Abstract
This study aimed to investigate the anti-quorum sensing (QS) activity of Artemisia argyi leaf extracts (AALE) towards Pseudomonas aeruginosa PAO1 as well as the underlying molecular mechanisms. Using a biosensor Chromobacterium violaceum CV026, AALE were found to have anti-QS activity as AALE treatment significantly inhibited the violacein production of C. violaceum CV026 while produced little effect on the cell growth. Beyond that a higher dosage of AALE inhibited cell growth, sub-MIC of AALE significantly reduced the production of QS-regulated virulence factors (pyocyanin, elastase, and rhamnolipid), biofilm formation, and the swarming and swimming motility in P. aeruginosa PAO1 with a dosage-dependent manner. Quantitative real-time PCR (qRT-PCR) analysis did not detect the direct inhibitory effect of AALE on the expression of QS genes (lasI, lasR, rhlI, and rhlR). By iTRAQ-based quantitative proteomic analysis, 129 proteins were found to be differentially expressed upon AALE treatment, with 85 upregulated and 44 downregulated proteins, respectively. Functional enrichment analysis of the differential proteins revealed that AALE exerted anti-QS activity towards P. aeruginosa PAO1 by upregulating the expression of the global regulator CsrA, inducing oxidative stress, and perturbing protein homeostasis. Moreover, the inhibitory effect of AALE on the virulence of P. aeruginosa PAO1 was likely to be achieved by attenuating the expression of QS-regulated genes instead of QS genes. Collectively, the results of this study provide a basis for the future use of AALE as a preservative in controlling food spoilage caused by P. aeruginosa.
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References
Abad MJ, Bedoya LM, Apaza L, Bermejo P (2012) The Artemisia L. Genus: a review of bioactive essential oils. Molecules 17:2542–2566. https://doi.org/10.3390/molecules17032542
Abdallah M, Benoliel C, Jama C, Drider D, Dhulster P, Chihib NE (2014) Thermodynamic prediction of growth temperature dependence in the adhesion of Pseudomonas aeruginosa and Staphylococcus aureus to stainless steel and polycarbonate. J Food Protect 77:1116–1126. https://doi.org/10.4315/0362-028x.Jfp-13-365
Abdallah M, Khelissa O, Ibrahim A, Benoliel C, Heliot L, Dhulster P, Chihib NE (2015) Impact of growth temperature and surface type on the resistance of Pseudomonas aeruginosa and Staphylococcus aureus biofilms to disinfectants. Int J Food Microbiol 214:38–47. https://doi.org/10.1016/j.ijfoodmicro.2015.07.022
Ahmed M, Ji M, Qin P, Gu Z, Liu Y, Sikandar A, Iqbal MF, Javeed A, Shafi J, Du Y (2019a) Determination of phytochemicals, antioxidant activity and biochemical composition of Chinese Mugwort (Artemisia Argyi L.) leaf extract from northeast China. Appl Ecol Environ Res 17:15349–15362. https://doi.org/10.15666/aeer/1706_1534915362
Ahmed SA, Rudden M, Smyth TJ, Dooley JS, Marchant R, Banat IM (2019b) Natural quorum sensing inhibitors effectively downregulate gene expression of Pseudomonas aeruginosa virulence factors. Appl Microbiol Biotechnol 103:3521–3535. https://doi.org/10.1007/s00253-019-09618-0
Arunkumar M, LewisOscar F, Thajuddin N, Pugazhendhi A, Nithya C (2020) In vitro and in vivo biofilm forming Vibrio spp: a significant threat in aquaculture. Process Biochem 94:213–223. https://doi.org/10.1016/j.procbio.2020.04.029
Banerjee M, Moulick S, Bhattacharya KK, Parai D, Chattopadhyay S, Mukherjee SK (2017) Attenuation of Pseudomonas aeruginosa quorum sensing, virulence and biofilm formation by extracts of Andrographis paniculata. Microb Pathogenesis 113:85–93. https://doi.org/10.1016/j.micpath.2017.10.023
