Unidad de Biología Integrativa, Centro de Investigación Científica de Yucatán, Mérida, Yucatán, Mexico.
Methods Mol Biol. 2024;2827:435-443. doi: 10.1007/978-1-0716-3954-2_29.
Protoplasts are plant cells from which the pectocellulosic cell wall has been removed, thus keeping the plasma membrane intact. For plant secondary metabolites research, this system is a powerful tool to study the metabolites' dynamics inside the cells, such as the subcellular localization of proteins, characterization of gene function, transcription factors involved in metabolite pathways, protein transport machinery, and to perform single-cell omics studies. Due to its lack of a cell wall, better images of the interior of the cell can be obtained compared to the whole tissue. This allows the identification of specific cell types involved in the accumulation of specialized metabolites, such as alkaloids, given their autofluorescence properties. Here is a simplified protocol to obtain protoplasts from leaves and in vitro cell cultures from Argemone mexicana, which produces the pharmacologically important alkaloids berberine and sanguinarine.
原生质体是去除了果胶纤维素细胞壁的植物细胞，从而保持质膜完整。对于植物次生代谢物的研究，该系统是研究细胞内代谢物动态的有力工具，例如蛋白质的亚细胞定位、基因功能的表征、参与代谢物途径的转录因子、蛋白质转运机制，并进行单细胞组学研究。由于缺乏细胞壁，可以获得比整个组织更好的细胞内部图像。这允许鉴定参与特殊代谢物积累的特定细胞类型，例如生物碱，因为它们具有自发荧光特性。这里是一个简化的方案，用于从墨西哥鸦胆子的叶片和体外细胞培养物中获得原生质体，该植物产生具有药理学重要性的生物碱小檗碱和血根碱。
