Transformation of Arabidopsis thaliana with Agrobacterium tumefaciens

• Published in: Science (American Association for the Advancement of Science) Vol. 234; no. 4775; pp. 464 - 466
• Main Authors: Lloyd, A.M, Barnason, A.R, Rogers, S.G, Byrne, M.C, Fraley, R.T, Horsch, R.B
• Format: Journal Article
• Language: English
• Published: Washington, DC The American Association for the Advancement of Science 24.10.1986; American Association for the Advancement of Science
• Subjects: AGROBACTERIUM; Agrobacterium tumefaciens; Agronomy. Soil science and plant productions; Arabidopsis thaliana; Biochemistry; Biological and medical sciences; Biotechnology; Callus; CRUCIFERAE; DNA; Flowers & plants; Fundamental and applied biological sciences. Psychology; Gap junctions; Genetic engineering; Genetic engineering applications; Genetic technics; Genetics; Genetics and breeding of economic plants; Genomes; Inoculation; Methods. Procedures. Technologies; NUCLEOTIDE; NUCLEOTIDES; NUCLEOTIDOS; Plant breeding: fundamental aspects and methodology; Plants; Plasmids; Seedlings; transformation; Transgenic plants; Vectors (cloning, transfer, expression). Insertion sequences and transposons; Vegetals; Genetic transformation; Agrobacterium tumefaciens; Genetic transfer; Arabidopsis thaliana; Cruciferae; Ti Plasmid; Dicotyledones; Angiospermae; Bacteria; Spermatophyta; Rhizobiaceae; Experimental plant; Hybrid gene
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.234.4775.464

Transformed Arabidopsis thaliana plants have been produced by a modified leaf disk transformation-regeneration method. Leaf pieces from sterilely grown plants were precultured for 2 days and inoculated with an Agrobacterium tumefaciens strain containing an avirulent Ti (tumor-inducing) plasmid with a chimeric gene encoding hygromycin resistance. After cocultivation for 2 days, the leaf pieces were placed on a medium that selects for hygromycin resistance. Shoots regenerated within 3 months and were excised, rooted, and transferred to soil. Transformation was confirmed by opine production, hygromycin resistance, and DNA blot hybridization of both primary transformants and progeny. This process for producing transgenic Arabidopsis plants should enhance the usefulness of the species for experimental biology.