Brencic A, Lory S (2009) Determination of the regulon and identification of novel mRNA targets of Pseudomonas aeruginosa RsmA. Mol Microbiol 72:612–632. https://doi.org/10.1111/j.1365-2958.2009.06670.x
Brindhadevi K, LewisOscar F, Mylonakis E, Shanmugam S, Verma TN, Pugazhendhi A (2020) Biofilm and quorum sensing mediated pathogenicity in Pseudomonas aeruginosa. Process Biochem 96:49–57. https://doi.org/10.1016/j.procbio.2020.06.001
Chen T, Sheng J, Fu Y, Li M, Wang J, Jia A (2017) 1H NMR-based global metabolic studies of Pseudomonas aeruginosa upon exposure of the quorum sensing inhibitor resveratrol. J Proteome Res 16:824–830. https://doi.org/10.1021/acs.jproteome.6b00800
Cornelis P (2020) Putting an end to the Pseudomonas aeruginosa IQS controversy. Microbiologyopen 9:e962. https://doi.org/10.1002/mbo3.962
Cosa S, Rakoma JR, Yusuf AA, Tshikalange TE (2020) Calpurnia aurea (Aiton) benth extracts reduce quorum sensing controlled virulence factors in Pseudomonas aeruginosa. Molecules. https://doi.org/10.3390/molecules25102283
Deng X, Weerapana E, Ulanovskaya O, Sun F, Liang H, Ji Q, Ye Y, Fu Y, Zhou L, Li J, Zhang H, Wang C, Alvarez S, Hicks LM, Lan L, Wu M, Cravatt BF, He C (2013) Proteome-wide quantification and characterization of oxidation-sensitive cysteines in pathogenic bacteria. Cell Host Microbe 13:358–370. https://doi.org/10.1016/j.chom.2013.02.004
Duran N, Justo GZ, Duran M, Brocchi M, Cordi L, Tasic L, Castro GR, Nakazato G (2016) Advances in Chromobacterium violaceum and properties of violacein—its main secondary metabolite: a review. Biotechnol Adv 34:1030–1045. https://doi.org/10.1016/j.biotechadv.2016.06.003
Garcia-Contreras R, Nunez-Lopez L, Jasso-Chavez R, Kwan BW, Belmont JA, Rangel-Vega A, Maeda T, Wood TK (2015) Quorum sensing enhancement of the stress response promotes resistance to quorum quenching and prevents social cheating. ISME J 9:115–125. https://doi.org/10.1038/ismej.2014.98
Garcia-Reyes S, Soberon-Chavez G, Cocotl-Yanez M (2020) The third quorum-sensing system of Pseudomonas aeruginosa: Pseudomonas quinolone signal and the enigmatic PqsE protein. J Med Microbiol 69:25–34. https://doi.org/10.1099/jmm.0.001116
Gebhardt MJ, Kambara TK, Ramsey KM, Dove SL (2020) Widespread targeting of nascent transcripts by RsmA in Pseudomonas aeruginosa. Proc Natl Acad Sci USA 117:10520–10529. https://doi.org/10.1073/pnas.1917587117
Johnson L, Mulcahy H, Kanevets U, Shi Y, Lewenza S (2012) Surface-localized spermidine protects the Pseudomonas aeruginosa outer membrane from antibiotic treatment and oxidative stress. J Bacteriol 194:813–826. https://doi.org/10.1128/JB.05230-11
Klopfenstein DV, Zhang LS, Pedersen BS, Ramirez F, Vesztrocy AW, Naldi A, Mungall CJ, Yunes JM, Botvinnik O, Weigel M, Dampier W, Dessimoz C, Flick P, Tang HB (2018) GOATOOLS: a Python library for gene ontology analyses. Sci Rep 8:10872. https://doi.org/10.1038/s41598-018-28948-z
Kong HS, Roberts DP, Patterson CD, Kuehne SA, Heeb S, Lakshman DK, Lydon J (2012) Effect of overexpressing rsmA from Pseudomonas aeruginosa on virulence of select phytotoxin-producing strains of P. syringae. Phytopathology 102:575–587. https://doi.org/10.1094/PHYTO-09-11-0267
Kostylev M, Kim DY, Smalley NE, Salukhe I, Greenberg EP, Dandekar AA (2019) Evolution of the Pseudomonas aeruginosa quorum-sensing hierarchy. Proc Natl Acad Sci USA 116:7027–7032. https://doi.org/10.1073/pnas.1819796116
Li T, Wang D, Liu N, Ma Y, Ding T, Mei Y, Li J (2018) Inhibition of quorum sensing-controlled virulence factors and biofilm formation in Pseudomonas fluorescens by cinnamaldehyde. Int J Food Microbiol 269:98–106. https://doi.org/10.1016/j.ijfoodmicro.2018.01.023
