Targeted subcellular localization of a novel fungicidal compound N-(naphthalen-1-yl) phenazine-1-carboxamide inhibiting Rhizoctonia solani

文献类型： 外文期刊

第一作者： Liu, Shuangqing

作者： Liu, Shuangqing;Wen, Ding;Jia, Wenhui;Liao, Xiaolan;Zhang, Ya;Wang, Chong;Pan, Hu

作者机构：

关键词： Rhizoctonia solani; N-(naphthalen-1-yl)phenazine-1-carboxamide; Polyclonal antibody; Subcellular localization

期刊名称：PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY （ 影响因子：4.0； 五年影响因子：4.5 ）

ISSN： 0048-3575

年卷期： 2025 年 213 卷

页码：

收录情况： SCI

摘要： N-(naphthalen-1-yl) phenazine-1-carboxamide (NNPCN) is a novel phenazine derivative that can inhibit Rhizoctonia solani. However, the molecular target of NNPCN is unclear. We constructed a fluorescent marker and a polyclonal antibody for NNPCN and used fluorescent labeling, immune fluorescence, and an immune colloidal gold technique to conduct subcellular localization of the target of NNPCN on R. solani. The results showed that NNPCN could be synthesized from N-(6-aminonaphthalen-1-yl) phenazine-1-carboxamide (NNPCN-NH2) and N-(6-(3-(3 ',6'-Dihydroxy-3-oxo-3 h-spiro [Isobenzofuran-1,9 '-Xanthen]-5-yl) thioureido) naphthalen-1-yl) phenazine-1-carboxamide (NNPPCN-NF), with respective molecular formulas and molecular weights of C23H16N4O, 365, and C44H27N5O6S, 753. The best solvent for NNPPCN-NF was acetone. The maximum excitation wavelength was 457.8 nm, and the emission wavelength was 557.8 nm. NNPCN-NF had an inhibitory effect on rice sheath blight; its median effect concentration (EC50) value of 15.71 mu g/mL was similar to that of the control agent. NNPCN-NF demonstrated specific binding within cells. Fluorescence labeling showed that NNPPCN-NF coincided with the fluorescence images from Mitro tracker red CmxRos; the Pearson's correlation coefficient (PCC) was 0.89, indicating that the compound acted on the mitochondria. A highly valent NNPCN polyclonal antibody was successfully prepared by immunizing rabbits. Immunofluorescence localization showed that Alexa fluor 488 tagged-goat anti-rabbit IgG coincided with the Mito tracker red CMXRos fluorescence images, and the PCC was 0.79, indicating that the target was the mitochondrion. Immunocolloidal gold co-localization showed that NNPCN-NF also acted on the cell wall. The results indicate that the targets of NNPCN in R. solani are the mitochondria and cell walls.

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