Pollen tetrad-based visual assay for meiotic recombination in Arabidopsis
Pollen tetrad-based visual assay for meiotic recombination in Arabidopsis
复制标题
DOI：
10.1073/pnas.0608936104
复制
发表时间：
2007-03-06
影响因子：
11.1
通讯作者：
Copenhaver, Gregory P.
中科院分区：
文献类型：
--
作者：
Francis, Kirk E.;Lam, Sandy Y.;Copenhaver, Gregory P.
重组以交叉（CO）和基因转换（GC）的形式出现，是从真菌到哺乳动物减数分裂的高度保守特征。重组有助于确保染色体分离并促进等位基因多样性。重组机制的损伤对于减数分裂通常是灾难性的，导致不育。我们开发了一种视觉检测方法，能够检测拟南芥中的 Cos 和 GC，并测量 CO 干扰。这种灵活的测定利用转基因构建体编码 qrt1 突变体背景中三种不同颜色的花粉表达荧光蛋白。通过观察 92,489 个花粉四分体中荧光等位基因的分离，我们证明了（i）发育位置与 CO 频率之间的相关性，（h）CO 频率的温度依赖性，（M）检测减数分裂 GC 事件的能力，以及（iv）快速评估 CO 干扰的能力。
Recombination, in the form of cross-overs (COs) and gene conversion (GC), is a highly conserved feature of meiosis from fungi to mammals. Recombination helps ensure chromosome segregation and promotes allelic diversity. Lesions in the recombination machinery are often catastrophic for meiosis, resulting in sterility. We have developed a visual assay capable of detecting Cos and GCs and measuring CO interference in Arabidopsis thaliana. This flexible assay utilizes transgene constructs encoding pollen-expressed fluorescent proteins of three different colors in the qrt1 mutant background. By observing the segregation of the fluorescent alleles in 92,489 pollen tetrads, we demonstrate (i) a correlation between developmental position and CO frequency, (h) a temperature dependence for CO frequency, (M) the ability to detect meiotic GC events, and (iv) the ability to rapidly assess CO interference.