Mechanical isolation and ultrastructural characterization of viable egg cells in Plumbago zeylanica
Mechanical isolation and ultrastructural characterization of viable egg cells in Plumbago zeylanica
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DOI：
10.1007/s004970050111
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发表时间：
1997-12-01
影响因子：
--
通讯作者：
Russell, SD
中科院分区：
文献类型：
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作者：
Cao, YJ;Russell, SD
报道了一种用机械解剖法分离白花丹卵的方法。分离的最佳溶液为0.8M甘露醇+10mM MOPS +10mM CaCl_2（pH 4.5-5.0），渗透压为860-940 mmol/kg。鸡蛋保持活力至少24小时。分离的卵是没有细胞壁的真正的原生质体，并且可以耐受437 mmol/kg至965 mmol/kg的渗透压。用透射电镜观察分离的卵细胞，结果表明，它们保存良好，并反映了生理活性细胞的超微结构，显示出与体内卵细胞相似的特征。值得注意的差异包括丝状器和致密颗粒的质体，这是最明显的卵细胞，在隔离过程中被损坏的积累的情况下。
A protocol for isolating viable eggs in Plumbago zeylanica by mechanical dissection is reported. The optimum solution for isolation was 0.8 M mannitol + 10 mM MOPS + 10 mM CaCl2, (pH 4.5-5.0) with an osmolality of 860-940 mmol/kg. Eggs retain their viability for at least 24 h. Isolated eggs were true protoplasts without cell walls and could tolerate osmolality of 437 mmol/kg to 965 mmol/kg. Observation of the isolated eggs using transmission electron microscopy indicated that they were well preserved and reflected the ultrastructure of physiologically active cells, displaying features similar to those of in vivo egg cells. Notable differences include the absence of a filiform apparatus and the accumulation of dense particles in the plastids, which was most conspicuous in egg cells that were damaged during isolation.