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. 2022 Nov;133(5):3161-3175.
doi: 10.1111/jam.15774. Epub 2022 Aug 22.

Virulence and biofilm inhibition of 3-methoxycinnamic acid against Agrobacterium tumefaciens

Deng Pan  1   2 , Xing-Yun Wang  1 , Jin-Wei Zhou  3 , Liang Yang  4 , Abbas Khan  5   6 , Dong-Qing Wei  5   6 , Jun-Jian Li  1   2 , Ai-Qun Jia  1   2   7
Affiliations

Virulence and biofilm inhibition of 3-methoxycinnamic acid against Agrobacterium tumefaciens

Deng Pan et al. J Appl Microbiol. 2022 Nov.

Abstract

Aims: In the current study the anti-virulence and anti-biofilm activities of the cinnamic acid derivative, 3-methoxycinnamic acid, was investigated against Agrobacterium tumefaciens.

Methods and results: Based on the disc diffusion test and β-galactosidase activity assay, 3-methoxycinnamic acid was shown to interfere with the quorum sensing (QS) system of A. tumefaciens. Crystal violet staining assay, phenol-sulfuric acid method, Bradford protein assay and confocal laser scanning microscopy (CLSM) revealed that the biofilm formation of A. tumefaciens was inhibited after the treatment of 3-methoxycinnamic acid. Employing high-performance liquid chromatography (HPLC) analysis of culture supernatant revealed that the production of 3-oxo-octanoylhomoserine lactone (3-oxo-C8-HSL) decreased concentration-dependently after treatment with 3-methoxycinnamic acid. Swimming and chemotaxis assays also indicated that 3-methoxycinnamic acid had a good effect on reducing the motility and chemotaxis of A. tumefaciens. In addition, the RT-qPCR, molecular docking and simulations further demonstrated that 3-methoxycinnamic acid could competitively inhibit the binding of 3-oxo-C8-HSL to TraR and down-regulate virulence-related genes.

Conclusions: 3-Methoxycinnamic acid is proved to have good anti-virulence and anti-biofilm activities against A. tumefaciens.

Significance and impact of the study: This is the first study that investigates the anti-virulence and anti-biofilm activities of 3-methoxycinnamic acid against A. tumefaciens. With its potential QS-related virulence and biofilm inhibitory activities, 3-methoxycinnamic acid is expected to be developed as a potent pesticide or adjuvant for the prevention and treatment of crown gall caused by A. tumefaciens.

Keywords: Agrobacterium tumefaciens; 3-methoxycinnamic acid; biofilm inhibition; quorum sensing; virulence.

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