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Showing 9 results for Tobacco
Bahareh Zakerghoran, Hamid Rajabi Memari, Daryoosh Nabati Ahmadi, Marzieh Siahmard,
Volume 1, Issue 1 (5-2014)
Abstract
The production of human gamma interferon in eukaryote expression systems refers as a therapeutic recombinant protein which has significant impact in medical studies. Unique com position of gamma interferon makes such protein as a suitable tool against cancer. It is documented that phosphinothricin (PPT) classified as non-selective herbicide group of bialaphos acts an inhibitor for glutamine synthetase. The bar gene is encoding the phosphinothricin-N-acetyltransferase (PAT) enzyme. This enzyme capable of boosting resistance against PPT herbicide, thus it can be selected as a selective marker within plant population. Then various colony PCR techniques, enzymatic digestion and sequencing were used to confirm the accuracy of fusion of IFNγ-bar gens within expression transporter. Using freezing and thawing method to transfer the pCAMBIA1305.1- IFNγ-bar construction into strain of LBA4404 of agrobacterium, then disc leaves was used to integrate into the genomic of tobacco plant. The transgenic plants were selected under selector condition which possess 30 mg/l of hygromycin. After the developed roots were transferred into soil, and PCR technique was used to confirm the presence of IFNγ-bar in the genomic of plants. Dot blot analysis was applied to detect IFNγ-bar protein in transgenic of to tobacco plants.
Marziyeh Shazdehahmadi, Mahin Kharrazi,
Volume 2, Issue 2 (3-2016)
Abstract
Determination of genetic diversity of breeding material is the first step in breeding programs. Evaluation of tobacco genetic diversity is essential for breeding programs and preservation of genetic resources. Genetic diversity level in tobacco genotypes, is very important for selection of parents in breeding programs. In this study, genetic diversity of 100 tobacco genotypes was evaluated using 25 ISSR markers. Banding pattern based on the presence or absence of the bands showed with 0 and 1, respectively. Out of 237 fragments produced in total cultivars, 195 bands were polymorphic and average of polymorphism ranged from 4 to 12 per primer. Average of polymorphism percentage was 94.10. To determine the efficiency of ISSR markers, PIC and their polymorphic percentage was calculated. UBC 818, UBC 812 and UBC 815 had the best marker parameters and were introduced as the best primers for assessment of genetic diversity. In order to evaluate the genetic similarity between cultivars, different similarity coefficient (SM, Dice and Jaccard) was calculated and Mantel corresponding test was performed. Finally, dendrogram was drawn based on SM similarity coefficient and UPGMA algorithm and the Cofenetic coefficient was calculated. All genotypes formed two distinct clusters indicating the high efficiency of used primers in amplification the approximate parts of the genome. The principle coordinate analysis showed that the first three components could explain 79.65 % of total variance. Totally, evaluation the tobacco genetic diversity using ISSR markers is suitable and ISSR marker can be used as appropriate marker system to identify the diversity and genetic relationship for breeding programs of this plant.
Reza Darvishzadeh, Mir Javad Mousavi Andazghi, Amir Fayyaz Moghaddam, Hossein Abbassi Holasou, Seyyed Reza Alavi,
Volume 3, Issue 2 (3-2017)
Abstract
In order to evaluate heritability and gene action for some of the important quantitative traits in oriental tobacco, two genotypes, Basma 16-10 and SPT406 were crossed with Basma S. 31 cultivar, separately in 2009-2010. Parents with F1, F2, BC1 and BC2 generations were planted in a randomized complete block design with three replications. Traits such as plant height, leaf length, leaf width, leaf number, internnode number, stem diameter and yield per plant were recorded. The results obtained from analysis of variance indicated that generations mean squares were statistically significant for all traits expect for stem diameter. Therefore, generation mean analysis was performed for significant triats to estimate gene actions using Chi-square and scaling tests. The Chi-square of simple three-parametric model (additive-dominance model) was significant for studied crosses, indicating the presence of non allelic-interactions in the inheritance of these traits in oriental tobacco. Both additive and dominance genetic effects were significant for plant height, leaf length, leaf width, leaf number and internnode number. In addition, presence of high amount of dominance effect and dominance × dominance interactions suggests the importance of non-additive genetic effects for these traits in oriental tobacco. Therefore, selection for these traits in early generations can not be successful. However, additive genetic effects play an important role in the inheritance of yield, and then selection for this trait is hopeful in early generations during tobacco breeding process.
