Department of Biology and Biotechnology, Worcester Polytechnic Institute, Worcester, MA.
Bioinformatics and Computational Biology Program, Worcester Polytechnic Institute, Worcester, MA.
PLoS Genet. 2018 May 10;14(5):e1007221. doi: 10.1371/journal.pgen.1007221. eCollection 2018 May.
Our ability to identify genes that participate in cell growth and division is limited because their loss often leads to lethality. A solution to this is to isolate conditional mutants where the phenotype is visible under restrictive conditions. Here, we capitalize on the haploid growth-phase of the moss Physcomitrella patens to identify conditional loss-of-growth (CLoG) mutants with impaired growth at high temperature. We used whole-genome sequencing of pooled segregants to pinpoint the lesion of one of these mutants (clog1) and validated the identified mutation by rescuing the conditional phenotype by homologous recombination. We found that CLoG1 is a novel and ancient gene conserved in plants. At the restrictive temperature, clog1 plants have smaller cells but can complete cell division, indicating an important role of CLoG1 in cell growth, but not an essential role in cell division. Fluorescent protein fusions of CLoG1 indicate it is localized to microtubules with a bias towards depolymerizing microtubule ends. Silencing CLoG1 decreases microtubule dynamics, suggesting that CLoG1 plays a critical role in regulating microtubule dynamics. By discovering a novel gene critical for plant growth, our work demonstrates that P. patens is an excellent genetic system to study genes with a fundamental role in plant cell growth.
我们识别参与细胞生长和分裂的基因的能力有限,因为它们的缺失往往会导致致命性。解决这个问题的方法是分离条件性突变体,在限制条件下可以观察到表型。在这里,我们利用苔藓植物Physcomitrella patens 的单倍体生长阶段来鉴定在高温下生长受损的条件性生长缺失(CLoG)突变体。我们对混合分离子代进行全基因组测序,以确定其中一个突变体(clog1)的病变,并通过同源重组恢复条件表型来验证鉴定出的突变。我们发现 CLoG1 是一种在植物中保守的新型古老基因。在限制温度下,clog1 植物的细胞较小,但可以完成细胞分裂,表明 CLoG1 在细胞生长中起重要作用,但在细胞分裂中不是必需的。CLoG1 的荧光蛋白融合表明它定位于微管上,偏向于解聚微管末端。沉默 CLoG1 会降低微管动力学,表明 CLoG1 在调节微管动力学中起关键作用。通过发现一个对植物生长至关重要的新基因,我们的工作表明 P. patens 是研究对植物细胞生长具有基本作用的基因的绝佳遗传系统。