A combination of overgrowth-control antibiotics improves Agrobacterium tumefaciens-mediated transformation efficiency for cultivated tomato (L. esculentum)

Authors: Hu, Wei; Phillips, Gregory

Source: In Vitro Cellular and Development Biology - Plant, Volume 37, Number 1, January 2001 , pp. 12-18(7)

Publisher: Springer

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Abstract:

The transformation efficiency of cultivated tomato (Lycopersicon esculentum cv. UC82) using Agrobacterium tumefaciens was improved from 14% in a previous report to 25% in the present study. Several variables potentially involved in the improvement of transformation efficiency were evaluated, including enhancements in the regeneration system, antibiotics used for Agrobacterium-overgrowth control, and method of applying kanamycin for selection. The most important variable identified was the influence of overgrowth-control antibiotics on both the regeneration response and transformation efficiency. The best transformant recovery and Agrobacterium-overgrowth control was obtained using 250 mg l⁻¹ claforan and 250 mg l⁻¹ ticareillin as the overgrowth-control antibiotics in the media. Selfed T₁ progeny plants showed Mendelian inheritance ratios in 77% of the independently transformed lines according to phenotype expression [β-glucuronidase (GUS) assay results], and confirmed by polymerase chain reaction amplification of the transgene in progeny.

Keywords: claforan; ticarcillin; vancomycin; transgene inheritance test

Document Type: Research article

DOI: http://dx.doi.org/10.1007/s11627-001-0003-4

Affiliations: 1: Email: grphilli@nmsu.edu

Publication date: 2001-01-01