Mohammad Reza Salavati Meybodi, Gholam Ali Ranjbar, Seyed Kamal Kazemitabar, Hamid Najafi Zarrini,
Volume 4, Issue 1 (9-2017)
Abstract
Water stress is one of the most common environmental stresses that limited tobacco production in IRAN. Identification of genetic diversity in tobacco germplasm and genetic relationships between traits help to improve tolerant varieties. In this study, the genetic diversity of 100 flue–cured tobacco (Nicotiana tabacum L.) cultivars was analyzed using 15 morpho-physiological traits and 25 ISSR primers. The cultivars were cultured in a simple lattice design (10×10) with two replications (with and without water stress) in Tirtash Tobacco Institute, Iran. Results showed relative water content (RWC) and cell membrane stability (CMS) in normal condition were significantly higher than stress condition. The estimated broadsense heritability was low for RWC and CMS that represents large environmental effects on these two traits. The results of genotypes clustering by UPGMA method with ISSR markers and by WARD method with morph-physiological traits did not match. Primer UBC814: (CT) 8A with 16 polymorphic bands of the 17 bands, had higher resolution than other primers and seems appropriate for molecular diversity studies in tobacco. The K394 genotype was identified as well as drought tolerant varieties. We can use results of this study for selecting genotypes with great genetic differences and chose the desirable traits and use in breeding programs for producing high heterosis hybrid with tolerance to drought stress in tobacco.
Somayeh Dadashi, Reza Darvishzadeh, Mojtaba Nouraein, Hamid Hatami Maleki,
Volume 6, Issue 1 (9-2019)
Abstract
For the purpose of graphical analysis and estimation of genetic parameters related to yield and its components, six tobacco genotypes was crossed in half -diallel mating design. In this study, the F2 progenies and their parents were evaluated in a randomized complete block design with three replications. The results of analysis of variance indicated a significant difference between genotypes for plant height, leaf number, leaf dry weight and leaf fresh weight. Hayman graphical method was utilized for analysis of data. Diallel analysis indicated existence of an additive and dominant actions in inheritance of all studied traits. Traits including plant height, leaf number, leaf length and width, stem diameter, internode distance and leaf dry weight possessed greater additive effects meanwhile in the leaf weight the dominance effect was greater. The additive gene action detected for leaf dry weight (yield) manifested the influence of selection methods in breeding of this trait. Due to the fact that the fresh weight of the leaf was controlled by dominance effects, so hybrid-based methods are effective in modifying this trait. Also, additive and dominance gene action were contributed in heritability of yield, plant height and number of leaves. Regarding the mean values of dominance degree and results of graphical analysis, the gene action for leaf fresh weight was over-domnance and so, the heterosis phenomenon could be used to increase and improve this trait. For the traits including plant height, leaf number and yield, the gene action type was relative dominance.
Maryam Tahmasbali, Reza Darvishzadeh, Amir Fayaz Moghaddam,
Volume 7, Issue 1 (9-2020)
Abstract
In a breeding program, it is important to find out information about the genes action, because knowledge in this field could help the researchers in their crossing programs and realizing effective selection. In this study, breeding values of different agronomic traits in oriental tobacco were predicted using the best linear unbiased prediction (BLUP) procedure. For this purpose, 89 tobacco genotypes were evaluated in a randomized complete block design with three replications under normal (without broomrape) and stress (with broomrape) conditions at Urmia Tobacco Research Centre, during two successive years. Broomrape stress was applied by mixing 0.06 gr broomrape seed with soil in pots. C.H.T.209.12e × F.K.40-1 genotype had high yield under both normal and broomrape stress conditions and was one of the desirable genotypes in terms of yield tolerance and stability index. The Rustica genotype was the best genotype in terms of the breeding value of most of studied traits in both normal and broomrape stress conditions. The result from cluster analysis based on the breeding values of the studied traits showed that, tobacco genotypes were divided into 6 and 5 groups in normal and broomrape stress conditions, respectively; but the distribution of genotypes within the groups was different depending on the conditions. The highest heritability was observed for root fresh weight under normal condition and for leaf fresh weight under broomrape stress conditionss. The results showed that a genotype with good phenotypic performance may have low breeding value. Therefore, considering breeding value information along with phenotypic mean of traits can increase the efficiency of breeding programs.
