Isolation of gametes and central cells from Oryza sativa L.
Isolation of gametes and central cells from Oryza sativa L.
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DOI：
10.1007/s00497-006-0020-x
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发表时间：
2006-02
影响因子：
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通讯作者：
T. Uchiumi;S. Komatsu;T. Koshiba;T. Okamoto
中科院分区：
文献类型：
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作者：
T. Uchiumi;S. Komatsu;T. Koshiba;T. Okamoto
利用玉米配子和中央细胞建立了高等植物的体外受精体系，可以产生胚和胚乳。本研究以水稻（Oryza sativa L.）CV.日本晴）为模式植物，建立水稻体外受精体系。卵细胞和中央细胞的酶处理未授粉胚珠的手工操作分离，并建立了一个替代的直接分离方法，不使用酶处理的卵细胞。荧光可视化的颗粒状结构在离体卵细胞的细胞质和核仁中的两个极核的离体中央细胞表明，这些细胞是可靠的配子和中央细胞。在精细胞分离中，通过渗透压诱导的花粉破裂释放花粉内容物。此外，成功地进行了与分离的配子的电融合。
In vitro fertilization system of higher plants has been well established using maize gametes and central cells, which can produce embryos and endosperms. In the present study, procedures for isolating gametes and central cells from rice (Oryza sativaL. cv. Nipponbare), a model plant, are reported with the goal of establishing rice in vitro fertilization system. Egg cells and central cells were isolated by manual manipulation of enzyme-treated unpollinated ovules, and an alternative direct isolation method for egg cells that does not use enzymatic treatment was also established. Fluorescent visualization of the granular structures in the cytoplasm of isolated egg cells and the nucleoli in two polar nuclei of isolated central cells suggest that these cells are reliable gametes and central cells. For sperm cell isolation, the contents of rice pollen grains were released by osmotic pressure-induced bursting of the grains. In addition, electrofusion with isolated gametes was successfully conducted.