Fungicide mode of action and resistance development in crop pathogenic fungi

杀菌剂的作用方式及作物病原真菌抗性发展

• 批准号: BB/P018335/1
• 负责人: Gero Steinberg
• 金额: $ 67.02万
• 依托单位: UNIVERSITY OF EXETER
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FP018335%2F1
• 关键词: Fungicide; mode; action; resistance; development

Continuous growth of the world population comes with increasing demand for food. As a consequence, agricultural practises have intensified. The large monocultures of important crop plant, such as maize and wheat, provide a rich food source for plant pathogenic fungi. In fact, fungi are the biggest challenge for our food security. Amongst the most devastating crop pathogens are the corn smut fungus (Ustilago maydis) and the Septoria tritici wheat blotch fungus (Zymoseptoria tritici/Mycosphaerella graminicola). Our farmers fight these fungi by spraying anti-fungal chemistries, so-called fungicides. These usually target the fungal cell, whilst showing little toxicity to the crop. To understand how a fungicide acts, detailed studies of the physiology of the fungal cell in the presence of the antifungal chemistry are required. The outcome is an understanding of the impact of the fungicide on the fungus, which describes is "mode of action" (MoA). In previous times, such studies were restricted by technical limitations. Consequently, the MoA of many fungicides is either not known or our knowledge is fragmentary. The recent development of live cell imaging techniques and tools for fungal pathogens allows visualisation of fungal cell in the presence of a fungicide. The PI's laboratory is world-leading in live cell imaging of fungal pathogens. The project aims to use this modern approach to monitor fungicide-induced changes in cells of U. maydis and Z. tritici. In a preliminary study, we provide a proof of principle study with the fungicide dodine, used to control fungal disease on apples. This revealed a novel MoA for dodine and illustrates the power of this approach. In the first part of the project, we will use the cell markers and live cell imaging to investigate the MoAs of 12 major fungicides that cover the most economically important fungicide groups in global use. Fungi have the ability to adapt to fungicides. Similar to bacteria, they can develop resistance, which is of high economic importance, as it renders the respective fungicide useless. Resistance can be achieved by modification of the protein that the fungicide binds to and inhibits. Alternatively, it can be achieved by other, much less understood ways, including an increased activity of cellular pumps that remove the fungicide from the fungal cell. Our understanding of the mechanism by which fungi develop resistance is limited to local changes in the genetic information of the pathogen. However, the ability to quickly sequence the entire genomic information of a resistant fungus opens the opportunity to look for all changes, accompanied by the appearance of fungicide resistance. We have developed techniques to generate fungicide resistant fungi in our laboratory (so far ~150 fungal cell lines (=strains), resistant against most of the 12 major fungicides). We have sequenced the genomic information of 15 of these fungal strains and already found strong indication for an unexpected and new mechanism conferring resistance in U. maydis. We aim to increase the number of resistant strains and extend the unbiased approach of sequencing entire genomes of resistant fungi. This, and the subsequent analysis of mutated genes, promises novel insight into the molecular adaptation of fungal pathogens to fungicides. It is increasingly difficult to control infections by the wheat pathogen Z. tritici. This is due to the appearance of resistance strains against the major fungicide classes. It was speculated that the ability to adapt to fungicides is due to the presence of 8 "dispensable" chromosomes. These are not essential for survival of the pathogen and, therefore, can be lost during cell division. In this part of the project, we will generate Z. tritici strains that are identical, but which lack individual dispensable chromosomes. We will expose these to fungicides and analyse the ability to develop resistance against the anti-fungal chemistries.

