Genetic Engineering Research Center, School of Life Sciences, Chongqing University, Chongqing, People's Republic of China.
Chongqing Engineering Research Center for Fungal Insecticide, Chongqing, People's Republic of China.
PLoS Pathog. 2024 Jul 30;20(7):e1012431. doi: 10.1371/journal.ppat.1012431. eCollection 2024 Jul.
Reactive carbonyl and oxygen species (RCS/ROS), often generated as metabolic byproducts, particularly under conditions of pathology, can cause direct damage to proteins, lipids, and nucleic acids. Glyoxal oxidases (Gloxs) oxidize aldehydes to carboxylic acids, generating hydrogen peroxide (H2O2). Although best characterized for their roles in lignin degradation, Glox in plant fungal pathogens are known to contribute to virulence, however, the mechanism underlying such effects are unclear. Here, we show that Glox in the insect pathogenic fungus, Metarhizium acridum, is highly expressed in mycelia and during formation of infection structures (appressoria), with the enzyme localizing to the cell membrane. MaGlox targeted gene disruption mutants showed RCS and ROS accumulation, resulting in cell toxicity, induction of apoptosis and increased autophagy, inhibiting normal fungal growth and development. The ability of the MaGlox mutant to scavenge RCS was significantly reduced, and the mutant exhibited increased susceptibility to aldehydes, oxidative and cell wall perturbing agents but not toward osmotic stress, with altered cell wall contents. The ΔMaGlox mutant was impaired in its ability to penetrate the host cuticle and evade host immune defense resulting in attenuated pathogenicity. Overexpression of MaGlox promoted fungal growth and conidial germination, increased tolerance to H2O2, but had little to other phenotypic effects. Transcriptomic analyses revealed downregulation of genes related to cell wall synthesis, conidiation, stress tolerance, and host cuticle penetration in the ΔMaGlox mutant. These findings demonstrate that MaGlox-mediated scavenging of RCS is required for virulence, and contributes to normal fungal growth and development, stress resistance.
活性羰基和氧自由基(RCS/ROS)通常作为代谢副产物产生,尤其是在病理条件下,会对蛋白质、脂质和核酸造成直接损伤。醛氧化酶(Gloxs)将醛氧化为羧酸,生成过氧化氢(H2O2)。尽管 Glox 最常被认为在木质素降解中发挥作用,但植物真菌病原体中的 Glox 被认为有助于毒力,然而,这种效应的机制尚不清楚。在这里,我们表明昆虫病原真菌绿僵菌中的 Glox 在菌丝体和侵染结构(附着胞)形成过程中高度表达,该酶定位于细胞膜上。MaGlox 靶向基因敲除突变体显示 RCS 和 ROS 积累,导致细胞毒性、细胞凋亡和自噬增加,抑制正常真菌生长和发育。MaGlox 突变体清除 RCS 的能力显著降低,突变体对醛类、氧化和细胞壁破坏剂的敏感性增加,但对渗透压胁迫的敏感性降低,细胞壁含量发生改变。ΔMaGlox 突变体穿透宿主表皮和逃避宿主免疫防御的能力受损,导致毒力减弱。MaGlox 的过表达促进了真菌的生长和分生孢子的萌发,增加了对 H2O2 的耐受性,但对其他表型效应影响不大。转录组分析显示,ΔMaGlox 突变体中与细胞壁合成、分生孢子形成、应激耐受和宿主表皮穿透相关的基因下调。这些发现表明,MaGlox 介导的 RCS 清除对于毒力是必需的,并有助于正常的真菌生长和发育、应激耐受。