蓝猪耳离体受精初试

Abstract

[中文文摘]用体内-体外方法分离了蓝猪耳精细胞。用酶解和解剖方法分离了其成熟卵细胞。分离的精、卵细胞用电融合介导尝试了体外诱导融合。在合适的渗透压(6%甘露醇)和合适的氯化钙(0.04%CaCl2·2H2O)溶液中,用交流电场为30～35V,10～25s使精、卵细胞排队;用直流电场400～600V,45～50μs的脉冲穿孔条件可诱导30%的精、卵细胞融合和70%以上的卵细胞之间的融合。尝试了人工合子的单细胞培养但未获成功。诱导蓝猪耳精、卵细胞融合的条件与玉米和水稻不同。[英文文摘]Sperm cells of Torenia fournieri were isolated from pollen tubes which were induced using an in vivo-in vitro method. Its egg cells were isolated by enzyme digesting followed by mechanical dissection. The electrical fusion method was applied to induce the fusion of the isolated sperm and egg cells. In a suitable solution containing 6% mannitol and 0.04% CaCl2 ·2H2O, 30% sperm-egg and over 70% egg-egg could be induced fusion using alternating current (30–35 V, 10–25 s) to align sperm-egg or egg-egg,following direct current (400–600 V, 45–50 μs) pulse to perforate the plasma member of both sperm and egg. The artifi cial zygotes were also tried to culture but no success. The conditions of inducing fusion of sperm cells and egg cells of Torenia fournieri are different with those of maize and rice.