Grapevine is among the most important economic plants that are susceptible to phytoplasma disease. Tools and procedures for genetic manipulation of grapevine are cumbersome and sometimes difficult to replicate; therefore, most of the research on plant–phytoplasma interactions focuses on species with more established transformation procedures instead, even though they might not be hosts of the studied phytoplasma. Studying the mechanisms of plant–phytoplasma interactions directly in the host plants would bring new and more accurate insights into the disease mechanisms. In this chapter, we present the protocols to obtain transformation-competent grapevine material from different tissues and the procedures for protoplast isolation and cultivation.