{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T03:02:56Z","timestamp":1789095776248,"version":"build-2803163510"},"reference-count":51,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,5]],"date-time":"2025-09-05T00:00:00Z","timestamp":1757030400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Key Research and Development Program of Guangxi","award":["AB18221047"],"award-info":[{"award-number":["AB18221047"]}]},{"name":"Key Research and Development Program of Guangxi","award":["GNKM202504-2"],"award-info":[{"award-number":["GNKM202504-2"]}]},{"name":"Guangxi Mushroom Science and Technology Vanguard","award":["AB18221047"],"award-info":[{"award-number":["AB18221047"]}]},{"name":"Guangxi Mushroom Science and Technology Vanguard","award":["GNKM202504-2"],"award-info":[{"award-number":["GNKM202504-2"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Bacterial diseases of Pleurotus pulmonarius, caused by diverse pathogens and associated with a range of symptoms, reduce its commercial value and lead to substantial economic losses. While most research has focused on Pseudomonas tolaasii and its non-volatile toxin tolaasin, little is known about other bacterial pathogens and their volatile metabolites. In this study, two bacterial pathogens were isolated from symptomatic P. pulmonarius fruiting bodies in Guangxi, China, and identified as Ewingella americana and Cedecea neteri. Using headspace solid-phase microextraction coupled with gas chromatography\u2013mass spectrometry (HS-SPME-GC-MS), we identified 16 volatile organic compounds (VOCs) produced by these two species, seven of which exhibited toxicity-inducing sunken lesions, discoloration, and inhibition of mycelial growth. Symptom severity was quantified by colorimetric analysis. Among the toxic VOCs, 2,4-di-tert-butylphenol was the most potent, inducing sunken lesions and slight discoloration at concentrations as low as 0.5 mg\/mL, and causing significant inhibition of mycelial growth at 5 \u03bcg\/L. The remaining VOCs also caused varying degrees of sunken lesions, yellowing or browning, and suppression of mycelial growth. This study is the first to demonstrate the pathogenic potential of VOCs produced by bacterial pathogens in P. pulmonarius, underscoring their role as important virulence factors and providing a foundation for further investigation into their mechanisms and control strategies.<\/jats:p>","DOI":"10.3390\/toxins17090449","type":"journal-article","created":{"date-parts":[[2025,9,5]],"date-time":"2025-09-05T12:18:18Z","timestamp":1757074698000},"page":"449","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Toxicity of Volatile Organic Compounds Produced by Pathogens Ewingella americana and Cedecea neteri Associated with Pleurotus pulmonarius"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-6614-861X","authenticated-orcid":false,"given":"Zhiyuan","family":"Wei","sequence":"first","affiliation":[{"name":"Institute of Applied Microbiology, College of Agriculture, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yifan","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Applied Microbiology, College of Agriculture, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jieheng","family":"Qiu","sequence":"additional","affiliation":[{"name":"Institute of Applied Microbiology, College of Agriculture, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yulu","family":"Nie","sequence":"additional","affiliation":[{"name":"Institute of Applied Microbiology, College of Agriculture, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lian","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Applied Microbiology, College of Agriculture, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9345-4674","authenticated-orcid":false,"given":"Bin","family":"Liu","sequence":"additional","affiliation":[{"name":"Institute of Applied Microbiology, College of Agriculture, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"113721","DOI":"10.1016\/j.scienta.2024.113721","article-title":"Transcriptome Sequencing Analysis of the Effects of 1-MCP and Ethephon Treatments on the Storage Quality of Pleurotus pulmonarius","volume":"338","author":"Chen","year":"2024","journal-title":"Sci. Hortic."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"5197","DOI":"10.1002\/ps.7725","article-title":"Ginger Blotches on Agaricus bisporus Due to Monoacetylphloroglucinol Production by the Pathogen Pseudomonas \u2018gingeri\u2019","volume":"79","author":"Huang","year":"2023","journal-title":"Pest Manag. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"03101","DOI":"10.1051\/e3sconf\/202338903101","article-title":"Harmful Competitors and Diseases of Pleurotus ostreatus and Their Control Measures","volume":"389","author":"Rakhmonov","year":"2023","journal-title":"E3S Web Conf."