a5b0a6395032ae9cc96cc1020d99759d jof-11-00852.pdf 27ed1d186fb8e78eed6cd39a8b4b852508fcdc35 jof-11-00852.pdf 63b964b4f3fbb04c075a8cd8fd6dcf39a657811c82433f98e84362eb763283b5 jof-11-00852.pdf Title: Environmental Compatibility of Penicillium rubens Strain 212: Impact on Indigenous Soil Fungal Community Dynamics Subject: Fusarium wilt causes substantial losses in many crops, and Penicillium rubens strain 212 (PO212) is a well-established biological control agent effective against several soil-borne pathogens, including the causal agents of Fusarium wilt. Before its widespread use, it is essential to assess whether applying PO212 may affect indigenous soil microbial communities. To address this, two open-field tomato trials were conducted to evaluate spatial and temporal changes in non-target soil fungal communities following the application of PO212. Fungal community profiles were monitored over one year using PCR–DGGE of fungal rDNA, and representative DGGE bands were sequenced for taxonomic confirmation. Community structure and variability were analysed using cluster analysis (UPGMA and Neighbor-Joining) and analysis of molecular variance (AMOVA) to determine the effects of treatment, sampling date, and soil depth. PO212 application did not significantly altered the composition or diversity of indigenous soil fungal communities. DGGE banding patterns and diversity indices were similar between treated and untreated soils throughout the study period. Observed community changes were driven primarily by temporal (seasonal) variation, with samples collected at 365 days clustering separately from earlier sampling dates for both treatments. AMOVA confirmed that sampling date, rather than PO212 treatment, explained most of the variance in community composition (p < 0.05). Although PO212 persisted in soil, fluctuations in other fungal populations were minor and within the range of natural seasonal variability. Overall, field application of PO212 did not disrupt indigenous soil fungal communities, supporting its environmental safety as a biocontrol agent for managing Fusarium wilt. Keywords: Penicillium rubens; PO212; biological control agent; Fusarium oxysporum; environmental risk assessment; soil fungal community Author: Belén Guijarro, Gema Vázquez, Antonieta De Cal, Paloma Melgarejo, Núria Gaju, Maira Martínez-Alonso and Inmaculada Larena Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Sat Nov 29 12:27:14 2025 CET ModDate: Sat Nov 29 12:46:46 2025 CET Custom Metadata: yes Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 18 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1966832 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- QKZKVH+VnURWPalladioL Type 1 Custom yes yes yes 106 0 IUICUF+URWPalladioL-Bold Type 1 Custom yes yes yes 107 0 YPJLJY+URWPalladioL-Roma Type 1 Custom yes yes yes 108 0 GHKUEP+URWPalladioL-Ital Type 1 Custom yes yes yes 109 0 BGUIAD+URWPalladioL-BoldItal Type 1 Custom yes yes yes 110 0 XMZODV+CMSY10 Type 1 Builtin yes yes yes 181 0 OVNEXW+EURM10 Type 1 Builtin yes yes yes 215 0 RFLZJB+CMR10 Type 1 Builtin yes yes yes 230 0 CIDFont+F1 CID TrueType Identity-H yes no yes 246 0 CIDFont+F2 CID TrueType Identity-H yes no yes 247 0 CIDFont+F3 CID TrueType Identity-H yes no yes 248 0 CIDFont+F3 CID TrueType Identity-H yes no yes 254 0 CIDFont+F1 CID TrueType Identity-H yes no yes 255 0 CIDFont+F2 CID TrueType Identity-H yes no yes 256 0 CIDFont+F1 CID TrueType Identity-H yes no yes 317 0 CIDFont+F2 CID TrueType Identity-H yes no yes 318 0 CIDFont+F2 CID TrueType Identity-H yes no yes 322 0 CIDFont+F1 CID TrueType Identity-H yes no yes 356 0 CIDFont+F2 CID TrueType Identity-H yes no yes 357 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2026-02-14 04:22:09 CET RepresentationInformation: jof-11-00852.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2026-02-13 05:01:19 CET Size: 1966832 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 516 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Item: Title: Materials and Methods Children: Item: Title: Culture Conditions and Plant Material Item: Title: Field Trials and Sampling Item: Title: DNA Extraction and PCR Amplification Item: Title: DGGE Profiling and Sequence Analysis Item: Title: Data Analysis Item: Title: Results Children: Item: Title: Fungal Community Profiles and Diversity Item: Title: Temporal and Spatial Dynamics vs. Treatment Effects Item: Title: Identification of Indigenous Fungi Item: Title: Discussion Item: Title: Conclusions Item: Title: References Info: Title: Environmental Compatibility of Penicillium rubens Strain 212: Impact on Indigenous Soil Fungal Community Dynamics Author: Belén Guijarro, Gema Vázquez, Antonieta De Cal, Paloma Melgarejo, Núria Gaju, Maira Martínez-Alonso and Inmaculada Larena Subject: Fusarium wilt causes substantial losses in many crops, and Penicillium rubens strain 212 (PO212) is a well-established biological control agent effective against several soil-borne pathogens, including the causal agents of Fusarium wilt. Before its widespread use, it is essential to assess whether applying PO212 may affect indigenous soil microbial communities. To address this, two open-field tomato trials were conducted to evaluate spatial and temporal changes in non-target soil fungal communities following the application of PO212. Fungal community profiles were monitored over one year using PCR–DGGE of fungal rDNA, and representative DGGE bands were sequenced for taxonomic confirmation. Community structure and variability were analysed using cluster analysis (UPGMA and Neighbor-Joining) and analysis of molecular variance (AMOVA) to determine the effects of treatment, sampling date, and soil depth. PO212 application did not significantly altered the composition or diversity of indigenous soil fungal communities. DGGE banding patterns and diversity indices were similar between treated and untreated soils throughout the study period. Observed community changes were driven primarily by temporal (seasonal) variation, with samples collected at 365 days clustering separately from earlier sampling dates for both treatments. AMOVA confirmed that sampling date, rather than PO212 treatment, explained most of the variance in community composition (p < 0.05). Although PO212 persisted in soil, fluctuations in other fungal populations were minor and within the range of natural seasonal variability. Overall, field application of PO212 did not disrupt indigenous soil fungal communities, supporting its environmental safety as a biocontrol agent for managing Fusarium wilt. 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