toxicity of antibiotics and herbicides on in vitro adventitious shoot formation on Pinus pinea L. cotyledons

• Published in: In vitro cellular & developmental biology. Plant 1999-07, Vol.35 (4), p.339-343
• Main Authors: Humara, J.M, Ordas, R.J
• Format: Article
• Language: English
• Subjects: adventitious shoots; Agrobacterium; Agrobacterium tumefaciens; Antibiotics; Biological and medical sciences; Biotechnology; Biotechnology/Genetic Transformation/Somatic Cell Genetics; cefotaxime; Cotyledons; Embryos; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; glufosinate; growth; Herbicides; In vitro propagation: entire plant regeneration from tissues and cell cultures; in vitro selection; kanamycin; Methods. Procedures. Technologies; phytotoxicity; Pinus pinea; Plant cells; Plant cells and fungal cells; plant development; Plant tissues; Plants; Regeneration; regenerative ability; ticarcillin; Transgenic plants; vancomycin
• ISSN: 1054-5476; 1475-2689
• DOI: 10.1007/s11627-999-0045-6

The toxicity of three antibiotics commonly used to eliminate Agrobacterium tumefaciens from plant tissue during transformation were tested to determine their effect on Pinus pinea L. morphogenesis. Cefotaxime and vancomycin at a dose of 250 μ g ml-1, as well as ticarcillin at$300 \mu g ml^{-1}$, were essentially nontoxic to the culture and significantly enhanced regeneration and shoot development. Kanamycin, a widely used selection agent for plant transformation, strongly inhibited regeneration even at very low doses. On the contrary, phosphinothricin and its ammonium salt glufosinate in the commercial formulation[REGISTERED SIGN] Finale proved to be the best selection agents because they were less toxic to stone pine cotyledons. Thus, they could be applied immediately after transformation to avoid escapes. Other schemes for selection and recovery of transgenic stone pine trees are discussed.