E-Book Overview
Agrobacterium Protocols offers beginning and experienced researchers the most comprehensive collection of step-by-step protocols for the genetic manipulation of plants using Agrobacterium. The topics range from the maintenance of bacterial culture collections to aspects of the metabolism and physiology of transformed tissues and transgenic plants. Drawing on the work of leading scientists from laboratories around the world, Agrobacterium Protocols provides a wealth of techniques for introducing specific DNA sequences into target plant species and discusses the environmental implications of genetically engineered plants. Its detailed procedures will facilitate rapid transfer of advanced techniques to other laboratories and their exploitation in fundamental and applied plant biology.
E-Book Content
CHAPTER1
Growth
and Storage Frauli$e
of Agrobacterium
M. van Asma
1. Introduction Proper preservation of strains is of great importance to all microbiologists. Working with contaminated or genetically changed cultures, or facing the loss of a strain are annoyances that often may be easily prevented. A reliable preservation method should fulfill the following criteria. First, the risk of introducing contaminants must be minimized. A method that invokes frequent subculturing is therefore considered less convenient. Such a method would also increase the risk of errors, including mislabeling and enrichment for mutants. Second, the cultures should be genetically stable during storage. Finally, loss of viability during preparation, storage, and reviving of the culture should be as low as possible. This guarantees a long shelf life and reduces the risk of selection for bacteria that are able to resist the exposure to stress conditions that occur during either processing or storage of cultures better. Genetic stability and viability levels durin