线粒体体征和亚细胞成像提供了深入了解卡巴龙对<i>革兰氏菌</i>的抗真菌机制的研究。<i>小麦</i>,Journal of Agricultural and Food Chemistry

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线粒体体征和亚细胞成像提供了深入了解卡巴龙对革兰氏菌的抗真菌机制的研究。小麦
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2017-12-20 00:00:00 , DOI: 10.1021/acs.jafc.7b03913
Lanying Wang _{1,

2} , Yunfei Zhang ₁ , Delong Wang ₁ , Mei Wang ₁ , Yong Wang ₁ , Juntao Feng _{1,

3}

Affiliation

Carabrone是一种植物性双环倍半萜内酯，具有广谱抗真菌活性，对毁灭性植物病原体Gaeumannomyces graminis var特别有效。小麦（Ggt）。但是，卡巴隆对Ggt的抗真菌机制仍不清楚。这项研究的主要目的是研究Carabrone在Ggt中的亚细胞定位，以更好地了解其作用机理。当Ggt暴露于carabrone（EC ₅₀值（28.45μg/ mL）持续7天，观察到线粒体浓度下降以及线粒体结构出现一些明显的变化，包括朦胧的轮廓，髓质过渡，不清楚的沉淀物的过多积累和液泡变性，表明碳纤维可能起作用直接在线粒体上。因此，设计并合成了荧光共轭物（TTY），作为碳水化合物的替代品，具有对Ggt具有相当的抗真菌活性（EC ₅₀33.68μg/ mL）。另外，还通过免疫小鼠制备了对甲萘醌特异且具有高滴度（256 000）的多克隆抗体。随后，应用了两种成像技术，即使用荧光偶联物（FC）和免疫荧光（IF）来确定甲萘醌的亚细胞定位。FC和IF荧光信号均显示其线粒体定位，FC的皮尔逊系数为0.83，IF的皮尔逊系数为0.86。这些结果表明，甲萘醌可通过干扰线粒体功能发挥其对Ggt的抗真菌活性。

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Mitochondrial Signs and Subcellular Imaging Provide Insight into the Antifungal Mechanism of Carabrone against Gaeumannomyces graminis var. tritici

Carabrone, a botanical bicyclic sesquiterpenic lactone, has broad-spectrum antifungal activity and is particularly efficient against the devastating phytopathogen Gaeumannomyces graminis var. tritici (Ggt). The antifungal mechanism of carabrone against Ggt, however, remains unclear. The main objective of this study was to investigate the subcellular localization of carabrone in Ggt to gain a better understanding of its mechanism of action. When Ggt was exposed to carabrone (EC₅₀ value of 28.45 μg/mL) for 7 days, a decline in mitochondrial concentration together with some obvious alternations in mitochondrial structure, including hazy outlines, medullary transitions, excess accumulation of unclear settlings, and vacuolar degeneration, were observed, indicating that carbrone may act on the mitochondria directly. A fluorescent conjugate (TTY) was thus designed and synthesized as a surrogate of carabrone that possessed comparable antifungal activity against Ggt (EC₅₀ of 33.68 μg/mL). Additionally, a polyclonal antibody specific to carabrone and with a high titer (256 000) was also prepared by immunizing mice. Subsequently, two imaging techniques, the use of the fluorescent conjugate (FC) and immunofluorescence (IF), were applied to determine the subcellular localization of carabrone. Both FC and IF fluorescent signals demonstrated its mitochondrial localization with a Pearson’s coefficient of 0.83 for FC and 0.86 for IF. These results imply that carabrone exerts its antifungal activity against Ggt by interfering with mitochondrial function.

更新日期：2017-12-20

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