不同电场强度和原生质体密度对马铃薯和薯蓣细胞融合效率的影响

EFFECTS OF ALTERNATING CURRENT FIELD AND PROTOPLAST DENSITY ON THE CELL FUSION FREQUENCY OF POTATO AND YAM

体细胞电融合技术因其融合效率高、对细胞无毒害作用而愈来愈普遍地用于细胞工程技术操作中。为了准确地确定实验参数，试验采用一个组装的电融合仪和自制平行电极，以马铃薯和薯蓣为材料，研究了在不同原生质体密度和不同电场强度条件下的细胞融合效率。试验结果显示，以交变电场频率1．0MHz、电压140Vpp．cm-处理原生质体20－40秒，尔后施加瞬时直流高压（120kv）脉冲1－2次，可以获得73．5％─76．7％的融合细胞，其中11．5％─14．4％的融合细胞可发生正常分裂。在本试验设计范围内，电场强度对细胞融合频率的影响主要表现在融合细胞能否发生分裂，而初始融合频率在各处理间无显著差异。

Electric fusion of somatic cells has been more widely used recently in cell engineering manipulation because of its high fusion frequency and non-toxic action to the cells. To decide optimum fusion parameters, the effects of alternating current (AC) field and protoplast density on the fusion frequency of potato and yam somatic cells were investisated. A combined cell hybrider and a lab-made paralIel electrode were used for the experiment. The results showed that fusion frequency of 73. 5%-76. 7% were obtained with treatments of AC 1, 0MHz. 140V for 20-40 seconds followed by 1-2 pulses of direct current (DC)voltage (120kV). Of the fusion cells. 11. 5%- 14. 4% had a normal cell division during subsequent culture. Among the treatments of this experiment, the optimum protoplast density was 1. 6-1. 8×105ml- 1. AC field had no remarkable effects on the fusion frequency but cell division rate of the fusion protoplasts.