Ramesar-Fortner Nicole S, Yeung Edward C
Department of Biological Sciences, University of Calgary, Calgary, AB, Canada.
Methods Mol Biol. 2011;710:29-37. doi: 10.1007/978-1-61737-988-8_3.
To further understand the events occurring during embryogenesis, it is imperative to have an experimental system. While somatic embryo is the system of choice due to the prolific number and success of protocols available, it is necessary to compare in vitro results with those in vivo. This process is often difficult due to the inaccessibility of the developing embryo and the complications of manipulating the embryo in vivo. The development of protocols that allow for manipulation and comparison of both somatic and zygotic embryos is key to elucidating the differences between the two types of embryos and determining if results observed using one system can be applied to the other. This chapter details a simple protocol for the culture of zygotic embryos of canola that allows for the processing of large numbers of embryos with little physical damage. Furthermore, this protocol allows for the experimental manipulation of zygotic embryos in vitro and comparisons to be made with a well-established microspore-derived embryo system.
为了进一步了解胚胎发生过程中发生的事件,拥有一个实验系统至关重要。虽然体细胞胚胎由于现有方案数量众多且成功率高而成为首选系统,但有必要将体外结果与体内结果进行比较。由于发育中的胚胎难以获取以及在体内操作胚胎存在复杂性,这个过程通常很困难。开发能够对体细胞胚胎和合子胚胎进行操作和比较的方案,对于阐明这两种胚胎类型之间的差异以及确定使用一种系统观察到的结果是否可以应用于另一种系统至关重要。本章详细介绍了一种用于油菜合子胚胎培养的简单方案,该方案允许处理大量胚胎且几乎不造成物理损伤。此外,该方案允许在体外对合子胚胎进行实验操作,并与成熟的小孢子衍生胚胎系统进行比较。