Research Progress on Preparation, Transformation and Application of Protoplasts Derived from Medicinal Plants

As unicellular systems, protoplasts derived from medicinal plant cells exhibit high totipotency and hold significant value in applications such as gene function analysis, genetic improvement, and cell engineering optimization. This review focuses on how protoplast technology addresses longstanding bottlenecks in medicinal plant research by systematically collating recent advances in the study of medicinal plant protoplasts. It explores the multifaceted factors influencing protoplast preparation, including the intrinsic and extrinsic properties of medicinal plant materials, pretreatment methods prior to enzymatic hydrolysis, the composition of enzymatic solutions, enzymatic hydrolysis parameters, external environmental conditions, and protoplast purification techniques. Additionally, the review summarizes the significance and value of medicinal plant protoplasts in gene function verification, gene editing, genetic transformation, single-cell sequencing, and cell fusion regulation. By comprehensively synthesizing the optimization of the preparation of medicinal plant protoplasts and their application in transient expression, gene function research, and plant regeneration, this work aims to provide critical guidance for subsequent research in genetic modification, germplasm resource breeding, spatiotemporal programming of active substances, and regulatory network analysis. Ultimately, it serves as a valuable reference for advancing research in the plant sciences.