Plant cell and tissue culture comprises a broad range of techniques of great value to research workers in the fields of cell and molecular biology, physiology, biochemistry, plant breeding and propagation and genetic engineering. This manual provides protocols for the major techniques in such a format that they can be followed step-by-step at the bench. Both applied and more fundamental uses of cell, tissue and organ culture are covered, and the ring-binder design allows the manual to be updated regularly with supplementary chapters. Each chapter, discussing a single technique or protocol, provides background information, references to the relevant literature and a guide to troubleshooting in addition to a thorough methodology. This second Supplement to the core text of the "Manual" provides important techniques to be added to the Sections on "Basic Techniques, Tissue Culture & Transformation of Crop Species and Reproductive Tissues".