世界人口的持续增长伴随着对粮食需求的增加。因此，农业的衰退加剧了。玉米、小麦等重要作物的大面积单作为植物病原真菌提供了丰富的食物来源。事实上，真菌是我们粮食安全面临的最大挑战。最具破坏性的农作物病原体是玉米黑穗病真菌（玉米黑粉菌）和小麦壳针孢小麦斑点真菌（小麦壳针孢菌/草生球腔菌）。我们的农民通过喷洒抗真菌化学物质，即所谓的杀真菌剂来对抗这些真菌。它们通常以真菌细胞为目标，同时对作物几乎没有毒性。为了了解杀真菌剂如何起作用，需要对真菌细胞在抗真菌化学物质存在下的生理学进行详细研究。结果是了解杀真菌剂对真菌的影响，其描述为“作用模式”（MoA）。在过去，这种研究受到技术限制。因此，许多杀真菌剂的MoA要么是未知的，要么是我们的知识是零碎的。真菌病原体的活细胞成像技术和工具的最新发展允许在杀真菌剂存在下可视化真菌细胞。PI的实验室在真菌病原体的活细胞成像方面处于世界领先地位。该项目旨在使用这种现代方法来监测杀菌剂诱导的U。maydis和玉米Z.三个月。在一项初步研究中，我们提供了一个证明的原则研究与杀菌剂多丁，用于控制苹果真菌病害。这揭示了dodine的一个新的MoA，并说明了这种方法的力量。在该项目的第一部分，我们将使用细胞标记物和活细胞成像来研究12种主要杀真菌剂的MoA，这些杀真菌剂涵盖了全球使用的最具经济意义的杀真菌剂类别。真菌具有适应杀真菌剂的能力。与细菌类似，它们可以产生抗性，这具有很高的经济重要性，因为它使相应的杀真菌剂无效。抗性可以通过修饰杀真菌剂结合并抑制的蛋白质来实现。或者，它可以通过其他不太了解的方式来实现，包括增加从真菌细胞中去除杀真菌剂的细胞泵的活性。我们对真菌产生耐药性的机制的理解仅限于病原体遗传信息的局部变化。然而，快速测序抗性真菌的整个基因组信息的能力为寻找所有变化提供了机会，伴随着杀菌剂抗性的出现。我们已经开发了在我们的实验室中产生抗杀真菌剂真菌的技术（迄今为止约150种真菌细胞系（=菌株），对12种主要杀真菌剂中的大多数具有抗性）。我们已经对其中15种真菌菌株的基因组信息进行了测序，并发现了一种意想不到的新机制，这种机制赋予了美国的耐药性。玉米粉我们的目标是增加耐药菌株的数量，并扩展对耐药真菌的整个基因组进行测序的无偏方法。这一点，以及随后对突变基因的分析，有望对真菌病原体对杀真菌剂的分子适应性有新的认识。控制小麦病原菌Z的侵染越来越困难。三个月。这是由于出现了对主要杀真菌剂类别的抗性菌株。据推测，这种对杀菌剂的适应能力是由于存在8条“”染色体。这些对于病原体的生存不是必需的，因此可以在细胞分裂期间丢失。在本项目的这一部分中，我们将生成Z。三倍体菌株是相同的，但缺乏单独的三倍体染色体。我们将这些暴露于杀真菌剂，并分析其对抗真菌化学物质产生抗性的能力。

项目成果

Alloxan disintegrates the plant cytoskeleton and suppresses mlo-mediated powdery mildew resistance.

• DOI: 10.1093/pcp/pcz216
• 发表时间: 2019-11
• 期刊: Plant & cell physiology
• 影响因子: 4.9
• 作者: Hongpo Wu;Weiwei Zhang;M. Schuster;M. Moch;R. Windoffer;G. Steinberg;C. Staiger;R. Panstruga

The fungicide dodine primarily inhibits mitochondrial respiration in Ustilago maydis, but also affects plasma membrane integrity and endocytosis, which is not found in Zymoseptoria tritici.

• DOI: 10.1016/j.fgb.2020.103414
• 发表时间: 2020-09
• 期刊: Fungal genetics and biology : FG & B
• 作者: Schuster M;Steinberg G
• 通讯作者: Steinberg G

Multi-site fungicides suppress banana Panama disease, caused by Fusarium oxysporum f. sp. cubense Tropical Race 4.

• DOI: 10.1371/journal.ppat.1010860
• 发表时间: 2022-10
• 期刊: PLOS PATHOGENS
• 影响因子: 6.7
• 作者: Cannon, Stuart;Kay, William;Kilaru, Sreedhar;Schuster, Martin;Gurr, Sarah Jane;Steinberg, Gero
• 通讯作者: Steinberg, Gero

Fungi, fungicide discovery and global food security.

• DOI: 10.1016/j.fgb.2020.103476
• 发表时间: 2020-11
• 期刊: Fungal genetics and biology : FG & B
• 作者: Steinberg G;Gurr SJ
• 通讯作者: Gurr SJ

Zymoseptoria tritici white-collar complex integrates light, temperature and plant cues to initiate dimorphism and pathogenesis.

• DOI: 10.1038/s41467-022-33183-2
• 发表时间: 2022-09-26
• 期刊: Nature communications
• 影响因子: 16.6