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Author:
Date:
2015-11-20
[Abstract] One of the major topics in plant and animal biology is sexual reproduction. It is, therefore, of great interest to isolate and study germ cells and accessory cells. The male gametophyte of the flowering plant Arabidopsis thaliana (A. thaliana), pollen, is the product of two post-meiotic mitotic divisions. Each mature pollen grain consists of two sperm cells contained within the vegetative cell, the non-reproductive companion cell. The tough pollen wall and its special nested structure make it difficult to study pollen cells separately. Here, we describe a simple and efficient method to fractionate A. thaliana sperm and vegetative cell nuclei by fluorescence activated cell sorting (FACS). Our protocol is based on differences in fluorescence intensity of sperm and ...
[摘要] 植物和动物生物学的一个主要议题是有性繁殖。因此，分离和研究生殖细胞和附属细胞是非常有意义的。开花植物拟南芥（Arabidopsis thaliana）（ A。thaliana ）的雄性配子体是两个减数分裂后有丝分裂的产物。每个成熟花粉粒由包含在营养细胞（非生殖伴侣细胞）内的两个精细胞组成。坚硬的花粉壁及其特殊的嵌套结构使得难以分别研究花粉细胞。在这里，我们描述一个简单而有效的方法分割。拟南芥精子和营养细胞核通过荧光激活细胞分选（FACS）。我们的方案基于用SYBR Green I染色的精子和营养细胞核的荧光强度的差异.100植物产生约1×10 6个精子和350,000个营养细胞核。该方法可用于纯化各种A的花粉核。拟南芥野生型种质和突变株系，并且原则上可适用于其它植物物种的花粉。
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