Regulation and function of the flavonoid-inducible efflux system, emrR-emrAB, in Agrobacterium tumefaciens C58

Khemthong, Sasimaporn; Nuonming, Puttamas; Dokpikul, Thanittra; Sukchawalit, Rojana; Mongkolsuk, Skorn

doi:10.1007/s00253-019-09899-5

Cited by 9 publications

(5 citation statements)

References 67 publications

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“…t -CNMA upregulated the gsp gene, which is associated with general stress, by 2.5-fold, and 4-nitro CNMA reduced the expression of a heat shock protein ( hspAT2 ). Also, some efflux pump genes, namely, emrA , norM , ifeA , and ifeR , were included ( Palumbo et al, 1998 ; Nuonming et al, 2018 ; Khemthong et al, 2019 ), but the only effect observed was that 4-nitro CNMA upregulated emrA by 3.7-fold ( Figure 8 ).…”

Section: Resultsmentioning

confidence: 99%

Inhibition of growth, biofilm formation, virulence, and surface attachment of Agrobacterium tumefaciens by cinnamaldehyde derivatives

et al. 2022

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Agrobacterium tumefaciens, a soil-borne, saprophytic plant pathogen that colonizes plant surfaces and induces tumors in a wide range of dicotyledonous plants by transferring and expressing its T-DNA genes. The limited availabilities and efficacies of current treatments necessitate the exploration of new anti-Agrobacterium agents. We examined the effects of trans-cinnamaldehyde (t-CNMA) and its derivatives on the cell surface hydrophobicity, exopolysaccharide and exo-protease production, swimming motility on agar, and biofilm forming ability of A. tumefaciens. Based on initial biofilm inhibition results and minimum inhibitory concentration (MIC) data, 4-nitro, 4-chloro, and 4-fluoro CNMAs were further tested. 4-Nitro, 4-chloro, and 4-fluoro CNMA at ≥150 μg/ml significantly inhibited biofilm formation by 94–99%. Similarly, biofilm formation on polystyrene or nylon was substantially reduced by 4-nitro and 4-chloro CNMAs as determined by optical microscopy and scanning electron microscopy (SEM) and 3-D spectrum plots. 4-Nitro and 4-chloro CNMAs induced cell shortening and concentration- and time-dependently reduced cell growth. Virulence factors were significantly and dose-dependently suppressed by 4-nitro and 4-chloro CNMAs (P ≤ 0.05). Gene expressional changes were greater after 4-nitro CNMA than t-CNMA treatment, as determined by qRT-PCR. Furthermore, some genes essential for biofilm formation, motility, and virulence genes significantly downregulated by 4-nitro CNMA. Seed germination of Raphanus sativus was not hindered by 4-nitro or 4-fluoro CNMA at concentrations ≤200 μg/ml, but root surface biofilm formation was severely inhibited. This study is the first to report the anti-Agrobacterium biofilm and anti-virulence effects of 4-nitro, 4-chloro, and 4-fluoro CNMAs and t-CNMA and indicates that they should be considered starting points for the development of anti-Agrobacterium agents.

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Section: Resultsmentioning

confidence: 99%

Inhibition of growth, biofilm formation, virulence, and surface attachment of Agrobacterium tumefaciens by cinnamaldehyde derivatives

et al. 2022

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“…Flavonoids like genistein, apigenin, luteolin, and naringenin were found to be the best inducers of the emrAB operon. These operons are responsible for the synthesis of bacterial cell wall inner and outer membrane proteins (Khemthong et al, 2019). Besides transformation, aztreonam treatment affected root morphology.…”

Section: Discussionmentioning

confidence: 99%

“…These operons synthesize the Agrobacterium inner and outer membrane fusion proteins. Among these flavonoids, quercetin and genistein were found to be more effective ( Khemthong et al, 2019 ). The luteolin accumulation during seed imbibition leads to nodulation in alfalfa by Rhizobium meliloti ( Hartwig and Phillips, 1991 ).…”

Section: Introductionmentioning

confidence: 99%

Aztreonam is a novel chemical inducer that promotes Agrobacteium transformation and lateral root development in soybean

Waqar Khan,

Yang,

et al. 2023

Front. Microbiol.

