This work describes the application of clearing on vibratome sections to study the embryo formation in cassava. This procedure provides high-resolution images and reduces significantly the number of sections that need to be analyzed per ovule. This methodology was instrumental for the development of the protocol for embryo rescue in cassava. It has been also applied to monitor the embryo formation response when optimizing seed setting from regular and broad crosses for cassava breeding. Broad crosses between cassava and castor bean (incompatible-euphorbiaceae species) were made aiming to induce doubled haploids through the elimination of the incompatible-male parent genome as done in cereals. Castor bean is widely available and provides continues supply of pollen. Our results suggest that this methodology is easy and effective to assess the response of hundreds of cassava ovules pollinated with castor bean pollen, allowing the identification of multicellular structures in the embryo sac without apparent formation of endosperm. The protocol is also useful when developing and optimizing a methodology to induce doubled haploids in cassava via gynogenesis or from ovules pollinated with irradiated cassava pollen.

这项工作描述了在玻璃纤维切片上进行清理的应用，以研究木薯中的胚胎形成。此过程可提供高分辨率图像，并显着减少了每个胚珠需要分析的切片数量。该方法学有助于开发木薯胚胎拯救方案。当用于常规和广泛杂交的木薯育种优化种子结实时，它也已用于监测胚胎形成反应。进行了木薯和蓖麻子（不相容大戟科物种）之间的广泛杂交，旨在通过消除谷物中不相容的雄性亲本基因组来诱导双倍单倍体。蓖麻子可广泛获得并且持续提供花粉。我们的结果表明，这种方法简便，有效，可以评估用蓖麻子花粉授粉的数百个木薯胚珠的反应，从而可以鉴定胚囊中的多细胞结构而没有明显的胚乳形成。该协议在开发和优化在木薯中诱导双倍单倍体的方法学时也很有用通过 雌核发育或胚珠经辐射的木薯花粉授粉。