Optimizing tissue culture and GUS gene transformation to shallots using Agrobacterium
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Abolfazl Hajiheidar , Masoud Tohidfar * , Mehdi Miri |
Shahid Beheshti University |
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Abstract: (5847 Views) |
Shallot (Allium stipitatum) is an edible vegetable that has important pharmacological properties. Therefore, optimization of an efficient in vitro regeneration and transformation system for shallot breeding through genetic engineering would be useful. Embryo, root and bulb explants of shallot were cultured in vitro on MS basal medium supplemented with different concentrations of growth regulators (NAA, 2,4-D and BA) for callus induction and plantlet regeneration. Explants were transformed using Agrobacterium tumefaciens strain LBA4404 and pBI121 plasmid carrying the gus reporter gene. The results indicated that bulb had a higher efficiency of callus induction and plantlet regeneration (100%) compared to embryo and root explants. The highest percentage of regeneration (100 %) was observed on MS medium supplemented with 5 mg.l-1 BA and 1 mg.l-1 NAA for bulb explants. Polymerase chain reaction (PCR) analysis of gus-positive transformants confirmed genetic transformation of the cultures. Furthermore, the lack of Agrobacterium-related infection was confirmed using virG-specific markers. In this study, the efficiency of transformation was 10 and 6.6% in embryo and bulb, respectively. |
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Keywords: Agrobacterium, GUS gene, Gene transformation, Shallot, Tissue culture |
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Full-Text [PDF 282 kb]
(3290 Downloads)
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Type of Study: Research |
Subject:
Animal Received: 2016/07/24 | Accepted: 2016/09/9 | Published: 2017/01/25
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