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"542","DOI":"10.1094\/PDIS-06-20-1260-RE","article-title":"Comparative Studies on the Disease Prevalence and Population Dynamics of Ginger Blotch and Brown Blotch Pathogens of Button Mushrooms","volume":"105","author":"Taparia","year":"2021","journal-title":"Plant Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3526","DOI":"10.1002\/ps.8057","article-title":"Elucidating the Role of Monoacetylphloroglucinol in the Pathogenicity of Pseudomonas \u2018gingeri\u2019 against Agaricus bisporus","volume":"80","author":"Huang","year":"2024","journal-title":"Pest Manag. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"113141","DOI":"10.1016\/j.scienta.2024.113141","article-title":"Differential Analyses of Morphology and Transcription from Oyster Mushroom Pleurotus ostreatus Response to Brown Blotch Disease","volume":"331","author":"Wang","year":"2024","journal-title":"Sci. Hortic."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1094\/PHYTO-12-20-0550-R","article-title":"Characterization, Pathogenicity, Phylogeny, and Comparative Genomic Analysis of Pseudomonas tolaasii Strains Isolated from Various Mushrooms in China","volume":"112","author":"Liu","year":"2022","journal-title":"Phytopathology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.cropro.2015.01.006","article-title":"Suppressiveness of White Vinegar and Steam-Exploded Liquid Waste against the Causal Agents of Pleurotus eryngii Yellowing","volume":"70","author":"Bruno","year":"2015","journal-title":"Crop Prot."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"110965","DOI":"10.1016\/j.scienta.2022.110965","article-title":"Serial Transcriptional Changes of Flammulina filiformis (Winter Mushroom) Mycelia Infected by Pseudomonas migulae","volume":"297","author":"Wang","year":"2022","journal-title":"Sci. Hortic."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1186\/s42483-024-00239-8","article-title":"Biological Characteristics of Two Pathogens Causing Brown Blotch in Agaricus bisporus and the Toxin Identification of Cedecea neteri","volume":"6","author":"Huang","year":"2024","journal-title":"Phytopathol. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1189","DOI":"10.1094\/PDIS-09-20-1886-PDN","article-title":"First Report of Cedecea neteri Causing Yellow Rot Disease in Pleurotus pulmonarius in China","volume":"105","author":"Liu","year":"2021","journal-title":"Plant Dis."},{"key":"ref_12","first-page":"1014","article-title":"First Report of Cedecea neteri Causing Yellow Sticky Disease in Flammulina velutipes in China","volume":"103","author":"Yan","year":"2019","journal-title":"Plant Dis."},{"key":"ref_13","unstructured":"Sugendra Kumar, V. (2021). Antimicrobial Activity of Silver Nanoparticles for the Control of Microbial Spoilage of Grey Oyster Mushroom (Pleurotus pulmonarius). [Ph.D. Dissertation, Universiti Malaya]. Available online: https:\/\/www.proquest.com\/dissertations-theses\/antimicrobial-activity-silver-nanoparticles\/docview\/3143984806\/se-2."},{"key":"ref_14","first-page":"216","article-title":"Diseases and Pests Noxious to Pleurotus spp. Mushroom Crops","volume":"50","author":"Bellettini","year":"2018","journal-title":"Rev. Argent. Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2513","DOI":"10.1094\/PDIS-10-22-2499-PDN","article-title":"First Report of Bacterial Brown Rot Disease Caused by Ewingella americana on Cultivated Naematelia aurantialba in China","volume":"107","author":"Yang","year":"2023","journal-title":"Plant Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1325","DOI":"10.1007\/s42161-021-00897-z","article-title":"First Report of Bacterial Brown Rot Disease on Shiitake Mushroom (Lentinula edodes) Caused by Ewingella americana in Korea","volume":"103","author":"Na","year":"2021","journal-title":"J. Plant Pathol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1007\/s42161-022-01244-6","article-title":"Brown Rot Caused by Ewingella americana on King Oyster Mushroom (Pleurotus eryngii) in Korea","volume":"105","author":"Kim","year":"2023","journal-title":"J. Plant Pathol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2633","DOI":"10.1094\/PDIS-02-18-0351-PDN","article-title":"First Report of Ewingella americana Causing Bacterial Brown Rot Disease on Cultivated Needle Mushroom (Flammulina velutipes) in China","volume":"102","author":"Liu","year":"2018","journal-title":"Plant Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1007\/s42114-022-00588-y","article-title":"Isolation, Identification, and Pathogenic Effects of Trichoderma spp. from Auricularia auricula","volume":"6","author":"Dang","year":"2023","journal-title":"Adv. Compos. Hybrid Mater."