Determination of physical membrane properties of plant cell protoplasts via the electrofusion technique: prediction of optimal fusion yields and protoplast viability
Determination of physical membrane properties of plant cell protoplasts via the electrofusion technique: prediction of optimal fusion yields and protoplast viability
复制标题
通过电融合技术测定植物细胞原生质体的物理膜特性：预测最佳融合产量和原生质体活力
作者：
W. Mehrle;B. Naton;R. Hampp
通过改变引起电融合和电穿孔的物理参数（电场强度、脉冲宽度），分析了不同类型植物细胞原生质体的质膜性质。较低的阈值，该领域的脉冲强度在膜击穿发生（记录为融合事件）取决于脉冲持续时间，原生质体的大小，和原生质体类型（烟草，燕麦;空泡，evacuolated）。植物原生质体的这种融合特性也可以作为膜的充电过程的量度，并且因此允许非侵入性地确定时间常数和比膜电容。虽然在脉冲持续时间/场强对下，例如，10 μs/1.5 kV*cm−1和200 μs/0.5 kV*cm−1时，杂交体存活率不高。烟草叶肉原生质体的细胞壁再生和细胞分裂的速率没有受到影响，但在短脉冲持续时间/高场强可能已经增加。与此相反，电镀效率显着下降，在低场强的脉冲持续时间较长。
By variation of physical parameters (field strength, pulse duration) which result in electrofusion and electroporation, properties of the plasma membrane of different types of plant cell protoplasts were analyzed. The lower threshold for that field pulse intensity at which membrane breakdown occurred (recorded as fusion event) depended on pulse duration, protoplast size, and protoplast type (tobacco, oat; vacuolated, evacuolated). This fusion characteristic of plant protoplasts can also be taken as a measure of the charging process of the membrane and allows thus a non-invasive determination of the time constant and the specific membrane capacitance. Although the fusion yield was comparable at pulse duration/field strength couples of, e.g., 10 μs/1.5 kV*cm−1 and 200 μs/0.5 kV*cm−1, hybrid viability was not. Rates of cell wall regeneration and cell division of tobacco mesophyll protoplasts were not affected but may have been increased at short pulse duration/high field strength. Plating efficiency, in contrast, was significantly decreased with longer pulse duration at low field strengths.