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Molecules 2015, 20(3), 3647-3666; doi:10.3390/molecules20033647

An Efficient Agrobacterium-Mediated Transformation of Strawberry cv. Camarosa by a Dual Plasmid System

1
Department of Agriculture Technology, Faculty of Agriculture, Universiti Putra Malaysia, Serdang, Selangor 43400, Malaysia
2
Laboratory of Plantation Crops, Institute of Tropical Agriculture, Universiti Putra Malaysia, Serdang, Selangor 43400, Malaysia
3
Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor 43400, Malaysia
4
Department of Biology, Faculty of Sciences, University of Zabol, Zabol 9861335856, Iran
*
Authors to whom correspondence should be addressed.
Academic Editor: Derek J. McPhee
Received: 24 October 2014 / Accepted: 25 November 2014 / Published: 23 February 2015
(This article belongs to the Section Natural Products)
View Full-Text   |   Download PDF [3433 KB, uploaded 23 February 2015]   |  

Abstract

An Agrobacterium-mediated transformation method was applied to introduce the luciferase reporter gene under the control of the CaMV35S promoter in the pGreen0049 binary vector into strawberry cv. Camarosa. The in vitro regeneration system of strawberry leaves to be used in the transformation was optimized using different TDZ concentrations in MS medium. TDZ at 16 µM showed the highest percentage (100%) of shoot formation and the highest mean number of shoots (24) produced per explant. Studies on the effects of different antibiotics, namely timentin, cefotaxime, carbenicillin and ampicillin, on shoot regeneration of strawberry leaf explants showed the best shoot regeneration in the presence of 300 mg/L timentin and 150 mg/L cefotaxime. Assessment of the different factors affecting Agrobacterium mediated-transformation of strawberry with the luciferase gene showed the highest efficiency of putative transformant production (86%) in the treatment with no preculture, bacterial OD600 of 0.6 and the addition of 150 mg/L cefotaxime in the pre-selection and selection media. The presence of the luciferase gene in the plant genome was verified by the luciferase reporter gene assay, nested PCR amplification and dot blot of genomic DNA isolated from the young leaves of each putatively transformed plantlet. View Full-Text
Keywords: stable transformation; Agrobacterium; strawberry camarosa; pGreen0049 binary vector; thidiazuron; timentin; cefotaxime stable transformation; Agrobacterium; strawberry camarosa; pGreen0049 binary vector; thidiazuron; timentin; cefotaxime
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Haddadi, F.; Abd Aziz, M.; Abdullah, S.N.A.; Tan, S.G.; Kamaladini, H. An Efficient Agrobacterium-Mediated Transformation of Strawberry cv. Camarosa by a Dual Plasmid System. Molecules 2015, 20, 3647-3666.

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