Instituto de Bioingeniería, Universidad Miguel Hernández, Campus de Elche, 03202, Elche, Spain.
Sci Rep. 2017 Dec 5;7(1):17010. doi: 10.1038/s41598-017-17218-z.
Several hundred genes are required for embryonic and gametophytic development in the model plant Arabidopsis thaliana, as inferred from the lethality of their mutations. Despite many of these genes are expressed throughout the plant life cycle, the corresponding mutants arrest at early stages, preventing the study of their post-embryonic functions by conventional methods. Clonal analysis represents an effective solution to this problem by uncovering the effects of embryo-lethal mutations in sectors of mutant cells within an otherwise normal adult plant. In this pilot study, we have evaluated the suitability of two sector induction methods for the large-scale study of the post-embryonic effects of embryo-lethal (emb) mutations in Arabidopsis. In line with the interests of our laboratory, we selected 24 emb mutations that damage genes that are expressed in wild-type vegetative leaves but whose effects on leaf development remain unknown. For the induction of mutant sectors in adult plants, we followed one approach based on the X-ray irradiation of 'cell autonomy' (CAUT) lines, and another based on the site-specific excision of transgenes mediated by Cre recombinase. We conclude that both methods are time-consuming and difficult to scale up, being better suited for the study of emb mutations on a case-by-case basis.
数百个基因对于拟南芥（Arabidopsis thaliana）胚胎和配子体发育是必需的，这可以从它们突变的致死性推断出来。尽管这些基因中的许多在整个植物生命周期中都有表达，但相应的突变体在早期就停止了生长，这使得传统方法无法研究它们的胚胎后功能。克隆分析通过揭示在正常成年植物的突变细胞的某些区域内的胚胎致死突变的影响，为解决这个问题提供了有效的解决方案。在这项初步研究中，我们评估了两种扇区诱导方法在大规模研究拟南芥胚胎致死（emb）突变的胚胎后效应中的适用性。根据我们实验室的兴趣，我们选择了 24 个 emb 突变，这些突变破坏了在野生型营养叶中表达的基因，但它们对叶片发育的影响尚不清楚。为了在成年植物中诱导突变扇区，我们采用了一种基于 X 射线照射“细胞自主性”（CAUT）系的方法，另一种基于 Cre 重组酶介导的转基因定点切除的方法。我们得出的结论是，这两种方法都很耗时，难以扩展，更适合逐个案例研究 emb 突变。