Machado I, Silva LR, Giaouris ED, Melo LF, Simoes M (2020) Quorum sensing in food spoilage and natural-based strategies for its inhibition. Food Res Int 127:108754. https://doi.org/10.1016/j.foodres.2019.108754
McLean RJ, Pierson LS 3rd, Fuqua C (2004) A simple screening protocol for the identification of quorum signal antagonists. J Microbiol Methods 58:351–360. https://doi.org/10.1016/j.mimet.2004.04.016
Pearson JP, Pesci EC, Iglewski BH (1997) Roles of Pseudomonas aeruginosa las and rhl quorum-sensing systems in control of elastase and rhamnolipid biosynthesis genes. J Bacteriol 179:5756–5767. https://doi.org/10.1128/jb.179.18.5756-5767.1997
Pessi G, Williams F, Hindle Z, Heurlier K, Holden MT, Camara M, Haas D, Williams P (2001) The global posttranscriptional regulator RsmA modulates production of virulence determinants and N-acylhomoserine lactones in Pseudomonas aeruginosa. J Bacteriol 183:6676–6683. https://doi.org/10.1128/JB.183.22.6676-6683.2001
Rahman MR, Lou Z, Zhang J, Yu F, Timilsena YP, Zhang C, Zhang Y, Bakry AM (2017) Star anise (Illicium verum Hook. f.) as quorum sensing and biofilm formation inhibitor on foodborne bacteria: study in milk. J Food Prot 80:645–653. https://doi.org/10.4315/0362-028X.JFP-16-294
Riggio GM, Wang Q, Kniel KE, Gibson KE (2019) Microgreens—a review of food safety considerations along the farm to fork continuum. Int J Food Microbiol 290:76–85. https://doi.org/10.1016/j.ijfoodmicro.2018.09.027
Sethupathy S, Prasath KG, Ananthi S, Mahalingam S, Balan SY, Pandian SK (2016) Proteomic analysis reveals modulation of iron homeostasis and oxidative stress response in Pseudomonas aeruginosa PAO1 by curcumin inhibiting quorum sensing regulated virulence factors and biofilm production. J Proteomics 145:112–126. https://doi.org/10.1016/j.jprot.2016.04.019
Shi G, Wang T, Wu S, Wang Y, Shao J, Zhou M, Wang C (2017) Activity of essential oil extracted from Artemisia argyi in inducing apoptosis of Candida albicans. China J Chin Mater Med 42:3572–3577. https://doi.org/10.19540/j.cnki.cjcmm.2017.0136
Song X, Wen X, He J, Zhao H, Li S, Wang M (2019) Phytochemical components and biological activities of Artemisia argyi. J Funct Foods 52:648–662. https://doi.org/10.1016/j.jff.2018.11.029
Sorensen HP, Madsen LS, Petersen J, Andersen JT, Hansen AM, Beck HC (2010) Oat (Avena sativa) seed extract as an antifungal food preservative through the catalytic activity of a highly abundant class I chitinase. Appl Biochem Biotechnol 160:1573–1584. https://doi.org/10.1007/s12010-009-8557-4
Tan SY, Chua SL, Chen Y, Rice SA, Kjelleberg S, Nielsen TE, Yang L, Givskov M (2013) Identification of five structurally unrelated quorum-sensing inhibitors of Pseudomonas aeruginosa from a natural-derivative database. Antimicrob Agents Chemother 57:5629–5641. https://doi.org/10.1128/AAC.00955-13
Vasavi HS, Arun AB, Rekha PD (2014) Anti-quorum sensing activity of Psidium guajava L. flavonoids against Chromobacterium violaceum and Pseudomonas aeruginosa PAO1. Microbiol Immunol 58:286–293. https://doi.org/10.1111/1348-0421.12150
Wang H, Chu W, Ye C, Gaeta B, Tao H, Wang M, Qiu Z (2019) Chlorogenic acid attenuates virulence factors and pathogenicity of Pseudomonas aeruginosa by regulating quorum sensing. Appl Microbiol Biotechnol 103:903–915. https://doi.org/10.1007/s00253-018-9482-7
Wang H, Zhang M, Ma Y, Wang B, Shao M, Huang H, Liu Y, Kang Z (2020) Selective inactivation of Gram-negative bacteria by carbon dots derived from natural biomass: Artemisia argyi leaves. J Mater Chem B 8:2666–2672. https://doi.org/10.1039/c9tb02735a
Wei Q, Bhasme P, Wang ZG, Wang L, Wang SW, Zeng YF, Wang Y, Ma LYZ, Li Y (2020) Chinese medicinal herb extract inhibits PQS-mediated quorum sensing system in Pseudomonas aeruginosa. J Ethnopharmacol 248:112272. https://doi.org/10.1016/j.jep.2019.112272
