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Showing 5 results for Amani

Mahdieh Sahshorpour, Jafar Amani, Mahyat Jafari, Ali Hatef Salmanian,
Volume 1, Issue 1 (5-2014)

One of the important pathogens which cause hemorrhagic colitis and hemolytic uremic syndrome in humans is enterohemorrhagic Escherichia coli (EHEC) O157:H7. Cattle are the most important reservoir of this bacterium. EspA and Intimin are two protein factors for bacteria colonization on intestinal epithelium and cause attaching/effacing lesion. The LEE pathogenicity islands code these proteins. EspA is part of type III secretion systems which delivers Tir to the host cell and integrate to membrane. Intimin encoded by eae gene and fused to Tir. In this research we supposed that chimeric recombinant form of EI gene containing EspA and Intimin were fused with a linker as an edible candidate vaccine would reduce colonization of E. coli O157:H7 in animal model. We constructed a synthetic gene EspA (E120) and intimin (Int282) fused by (EAAAK)4 sequence. The synthetic gene (EI) was codon optimized and subcloned into plant expression vector (PBI121) under CaMV35S promoter and then transferred to tobacco plant by agrobacterium mediated protocol. The presence of inserted gene in plant genome was documented by PCR and RT-PCR methods. The amount of EI protein in transgenic tobacco leaves were estimated 0.1% of the total soluble protein (TSP) by ELISA method.
Shahin Jahangirzadeh Khiavi, Zabihollah Zamani, Mohamadreza Fatahi, Masoumeh Ashourpour,
Volume 5, Issue 1 (9-2018)

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.

Negin Eslahi, Mojegan Kowsari, Mostafa Motallebi#, Mohammad Reza Zamani, Sepideh Akbari,
Volume 6, Issue 1 (9-2019)

The transition from the vegetative phase to reproductive phase is the most important event in production and genetic innovation. This phenomenon is influenced by many genetic and environmental factors in plants. According to studies carried out in this field, one of the environmental factors affects the reproductive and flowering process is Trichoderma species, which is abundant in soil. This study was carried out to evaluate the ability of two recombinant Trichoderma harzianum strains containing chimeric chit 42 (with CHBD domain) and wild-type strain to promote common bean flowering and yield increase in vivo condition. To do this, flowering parameters such as a number of flowers, flowering time and effective parameters in yield were evaluated. Also, expression level of some flowering-related genes such as FT and SOC1 were measured using real-time PCR. The results showed that the bean plants treated with recombinant strains had a significantly increased number of flowers and earlier flowering compared to the control and wild type Trichoderma. Also, plants treated with recombinant strains showed a significant difference in the number and weight of the pod compared to the plant treated with wild type strain and non-treated plants. In addition, the plants treated with T13 strain showed more expression levels of the FT and SOC1 genes (with ratio of 3.42 and 3.41 fold respectively) compared to other treatments and control plant. Finally, T13 recombinant strain exhibited a better performance compared to the other strains through a positive effect on flowering and then increased the crop yield.

Raheleh Aziznia, Hedieh Badakhshan, Taimoor Javadi, Soma Zamani,
Volume 6, Issue 2 (3-2020)

In this study, variation of beta-glucan content was assessed in 20 barley line and cultivars based on complete block design with three replications. Genetic diversity of these genotypes was also evaluated using ISSR markers. Beta-glucan extracted by an enzymatic method. Significant differences were found at the level of 1% among barley genotypes for beta-glucan content. The beta glucan content was variable from 7.21 to 12.48 and, the Yosef, E94B3 and E94B17 genotypes hold the highest content of the beta-glucan. ISSR primers with average polymorphism of 66.79%, genetic diversity of 0.25 and Shannon index of 0.37 were determined as efficient markers for studying genetic diversity. The barley lines and cultivars were assigned in two distinct groups according to their genetic pedigree. On the basis of non-parametric Kruskal-Wallis, Spearman correlation, and stepwise regression analysis, nine informative primers were detected explaining highest seed’s beta-glucan content variations ranging from 24.3 to 42.4 percent. The ISSR6 (700), the combination of ISSR1+ISSR4 (1400) and IS2+ ISSR2 (1400) primers were the most informative primers for the beta-glucan content. The informative markers provide possible functional and efficient marker based selection method and, screening the barley germplasms for the highest beta-glucan content.

Sahar Dashchi, Hassan Rahnama, Kianoosh Cheghamirza, Katayun Zamani,
Volume 7, Issue 2 (3-2021)

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.

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