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Optimization of callus induction and Agro-transformation on some Iranian Rice cultivars
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Afsaneh Ebrahimi   , Ebrahim Dorani * , Elaheh Bina , Behzad Ghareyazi |
| Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Tabriz, Iran , uliaie@tabrizu.ac.ir |
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Abstract: (3915 Views) |
| Improving new high performance cultivars, with better quality and resistance to biotic and abiotic stresses are important objectives of rice (oryza sativa L.) breeding through genetic engineering. This study, have been done to optimization of callus induction and, transformation protocol for some cultivar of Iranian Rice. In first experiment, effect of plant growth regulators (NAA, 2,4-D, Kin), proline (200, 400 and 600 mg/L) and Rice genotypes (including Moosa tarom, Nemat, Dom sorkh, Garib, Ali kazemi and Derafk) in callus induction, bacterial concentrations (OD600 = 0.2, 0.6 and 1) and strain types (LBA4404, EHA105, AGL1) on selected cultivar transformation were evaluated. The results were analyzed in a factorial experiment based on a completely randomized design. Analyses of variance were showed significant difference between plant growth regulator and genotype in response to callus induction. Almost all of the cultivars induced callus up to 80% in majority of PGRs combinations. HasanSaraei, had the higher callus growth rate (over 0.49 g) in the medium supplemented with 2 mg/l 2,4-D, and DomSorkh and Garib had the highest callus fresh weight (above 0.4 g) in media containing 2 mg/l 2,4-D plus 2 mg/l NAA. Using prolin in callus induction medium decreased callus growth. Lowest callus biomass (0.138 g) produced in medium containing 600 mg/L prolin. Among the three evaluated strain of Agrobacterium tumefaciens, only the EHA105 produced transgenic calli. Analysis of variance showed there was no significant difference between bacterial OD600 on transformation efficiency, but OD600=1 produced more transgenic calli. According to the results, the DomSorkh cultivar is more capable of callus formation in 2,4-D medium and can be used for gene transfer for different purposes. |
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| Keywords: Prolin, 2, 4-D, Callus induction, Transformation, DomSorkh, EHA105 |
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Full-Text [PDF 811 kb]
(1585 Downloads)
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Type of Study: Research |
Subject:
Plant
Received: 2019/06/9 | Accepted: 2019/10/3 | Published: 2019/10/8
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