Unité Mixte de Recherches Interactions Biotiques et Santé Végétale, INRA-CNRS-UNS, Sophia Antipolis, France.
PLoS Pathog. 2011 Dec;7(12):e1002343. doi: 10.1371/journal.ppat.1002343. Epub 2011 Dec 1.
Reorganization of the microtubule network is important for the fast isodiametric expansion of giant-feeding cells induced by root-knot nematodes. The efficiency of microtubule reorganization depends on the nucleation of new microtubules, their elongation rate and activity of microtubule severing factors. New microtubules in plants are nucleated by cytoplasmic or microtubule-bound γ-tubulin ring complexes. Here we investigate the requirement of γ-tubulin complexes for giant feeding cells development using the interaction between Arabidopsis and Meloidogyne spp. as a model system. Immunocytochemical analyses demonstrate that γ-tubulin localizes to both cortical cytoplasm and mitotic microtubule arrays of the giant cells where it can associate with microtubules. The transcripts of two Arabidopsis γ-tubulin (TUBG1 and TUBG2) and two γ-tubulin complex proteins genes (GCP3 and GCP4) are upregulated in galls. Electron microscopy demonstrates association of GCP3 and γ-tubulin as part of a complex in the cytoplasm of giant cells. Knockout of either or both γ-tubulin genes results in the gene dose-dependent alteration of the morphology of feeding site and failure of nematode life cycle completion. We conclude that the γ-tubulin complex is essential for the control of microtubular network remodelling in the course of initiation and development of giant-feeding cells, and for the successful reproduction of nematodes in their plant hosts.
微管网络的重组成因根结线虫诱导的巨型取食细胞的快速等径扩张非常重要。微管重排的效率取决于新微管的成核、它们的延伸速度和微管切割因子的活性。植物中的新微管由细胞质或微管结合的γ-微管蛋白环复合物引发。在这里,我们使用拟南芥和根结线虫属之间的相互作用作为模型系统,研究了γ-微管蛋白复合物对巨型取食细胞发育的要求。免疫细胞化学分析表明,γ-微管蛋白定位于巨型细胞的皮质细胞质和有丝分裂微管阵列中,在那里它可以与微管结合。两种拟南芥γ-微管蛋白(TUBG1 和 TUBG2)和两种γ-微管蛋白复合物蛋白基因(GCP3 和 GCP4)的转录物在虫瘿中上调。电子显微镜证明 GCP3 和 γ-微管蛋白作为复合体的一部分存在于巨型细胞的细胞质中。敲除一个或两个γ-微管蛋白基因会导致取食部位形态的基因剂量依赖性改变,并且线虫生命周期的完成失败。我们得出结论,γ-微管蛋白复合物对于控制巨型取食细胞起始和发育过程中的微管网络重塑以及线虫在其植物宿主中的成功繁殖是必不可少的。