Sahar Dashchi, Hassan Rahnama, Kianoosh Cheghamirza, Katayun Zamani,
Volume 7, Issue 2 (3-2021)
Abstract
In oilseed crops, a number of genes involved in the production of triacylglycerol have been identified that changes in their expression have increase the seed oil content. WRI1 and LPAAT are key genes in this synthetic pathway that their overexpression can increase the oil content. In this study, the expression vectors carrying WRI1 and LPAAT genes were designed and constructed for genetic transformation of tobacco (Nicotiana tabaccum) plants. The synthetic WRI1 and LPAAT genes were isolated from the PGH.WRI1 and PGH.LPAAT cloning vector using specific restriction enzymes and then cloned in the PGH.O3.2.2 intermediate vector under the control of SBP and Napin promoters and E9 terminator. Finally, the genetic cassettes were transferred to the plant transformation pBin19 binary vector. The resulting constructs were transferred to Agrobacterium tumefacience strain EHA105 and were used for genetic transformation of tobacco plants. Molecular analysis of transgenic plants confirmed the presence and activity of WRI1 and LPAAT genes. Seeds from transgenic plants were selected on the medium containing kanamycin and developed strong and healthy seedlings.
Maryam Ehsasatvatan, Dr Bahram Baghban Kohnehrouz,
Volume 9, Issue 2 (3-2023)
Abstract
Plastid engineering gives numerous benefits for the next generation of transgenic technology, consisting of the convenient use of transgene stacking and the production of high expression levels of recombinant proteins. Designed ankyrin repeat proteins (DARPin) are relatively small non-immunoglobulin scaffold proteins that bind to their specific target with high affinity. The G3 is a type of DARPin designed to bind to the HER2 tyrosine kinase receptor (human epidermal growth factor receptor 2). We previously developed a bioprocess for the production of DARPin G3 in tobacco chloroplasts as an imaging agent in HER2 over-expressed cancers. In this study, we analyzed the expression and homoplasmic stability of DARPin G3 gene in vegetative and generative T1 generation of transplastomic tobacco plants. The presence of DARPin G3 gene in the next generation of transplastomic plants was confirmed with specific primers by PCR analysis. Southern blot analysis confirmed the homoplasmic status of transplastomic plants. The western blot analysis confirmed the accumulation of the DARPin G3 in the chloroplasts of next generation of transplastomic plants. The DARPin G3 protein content was estimated around 33% by ELISA in chloroplast total soluble protein (TSP) of the transplastomic plants. Results confirmed that the DARPin G3 gene in vegetative and generative T1 generation of transplastomic tobacco plants was stably and highly expressed.
Maryam Ehsasatvatan, Bahram Baghban Kohnehrouz,
Volume 10, Issue 2 (2-2024)
Abstract
The global prevalence of type 2 diabetes mellitus is continuously increasing, and there is currently no definitive cure for type 2 diabetes. The potent glucagon-like peptide 1 (GLP-1), a natural small incretin hormone, enhances insulin secretion in a glucose-dependent manner. However, the exceedingly short half-life of GLP-1 limits its therapeutic applications. Albumin-binding DARPin can be used to increase the serum half-lives of therapeutic proteins, peptides, and small compounds. In this study, a long-acting GLP-1 agonist with oral delivery potential containing a protease-resistant GLP-1, an albumin-binding DARPin, and Penetratin as a fusion protein was expressed in a bioencapsulated form within tobacco chloroplasts to confer digestive system protection in plant cells. The successful transformation of tobacco chloroplasts with trivalent fusion protein-coding genes was conducted using a pPRV111A chloroplastic expression vector and a gene gun. Homoplasmic transplastomic plants were obtained after three rounds of selection in selection medium containing 500 mg/L spectinomycin and streptomycin. Transgene integration and homoplasmic status in the transplastomic plants were confirmed by PCR and Southern blot analyses. Western blot analysis confirmed the accumulation of the mGLP1-DARPin-Pen fusion protein in the chloroplasts of the transplastomic plants. The fusion protein content estimated by ELISA was 21.8% of the total soluble protein content in the transplastomic plants. The successful expression of the designed fusion protein indicated that the production of functional GLP-1 in plants may facilitate the development of a low-cost, orally deliverable form of this protein for the treatment of type 2 diabetes.
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