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Showing 6 results for Chloroplast
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Volume 4, Issue 1 (9-2017)
Abstract
Chrysanthemum is one of the world's most important cut flower crops. The genetic diversity assessment among present Iranian Chrysanthemum cultivars are needed for future Chrysanthemum breeding programs. In an attempt to reveal the genetic variation and relationship among 30 Chrysanthemum cultivars, some morphological and a chloroplastic gene (rpoC) DNA molecular markers were employed. Chrysanthemum cultivars were cultivated in a Randomized Complete Block design (RCBD) with 3 replications in the field. Meanwhile, a phylogenetic tree was constructed based on chloroplast rpoC gene sequence. Analysis of variance (ANOVA) indicated that a significant variation exists among the Chrysanthemum cultivars for almost all the traits under study. In general, phenotypic coefficient of variation was higher than the genotypic coefficient of variation indicating the predominant role of environment effects. Ward Cluster analysis divided cultivars in two main clusters with highest number of cultivars falling under cluster I. Phylogenetic relationships among Chrysanthemum cultivars based on Maximum likelihood method showed that Iranian cultivars belong to Chrysanthemum morifolium, suggesting that these cultivars have presumably been bred from the same set of parents. Furthermore, genetic distance between cultivars ranged from 0.4 to 2.9, indicating sufficient genetic diversity among cultivars for crossing and selecting the most appropriate parents for Chrysanthemum morifolium breeding programs.
Shahin Jahangirzadeh Khiavi, Zabihollah Zamani, Mohamadreza Fatahi, Masoumeh Ashourpour,
Volume 5, Issue 1 (9-2018)
Abstract
The chloroplastic DNA of 64 accessions of apple (Malus spp.) (54 Iranian genotypes, five commercial cultivars and five rootstocks) were analyzed to reveal their haplotypes by using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) method. Approximately 4320 bp of the chloroplast genome was analyzed, by using two chloroplast universal primer pairs and two restriction enzymes (EcoRI and MseI). All the mutations detected were insertion-deletions been in the range between 10-40 bp. The combination of all the mutations resulted to six haplotypes (H1, H2, H3, H4, H5 and H6) in the studied apple accessions. It was defined that the Iranian apple rootstock 'Gami-Almasi' had a specific haplotype form all the other studied accessions. It was approved that cpDNA diversity data can be considered for phylogenetic studies in this plant and the polymorphism determined in cytoplasmic genome by PCR-RFLP method can help to understand the maternal inheritance in apple.
Seyedeh Zahra Hosseini, Behroz Shiran, Mohammad Ali Ebrahimi,
Volume 5, Issue 2 (3-2019)
Abstract
Phylogenetic relations among 12 wild species of almonds, one cultivated almond and one species of peach were investigated by using of ITS1-5.8S rDNA-ITS2 sequences and trnL region of chloroplast DNA. To do this, maximum-parsimony and neighbor joining analysis adopted. Results of ITS data showed that studied species of Prunus only divided in two groups but incapable to separate different sections. P. tenella showed more diverse genetic distance in compare to other studied species and it seems that this species does not belong to Amygdalus. Also, by using the ITS data it can be reported that Prunus is monophyletic. In this research, the genetic distances for each pair of species were determined and the average genetic distance between species shows only the lowest genetic distance within the genus. Therefore, Prunus is a single genus. Regarding its high similarity of trnL region in wild almond species, it can be reported that maternal ancestor of Prunus is the same and trnL is not optimum marker to separate species of almond.
Halime Arbabi, Mojtaba Keykhasaber, Leila Fahmideh, Valiollah Ghasemi Omran,
Volume 9, Issue 2 (3-2023)
Abstract
Luffa (Luffa cylindrica) is a plant from the Cucurbitaceae family that grows mostly in tropical and subtropical regions, as well as in most regions of Iran. In this research, the genetic diversity of nine native and non-native genotypes of L. cylindrica was investigated through the evaluation of the chloroplast trnH-psbA intergenic region (IGS). After sampling the young leaves, DNA extraction was performed by using the Dellaporta method, and PCR was conducted by using IGS intergenic region primers. After sequencing of the amplified products, their quality was determined using Chromas software and then aligned using ClustalW method by BioEdit and MEGA7 softwares. Next, the dendrogram of phylogenic relationships was drawn and the matrix of the difference and similarity of the sequences were determined. In the present research, by analyzing the relationships between studied samples, based on the trnH-psbA (IGS) marker, a strong intraspecies variation was observed in native and non-native L. cylindrical genotypes. The genetic distance matrix between the samples examined in this research ranged from 0 to 6.865 with an overall average distance of 2.53. The average value of synonymous and non-synonymous substitutions (dN/dS) for the IGS sequence was ds/dn = 0.68, which indicates positive and pure line selections in the process of natural selection of studied genotypes. The results of this research showed that trnH-psbA is a suitable marker for evaluation of the intraspecific diversity of luffa species.
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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