This second edition volume expands on the previous edition with updated chapters in model systems and new crops. The book contains protocols for plastid engineering in leaves, tissue culture cells, and the shoot apex of plants, as well as for marker excision from the plastid genome and engineering Rubisco, the key enzyme of photosynthesis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and practical, Chloroplast Biotechnology: Methods and Protocols, Second Edition is a valuable resource for researchers who wish to enter the field, and for practitioners looking for insights of applications in agriculture, industrial biotechnology and healthcare.