Niño-Sánchez Jonathan, Casado-Del Castillo Virginia, Tello Vega, De Vega-Bartol José J, Ramos Brisa, Sukno Serenella A, Díaz Mínguez José María
Instituto Hispano-Luso de Investigaciones Agrarias (Ciale), Departamento de Microbiología y Genética, Universidad de Salamanca, C/Río Duero 12, Villamayor, 37185, Salamanca, Spain.
Instituto de Biologia Experimental e Tecnológica/Instituto de Tecnologia Química e Biológica-Universidade Nova de Lisboa (IBET/ITQB-UNL), Av. República, 2784-505, Oeiras, Portugal.
Mol Plant Pathol. 2016 Sep;17(7):1124-39. doi: 10.1111/mpp.12373. Epub 2016 Apr 12.
The FTF (Fusarium transcription factor) gene family comprises a single copy gene, FTF2, which is present in all the filamentous ascomycetes analysed, and several copies of a close relative, FTF1, which is exclusive to Fusarium oxysporum. An RNA-mediated gene silencing system was developed to target mRNA produced by all the FTF genes, and tested in two formae speciales: F. oxysporum f. sp. phaseoli (whose host is common bean) and F. oxysporum f. sp. lycopersici (whose host is tomato). Quantification of the mRNA levels showed knockdown of FTF1 and FTF2 in randomly isolated transformants of both formae speciales. The attenuation of FTF expression resulted in a marked reduction in virulence, a reduced expression of several SIX (Secreted In Xylem) genes, the best studied family of effectors in F. oxysporum, and lower levels of SGE1 (Six Gene Expression 1) mRNA, the presumptive regulator of SIX expression. Moreover, the knockdown mutants showed a pattern of colonization of the host plant similar to that displayed by strains devoid of FTF1 copies (weakly virulent strains). Gene knockout of FTF2 also resulted in a reduction in virulence, but to a lesser extent. These results demonstrate the role of the FTF gene expansion, mostly the FTF1 paralogues, as a regulator of virulence in F. oxysporum and suggest that the control of effector expression is the mechanism involved.
FTF(镰刀菌转录因子)基因家族包含一个单拷贝基因FTF2,该基因存在于所有已分析的丝状子囊菌中,以及几个近亲拷贝FTF1,它是尖孢镰刀菌特有的。开发了一种RNA介导的基因沉默系统,以靶向所有FTF基因产生的mRNA,并在两个专化型中进行了测试:尖孢镰刀菌菜豆专化型(其寄主是菜豆)和尖孢镰刀菌番茄专化型(其寄主是番茄)。mRNA水平的定量分析表明,在这两个专化型的随机分离转化体中,FTF1和FTF2的表达均被敲低。FTF表达的减弱导致毒力显著降低,几个SIX(木质部分泌蛋白)基因的表达减少,SIX基因是尖孢镰刀菌中研究最深入的效应子家族,同时SGE1(Six基因表达1)mRNA水平降低,SGE1是SIX表达的假定调节因子。此外,敲低突变体显示出与缺乏FTF1拷贝的菌株(弱毒力菌株)相似的寄主植物定殖模式。FTF2的基因敲除也导致毒力降低,但程度较小。这些结果证明了FTF基因扩展,主要是FTF1旁系同源物,在尖孢镰刀菌毒力调节中的作用,并表明效应子表达的调控是其中涉及的机制。