Caracuel Zaira, Martínez-Rocha Ana Lilia, Di Pietro Antonio, Madrid Marta P, Roncero M Isabel G
Departamento de Genetica, Universidad de Córdoba, Campus de Rabanales, Spain.
Mol Plant Microbe Interact. 2005 Nov;18(11):1140-7. doi: 10.1094/MPMI-18-1140.
Glycosylphosphatidylinositol-anchored (beta)-1,3-glucanosyltransferases play active roles in fungal cell wall biosynthesis and morphogenesis and have been implicated in virulence on mammals. The role of beta-1,3-glucanosyltransferases in pathogenesis to plants has not been explored so far. Here, we report the cloning and mutational analysis of the gas1 gene encoding a putative beta-1,3-glucanosyltransferase from the vascular wilt fungus Fusarium oxysporum. In contrast to Candida albicans, expression of gas1 in F. oxysporum was independent of ambient pH and of the pH response transcription factor PacC. Gene knockout mutants lacking a functional gas1 allele grew in a way similar to the wildtype strain in submerged culture but exhibited restricted colony growth on solid substrates. The restricted growth phenotype was relieved by the osmotic stabilizer sorbitol, indicating that it may be related to structural alterations in the cell wall. Consistent with this hypothesis, deltagas1 mutants exhibited enhanced resistance to cell wall-degrading enzymes and increased transcript levels of chsV and rho1, encoding a class V chitin synthase and a small monomeric G protein, respectively. The deltagas1 mutants showed dramatically reduced virulence on tomato, both in a root infection assay and in a fruit tissue-invasion model, thus providing the first evidence for an essential role of fungal beta-1,3-glucanosyltransferases during plant infection.
糖基磷脂酰肌醇锚定的β-1,3-葡聚糖基转移酶在真菌细胞壁生物合成和形态发生中发挥着积极作用,并与对哺乳动物的毒力有关。迄今为止,β-1,3-葡聚糖基转移酶在植物致病过程中的作用尚未得到探索。在此,我们报告了来自维管束萎蔫真菌尖孢镰刀菌的假定β-1,3-葡聚糖基转移酶编码基因gas1的克隆和突变分析。与白色念珠菌不同,gas1在尖孢镰刀菌中的表达与环境pH值和pH响应转录因子PacC无关。缺乏功能性gas1等位基因的基因敲除突变体在液体培养中的生长方式与野生型菌株相似,但在固体培养基上菌落生长受限。渗透稳定剂山梨醇可缓解这种生长受限的表型,表明这可能与细胞壁的结构改变有关。与此假设一致,gas1缺失突变体对细胞壁降解酶的抗性增强,并且分别编码V类几丁质合酶和小的单体G蛋白的chsV和rho1的转录水平增加。在根部感染试验和果实组织侵染模型中,gas1缺失突变体在番茄上的毒力均显著降低,从而为真菌β-1,3-葡聚糖基转移酶在植物感染过程中的重要作用提供了首个证据。