Integrated Transcriptome and Proteome Analysis Reveals that the Antimicrobial Griseofulvin Targets Didymella segeticola Beta-Tubulin to Control Tea Leaf Spot

Author:

Huang Hongke12,Li Dongxue1,Jiang Shilong13,Yang Rui13,Yang Yuqing12,Xia Zhongqiu12,Jiang Xinyue1,Zhao Yongtian14,Wang Delu5,Song Baoan1,Chen Zhuo1ORCID

Affiliation:

1. Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Guiyang, Guizhou, P.R. China

2. College of Tea Science, Guizhou University, Guiyang, Guizhou, P.R. China

3. Agricultural College, Guizhou University, Guiyang, Guizhou, P.R. China

4. College of Life Science and Agriculture, Qiannan Normal University for Nationalities, Duyun, Guizhou, P.R. China

5. College of Forestry, Guizhou University, Guiyang, Guizhou, P.R. China

Abstract

Because effective control measures are lacking, tea leaf spot caused by Didymella segeticola results in huge tea ( Camellia sinensis) production losses on tea plantations in Guizhou Province, southwestern China. Screening for natural antimicrobial agents with higher control effects against this pathogen and studying their modes of action may contribute to disease management. Here, Penicillium griseofulvum-derived antimicrobial griseofulvin (GSF) can inhibit the hyphal growth of D. segeticola strain GZSQ-4, with a half-maximal effective concentration of 0.37 μg/ml in vitro and a higher curative efficacy at a lower dose of 25 μg/ml for detached tea twigs. GSF induces deformed and slightly curly hyphae with enlarged ends, with protoplasts agglutinated in the hyphae, and higher numbers of hyphal protuberances. GSF alters hyphal morphology and the subcellular structure's order. The integrated transcriptome and proteome data revealed that the transport of materials in cells, cellular movement, and mitosis were modulated by GSF. Molecular docking indicated that beta-tubulin was the most potent target of GSF, with a binding free energy of −13.59 kcal/mol, and microscale thermophoresis indicated that the dissociation constant (Kd) value of GSF binding to beta-tubulin 1, compared with beta-tubulin 2, was significantly lower. Thus, GSF potentially targets beta-tubulin 1 to disturb the chromosomal separation and fungal mitosis, thereby inhibiting hyphal growth.

Funder

National Key Research and Development Program of China

National Natural Science Foundation of China

China Agriculture Research System

Program of Introducing Talents to Chinese Universities

Publisher

Scientific Societies

Subject

Plant Science,Agronomy and Crop Science

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