Developing a toolbox of Tier I tests to assess pesticides toxicity on the asymbiotic and symbiotic phases of arbuscular mycorrhizal fungi

Soil microorganisms are a key protection goal in the European Union (EU) pesticide regulatory framework. Arbuscular mycorrhizal fungi (AMF) were identified as good proxies for assessing pesticides toxicity on the soil microbiota. This could involve ecotoxicity testing at the different life stages of...

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Main Authors: Christos Papadopoulos, Marjan Roshanfekrrad, Daniela Tsikou, Kalliope K. Papadopoulou, Maryline Calonne-Salmon, Stephan Declerck, Dimitrios G. Karpouzas
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:Ecotoxicology and Environmental Safety
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Online Access:http://www.sciencedirect.com/science/article/pii/S0147651325012370
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author Christos Papadopoulos
Marjan Roshanfekrrad
Daniela Tsikou
Kalliope K. Papadopoulou
Maryline Calonne-Salmon
Stephan Declerck
Dimitrios G. Karpouzas
author_facet Christos Papadopoulos
Marjan Roshanfekrrad
Daniela Tsikou
Kalliope K. Papadopoulou
Maryline Calonne-Salmon
Stephan Declerck
Dimitrios G. Karpouzas
author_sort Christos Papadopoulos
collection DOAJ
description Soil microorganisms are a key protection goal in the European Union (EU) pesticide regulatory framework. Arbuscular mycorrhizal fungi (AMF) were identified as good proxies for assessing pesticides toxicity on the soil microbiota. This could involve ecotoxicity testing at the different life stages of AMF. We evaluated the effects of five pesticides (pyraclostrobin, fludioxonil, hymexazol, etridiazole, glyphosate) and a transformation product (AMPA), with distinct mode of action, on the development and functionality of Rhizophagus irregularis at the asymbiotic and symbiotic phase using a spore germination assay and a gnotobiotic AMF-host plant system (AMF-sandwich test), respectively. Based on arbuscular colonization in the AMF-sandwich test, fludioxonil was the most toxic (EC50 0.085 mg/L) followed by glyphosate (EC50 2.58 mg/L) and pyraclostrobin (EC50 9.22 mg/L), while etridiazole, hymexazol, and AMPA showed EC50 values higher than the highest tested concentration. However, for glyphosate and pyraclostrobin negative effects on symbiosis functioning were observed at lower concentrations than for colonization, as depicted by the expression of plant marker genes and/or P-uptake, suggesting the establishment of non-functional arbuscular symbiosis. The high toxicity of fludioxonil (EC50 0.03 mg/L) and the low toxicity of AMPA (EC50 > 432 mg/L) on R. irregularis was verified also for the asymbiotic phase via spore germination assay. Comparative tests showed differences in the toxicity of pure active substances and commercial formulations of fludioxonil and pyraclostrobin on the AMF-sandwich test. We propose that the AMF-sandwich system together with the spore germination test could be used as a toolbox for Tier-I assessment of pesticides toxicity on AMF.
