The role of oxalic acid in Clarireedia jacksonii virulence and development on creeping bentgrass.

IF 2.6 2区农林科学 Q2 PLANT SCIENCES Phytopathology Pub Date : 2024-08-15 DOI:10.1094/PHYTO-03-24-0094-R

Daowen Huo, Nathaniel M Westrick, Ashley Nelson, Mehdi Kabbage, Paul Koch

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Abstract

Dollar spot is a destructive foliar disease of amenity turfgrass caused by the fungus Clarireedia spp., and mainly Clarireedia jacksonii on the northern US region's cool-season grass. Oxalic acid (OA) is an important pathogenicity factor in related fungal plant pathogens such as Sclerotinia sclerotiorum, however, the role of OA in the pathogenic development of C. jacksonii remains unclear due to its recalcitrance to genetic manipulation. To overcome these challenges, a CRISPR/Cas9-mediated homologous recombination approach was developed. Using this novel approach, the oxaloacetate acetylhydrolase (Oah) gene that is required for the biosynthesis of OA was deleted from C. jacksonii wild-type stain. Two independent knockout mutants, ΔCjoah-1 and ΔCjoah-2, were generated and inoculated on potted creeping bentgrass along with a wild-type isolate (WT) and a genome sequenced isolate LWC-10. After 12 days, bentgrass inoculated with the mutants ΔCjoah-1 and ΔCjoah-2 exhibited 59.41% lower dollar spot severity compared to the WT and LWC-10 isolates. Oxalic acid production and environmental acidification were significantly reduced in both mutants when compared to the WT and LWC-10. Surprisingly, stromal formation was also severely undermined in the mutants in vitro, suggesting a critical developmental role of OA independent of plant infection. These results demonstrate that OA plays a significant role in C. jacksonii virulence and provide novel directions for future management of dollar spot.

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草酸在 Clarireedia jacksonii 对匍匐翦股颖的毒力和发育过程中的作用。

美元斑是由真菌 Clarireedia spp.（主要是 Clarireedia jacksonii）在美国北部冷季型草坪上引起的一种破坏性草坪叶面病害。草酸（OA）是相关真菌植物病原体（如硬皮病菌）的重要致病因子，但由于其对遗传操作的顽固性，OA 在 C. jacksonii 的致病发展中的作用仍不清楚。为了克服这些挑战，我们开发了一种 CRISPR/Cas9 介导的同源重组方法。利用这种新方法，从 C. jacksonii 野生型染色中删除了 OA 生物合成所需的草酰乙酸乙酰水解酶（Oah）基因。产生了两个独立的基因敲除突变体：ΔCjoah-1 和 ΔCjoah-2，并将其与野生型分离株（WT）和基因组测序分离株 LWC-10 一起接种到盆栽匍匐翦股颖上。12 天后，接种了突变体 ΔCjoah-1 和 ΔCjoah-2 的翦股颖与 WT 和 LWC-10 株系相比，美元斑的严重程度降低了 59.41%。与 WT 和 LWC-10 相比，这两个突变体的草酸产量和环境酸化程度都明显降低。令人惊讶的是，突变体在体外的基质形成也受到了严重破坏，这表明 OA 在植物感染之外还起着关键的发育作用。这些结果表明，OA 在 C. jacksonii 的毒力中起着重要作用，并为未来美元斑的管理提供了新的方向。

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来源期刊

Phytopathology 生物-植物科学

CiteScore

5.90

自引率

9.40%

发文量

505

审稿时长

4-8 weeks

期刊介绍： Phytopathology publishes articles on fundamental research that advances understanding of the nature of plant diseases, the agents that cause them, their spread, the losses they cause, and measures that can be used to control them. Phytopathology considers manuscripts covering all aspects of plant diseases including bacteriology, host-parasite biochemistry and cell biology, biological control, disease control and pest management, description of new pathogen species description of new pathogen species, ecology and population biology, epidemiology, disease etiology, host genetics and resistance, mycology, nematology, plant stress and abiotic disorders, postharvest pathology and mycotoxins, and virology. Papers dealing mainly with taxonomy, such as descriptions of new plant pathogen taxa are acceptable if they include plant disease research results such as pathogenicity, host range, etc. Taxonomic papers that focus on classification, identification, and nomenclature below the subspecies level may also be submitted to Phytopathology.