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Agrobacterium-mediated soybean transformation is the simplest method of gene transfer. However, the low transformation due to the intractable nature of soybean genotypes hinders this process. The use of biochemicals (acetosyringone, cinnamic acid, flavonoids, etc.) plays an important role in increasing soybean transformation. These biochemicals induce chemotaxis and virulence gene activation during the infection process. Here we identified a biochemical, aztreonam (a monobactam), for high agrobacterium-mediated transformation in soybean. The soybean explants from three genotypes were inoculated with A. tumefaciens (GV3101) harboring the pMDC32 vector containing hpt or the GmUbi-35S-GUS vector containing the GUS gene during two separate events. High transient GUS expression was obtained during cotyledon explant culture on MS media supplemented with 2.5 mg/L aztreonam. The aztreonam-treated explants showed high efficiency in transient and stable transformation as compared to the untreated control. The transformation of aztreonam-treated explants during seed imbibition resulted in an average of 21.1% as compared to 13.2% in control by using the pMDC32 vector and 28.5 and 20.7% while using the GUS gene cassette, respectively. Based on these findings, the metabolic analysis of the explant after aztreonam treatment was assessed. The high accumulation of flavonoids was identified during an untargeted metabolic analysis. The quantification results showed a significantly high accumulation of the four compounds, i.e., genistein, apigenin, naringenin, and genistin, in cotyledon explants after 18 hours of aztreonam treatment. Alongside this, aztreonam also had some surprising effects on root elongation and lateral root formation when compared to indole-3-butyric acid (IBA). Our findings were limited to soybeans. However, the discovery of aztreonam and its effect on triggering flavonoids could lead to the potential role of aztreonam in the agrobacterium-mediated transformation of different crops.

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“…Since the operon is capable of handling structurally unrelated drugs/compounds, their intrinsic function was suggested to be the efflux of these chemicals in E. coli (Lomovskaya and Lewis, 1995). Proper regulation of emrR-emrAB operon is essential for the survival of Agrobacterium tumefaciens on exposure to toxins (Khemthong et al, 2019). Hence, it can be concluded that emr locus essentially protects and responsible for the high tolerance of E .coli to chlorpyrifos.…”

Section: Global Comparison Of Differential Gene Expression and Protein Expression On Exposure To Chlorpyrifosmentioning

confidence: 99%

Efflux mediated chlorpyrifos tolerance inEscherichia coliBL21(DE3)

Aswathi

Pandey

Sukumaran

2020

Preprint

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Bacteria are continually challenged with variety of synthetic chemicals/xenobiotics in their immediate surroundings, including pesticides. Chlorpyrifos is one of the most commonly used organophosphate pesticides in the world. The non-environmental strain of Escherichia coli, BL21 (DE3) displayed high tolerance to chlorpyrifos but with no/negligible degradation. The intrinsic resistance mechanisms that aid the organism in its high tolerance are probed. Efflux pumps being ubiquitous in nature and capable of conferring resistance against wide variety of xenobiotics were found to be over-expressed in the presence of CP. Also, an efflux pump inhibitor PAβN increased the susceptibility of E. coli to chlorpyrifos due to the intracellular accumulation of CP. The tripartite efflux pump EmrAB-TolC with increased expression in both transcript and protein on CP exposure, might play a major role in CP tolerance. The transcriptional regulators involved in multidrug resistance along with transporters belonging to all the major families conferring antimicrobial resistance were up-regulated. Also up-regulated were the genes involved in phopshonate metabolism and all the genes in the copper or silver export system. The common resistance mechanisms i.e, activation of efflux pumps between CP, antibacterial metals and antibiotics resistance might result in cross-resistance, ultimately increasing the prevalence of multidrug resistant strains, making infections hard to treat.

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Regulation and function of the flavonoid-inducible efflux system, emrR-emrAB, in Agrobacterium tumefaciens C58

Cited by 9 publications

References 67 publications

Inhibition of growth, biofilm formation, virulence, and surface attachment of Agrobacterium tumefaciens by cinnamaldehyde derivatives

Inhibition of growth, biofilm formation, virulence, and surface attachment of Agrobacterium tumefaciens by cinnamaldehyde derivatives

Aztreonam is a novel chemical inducer that promotes Agrobacteium transformation and lateral root development in soybean

Efflux mediated chlorpyrifos tolerance inEscherichia coliBL21(DE3)

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