Pastuglia Martine, Azimzadeh Juliette, Goussot Magali, Camilleri Christine, Belcram Katia, Evrard Jean-Luc, Schmit Anne-Catherine, Guerche Philippe, Bouchez David
Station de Génétique et d'Amélioration des Plantes, Institut National de la Recherche Agronomique, 78026 Versailles Cedex, France.
Plant Cell. 2006 Jun;18(6):1412-25. doi: 10.1105/tpc.105.039644. Epub 2006 May 12.
The process of microtubule nucleation in plant cells is still a major question in plant cell biology. gamma-Tubulin is known as one of the key molecular players for microtubule nucleation in animal and fungal cells. Here, we provide genetic evidence that in Arabidopsis thaliana, gamma-tubulin is required for the formation of spindle, phragmoplast, and cortical microtubule arrays. We used a reverse genetics approach to investigate the role of the two Arabidopsis gamma-tubulin genes in plant development and in the formation of microtubule arrays. Isolation of mutants in each gene and analysis of two combinations of gamma-tubulin double mutants showed that the two genes have redundant functions. The first combination is lethal at the gametophytic stage. Disruption of both gamma-tubulin genes causes aberrant spindle and phragmoplast structures and alters nuclear division in gametophytes. The second combination of gamma-tubulin alleles affects late seedling development, ultimately leading to lethality 3 weeks after germination. This partially viable mutant combination enabled us to follow dynamically the effects of gamma-tubulin depletion on microtubule arrays in dividing cells using a green fluorescent protein marker. These results establish the central role of gamma-tubulin in the formation and organization of microtubule arrays in Arabidopsis.
植物细胞中微管成核的过程仍是植物细胞生物学中的一个主要问题。γ-微管蛋白是动物和真菌细胞中微管成核的关键分子参与者之一。在此,我们提供了遗传学证据,表明在拟南芥中,γ-微管蛋白是纺锤体、成膜体和皮层微管阵列形成所必需的。我们采用反向遗传学方法来研究拟南芥中两个γ-微管蛋白基因在植物发育和微管阵列形成中的作用。对每个基因的突变体进行分离,并对γ-微管蛋白双突变体的两种组合进行分析,结果表明这两个基因具有冗余功能。第一种组合在配子体阶段是致死的。两个γ-微管蛋白基因的破坏会导致纺锤体和成膜体结构异常,并改变配子体中的核分裂。γ-微管蛋白等位基因的第二种组合影响幼苗后期发育,最终导致萌发后3周死亡。这种部分存活的突变体组合使我们能够使用绿色荧光蛋白标记动态跟踪γ-微管蛋白缺失对分裂细胞中微管阵列的影响。这些结果确立了γ-微管蛋白在拟南芥微管阵列形成和组织中的核心作用。