Isolation of two populations of sperm cells and microelectrophoresis of pairs of sperm cells from pollen tubes of tobacco (Nicotiana tabacum)
Isolation of two populations of sperm cells and microelectrophoresis of pairs of sperm cells from pollen tubes of tobacco (Nicotiana tabacum)
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DOI：
10.1007/s00497-005-0248-x
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发表时间：
2005-06
影响因子：
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通讯作者：
Yh Yang;Yl Qiu;Chao Tian Xie;Hui-Qiao Tian;Zhaojie Zhang;Scott D. Russell
中科院分区：
文献类型：
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作者：
Yh Yang;Yl Qiu;Chao Tian Xie;Hui-Qiao Tian;Zhaojie Zhang;Scott D. Russell
先前的研究表明，烟草的两个精细胞是二态的，这表明它们可能在与卵细胞和中央细胞的体内融合过程中参与优先受精。利用现代敏感的分子生物学技术来探讨这种植物潜在的优先受精机制是十分必要的。为了这个目的，两个单独的人口的两个精细胞，构成Svn（与营养核）和Sua（与营养核无关），分离在成千上万的烟草花粉管与显微操作器作为初步的一步研究配子识别使用分子技术。对单个花粉管中不同发育时期的成对精细胞进行了显微电泳。从1-，2-，3-和4-cm的花柱长度分离的精子细胞迁移到负极，与苏阿显示显着更大的电泳迁移率比Svn，反映了更多的正电荷的苏阿细胞表面。从1cm花柱中分离的精细胞对电泳场中的电子电位非常敏感，这可能反映了它们仍处于年轻状态。Sua和Svn细胞表面电荷的差异可能与受精过程中的细胞命运有关。
Prior research has indicated that the two sperm cells ofNicotiana tabacumare dimorphic, suggesting that they may participate in preferential fertilization during in vivo fusion with the egg and central cells. To probe the mechanism of potential preferential fertilization in this plant, it will be necessary to use modern sensitive molecular techniques. For this purpose, two individual populations of two sperm cells, constituting the Svn (associated with the vegetative nucleus) and Sua (unassociated with the vegetative nucleus), were isolated in the thousands from tobacco pollen tubes with a micromanipulator as a preliminary step toward research on gametic recognition using molecular techniques. Microelectrophoresis of paired sperm cells from a single pollen tube was conducted at different developmental stages. Sperm cells isolated from 1-, 2-, 3- and 4-cm stylar lengths migrated to the negative pole, with the Sua displaying significantly greater electrophoretic mobility than the Svn, reflecting a more positively charged cell surface on the Sua. The sperm cells isolated from 1-cm style are very sensitive to electron potential in an electrophoretic field, presumably reflecting that they are still in a young state. Differences in cell surface charge between the Sua and Svn may be related with cell fate during fertilization.