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Lo Cantore, P., Giorgio, A., and Iacobellis, N.S. (2015). Bioactivity of Volatile Organic Compounds Produced by Pseudomonas tolaasii. Front. Microbiol., 6.","DOI":"10.3389\/fmicb.2015.01082"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1007\/s10526-020-10071-6","article-title":"Response of Pseudomonas tolaasii, the Causal Agent of Mushroom Brown Blotch Disease, to the Volatile Compounds Produced by Endofungal Bacteria","volume":"66","author":"Ghasemi","year":"2021","journal-title":"BioControl"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1007\/BF00314581","article-title":"Isolation of Ewingella americana from Mollusks","volume":"31","author":"Fanning","year":"1995","journal-title":"Curr. Microbiol."},{"key":"ref_23","first-page":"92","article-title":"Identification and Pollution Source Analysis of Pleurotus pulmonarius Blight Disease Caused by Ewingella americana","volume":"30","author":"Lu","year":"2023","journal-title":"Acta Edulis Fungi"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"334","DOI":"10.1007\/s002849900124","article-title":"Isolation of Ewingella americana from the Cultivated Mushroom, Agaricus bisporus","volume":"33","author":"Inglis","year":"1996","journal-title":"Curr. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/S0168-6496(03)00283-6","article-title":"Prevalence of Ewingella americana in Retail Fresh Cultivated Mushrooms (Agaricus bisporus, Lentinula edodes, and Pleurotus ostreatus) in Zaragoza (Spain)","volume":"47","author":"Reyes","year":"2004","journal-title":"FEMS Microbiol. Ecol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1094\/MPMI-19-1113","article-title":"Biological Characterization of White Line-Inducing Principle (WLIP) Produced by Pseudomonas reactans NCPPB1311","volume":"19","author":"Lazzaroni","year":"2006","journal-title":"Mol. Plant Microbe Interact."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"185","DOI":"10.3186\/jjphytopath.62.185","article-title":"Production of Volatile Components by Pseudomonas tolaasii and Their Toxic Activity","volume":"62","author":"Shirata","year":"1996","journal-title":"Jpn. J. Phytopathol."},{"key":"ref_28","first-page":"692","article-title":"Volatile Metabolites and Toxin Activity of Valsa mali","volume":"27","author":"Li","year":"2018","journal-title":"Acta Agric. Boreali-Occident. Sin."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1006\/pmpp.1999.0200","article-title":"The Effects of Tolaasin, the Toxin Produced by Pseudomonas tolaasii, on Tyrosinase Activities and the Induction of Browning in Agaricus bisporus Fruiting Bodies","volume":"55","author":"Arpin","year":"1999","journal-title":"Physiol. Mol. Plant Pathol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"493","DOI":"10.3186\/jjphytopath.61.493","article-title":"Isolation and Biological Activity of Toxins Produced by a Japanese Strain of Pseudomonas tolaasii, the Pathogen of Bacterial Rot of Cultivated Oyster Mushroom","volume":"61","author":"Shirata","year":"1995","journal-title":"Jpn. J. Phytopathol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Zhao, F.Q., Wang, P., Lucardi, R.D., Su, Z.S., and Li, S.Y. (2020). Natural Sources and Bioactivities of 2,4-Di-Tert-Butylphenol and Its Analogs. Toxins, 12.","DOI":"10.3390\/toxins12010035"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"6138","DOI":"10.1021\/jf5001138","article-title":"Purification, Characterization, and In Vitro Activity of 2,4-Di-tert-butylphenol from Pseudomonas monteilii PsF84: Conformational and Molecular Docking Studies","volume":"62","author":"Dharni","year":"2014","journal-title":"J. Agric. Food Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"17723","DOI":"10.1021\/acs.jafc.3c05157","article-title":"Antifungal Activity and Action Mechanisms of 2,4-Di-tert-butylphenol against Ustilaginoidea virens","volume":"71","author":"Fan","year":"2023","journal-title":"J. Agric. Food Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3785","DOI":"10.3390\/toxins7093785","article-title":"Are Some Fungal Volatile Organic Compounds (VOCs) Mycotoxins?","volume":"7","author":"Bennett","year":"2015","journal-title":"Toxins"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Sheikh, T.M.M., Zhou, D.M., Haider, M.S., Hussain, S., Wang, N., Chen, S.Q., Zhao, Y.S., Wen, X., Feng, H., and Wang, X.Y. (2023). Volatile Organic Compounds from Pythium oligandrum Play a Role in Its Parasitism on Plant-Pathogenic Pythium myriotylum. Appl. Environ. Microbiol., 89.","DOI":"10.1128\/aem.02036-22"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"101160","DOI":"10.1016\/j.fpsl.2023.101160","article-title":"Antifungal Effect of Volatile Organic Compounds (VOCs) Released from Antarctic Bacteria under Postharvest Conditions","volume":"39","author":"Ulloa","year":"2023","journal-title":"Food Packag. Shelf Life"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Corre, M.