Strategic Innovation Platform, International Rice Research Institute, Manila, Philippines.
Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan.
Methods Mol Biol. 2021;2238:313-324. doi: 10.1007/978-1-0716-1068-8_21.
Protoplasts are a versatile and powerful cell-based system to study different plant processes in vivo, due to their ability to maintain cell identity and carry out reactions and metabolic processes similar to intact plants. In rice, despite numerous reports, difficulties are encountered in protoplast isolation and transfection. These include insufficient numbers of protoplasts isolated and inefficient transfection. Such difficulties limit the use of this simple yet useful technology. The need to use protoplasts is particularly important when similar experiments may not work in yeast or Pichia, due to differences in functionally essential protein post-translation modifications. In this chapter, we describe a rice protoplast isolation and transfection method.
原生质体是一种通用且强大的基于细胞的系统，可用于研究体内不同的植物过程，因为它们能够维持细胞特性，并进行类似于完整植物的反应和代谢过程。在水稻中，尽管有许多报道，但在原生质体分离和转染方面仍存在困难。这些困难包括分离的原生质体数量不足和转染效率低下。这些困难限制了这项简单而有用的技术的应用。当在酵母或毕赤酵母中进行类似的实验可能无法进行时，使用原生质体的需求尤为重要，因为功能必需的蛋白质翻译后修饰存在差异。在本章中，我们描述了一种水稻原生质体分离和转染方法。
