ali akbar habashi

This study was aimed to eradicate three viruses, Apple chlorotic leaf spot virus (ACLSV),Apple stem grooving virus (ASGV) and Apple stem pitting virus (ASPV) from seven pear cultivars including Abate Fetel, Beiruti, Dargazi, Coscia, Louise Bonne, Mellina, Spadona. Experiments were performed by evaluation of the effectiveness of thermotherapy duration including 0, 7, 14 and 21 days at 38 °C and meristem culturing in size of less than 0.2 mm on virus eradication rates of explants. At first mother samples were tested for ACLSV, ASGV and ASPV by and RT-PCR and all samples were infected by all three viruses, except Abate Fetel and Beiruti samples, that were free of ASP virus. Thermotherapy and meristem culturing were performed in vitro</em>. Explants from thermotherapy and meristem culturing were tested by RT-PCR for all three viruses and the results showed that eradication rates of ACLSV, ASGV and ASPV, respectively 78.46, 26.63 and 35.5 percent were different in the different pear cultivars. The highest rate of virus eradication was related to Coscia and the lowest to Spadona cultivars. There was a direct relationship between increasing the duration of thermotherapy and virus elimination of explants, but this increased period of time up to 21 days reduced the growth and multiplication and even destroyed the explants. Therefore, 14 days of thermotherapy was the most effective treatment for elimination of ASGV, ASPV and ACLSV, respectively 61.4, 100 and 45.5 percent, from pear explants. At the end of the experiment, samples that were diagnosed virus free using RT-PCR, were proliferated, rooted and transferred to greenhouse condition for acclimation stage.

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.

This study was aimed to production of healthy primary pear plants for propagation and establishment of a high health maternal orchard in 2014 at Agriculture Biotechnology Research Institute of Iran (ABRII). The experiments were conducted by evaluate the effectiveness of the chemotherapy treatments (Ribavirin (0, 20, 40 and 80 mg/l) and the sizes of apical meristems culturing (less than 0.2 mm, between 0.2 to 0.7 mm and larger than 0.7 mm) on virus eradicate (ACLSV, ASPV, ASGV) from seven pear cultivars ("Abate Fetal", "Beiruti", "Coscia", "Dargazi", "Louise Bonne", "Mellina" and "Spadona"). At first maternal samples were tested for ACLSV, ASGV and ASPV by RT-PCR and all explants had been infected with all three viruses except Fettel and Beirut that were free from ASPV. We performed the chemotherapy treatments then meristems were cultured and grown in vitro. Regenerated shoots from meristem were tested by RT-PCR for all three viruses. The results showed that increasing the concentration of ribavirin and reducing the size of cultivated meristem was effective on all the three viruses’ elimination from explants. Also the responses of studied cultivars and virus varieties were different in treatments of this research. Samples that were diagnosed virus free by using RT-PCR, were proliferated, rooted and transferred into the pots to be used for later propagation and establishment of maternal orchard.

Promoter sequences are one of the key factors in directed gene expression. Different types of promoters can be used according to the type of tissue or expression levels desired. β-Conglycinin is one of the major storage proteins in soybean seed embryos and is produced in the seed development stage. The synthesis of storage proteins is primarily controlled at the transcriptional level and seed-specific expression has been shown to be conferred upon the promoter regions of many storage protein genes. In this research, a seed specific promoter (β-Conglycinin) of Iranian Glycin max was isolated from genomic DNA. Then, the promoter was cloned into pTZ57R/T and sequenced. Sequence analysis showed that the cloned promoter contained all of the typical conserved motifs such as TATA box, CAAT box as well as other previously identified seed-specific motifs such as SKn-1, RY repeats, G box. Finally, the β-Conglycinin promoter was cloned into pBI121 binary vector for further research.

Conventional methods of pear breeding, largely based on intra- and inter-specific hybridization, are difficult because pear is highly heterozygous, polygenic and has a long juvenile period. Genetic improvements of pear cultivars are possible through induction of mutations and gene transfer by genetic engineering. A general prerequisite for these approaches is to establish an efficient plant regeneration system. In the present study, the effect of two basal media (MS and NN) and different concentrations of TDZ (0, 2.5, 5, 7.5 μM) or BAP (0, 4, 8, 16 μM) in combination with NAA (1 μM) on direct shoot regeneration of two pear (Pyrus communis L.) genotypes 'Bartlett' and 'Dargazi' was investigated. The obtained results showed that 'Dargazi' had higher rates of shoot regeneration than 'Bartlett' and in both cultivars the highest percent of shoot regeneration was observed from lower sections of the leaves. Although the highest percent of shoot regeneration in 'Bartlett' (38%) was attained in the NN medium containing 2.5 µM TDZ and 1 µM NAA, the differences in shoot regeneration between this medium and the NN media containing 5 or 7.5 µM TDZ and 1 µM NAA were not significant. The highest percent of shoot regeneration in 'Dargazi' (56%) was obtained in NN medium containing 7.5 µm TDZ and 1 µm NAA. It can be concluded that genotypes, explant types and culture media composition could effect on direct shoot regeneration of pear.

Human granulocyte colony-stimulating factor (hG-CSF) can serve as valuable biopharmaceutical for research and treatment of the human blood cancer. Transplastomic plants have been emerged as a new and high potential candidate for production of recombinant biopharmaceutical proteins in comparison with transgenic plants due to extremely high level expression، biosafety and many other advantages. hG-CSF gene was cloned into pCL vector between prrn16S promoter and TpsbA terminator. The recombinant vector was coated on nanogold particles and transformed to lettuce chloroplasts through biolistic method. Callogenesis and regeneration of cotyledonary explants were obtained by Murashige and Skoog media containing 6-benzylaminopurine and 1-naphthaleneacetic acid hormones. The presence of hG-CSF gene in plastome was studied with four specific PCR primers and expression by Western immunoblotting. hG-CSF gene cloning was confirmed by digestion and sequencing. Transplastomic lettuce lines were regenerated and subjected to molecular analysis. The presence of hG-CSF in plastome was confirmed by PCR using specific primers designed from the plastid genome. Western immunoblotting of extracted protein from transplastomic plants showed a 20-kDa band، which verified the expression of recombinant protein in lettuce chloroplasts. This study is the first report that successfully express hG-CSF gene in lettuce chloroplast. The lettuce plastome can provide a cheap and safe expression platform for producing valuable biopharmaceuticals for research and treatment.