Studies of surfactants by employing the protoplast system.

本研究旨在检测原生质体系统是否适用于检测不同表面活性剂对活细胞的影响。用4种非离子表面活性剂和2种阴离子表面活性剂对黄瓜炭疽菌(Colletotrichum Lagenarium)菌丝和叶片原生质体进行处理。根据非离子表面活性剂的作用效果，将其分为2组，其中2组对黄瓜和真菌原生质体均无明显影响，另2组引起原生质体肿胀，继而破裂。因此，后两者可能干扰了质膜的通透性。两种阴离子表面活性剂在处理开始后立即导致原生质体突然破裂，表明它们以某种方式直接破坏质膜结构。不同的细胞器对膜的作用不同，尤其是线粒体膜和叶绿体颗粒膜对某些表面活性剂敏感。用不同表面活性剂处理黄瓜子叶，发现覆盖在子叶表面的角质层对某些表面活性剂的透性有抑制作用。因此，当完整的子叶用各自的表面活性剂处理时，子叶细胞不受大多数表面活性剂的影响。然而，当剥离的没有角质层的表皮的下表面被处理时，它们的细胞反应类似于原生质体。用表面活性剂处理真菌孢子也得到了类似的结果。这些结果表明，细胞壁的存在可能不是表面活性剂渗透性的负面因素。总而言之，尽管制备原生质体需要时间和人力，但原生质体系统是分析表面活性剂对活细胞的直接细胞学影响的一种适当敏感的系统。然而，表面活性剂通过角质层的渗透性应该用单独的方法来检查。

The present study aimed to examine the suitability of the protoplast system for testing effects of various surfactants on living cells. Protoplasts prepared from cucumber leaves and hyphae of cucumber anthracnose fungus, Colletotrichum lagenarium, were treated with 4 nonionic and 2 anionic surfactants. Test nonionic surfactants were divided into 2 groups on the basis of their effects: 2 of them did not give any detectable effects to both cucumber and fungus protoplasts, while other 2 caused swelling of protoplasts, followed by their bursting. Thus, the latter 2 perhaps interfere with permeability of plasma membrane. Two anionic surfactants caused the sudden bursting of protoplasts immediately after the onset of treatment, suggesting that they way directly damage the structure of plasma membrane. The effects of the respective surfactants on membranes varied among organelles: especially membranes of mitochondrial cristae and chloroplast granae were sensitive to some surfactants . The treatment of intact cotyledons of cucumber with the respective surfactants revealed that the cuticle layer covering the cotyledon surfaces suppressed the permeability of some surfactants. Therefore, when intact cotyledons were treated with the respective surfactants, cotyledon cells were not affected by most surfactants. However, when the lower surfaces of stripped epidermises which had no cuticle were treated, their cells responded similarly as protoplasts. Similar results were obtained when spores of the fungus were treated with the surfactants. These results suggest that the presence of cell walls may not be a negative factor for permeability of the surfactants. In conclusion, the protoplast system is a propers sensitive system for analyzing the direct, cytological effects of surfactants on living cells, although time and labor are required for preparing protoplasts. However, the permeability of surfactants through cuticles should be examined by a separate method.