A collection of the most frequently used techniques for plant cell and tissue culture. The readily reproducible methods range from general methodologies, such as culture induction, growth and viability evaluation, and contamination control, to such highly specialized techniques as chloroplast transformation involving the laborious process of protoplast isolation and culture. Most of the protocols are currently used in the research programs of the authors or represent important parts of business projects aimed at the generation of improved plant materials. Two new appendices explain the principles for formulating culture media and the composition of the eight most commonly used media formulations, and list more than 100 very useful internet sites.
A collection of readily reproducible techniques for the genetic transformation and regeneration of plants. The authors cover the most commonly used transformation systems for the regeneration of whole transgenic plants. Among the techniques discussed for the detection of transgenes are quantitative real-time PCR, RT-PCR, in situ hybridization, FISH, and TAIL-PCR. Risk assessment methods offer the opportunity to study Agrobacterium persistence in plant tissues and to investigate the possibility of transgene dispersal. Each protocol includes step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls
