Fertilization in Arabidopsis thaliana wild type:: Developmental stages and time course
Fertilization in Arabidopsis thaliana wild type:: Developmental stages and time course
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DOI：
10.1046/j.1365-313x.2002.01305.x
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发表时间：
2002-05-01
期刊：
影响因子：
7.2
通讯作者：
Dumas, C
中科院分区：
文献类型：
--
作者：
Faure, JE;Rotman, N;Dumas, C
我们描述了一些以前未描述的阶段，在拟南芥受精，并提供了第一次精确的时间过程中的受精过程。我们用野生型花粉（哥伦比亚生态型）人工授粉野生型雌蕊，在授粉后的不同时间将它们固定，并使用共聚焦激光扫描显微镜分析了600个胚囊。其中一个助细胞的退化开始于花粉后5小时（HAP）。助细胞退化后，卵的极性迅速改变。然后通过在卵和中央细胞核中存在两个不同直径的核仁来检测核配，7-8 HAP。在接下来的一个小时内，第一次核分裂发生在受精的中央细胞中，然后可以在每个产生的核中短暂地看到两个核仁。在第二组实验中，我们用来自转基因promLAT 52：：EGFP系的花粉人工授粉野生型雌蕊，所述转基因promLAT 52：：EGFP系在其花粉营养细胞中表达EGFP。在活体材料中可检测到花粉管内容物释放到助细胞中。我们发现，助细胞退化和花粉管释放的时间相关性很好，这表明无论是助细胞退化的时候，花粉管放电或之前不久it. These观察和协议构成了一个重要的基础，为进一步的表型分析受影响的突变体受精。
We describe some previously uncharacterised stages of fertilization in Arabidopsis thaliana and provide for the first time a precise time course of the fertilization process. We hand-pollinated wild type pistils with wild type pollen (Columbia ecotype), fixed them at various times after pollination, and analysed 600 embryo sacs using Confocal Laser Scanning Microscopy. Degeneration of one of the synergid cells starts at 5 Hours After Pollination (HAP). Polarity of the egg changes rapidly after this synergid degeneration. Karyogamy is then detected by the presence of two nucleoli of different diameters in both the egg and central cell nuclei, 7-8 HAP. Within the next hour, first nuclear division takes place in the fertilized central cell and two nucleoli can then be seen transiently in each nucleus produced. In a second set of experiments, we hand-pollinated wild type pistils with pollen from a transgenic promLAT52::EGFP line that expresses EGFP in its pollen vegetative cell. Release of the pollen tube contents into the synergid cell could be detected in living material. We show that the timing of synergid degeneration and pollen tube release correlate well, suggesting that either the synergid cell degenerates at the time of pollen tube discharge or very shortly before it. These observations and protocols constitute an important basis for the further phenotypic analysis of mutants affected in fertilization.