Fungicide-driven evolution and molecular basis of multidrug resistance in field populations of the grey mould fungus Botrytis cinerea.
Fungicide-driven evolution and molecular basis of multidrug resistance in field populations of the grey mould fungus Botrytis cinerea.
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DOI：
10.1371/journal.ppat.1000696
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发表时间：
2009-12
期刊：
影响因子：
6.7
通讯作者：
Hahn M
中科院分区：
文献类型：
--
作者：
Kretschmer M;Leroch M;Mosbach A;Walker AS;Fillinger S;Mernke D;Schoonbeek HJ;Pradier JM;Leroux P;De Waard MA;Hahn M
灰霉菌导致世界范围内重要的商业水果、蔬菜和观赏植物的损失。杀菌剂治疗对疾病控制是有效的，但承担着产生抗药性的风险。真菌的主要耐药机制是靶蛋白修饰导致药物结合力降低。由外排活性增加引起的多重耐药(MDR)在人类病原菌中很常见，但在植物病原菌中却很少被报道。对法国和德国经杀菌剂处理的葡萄园的田间分离物的抗药性年度监测显示，具有三种不同MDR表型的灰霉病菌田间种群迅速增加。所有MDR菌株都表现出杀菌剂外排活性增加和外排转运蛋白基因过表达。与由于转录因子突变而导致的念珠菌临床多药耐药菌株相似，所有MDR1菌株都被证明在控制ABC转运蛋白AtrB基因的转录因子(MRr1)中存在激活突变。MDR2菌株通过插入反转录转座子衍生序列，在主要促进因子超家族转运蛋白基因mfsM2的启动子区域发生了独特的重排。携带相同重排mfsM2等位基因的MDR2菌株可能已经从法国葡萄酒产区迁徙到德国。基因敲除突变体和过表达突变体的表型证实了atrB、mrr1和mfsM2的作用。有性杂交证实，mrr1和mfsM2突变的组合导致MDR3菌株具有更高的广谱耐药性。田间试验表明，一株MDR3菌株可通过杀菌剂处理对敏感菌株进行筛选。我们的数据首次证明，在农业环境中的一种主要植物病原体中，MDR种群的流行率、传播和分子基础不断上升。这些种群将增加灰霉菌腐烂的风险，并阻碍当前杀菌剂抗性管理战略的有效性。细菌和真菌病原体在人类和植物中都会引起疾病。抗生素和杀菌剂被用于疾病控制，但微生物能够通过突变迅速适应这些药物。多重耐药(MDR)在人类病原体中得到了很好的研究，并导致了越来越多的抗生素治疗问题。在商业葡萄园持续使用杀菌剂的推动下，自20世纪90年代中期以来，法国葡萄园出现了三种快速增长的灰霉菌灰霉菌多药抗药性种群。结合生理学、分子和遗传学技术，我们证明了这些多药耐药表型与药物外排活性增加和两个外排转运体的过度表达相关。仅两种类型的突变，一种是控制药物外流的调节蛋白，另一种是外流转运体本身的基因，就足以解释三种MDR表型。我们还提供了证据表明，法国耐多药菌株的一个亚群已经向东迁移到德国葡萄酒产区。我们预计，随着多重抗性菌株的不断选育，田间灰霉病的化学防治将变得越来越困难。
The grey mould fungus Botrytis cinerea causes losses of commercially important fruits, vegetables and ornamentals worldwide. Fungicide treatments are effective for disease control, but bear the risk of resistance development. The major resistance mechanism in fungi is target protein modification resulting in reduced drug binding. Multiple drug resistance (MDR) caused by increased efflux activity is common in human pathogenic microbes, but rarely described for plant pathogens. Annual monitoring for fungicide resistance in field isolates from fungicide-treated vineyards in France and Germany revealed a rapidly increasing appearance of B. cinerea field populations with three distinct MDR phenotypes. All MDR strains showed increased fungicide efflux activity and overexpression of efflux transporter genes. Similar to clinical MDR isolates of Candida yeasts that are due to transcription factor mutations, all MDR1 strains were shown to harbor activating mutations in a transcription factor (Mrr1) that controls the gene encoding ABC transporter AtrB. MDR2 strains had undergone a unique rearrangement in the promoter region of the major facilitator superfamily transporter gene mfsM2, induced by insertion of a retrotransposon-derived sequence. MDR2 strains carrying the same rearranged mfsM2 allele have probably migrated from French to German wine-growing regions. The roles of atrB, mrr1 and mfsM2 were proven by the phenotypes of knock-out and overexpression mutants. As confirmed by sexual crosses, combinations of mrr1 and mfsM2 mutations lead to MDR3 strains with higher broad-spectrum resistance. An MDR3 strain was shown in field experiments to be selected against sensitive strains by fungicide treatments. Our data document for the first time the rising prevalence, spread and molecular basis of MDR populations in a major plant pathogen in agricultural environments. These populations will increase the risk of grey mould rot and hamper the effectiveness of current strategies for fungicide resistance management. Bacterial and fungal pathogens cause diseases in humans and plants alike. Antibiotics and fungicides are used for disease control, but the microbes are able to adapt quickly to these drugs by mutation. Multiple drug resistance (MDR) is well investigated in human pathogens and causes increasing problems with antibiotic therapy. Driven by the continuous use of fungicides in commercial vineyards, three types of rapidly increasing multidrug resistant populations of the grey mould fungus Botrytis cinerea have appeared in French vineyards since the mid 1990s. Using a combination of physiological, molecular and genetic techniques, we demonstrate that these MDR phenotypes are correlated with increased drug efflux activity and overexpression of two efflux transporters. Just two types of mutations, one in a regulatory protein that controls drug efflux, and the other in the gene for an efflux transporter itself, are sufficient to explain the three MDR phenotypes. We also provide evidence that a subpopulation of the French MDR strains has migrated eastward into German wine-growing regions. We anticipate that by continuous selection of multi-resistant strains, chemical control of grey mould in the field will become increasingly difficult.
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DOI：
10.1007/s004380051210
发表时间：
2000-05-01
期刊：
MOLECULAR AND GENERAL GENETICS
影响因子：
--
作者：
Kaneko, I;Tanaka, A;Tsuge, T
通讯作者：
Tsuge, T
影响因子：
2
作者：
Kretschmer, M.;Hahn, M.
通讯作者：
Hahn, M.
影响因子：
2.8
作者：
Coste, Alix T.;Ramsdale, Mark;Sanglard, Dominique
通讯作者：
Sanglard, Dominique
影响因子：
4.1
作者：
Leroux, P;Fritz, R;Chapeland, F
通讯作者：
Chapeland, F
影响因子：
4.4
作者：
Hayashi, K;Schoonbeek, HJ;De Waard, MA
通讯作者：
De Waard, MA