA-AFLP technique was used to compare the expression profiles of KCl stress at three levels: ...
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Soil salinity limits crop production by creating osmotic stress and disruption of ion homeostasis, leads to damage at the molecular level and finally cell death. In this study, gene expression analysis based on cDNA-AFLP technique was used to compare the expression profiles of KCl stress at three levels: 0 (control), 200mM and 400mM, in Aeluropus littoralis which is the closest family to cereal. Among 34 isolated ESTs, 27 ESTs were obtained with the average length of 280 bp. The nucleotide sequences were compared with those in the GenBank database. Approximately 80% of the ESTs show homology to nucleotide or amino acid sequences in the GenBank database and 6 ESTs show no significant similarity in the GenBank database which considered as novel genes. Finally, 27 ESTs were recorded in NCBI database which are included potassium transporter, ribosomal protein, NADH dehydrogenase and golgin. The result of this research is very important to understand molecular basis and resistance mechanisms of drought stress for breeding and genetic engineering to improve crop resistance against stress and the production of resistant plants. EST classification based on responses to stress, will facilitate performance analysis, characterization of responsive genes in plant roots of Aeluropus littoralis to stress in future studies on this herb that is a member of the Poaceae family.
Molecular Genetics and Genetic Engineering
Somayeh Khosrojerdi; Maryam Hashemi; Maryam Mousivand
Volume 4, Issue 6 , October 2014, , Pages 95-105
Abstract
Cellulases can hydrolysis cellulose into glucose units which act in a synergistic manner of three enzymes endo-beta -1, 4-glucanase, cellobiohydrolase and beta-glucosidase. Nowadays, cellulases are used in various industries including animal feeds, textile, sewage treatment, brewery industries and biofuel ...
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Cellulases can hydrolysis cellulose into glucose units which act in a synergistic manner of three enzymes endo-beta -1, 4-glucanase, cellobiohydrolase and beta-glucosidase. Nowadays, cellulases are used in various industries including animal feeds, textile, sewage treatment, brewery industries and biofuel production. Demand for cellulolytic enzymes with unique catalytic properties is growing more rapidly than ever before. In this study, Bacillus subtilis A14h with cellulase production potential was identified using biochemical and molecular (16SrDNA gene) methods.The optimum conditions of B. subtilis A14h cellulase activity was evaluated in the presence of two type of substrates (carboxymethyl cellulose and beta-glucan), temperatures range of 30-70°C,and pH 4.6. Also, endo 1, 4-beta glucanase gene amplified using specific primers, sequencing outcome was recorded in the NCBI database. Sequence analysis and construction of phylogenetic trees was performed using vector NTI and Mega.4 softwares.The results showed that the isolate belongs to the Bacillus subtilis species. The phylogenetic analysis and comparison of the amino acid sequence of endo -1, 4-beta glucanase gene showed that the enzyme catalytic region belongs to glycosyl hydrolases family 5 and the non catalytic region was placed in the CBM3 family. The B. subtilis A14h cellulase was most closely related to the cellulase of B.subtilis strains. The results showed that the highest enzyme activity equal to 1464.25 U/ml was obtained in the presence of beta-glucan substrate at 55° C.
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.
Molecular Plant Breeding
Masume Rajabi Hashjin; Mostafa Agahee Sarbrzeh; Mohammad Hossein Fotokian; Mohsen Mohammadi
Volume 3, Issue 4 , September 2013, , Pages 33-41
Abstract
In order to evaluate the quality characteristics and seed storage proteins, 11 durum wheat genotypes as well as Eagle, Falat and Verinac genotypes were Chosen. Eight quality traits including hardness, grain quality, Zeleny number, dry gluten, wet gluten, bread mass, grain moisture, water absorption and ...
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In order to evaluate the quality characteristics and seed storage proteins, 11 durum wheat genotypes as well as Eagle, Falat and Verinac genotypes were Chosen. Eight quality traits including hardness, grain quality, Zeleny number, dry gluten, wet gluten, bread mass, grain moisture, water absorption and protein content were measured. For separation of glutenin subunits SDS-PAGE method was used. The correlations between protein content and zeleny number, hardness, dry gluten and wet gluten were significant and positive. On the other hand, there were significant and negative correlation between protein content and bread mass and moisture content. The stepwise regression results indicated that dry gluten was the trait most related to protein content since it explained 90% of protein variation. The high molecular weight Glutenin subunits (HMW-GS) at the Glu-A1 locus, the 2* allele was observed in the Eagle, Falat and Verinac and three genotypes (Kc-525, Wc-3122, Wc-45505). At the Glu-B1 locus, the 7+9 was observed in Falat cultivar. The 17+18 allele was presented in Eagle, Verinac and 6 genotypes (TN-12590, Kc-3638, TN-12501, Kc-525, Wc-3122, Wc-45505). The 7+8 allele also was observed in two genotypes (TN-12595, TN-12567). Cluster analyses of qualitative traits, the genotypes were divided into 3 groups. The findings of the present study could be utilized in breeding programs aimed at improving durum wheat.
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
Meysam Alizadeh; Ghorban Ali Nematzadeh; Mohammad Ali Ebrahimi; Hamidreza Hashemi
Volume 3, Issue 4 , September 2013, , Pages 53-60
Abstract
Rice is the main source of carbohydrates for more than a third of the world's population. It is also a principal food stuff for the Iranian people. In this study, AFLP markers were used to evaluate and fingerprint a number of rice (Oryza sativa L.) genotypes, including 28 Iranian local varieties and ...
