Agrobacterium Protocols (2^nd Ed., Softcover reprint of hardcover 2nd ed. 2006)
Volume I
Methods in Molecular Biology Series, Vol. 343

Agrobacterium tumefaciens is a soil bacterium that for more than a century has been known as a pathogen causing the plant crown gall disease. Unlike many other pathogens, Agrobacterium has the ability to deliver DNA to plant cells and permanently alter the plant genome. The discovery of this unique feature 30 years ago has provided plant scientists with a powerful tool to genetically transform plants for both basic research purposes and for agricultural development. Compared to physical transformation methods such as particle bomba- ment or electroporation, Agrobacterium-mediated DNA delivery has a number of advantages. One of the features is its propensity to generate a single or a low copy number of integrated transgenes with defined ends. Integration of a single transgene copy into the plant genome is less likely to trigger ?gene silencing? often associated with multiple gene insertions. When the first edition of Agrobacterium Protocols was published in 1995, only a handful of plants could be routinely transformed using Agrobacterium. Agrobacterium-mediated transformation is now commonly used to introduce DNA into many plant species, including monocotyledon crop species that were previously considered non-hosts for Agrobacterium. Most remarkable are recent developments indicating that Agrobacterium can also be used to deliver DNA to non-plant species including bacteria, fungi, and even mammalian cells.

AGROBACTERIUM HANDLING.- Culture and Maintenance of Agrobacterium Strains.- Binary Vectors and Super-binary Vectors.- Three Methods for the Introduction of Foreign DNA into Agrobacterium.- Integration of Genes into the Chromosome of Agrobacterium tumefaciens C58.- Nucleic Acid Extraction from Agrobacterium Strains.- Agrobacterium Virulence Gene Induction.- Model Plants.- Arabidopsis thaliana Floral Dip Transformation Method.- Agrobacterium Transformation of Arabidopsis thaliana Roots.- Medicago truncatula Transformation Using Leaf Explants.- Medicago truncatula Transformation Using Cotyledon Explants.- Medicago truncatula Transformation Using Root Explants.- Nicotiana (Nicotiana tobaccum, Nicotiana benthamiana).- Generation of Composite Plants Using Agrobacterium rhizogenes.- Cereal Crops.- Barley (Hordeum vulgare L.).- Maize (Zea mays L.).- Indica Rice (Oryza sativa, BR29 and IR64).- Japonica Rice Varieties (Oryza sativa, Nipponbare, and Others).- Rye (Secale cereale L.).- Sorghum (Sorghum bicolor L.).- Wheat (Triticum aestivum L.).- Industrial Plants.- Canola (Brassica napus L.).- Cotton (Gossypium hirsutum L.).- Indian Mustard [Brassica juncea (L.) Czern.].- Sunflower (Helianthus annuus L.).- Legume Plants.- Alfalfa (Medicago sativa L.).- Chickpea (Cicer arietinum L.).- Clovers (Trifolium spp.).- Peas (Pisum sativum L.).- Peanut (Arachis hypogaea L.).- Pigeonpea (Cajanus cajn (L.) Millsp.).- Red Clover (Trifolium pratense).- Soybean (Glycine max) Transformation Using Mature Cotyledonary Node Explants.- Soybean (Glycine max) Transformation Using Immature Cotyledon Explants.- Tepary Bean (Phaseolus acutifolius).- Vegetable Plants.- Brassica oleracea.- Cucumber (Cucumis sativus L.).- Eggplant (Solanum melongena L.).- Lettuce (Lactuca sativa L.).- Tomato (Lycopersicum esculentum).

Includes supplementary material: sn.pub/extras