Edwards G E, Black C C
Department of Biochemistry, University of Georgia, Athens, Georgia 30601.
Plant Physiol. 1971 Jan;47(1):149-56. doi: 10.1104/pp.47.1.149.
A technique is described for the separation of mesophyll and bundle sheath cells from Digitaria sanguinalis leaves and evidence for separation is given with light and scanning electron micrographs. Gentle grinding of fully differentiated leaves in a mortar releases mesophyll cells which are isolated on nylon nets by filtration. More extensive grinding of the remaining tissue yields bundle sheath strands which are isolated by filtration with stainless steel sieves and nylon nets. Further grinding of bundle sheath strands in a tissue homogenizer releases bundle sheath cells which are collected on nylon nets. Percentage of purity derived from cell counts and yield data on a chlorophyll basis are given.The internal leaf cell morphology is presented in scanning electron micrographs and compared with light micrographs of fully-differentiated D. sanguinalis leaves. In leaves of plants which possess the C(4)-dicarboxylic acid cycle of photosynthesis, the relationship of leaf morphology to photosynthesis in mesophyll and bundle sheath cells is considered, and the hypothesis is presented that as atmospheric CO(2) enters a leaf about 85% is fixed by the C(4)-dicarboxylic acid cycle in the mesophyll cells and 10 to 15% is fixed by the reductive pentose phosphate cycle in the bundle sheath cells.A technique also is given for the isolation of mesophyll cells from spinach leaves.
本文描述了一种从马唐叶片中分离叶肉细胞和维管束鞘细胞的技术，并通过光学显微镜和扫描电子显微镜照片给出了分离的证据。在研钵中对完全分化的叶片进行轻柔研磨可释放叶肉细胞，通过过滤将其分离在尼龙网上。对剩余组织进行更广泛的研磨可得到维管束鞘束，通过不锈钢筛网和尼龙网过滤将其分离。在组织匀浆器中对维管束鞘束进行进一步研磨可释放维管束鞘细胞，这些细胞收集在尼龙网上。给出了基于细胞计数的纯度百分比以及以叶绿素为基础的产量数据。扫描电子显微镜照片展示了叶片内部细胞形态，并与完全分化的马唐叶片的光学显微镜照片进行了比较。在具有光合作用C(4)-二羧酸循环的植物叶片中，考虑了叶肉细胞和维管束鞘细胞中叶形态与光合作用的关系，并提出了一个假设，即当大气中的二氧化碳进入叶片时，约85%由叶肉细胞中的C(4)-二羧酸循环固定，10%至15%由维管束鞘细胞中的还原戊糖磷酸循环固定。还给出了一种从菠菜叶片中分离叶肉细胞的技术。
