An efficient protocol for sugarcane (Saccharum spp. L.) transformation mediated by Agrobacterium tumefaciens |
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Authors: | Arencibia Ariel D. Carmona Elva R. Tellez Pilar Chan Ming-Tsair Yu Su-May Trujillo Luis E. Oramas Pedro |
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Affiliation: | (1) Centre for Genetic Engineering and Biotechnology, PO Box 6162, Cuba;(2) Institute of BiAgriculture Science, Academia Sinica Nankang, Taipei, Taiwan |
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Abstract: | This is the first successful report of the recovery of morphologically normal transgenic sugarcane plants from co-cultivation of calluses with Agrobacterium tumefaciens. Transformation frequencies (total of transgenic plants/number of cell clusters) were between 9.4 × 10–3 and 1.15 × 10–2. In our experiments, both LBA4404 (pTOK233) and EHA101 (pMTCA3IG), carrying a super-binary vector or supervirulent strain, respectively, were successful for sugarcane transformation. We found that three main factors: (1) the use of young regenerable calluses as target explants; (2) induction and/or improvement of the A. tumefaciens virulence system with sugarcane cell cultures and (3) pre-induction of organogenesis or somatic-embryogenesis-like sexual embryos, seem to be crucial in order to increase the cells competence for T-DNA transfer process. Patterns generated by Southern hybridization confirmed that T-DNAs were randomly integrated into sugarcane genome without th e persistence of A. tumefaciens in the transgenic plants |
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Keywords: | Saccharum Agrobacterium transformation |
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