Department of Botany, University of Georgia, 30602, Athens, GA, USA.
Planta. 1980 Jan;149(4):389-401. doi: 10.1007/BF00571175.
Protoplasts were prepared from the guard cells ofA. cepa. Epidermal peels taken from expanding green leaves and largely free of mesophyll were treated with Cellulysin, and protoplasts were harvested after 18 h of digestion. That the protoplasts were derived from guard cells was ascertained from their characteristic vacuolar autofluorescence and from observations showing that all other epidermal cells are killed in the peeling procedure. The protoplasts proved to be a good system with which to view the cell cortex and inner surface of the plasmalemma. The lysis of cells adhering to polylysine-treated, Formvar-coated grids, followed by negative staining in uranyl acetate, showed that many microtubules normally present in ordered arrays in situ remain closely applied to the inner surface of the plasmalemma in protoplasts. In addition, numerous vesiculate elements including coated vesicles and/or pits are present amongst the microtubules. Similar vesicles are evident in thin sections of fixed, embedded guard cells and protoplasts. The significance of these structures in the cell cortex is discussed.
从洋葱的保卫细胞中制备原生质体。从展开的绿叶中提取的表皮薄片，基本上不含叶绿体，用纤维素酶处理，消化 18 小时后收获原生质体。从原生质体特有的液泡自发荧光和观察结果可以确定，在去皮过程中所有其他表皮细胞都被杀死。原生质体被证明是一个很好的系统，可以观察细胞皮层和质膜的内表面。用多聚赖氨酸处理的 Formvar 涂层网格上的细胞裂解，然后用醋酸铀负染色，表明许多在原位以有序排列存在的微管在质膜的内表面紧密附着。此外，在微管之间存在许多囊泡状元件，包括有被囊泡和/或陷窝。在固定、包埋的保卫细胞和原生质体的薄片中也可以看到类似的囊泡。本文讨论了这些结构在细胞皮层中的意义。
