A functional analysis of mitochondrial respiratory chain cytochrome bc1 complex in Gaeumannomyces tritici by RNA silencing as a possible target of carabrone,Molecular Plant Pathology

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A functional analysis of mitochondrial respiratory chain cytochrome bc1 complex in Gaeumannomyces tritici by RNA silencing as a possible target of carabrone
Molecular Plant Pathology ( IF 4.8 ) Pub Date : 2020-09-30 , DOI: 10.1111/mpp.12993
Mei Wang _{1,

2} , Xingyu Ren ₁ , Lanying Wang _{1,

3} , Xiang Lu ₁ , Lirong Han ₁ , Xing Zhang _{1,

4} , Juntao Feng _{1,

4}

Affiliation

Gaeumannomyces tritici, an ascomycete soilborne fungus, causes a devastating root disease in wheat. Carabrone, a botanical bicyclic sesquiterpenic lactone, is a promising fungicidal agent that can effectively control G. tritici. However, the mechanism of action of carabrone against G. tritici remains largely unclear. Here, we used immunogold for subcellular localization of carabrone and the results showed that carabrone is subcellularly localized in the mitochondria of G. tritici. We then explored the functional analysis of genes GtCytc₁, GtCytb, and GtIsp of the mitochondrial respiratory chain cytochrome bc₁ complex in G. tritici by RNA silencing as a possible target of carabrone. The results showed that the silenced mutant ∆GtIsp is less sensitive to carabrone compared to ∆GtCytc₁ and ∆GtCytb. Compared with the control, the activities of complex III in all the strains, except ∆GtIsp and carabrone‐resistant isolate 24‐HN‐1, were significantly decreased following treatment with carabrone at EC₂₀ and EC₈₀ in vitro (40%–50% and 70%–80%, respectively). The activities of mitochondrial respiratory chain complex III and the mitochondrial respiration oxygen consumption rates in all the strains, except ∆GtIsp and 24‐HN‐1, were higher with respect to the control when treated with carabrone at EC₂₀ in vivo. The rates of mitochondrial respiration of all strains, except ∆GtIsp, were significantly inhibited following treatment with carabrone at EC₈₀ (ranging from 57% to 81%). This study reveals that the targeting of the iron–sulphur protein encoded by GtIsp is highly sensitive to carabrone and provides a direction for the research of carabrone's target.

中文翻译：

通过 RNA 沉默作为卡拉布隆的可能靶标对小麦全麦酵母线粒体呼吸链细胞色素 bc1 复合物进行功能分析

小麦全芽孢 (Gaeumannomyces tritici ) 是一种子囊菌土传真菌，可引起小麦毁灭性的根部病害。 Carabrone 是一种植物双环倍半萜内酯，是一种很有前景的杀菌剂，可以有效控制小麦小麦。然而，卡拉布隆对抗小麦小麦的作用机制仍不清楚。在这里，我们使用免疫金对 carabrone 进行亚细胞定位，结果表明 carabrone 亚细胞定位于小麦小麦的线粒体中。然后，我们通过 RNA 沉默作为 carabrone 的可能靶点，探索了对小麦线粒体呼吸链细胞色素bc ₁复合物的基因GtCytc ₁ 、 GtCytb和GtIsp的功能分析。结果表明，与 ΔGtCytc ₁和 ΔGtCytb 相比，沉默突变体 ΔGtIsp 对卡拉布朗的敏感性较低。与对照相比，在体外 EC ₂₀和 EC ₈₀条件下用 Carabrone 处理后，除 ΔGtIsp 和耐 Carabrone 分离株 24-HN-1 外，所有菌株中复合物 III 的活性均显着降低（40%–50%）。和 70%–80%）。当在 EC ₂₀体内用卡拉布隆处理时，除 ΔGtIsp 和 24-HN-1 外，所有菌株的线粒体呼吸链复合物 III 的活性和线粒体呼吸耗氧率均高于对照。除 ΔGtIsp 外，所有菌株的线粒体呼吸速率在 EC ₈₀下用卡拉布隆处理后均受到显着抑制（范围为 57% 至 81%）。该研究揭示了GtIsp编码的铁硫蛋白的靶向性对carabrone高度敏感，为carabrone靶点的研究提供了方向。

更新日期：2020-11-27

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