Molecular biology and (trans) gene expression technology in plant breeding
Meysam Bastami; Hosseini Ramin
Abstract
The regulatory sequence of rice alpha amylase 3D gene (Ramy3D) is amongst the most successful expression systems used for recombinant protein expression in plants. In the current study a 995 bp fragment consisting of Ramy3D promoter and its 5′ untranslated region was amplified from the genomic ...
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The regulatory sequence of rice alpha amylase 3D gene (Ramy3D) is amongst the most successful expression systems used for recombinant protein expression in plants. In the current study a 995 bp fragment consisting of Ramy3D promoter and its 5′ untranslated region was amplified from the genomic DNA of an Iranian rice cultivar ″Nemat″, using polymerase chain reaction. The amplified fragment was ligated into the pTG19-T vector and the cloned fragment was sequenced. For in silico characterization, the rice specific consensus sequences of TATA-box and YR Rule motifs were scanned against the cloned fragment sequence using FIMO program and the cis acting elements existing in the promoter region were investigated using PlantCare database. A TATA-box motif with the rice specific pattern was identified at upstream position of the transcription start site. The identification of TATA-box in Ramy3D promoter is consistent with its metabolic and tissue specific regulation manner. Several cis regulatory motifs responsible for the metabolic and hormonal regulation of Ramy3D gene were identified including ABRE, G-Box, GC-box, GATA motif and TATCCA T/C motif. In addition, several motifs involved in response to various stimuli such as plant hormones, light and biotic and abiotic stresses were identified which include circadian motif, as-2-box, WUN-motif, TGACG-motif, Skn-1 motif, O2-site, MBS, LAMP-element, I-box, HSE, GCC Box, GATT motif, CGTCA-motif and GAG-motif.
Plant molecular physiology & breeding
Ghasemali Garoosi; Moharram-ali Delijam; Esmaeil Nezami-Alanagh; Ramin Hosseini
Abstract
A complete micropropagation protocol was developed for Pistacia vera cv. Ghazvini, an important rootstock in Pistachio orchards in Iran. In present study, the efficiency of a new medium called GNH (Garoosi, Nezami and Haddad) was investigated against some standard media. Different vitamins, calcium (Ca) ...
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A complete micropropagation protocol was developed for Pistacia vera cv. Ghazvini, an important rootstock in Pistachio orchards in Iran. In present study, the efficiency of a new medium called GNH (Garoosi, Nezami and Haddad) was investigated against some standard media. Different vitamins, calcium (Ca) sources and plant growth regulators (PGRs) were tested on in vitro shoot multiplication and root induction. Results indicated that mean number of shoots (4.25 ± 0.25), and productivity (69.87 ± 9.19 mm) increased significantly when GNH medium formed the basal medium, compared to the standard media including Murashige and Skoog medium (MS), Juglans Medium (DKW), and McCown Woody Plant Medium (WPM). The most suitable concentration Ca and vitamin sources for shoot multiplication were 3.0 mM Ca gluconate and DKW-vitamins. The most suitable PGRs were a combination of 0.5 or 1.0 mg l-1 BAP and 0.1 mg l-1 IBA. The highest rooting parameters were obtained when 3.0 mM Ca gluconate or Fe-EDDHA (with 0.2 mM Fe) were incorporated into the GNH medium containing 2.0 mg l-1 α-naphthalenacetic acid (NAA). Finally, nearly 70% of the plantlets survived acclimatization in the greenhouse. The results suggested the GNH medium (supplemented with Ca gluconate and DKW-vitamins), as a considerable and specific medium for the rapid micropropagation of Pistacia vera cv. Ghazvini.