Xia K, Zang N, Zhang J, Zhang H, Li Y, Liu Y, Feng W, Liang X (2016) New insights into the mechanisms of acetic acid resistance in Acetobacter pasteurianus using iTRAQ-dependent quantitative proteomic analysis. Int J Food Microbiol 238:241–251. https://doi.org/10.1016/j.ijfoodmicro.2016.09.016
Xiao J, Liu W, Sun H, Li W, Koike K, Kikuchi T, Yamada T, Li D, Feng F, Zhang J (2019) Bioactivity-based analysis and chemical characterization of hypoglycemic and antioxidant components from Artemisia argyi. Bioorg Chem. https://doi.org/10.1016/j.bioorg.2019.103268
Xie C, Mao X, Huang J, Ding Y, Wu J, Dong S, Kong L, Gao G, Li C, Wei L (2011) KOBAS 2.0: a web server for annotation and identification of enriched pathways and diseases. Nucleic Acids Res 39:W316–W322. https://doi.org/10.1093/nar/gkr483
Xu J, Xia K, Li P, Qian C, Li Y, Liang X (2020) Functional investigation of the chromosomal ccdAB and hipAB operon in Escherichia coli Nissle 1917. Appl Microbiol Biotechnol 104:6731–6747. https://doi.org/10.1007/s00253-020-10733-6
Yan X, He B, Liu L, Qu G, Shi J, Hu L, Jiang G (2018) Antibacterial mechanism of silver nanoparticles in Pseudomonas aeruginosa: proteomics approach. Metallomics 10:557–564. https://doi.org/10.1039/c7mt00328e
Yao X, He W, Lu CD (2011) Functional characterization of seven gamma-glutamylpolyamine synthetase genes and the bauRABCD locus for polyamine and beta-Alanine utilization in Pseudomonas aeruginosa PAO1. J Bacteriol 193:3923–3930. https://doi.org/10.1128/JB.05105-11
Yi S, Zhu J, Fu L, Li J (2010) Tea polyphenols inhibit Pseudomonas aeruginosa through damage to the cell membrane. Int J Food Microbiol 144:111–117. https://doi.org/10.1016/j.ijfoodmicro.2010.09.005
Zhang Y, Wei J, Yuan Y, Yue T (2019) Diversity and characterization of spoilage-associated psychrotrophs in food in cold chain. Int J Food Microbiol 290:86–95. https://doi.org/10.1016/j.ijfoodmicro.2018.09.026
Zhang Y, Pan X, Liao S, Jiang C, Wang L, Tang Y, Wu G, Dai G, Chen L (2020) Quantitative proteomics reveals the mechanism of silver nanoparticles against multidrug-resistant Pseudomonas aeruginosa biofilms. J Proteome Res 19:3109–3122. https://doi.org/10.1021/acs.jproteome.0c00114
Zhou J, Muhammad J, Sun B, Yang R, Wadood A, Wang J, Jia A (2019) Metabolomic analysis of quorum sensing inhibitor hordenine on Pseudomonas aeruginosa. Appl Microbiol Biotechnol 103:6271–6285. https://doi.org/10.1007/s00253-019-09878-w
Zhu J, Huang X, Zhang F, Feng L, Li J (2015) Inhibition of quorum sensing, biofilm, and spoilage potential in Shewanella baltica by green tea polyphenols. J Microbiol 53:829–836. https://doi.org/10.1007/s12275-015-5123-3
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The work was financially supported by Grants from the Natural Science Foundation of Zhejiang Province (LY19C200002 and LGN19C160001).
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JK and YW conducted lab work and data analysis. KX interpreted the data, designed the experiments, and drafted the manuscript. ZZ and HZ conducted data analysis. XL conceived of the study, performed data review, and contributed to the paper writing. All authors read and approved the final manuscript.
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Kong, J., Wang, Y., Xia, K. et al. New insights into the antibacterial and quorum sensing inhibition mechanism of Artemisia argyi leaf extracts towards Pseudomonas aeruginosa PAO1. 3 Biotech 11, 97 (2021). https://doi.org/10.1007/s13205-021-02663-5
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DOI: https://doi.org/10.1007/s13205-021-02663-5