Sella Luca, Castiglioni Carla, Paccanaro Maria Chiara, Janni Michela, Schäfer Wilhelm, D'Ovidio Renato, Favaron Francesco
1 Dipartimento Territorio e Sistemi Agro-Forestali (TeSAF), Research group in Plant Pathology, Università di Padova, Viale dell'Università 16, 35020 Legnaro (PD), Italy;
2 Dipartimento di Scienze e tecnologie per l'Agricoltura, le Foreste, la Natura e l'Energia, (DAFNE), Università della Tuscia, Via S. Camillo de Lellis snc, 01100 Viterbo, Italy; and.
Mol Plant Microbe Interact. 2016 Apr;29(4):258-67. doi: 10.1094/MPMI-07-15-0174-R. Epub 2016 Mar 9.
The genome of Fusarium graminearum, the causal agent of Fusarium head blight of wheat, contains two putative pectin methylesterase (PME)-encoding genes. However, when grown in liquid culture containing pectin, F. graminearum produces only a single PME, which was purified and identified. Its encoding gene, expressed during wheat spike infection, was disrupted by targeted homologous recombination. Two Δpme mutant strains lacked PME activity but were still able to grow on highly methyl-esterified pectin even though their polygalacturonase (PG) activity showed a reduced capacity to depolymerize this substrate. The enzymatic assays performed with purified F. graminearum PG and PME demonstrated an increase in PG activity in the presence of PME on highly methyl-esterified pectin. The virulence of the mutant strains was tested on Triticum aestivum and Triticum durum spikes, and a significant reduction in the percentage of symptomatic spikelets was observed between 7 and 12 days postinfection compared with wild type, demonstrating that the F. graminearum PME contributes to fungal virulence on wheat by promoting spike colonization in the initial and middle stages of infection. In contrast, transgenic wheat plants with increased levels of pectin methyl esterification did not show any increase in resistance to the Δpme mutant, indicating that the infectivity of the fungus relies only to a certain degree on pectin degradation.
禾谷镰刀菌是小麦赤霉病的病原菌,其基因组包含两个假定的果胶甲基酯酶(PME)编码基因。然而,当在含有果胶的液体培养基中生长时,禾谷镰刀菌仅产生一种PME,该酶已被纯化和鉴定。其编码基因在小麦穗感染期间表达,通过靶向同源重组被破坏。两个Δpme突变菌株缺乏PME活性,但仍能够在高度甲酯化的果胶上生长,尽管它们的多聚半乳糖醛酸酶(PG)活性显示出对该底物解聚能力的降低。用纯化的禾谷镰刀菌PG和PME进行的酶活性测定表明,在高度甲酯化的果胶上,PME存在时PG活性增加。在普通小麦和硬粒小麦穗上测试了突变菌株的毒力,与野生型相比,在感染后7至12天观察到有症状小穗的百分比显著降低,这表明禾谷镰刀菌PME通过在感染的初始和中期促进穗定殖,对小麦上的真菌毒力有贡献。相反,果胶甲酯化水平增加的转基因小麦植株对Δpme突变体没有表现出任何抗性增加,这表明真菌的感染性仅在一定程度上依赖于果胶降解。