An optimized Agrobacterium-mediated transformation procedure for Phaseolus acutifolius A. Gray

• Published in: Plant cell reports Vol. 21; no. 4; pp. 333 - 340
• Main Authors: DE CLERCQ, J, ZAMBRE, M, VAN MONTAGU, M, DILLEN, W, ANGENON, G
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.11.2002; Springer Nature B.V
• Subjects: Agronomy. Soil science and plant productions; Bacteria; Biological and medical sciences; Biotechnology; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic engineering applications; Genetic technics; Genetics and breeding of economic plants; Methods. Procedures. Technologies; Plant breeding: fundamental aspects and methodology; Transgenic animals and transgenic plants; Transgenic plants; Callus; Temperature; Transformation; Plant pathogen; Agrobacterium tumefaciens; Environmental factor; Phaseolus; Optimization; Agrobacterium; Dicotyledones; Angiospermae; Bacteria; Transgenic plant; Developmental stage; Intraspecific comparison; Tepary bean; Stability; Genetic transformation; Treatment efficiency; Photoperiod; Phaseolus acutifolius; Experimental protocol; Infection; Pathogenicity; Grain legume; Opine; Leguminosae; Phenols; Spermatophyta; Rhizobiaceae; Inheritance(genetics)
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-002-0518-0

To improve Agrobacterium tumefaciens-mediated transformation of Phaseolus acutifolius, we examined the effect of different factors on T-DNA transfer by measuring transient expression levels of an intron-containing β-glucuronidase gene. Improved transformation frequencies were obtained with an A. tumefaciens strain carrying nopaline-type virulence genes and when calli were infected with Agrobacterium cells in the early-log growth phase. Optimized co-cultivation was performed at 22°C under a 16/8-h (day/night) photoperiod in an acidic medium (pH 5.5) in the presence of 200 µM acetosyringone. By combining the best treatments, an efficient and reproducible transformation procedure was established for the P. acutifolius genotype NI576. Southern and immunoblot analyses confirmed the stable integration and expression of the transgenes in the primary transgenic plants and their progeny.[PUBLICATION ABSTRACT]