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首页> 外文期刊>Journal of Experimental Botany >Novel features of Brassica napus embryogenic microspores revealed by high pressure freezing and freeze substitution: evidence for massive autophagy and excretion-based cytoplasmic cleaning.
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Novel features of Brassica napus embryogenic microspores revealed by high pressure freezing and freeze substitution: evidence for massive autophagy and excretion-based cytoplasmic cleaning.

机译：高压冷冻和冷冻替代揭示了甘蓝型油菜胚性小孢子的新特征：大量自噬和基于排泄的细胞质清洁的证据。

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Induction of embryogenesis from isolated microspore cultures is a complex experimental system where microspores undergo dramatic changes in developmental fate. After ~40 years of application of electron microscopy to the study of the ultrastructural changes undergone by the induced microspore, there is still room for new discoveries. In this work, high pressure freezing and freeze substitution (HPF/FS), the best procedures known to date for ultrastructural preservation, were used to process Brassica napus microspore cultures covering all the stages of microspore embryogenesis. Analysis of these cultures by electron microscopy revealed massive processes of autophagy exclusively in embryogenic microspores, but not in other microspore-derived structures also present in cultures. However, a significant part of the autophagosomal cargo was not recycled. Instead, it was transported out of the cell, producing numerous deposits of extracytoplasmic fibrillar and membranous material. It was shown that commitment of microspores to embryogenesis is associated with both massive autophagy and excretion of the removed material. It is hypothesized that autophagy would be related to the need for a profound cytoplasmic cleaning, and excretion would be a mechanism to avoid excessive growth of the vacuolar system. Together, the results also demonstrate that the application of HPF/FS to the study of the androgenic switch is the best option currently available to identify the complex and dramatic ultrastructural changes undergone by the induced microspore. In addition, they provide significant insights to understand the cellular basis of induction of microspore embryogenesis, and open a new door for the investigation of this intriguing developmental pathway.
机译：从分离的小孢子培养物中诱导胚胎发生是一个复杂的实验系统，在该系统中，小孢子的发育命运发生了巨大变化。在将电子显微镜应用于诱导小孢子所经历的超微结构变化的研究中，经过约40年的研究，仍有新发现的空间。在这项工作中，高压冷冻和冷冻替代（HPF / FS）是迄今为止已知的用于超微结构保存的最佳方法，用于处理覆盖小孢子胚胎发生所有阶段的甘蓝型油菜小孢子培养物。通过电子显微镜对这些培养物的分析显示，自噬的大量自噬过程仅在胚胎发生的小孢子中发生，而在培养物中也存在的其他小孢子衍生的结构中则没有。但是，自噬体货物的很大一部分没有回收。取而代之的是，它被转运出细胞，产生了大量胞浆外纤维状和膜状物质的沉积物。结果表明，小孢子对胚胎发生的承诺与大量自噬和所去除物质的排泄有关。据推测，自噬与彻底清洗细胞质有关，排泄是避免液泡系统过度生长的一种机制。总之，结果还表明，将HPF / FS应用于雄性激素转换的研究是目前可用于鉴定诱导的小孢子经历的复杂而剧烈的超微结构变化的最佳选择。此外，他们提供了重要的见识，以了解诱导小孢子胚胎发生的细胞基础，并为研究这种有趣的发育途径打开了新的大门。

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