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Rice is the main source of carbohydrates for more than a third of the world's population. It is also a principal food stuff for the Iranian people. In this study, AFLP markers were used to evaluate and fingerprint a number of rice (Oryza sativa L.) genotypes, including 28 Iranian local varieties and 19 exotic varieties. Using 10 primer combinations, 675 bands were obtained out of which 429 bands (63.5%) showed polymorphism. Among the used primers, E-TTG, M-CAT with107 bands and E-AGG, M-CTG with 34 bands had the highest and the lowest band numbers, respectively. Average polymorphism information content (PIC) was at 0.34. The best primer combination to differentiate rice samples were E-TTG , M-CAT with the marker index of 24.1. The average genetic similarity based on Nei coefficient, was estimated at 0.67 (0.97 to 0.40). The dendrogram obtained by using the UPGMA determined three main groups among the investigated varieties which was in concordance with the PCR analysis results.In conclusion, AFLP markers could provide unique fingerprinting patterns for all the studied varieties. The finding of this study could be used in inbreeding programs to produce hybrid and promising varieties based on the genetic distances and the identification of heterotic groups.
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.
Tissue culture and Micropropagation
Hamed Ebrahimizadeh; Mahmoud Lotfi; Shiva Azizinia; Farangis Ghanavati
Volume 3, Issue 4 , September 2013, , Pages 99-108
Abstract
Production of haploid plants was studied in summer squash via induction of parthenogenic embryos. Female flowers were pollinated with anthers irradiated by different doses of gamma ray (25, 50, 75, 100 and 200 Gray) and induced embryos were rescued and cultured in specific medium. The results achieved ...
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Production of haploid plants was studied in summer squash via induction of parthenogenic embryos. Female flowers were pollinated with anthers irradiated by different doses of gamma ray (25, 50, 75, 100 and 200 Gray) and induced embryos were rescued and cultured in specific medium. The results achieved showed that the highest number of embryos was obtained in 50 and 75 Gray doses (82%), however, no embryo was produced at 200 Gray. Gamma ray doses and embryo stage had significant effect on frequency of embryo and plant regeneration. Highest embryo regeneration and haploid formation (27 plant regeneration and 11 haploids formation, respectively) were recorded at 50 Gray. All amorphous embryos produced only diploid plant, though 29.17, 33.33, 57.14, 66.67, 100 and 100 percent of plants derived from cotyledon, heart, torpedo, arrow-tip, torpedo-tip and globular embryos respectively, were haploid. Based on this study, out of the 7744 extracted seeds, 127 embryos and 44 plants were regenerated; among those 17 plants were identified as haploid plants using chloroplast counting, Flow-Cytometry technique and morphologic traits evaluations.
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 Genetics and Genetic Engineering
Fatemeh Pirasteh Broujeni; Mohammad Reza Naghavi; Alireza Abbasi; Hasan Soltanloo; Mojtaba Ranjbar; Sara Raeisy
Volume 3, Issue 5 , February 2013, , Pages 23-31
Abstract
Artemisia species due to have different compounds of terpenoids are considered the most important medical plants. Artemisinin, a sesquiterpene with antimalarial and anticancer properties, and tritenpenes, squalene and β-amyrin, are important medicinal compounds which are produced by Artemisia species. ...
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Artemisia species due to have different compounds of terpenoids are considered the most important medical plants. Artemisinin, a sesquiterpene with antimalarial and anticancer properties, and tritenpenes, squalene and β-amyrin, are important medicinal compounds which are produced by Artemisia species. Since farnesyl diphosphate is the precursor of all tri- and sesquiterpenes, expression of farnesyl diphosphate synthase (FDS), squalene synthase (SQS) and β-amyrin synthase in three developmental stages are studied in seven Artemisia species native of Iran by real-time PCR. Furthermore, artemisinin content was determined by HPLC. Our results showed A. annua has maximum artemisinin content in budding stage and A. diffusa and A. spicigeria have minimum artemisinin content in vegetative stage. In this manner expression of FDS has no difference between the species and although its effective role in biosynthesis of artemisinin, it is not useful to manipulate for increase of artemisinin. Also lower expression of SQS means we will have higher artemisinin but the revers is not true. Also A. scoparia in flowering stage is the best source to access of squalene and β-amyrin.
Bioinformatics
Reyhaneh Ebrahimi; Behrouz Shiran; Esmaeil Ebrahimi; Hossein Fallahi
Volume 3, Issue 5 , February 2013, , Pages 49-51
Abstract
microRNAs are small non-coding RNAs approximately 21-22 nucleotides long and have been identified as negative regulators of gene expression in the post-transcriptional level in plants and animals. Temperature is one of the most important climate change factors and higher temperatures affect agriculture ...
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microRNAs are small non-coding RNAs approximately 21-22 nucleotides long and have been identified as negative regulators of gene expression in the post-transcriptional level in plants and animals. Temperature is one of the most important climate change factors and higher temperatures affect agriculture and crop production adversely. In this study, the expression pattern of miR398 and its target gene NtGT5b were measured in root and leaf tissues of sunflower (Helianthus annuus) in various heat stress conditions (0h, 1.5h, 3h , 6h) at 42 °C by qRT-PCR. The results showed different pattern of miRNA expression in both root and leaf tissues during mild, moderate and severe heat stress, respectively. miR398 was highly up regulated and its target gene down regulated in leaf tissue that showed thermo tolerance of this tissue in term of heat stress. There is a different miRNA expression patterns in leaf and root tissues under various stress conditions which is an indication of induced signal transmission after heat stress. Investigating miR398 network shows that this miRNA through presences of important genes and transcription factors such as AFO and INO has the ability to change the pattern of gene expression of many processes such as growth, development and response to stress.
