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Synergistic antimicrobial effect and mode of action of palmarosa oil-loaded nanoemulsion and citric acid against Pectobacterium carotovorum.

ABSTRACT: The synergistic antimicrobial activity of palmarosa oil (Cymbopogon martini, PO)-loaded nanoemulsion (PO-NE) and citric acid (CA) against Pectobacterium, the major pathogen for soft-rot disease, was evaluated. The combination of PO-NE and CA (PO-NE + CA) significantly improved the storage stability of PO-NE at 30 °C. Compared to the anti-Pectobacterium activity of alone, PO-NE + CA reduced the minimum inhibitory concentration (MIC) by 1/4 and 1/2, respectively. Bactericidal efficacy of PO-NE + CA against P. carotovorum PCC3 was similar of PO-NE alone in the MIC in time-kill kinetic assay. PO-NE treatment mainly influenced membrane integrity, while CA treatment strongly stimulated intracellular ATP depletion. This synergistic combination effectively reduced the use of PO-NE, imparting a strong flavor note without sacrificing the antimicrobial efficacy against Pectobacterium.

Supplementary information

The online version contains supplementary material available at 10.1007/s10068-022-01217-6.

SUBMITTER: Park JH

PROVIDER: S-EPMC10082693 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature

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Synergistic antimicrobial effect and mode of action of palmarosa oil-loaded nanoemulsion and citric acid against Pectobacterium carotovorum.

Park Ji-Hwa JH Kim Sangbin S Chang Yoonjee Y Imm Jee-Young JY

Food science and biotechnology 20221208 6

The synergistic antimicrobial activity of palmarosa oil (Cymbopogon martini, PO)-loaded nanoemulsion (PO-NE) and citric acid (CA) against Pectobacterium, the major pathogen for soft-rot disease, was evaluated. The combination of PO-NE and CA (PO-NE + CA) significantly improved the storage stability of PO-NE at 30 °C. Compared to the anti-Pectobacterium activity of alone, PO-NE + CA reduced the minimum inhibitory concentration (MIC) by 1/4 and 1/2, respectively. Bactericidal ...[more]

PMID: 37041810

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Project description:Pectobacterium carotovorum subsp. carotovorum is a necrotrophic plant pathogen that secretes plant cell wall-degrading enzymes (PCWDEs) that cause soft rot disease in various crops. Bacteriophages have been under consideration as harmless antibacterial agents to replace antibiotics and copper-based pesticides. However, the emergence of bacteriophage resistance is one of the main concerns that should be resolved for practical phage applications. In this study, we developed a phage cocktail with three lytic phages that recognize colanic acid (phage POP12) or flagella (phages POP15 and POP17) as phage receptors to minimize phage resistance. The phage cocktail effectively suppressed the emergence of phage-resistant P. carotovorum subsp. carotovorum compared with single phages in in vitro challenge assays. The application of the phage cocktail to napa cabbage (Brassica rapa subsp. pekinensis) resulted in significant growth retardation of P. carotovorum subsp. carotovorum (P < 0.05) and prevented the symptoms of soft rot disease. Furthermore, phage cocktail treatments of young napa cabbage leaves in a greenhouse environment indicated effective prevention of soft rot disease compared to that in the nonphage negative control. We isolated 15 phage-resistant mutants after a phage cocktail treatment to assess the virulence-associated phenotypes compared to those of wild-type (WT) strain Pcc27. All mutants showed reduced production of four different PCWDEs, leading to lower levels of tissue softening. Ten of the 15 phage-resistant mutants additionally exhibited decreased swimming motility. Taken together, these results show that the phage cocktail developed here, which targets two different types of phage receptors, provides an effective strategy for controlling P. carotovorum subsp. carotovorum in agricultural products, with a potential ability to attenuate P. carotovorum subsp. carotovorum virulence. IMPORTANCE Pectobacterium carotovorum subsp. carotovorum is a phytopathogen that causes soft rot disease in various crops by producing plant cell wall-degrading enzymes (PCWDEs). Although antibiotics and copper-based pesticides have been extensively applied to inhibit P. carotovorum subsp. carotovorum, the emergence of antibiotic-resistant bacteria and demand for harmless antimicrobial products have emphasized the necessity of finding alternative therapeutic strategies. To address this problem, we developed a phage cocktail consisting of three P. carotovorum subsp. carotovorum-specific phages that recognize colanic acids and flagella of P. carotovorum subsp. carotovorum. The phage cocktail treatments significantly decreased P. carotovorum subsp. carotovorum populations, as well as soft rot symptoms in napa cabbage. Simultaneously, they resulted in virulence attenuation in phage-resistant P. carotovorum subsp. carotovorum, which was represented by decreased PCWDE production and decreased flagellum-mediated swimming motility. These results suggested that preparations of phage cocktails targeting multiple receptors would be an effective approach to biocontrol of P. carotovorum subsp. carotovorum in crops.

Dataset Information

Synergistic antimicrobial effect and mode of action of palmarosa oil-loaded nanoemulsion and citric acid against Pectobacterium carotovorum.

Supplementary information

Publications

Synergistic antimicrobial effect and mode of action of palmarosa oil-loaded nanoemulsion and citric acid against <i>Pectobacterium carotovorum</i>.

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