-H., Mercier, A., Bouteiller, M., Khalil, A., Ginevra, C., Depayras, S., Dupont, C., Rouxel, M., Gallique, M., and Grac, L. (2021). Bacterial Long-Range Warfare: Aerial Killing of Legionella pneumophila by Pseudomonas fluorescens. Microbiol. Spectr., 9.","DOI":"10.1128\/Spectrum.00404-21"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"14468","DOI":"10.1021\/acs.jafc.2c06187","article-title":"ROS Stress and Cell Membrane Disruption Are the Main Antifungal Mechanisms of 2-Phenylethanol Against Botrytis cinerea","volume":"70","author":"Zou","year":"2022","journal-title":"J. Agric. Food Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"111359","DOI":"10.1016\/j.foodcont.2025.111359","article-title":"Biocontrol of Postharvest Soft Rot Caused by Rhizopus stolonifer in Sweet Potatoes Using Volatile Organic Compounds from Actinomycete Nocardiopsis dassonvillei MI-S24","volume":"111","author":"Gomaa","year":"2025","journal-title":"Food Control"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1007\/s42161-022-01056-8","article-title":"Antifungal Activity and Control Efficiency of Endophytic Bacillus velezensis ZJ1 Strain and Its Volatile Compounds Against Alternaria solani and Botrytis cinerea","volume":"104","author":"Ren","year":"2022","journal-title":"J. Plant Pathol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Delgado, N., Olivera, M., C\u00e1diz, F., Bravo, G., Montenegro, I., Madrid, A., Fuentealba, C., Pedreschi, R., Salgado, E., and Besoain, X. (2021). Volatile Organic Compounds (VOCs) Produced by Gluconobacter cerinus and Hanseniaspora osmophila Displaying Control Effect against Table Grape-Rot Pathogens. Antibiotics, 10.","DOI":"10.3390\/antibiotics10060663"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Rey-Serra, P., Mnejja, M., and Monfort, A. (2022). Inheritance of Esters and Other Volatile Compounds Responsible for the Fruity Aroma in Strawberry. Front. Plant Sci., 13.","DOI":"10.3389\/fpls.2022.959155"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1007\/s10661-008-0589-7","article-title":"Study of Odor Compounds in Gaseous Effluents Generated During Production of Poultry Feather and Viscera Meal Using Headspace Solid Phase Microextraction","volume":"158","author":"Dallago","year":"2009","journal-title":"Environ. Monit. Assess."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2311","DOI":"10.1021\/ac9711394","article-title":"Sampling and Determination of Formaldehyde Using Solid-Phase Microextraction with On-Fiber Derivatization","volume":"70","author":"Martos","year":"1998","journal-title":"Anal. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2409","DOI":"10.5194\/amt-8-2409-2015","article-title":"Characterisation and Optimisation of a Method for the Detection and Quantification of Atmospherically Relevant Carbonyl Compounds in Aqueous Medium","volume":"8","author":"Rodigast","year":"2015","journal-title":"Atmos. Meas. Tech. Discuss."},{"key":"ref_46","first-page":"101224","article-title":"Effect of Different Isolation Sources of Lactococcus lactis subsp. lactis on Volatile Metabolites in Fermented Milk","volume":"21","author":"Yu","year":"2024","journal-title":"Food Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"967","DOI":"10.12688\/f1000research.11823.1","article-title":"Analytical Challenges of Untargeted GC-MS-Based Metabolomics and the Critical Issues in Selecting the Data Processing Strategy","volume":"6","author":"Han","year":"2017","journal-title":"F1000Research"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"7828","DOI":"10.1128\/AEM.01303-08","article-title":"Cross-kingdom amplification using bacteria-specific primers: Complications for studies of coral microbial ecology","volume":"74","author":"Galkiewicz","year":"2008","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/j.diagmicrobio.2006.12.019","article-title":"Phylogeny and species identification of the family Enterobacteriaceae based on dnaJ sequences","volume":"58","author":"Pham","year":"2007","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1104","DOI":"10.1128\/aem.61.3.1104-1109.1995","article-title":"PCR amplification and direct sequencing of gyrB genes with universal primers and their application to the detection and taxonomic analysis of Pseudomonas putida strains","volume":"61","author":"Yamamoto","year":"1995","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"D1692","DOI":"10.1093\/nar\/gkae961","article-title":"mVOC 4.0: A Database of Microbial Volatiles","volume":"53","author":"Kemmler","year":"2025","journal-title":"Nucleic Acids Res."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/17\/9\/449\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:40:13Z","timestamp":1760035213000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/17\/9\/449"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,5]]},"references-count":51,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["toxins17090449"],"URL":"https:\/\/doi.org\/10.3390\/toxins17090449","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,5]]}}}