Journal
PLANT CELL TISSUE AND ORGAN CULTURE
Volume 76, Issue 3, Pages 269-275Publisher
SPRINGER
DOI: 10.1023/B:TICU.0000009249.14051.77
Keywords
acetosyringone; genetic transformation; Lycopersicon esculentum; tissue culture; transgenic plants; zeatin riboside
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Tomato transformation and regeneration were analysed and optimized. Cotyledon explants from Lycopersicon esculentum cv. UC82B, were infected by Agrobacterium tumefaciens strain LBA4404 harbouring the neomycin phosphotransferase (NPTII) reporter gene. The effects of phenolic compounds, vitamins and growth regulators on plant transformation and regeneration were studied. Increasing the vitamin thiamine concentration from 0.1 mg l(-1) in standard medium to 0.4 mg l(-1) decreased the chlorophyll lost that accompanied the expansion of necrotic areas in cotyledon explants. Optimal shoot regeneration rate was obtained with a balanced concentration of 0.5 mg l(-1) auxin indolelacetic acid (IAA) and 0.5 mg l(-1) cytokinin zeatin riboside. Finally, when the phenolic acetosyringone was present in the co-culture medium at 200 muM, confirmed transgenic lines reached 50% of antibiotic resistant shoots. Under the above conditions, the transformation efficiency reached 12.5%.
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