Peng Xiong Bo, Sun Meng Xiang, Yang Hong Yuan
Key Laboratory of Ministry of Education for Plant Developmental Biology, College of Life Sciences, Wuhan University, China.
Cell Res. 2005 Sep;15(9):734-8. doi: 10.1038/sj.cr.7290343.
Various systems by using electric pulse, calcium, or polyethylene glycol have been developed in the past decade for the in vitro fusion of plant gametes. These in vitro systems provide a new way to study the fertilization mechanisms of plants. In this study, we developed a bovine serum albumin (BSA)-mediated fusion system for the in vitro fusion of maize gametes. The in vitro fusion of the isolated single egg cell and sperm cell of maize was observed microscopically in the BSA solution and the fertilized egg cell showed normal cell wall regeneration and nuclear division. The effects of the BSA concentration, pH value and calcium level on the efficiency of the maize gamete fusion were also assessed. BSA concentration and pH value did significantly affect the efficiency of the gamete fusion. Calcium was not necessary for the gamete fusion when BSA was present. The optimal solution for the gamete fusion contained 0.1% BSA, pH 6.0. The fusion frequency was as high as 96.7% in that optimal solution. This new in vitro fertilization system offers an alternative tool for the in vitro study of fertilization mechanisms with much simpler manipulating procedure than PEG system, and it will be especially useful for the in vitro study of the calcium dynamics during plant fertilization.
在过去十年中，人们开发了各种利用电脉冲、钙或聚乙二醇的系统来实现植物配子的体外融合。这些体外系统为研究植物的受精机制提供了一种新方法。在本研究中，我们开发了一种用于玉米配子体外融合的牛血清白蛋白（BSA）介导的融合系统。在BSA溶液中通过显微镜观察到了分离的玉米单个卵细胞和精子细胞的体外融合，并且受精卵细胞显示出正常的细胞壁再生和核分裂。还评估了BSA浓度、pH值和钙水平对玉米配子融合效率的影响。BSA浓度和pH值确实显著影响配子融合效率。当存在BSA时，钙对于配子融合不是必需的。配子融合的最佳溶液含有0.1% BSA，pH值为6.0。在该最佳溶液中融合频率高达96.7%。这种新的体外受精系统为受精机制的体外研究提供了一种替代工具，其操作程序比PEG系统简单得多，并且对于植物受精过程中钙动态的体外研究将特别有用。
