Institute of Microbiology, College of Life Science, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
PLoS One. 2013 Apr 15;8(4):e62179. doi: 10.1371/journal.pone.0062179. Print 2013.
Multidrug resistance (MDR) confers agrochemical compatibility to fungal cells-based mycoinsecticdes but mechanisms involved in MDR remain poorly understood for entomopathogenic fungi, which have been widely applied as biocontrol agents against arthropod pests. Here we characterized the functions of five ATP-binding cassette (ABC) transporters, which were classified to the subfamilies ABC-B (Mdr1), ABC-C (Mrp1) and ABC-G (Pdr1, Pdr2 and Pdr5) and selected from 54 full-size ABC proteins of Beauveria bassiana based on their main domain architecture, membrane topology and transcriptional responses to three antifungal inducers. Disruption of each transporter gene resulted in significant reduction in resistance to four to six of eight fungicides or antifungal drugs tested due to their differences in structure and function. Compared with wild-type and complemented (control) strains, disruption mutants of all the five transporter genes became significantly less tolerant to the oxidants menadione and H₂O₂ based on 22-41% and 10-31% reductions of their effective concentrations required for the suppression of 50% colony growth at 25°C. Under a standardized spray, the killing actions of ΔPdr5 and ΔMrp1 mutants against Spodoptera litura second-instar larvae were delayed by 59% and 33% respectively. However, no significant virulence change was observed in three other delta mutants. Taken together, the examined five ABC transporters contribute differentially to not only the fungal MDR but antioxidant capability, a phenotype rarely associated with ABC efflux pumps in previous reports; at least some of them are required for the full virulence of B. bassiana, thereby affecting the fungal biocontrol potential. Our results indicate that ABC pump-dependent MDR mechanisms exist in entomopathogenic fungi as do in yeasts and human and plant pathogenic fungi.
多药耐药性 (MDR) 使真菌细胞基生虫剂具有农用化学品相容性,但昆虫病原真菌的 MDR 相关机制仍知之甚少,这些真菌已被广泛用作防治节肢动物害虫的生物防治剂。在这里,我们对五个 ABC 转运蛋白的功能进行了表征,这些蛋白根据其主要结构域结构、膜拓扑结构和对三种抗真菌诱导剂的转录响应,分为 ABC-B(Mdr1)、ABC-C(Mrp1)和 ABC-G(Pdr1、Pdr2 和 Pdr5)亚家族。每个转运蛋白基因的破坏导致对八种杀真菌剂或抗真菌药物中的四种至六种的耐药性显著降低,这是由于它们在结构和功能上的差异。与野生型和互补(对照)菌株相比,所有五个转运蛋白基因的敲除突变体对氧化剂甲萘醌和 H₂O₂的耐受性显著降低,这两种氧化剂的有效浓度分别降低了 22-41%和 10-31%,以抑制 25°C 下 50%菌落生长所需的浓度。在标准化喷雾下,ΔPdr5 和 ΔMrp1 突变体对斜纹夜蛾二龄幼虫的杀伤作用分别延迟了 59%和 33%。然而,在其他三个 delta 突变体中没有观察到明显的毒力变化。总之,研究的五个 ABC 转运蛋白不仅对真菌的 MDR,而且对抗氧化能力有不同的贡献,这是以前的报道中很少与 ABC 外排泵相关的表型;至少其中一些是白僵菌完全毒力所必需的,从而影响了真菌的生物防治潜力。我们的结果表明,与酵母和人类及植物病原真菌一样,昆虫病原真菌中也存在 ABC 泵依赖性 MDR 机制。