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spelling doaj-art-1175c3cf61a045658431e5e5b0a4af332025-08-20T05:04:11ZengElsevierEcotoxicology and Environmental Safety0147-65132025-09-0130311889210.1016/j.ecoenv.2025.118892Developing a toolbox of Tier I tests to assess pesticides toxicity on the asymbiotic and symbiotic phases of arbuscular mycorrhizal fungiChristos Papadopoulos0Marjan Roshanfekrrad1Daniela Tsikou2Kalliope K. Papadopoulou3Maryline Calonne-Salmon4Stephan Declerck5Dimitrios G. Karpouzas6Laboratory of Plant and Environmental Biotechnology, Department of Biochemistry and Biotechnology, University of Thessaly, Biopolis Campus, Larissa 41500, GreeceUniversite Catholique de Louvain, Earth and Life Institute, Mycology, Croix du Sud 2, Box L7.05.06, Louvain‑La‑Neuve 1348, BelgiumLaboratory of Plant and Environmental Biotechnology, Department of Biochemistry and Biotechnology, University of Thessaly, Biopolis Campus, Larissa 41500, GreeceLaboratory of Plant and Environmental Biotechnology, Department of Biochemistry and Biotechnology, University of Thessaly, Biopolis Campus, Larissa 41500, GreeceUniversite Catholique de Louvain, Earth and Life Institute, Mycology, Croix du Sud 2, Box L7.05.06, Louvain‑La‑Neuve 1348, BelgiumUniversite Catholique de Louvain, Earth and Life Institute, Mycology, Croix du Sud 2, Box L7.05.06, Louvain‑La‑Neuve 1348, BelgiumLaboratory of Plant and Environmental Biotechnology, Department of Biochemistry and Biotechnology, University of Thessaly, Biopolis Campus, Larissa 41500, Greece; Corresponding author.Soil microorganisms are a key protection goal in the European Union (EU) pesticide regulatory framework. Arbuscular mycorrhizal fungi (AMF) were identified as good proxies for assessing pesticides toxicity on the soil microbiota. This could involve ecotoxicity testing at the different life stages of AMF. We evaluated the effects of five pesticides (pyraclostrobin, fludioxonil, hymexazol, etridiazole, glyphosate) and a transformation product (AMPA), with distinct mode of action, on the development and functionality of Rhizophagus irregularis at the asymbiotic and symbiotic phase using a spore germination assay and a gnotobiotic AMF-host plant system (AMF-sandwich test), respectively. Based on arbuscular colonization in the AMF-sandwich test, fludioxonil was the most toxic (EC50 0.085 mg/L) followed by glyphosate (EC50 2.58 mg/L) and pyraclostrobin (EC50 9.22 mg/L), while etridiazole, hymexazol, and AMPA showed EC50 values higher than the highest tested concentration. However, for glyphosate and pyraclostrobin negative effects on symbiosis functioning were observed at lower concentrations than for colonization, as depicted by the expression of plant marker genes and/or P-uptake, suggesting the establishment of non-functional arbuscular symbiosis. The high toxicity of fludioxonil (EC50 0.03 mg/L) and the low toxicity of AMPA (EC50 > 432 mg/L) on R. irregularis was verified also for the asymbiotic phase via spore germination assay. Comparative tests showed differences in the toxicity of pure active substances and commercial formulations of fludioxonil and pyraclostrobin on the AMF-sandwich test. We propose that the AMF-sandwich system together with the spore germination test could be used as a toolbox for Tier-I assessment of pesticides toxicity on AMF.http://www.sciencedirect.com/science/article/pii/S0147651325012370Arbuscular mycorrhizal fungiPesticidesEcotoxicityAMF-sandwich testSpore gemination test
spellingShingle Christos Papadopoulos
Marjan Roshanfekrrad
Daniela Tsikou
Kalliope K. Papadopoulou
Maryline Calonne-Salmon
Stephan Declerck
Dimitrios G. Karpouzas
Developing a toolbox of Tier I tests to assess pesticides toxicity on the asymbiotic and symbiotic phases of arbuscular mycorrhizal fungi
Ecotoxicology and Environmental Safety
Arbuscular mycorrhizal fungi
Pesticides
Ecotoxicity
AMF-sandwich test
Spore gemination test
title Developing a toolbox of Tier I tests to assess pesticides toxicity on the asymbiotic and symbiotic phases of arbuscular mycorrhizal fungi
title_full Developing a toolbox of Tier I tests to assess pesticides toxicity on the asymbiotic and symbiotic phases of arbuscular mycorrhizal fungi
title_fullStr Developing a toolbox of Tier I tests to assess pesticides toxicity on the asymbiotic and symbiotic phases of arbuscular mycorrhizal fungi
title_full_unstemmed Developing a toolbox of Tier I tests to assess pesticides toxicity on the asymbiotic and symbiotic phases of arbuscular mycorrhizal fungi
title_short Developing a toolbox of Tier I tests to assess pesticides toxicity on the asymbiotic and symbiotic phases of arbuscular mycorrhizal fungi
title_sort developing a toolbox of tier i tests to assess pesticides toxicity on the asymbiotic and symbiotic phases of arbuscular mycorrhizal fungi
topic Arbuscular mycorrhizal fungi
Pesticides
Ecotoxicity
AMF-sandwich test
Spore gemination test
url http://www.sciencedirect.com/science/article/pii/S0147651325012370
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