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Chemical control suffers from the loss of available conventional active ingredients due to strict environmental safety regulations which, combined with the loss of fungicide efficacy due to resistance development, constitute major problems of contemporary crop protection. Metal-containing nanoparticles (MNPs) appear to have all the credentials to be next-generation, eco-compatible fungicide alternatives and a valuable anti-resistance management tool. Could the introduction of MNPs as nano-fungicides be the answer to both reducing the environmental footprint of xenobiotics and dealing with fungicide resistance? The potential of MNPs to be utilized as nano-fungicides, both as alternatives to conventional fungicides or/and as partners in combating fungicide resistance, is discussed in terms of effectiveness, potential antimicrobial mechanisms as well as synergy profiles with conventional fungicides. However, their "golden" potential to be used both as alternatives and partners of conventional fungicides to combat resistance and reduce environmental pollution is challenged by undesirable effects towards non-target organisms such as phytotoxicity, toxicity to humans and environmental ecotoxicity, constituting risks that should be considered before their commercial introduction as nano-pesticides at a large scale. © 2022 Society of Chemical Industry., (© 2022 Society of Chemical Industry.)

The antifungal potential of ZnO-NPs against Alternaria alternata isolates with reduced sensitivity to the succinate dehydrogenase inhibitor (SDHI) boscalid, resulting from target site modifications, was evaluated in vitro and in vivo. ZnO-NPs could effectively inhibit mycelial growth in a dose-dependent way in both boscalid (BOSC) sensitive (BOSC-S) and resistant (BOSC-R) isolates. The fungitoxic effect of ZnO-NPs against the pathogen was significantly enhanced when combined with boscalid compared to the individual treatments in all phenotype cases (BOSC-S/R) both in vitro and in vivo. Fungitoxic effect of ZnO-NPs could be, at least partly, attributed to zinc ion release as indicated by the positive correlation between sensitivities to the nanoparticles and their ionic counterpart ZnSO 4 and the alleviation of the ZnO-NPs fungitoxic action in the presence of the strong chelating agent EDTA. The superior effectiveness of ZnO-NPs against A. alternata, compared to ZnSO 4, could be due to nanoparticle properties interfering with cellular ion homeostasis mechanisms. The observed additive action of the oxidative phosphorylation-uncoupler fluazinam (FM) against all phenotypes indicates a possible role of ATP-dependent ion efflux mechanism in the mode of action of ZnO-NPs. A potential role of ROS production in the fungitoxic action of ZnO-NPs was evident by the additive/synergistic action of salicylhydroxamate (SHAM), which blocks the alternative oxidase antioxidant action. Mixture of ZnO-NPs and boscalid, resulting in a "capping" effect for the nanoparticles and significantly reducing their mean size, probably accounted for the synergistic effect of the mixture against both sensitive and resistant A. alternata isolates. Summarizing, results indicated that ZnO-NPs can be effectively used against A. alternata both alone or in combination with boscalid, providing an effective tool for combating SDHI-resistance and reducing the environmental fingerprint of synthetic fungicides., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The effect of copper (Cu-NPs, CuO-NPs), silver (Ag-NPs) and zinc oxide (ZnO-NPs) nanoparticles (NPs) on plant growth, physiological properties of tomato plants and their symbiotic relationships with the endophytic Fusarium solani FsK strain was investigated. Fungitoxicity tests revealed that the FsK strain was significantly more sensitive to Cu-NPs and ZnO-NPs than CuO-NPs and Ag-NPs both in terms of mycelial growth and spore germination. All NPs were more toxic to FsK compared to their bulk counterparts except for AgNO 3 , which was 8 to 9-fold more toxic than Ag-NPs. Apart from AgNO 3 , NPs and bulk counterparts did not affect the number of germinated tomato seeds even in higher concentrations, while root length was significantly reduced in a dose dependent way in most cases. Dry weight of tomato plants was also significantly reduced upon treatment with NPs and counterparts with most pronounced effects in the cases of AgNO 3 , Cu-NPs, ZnO-NPs, and ZnSO 4 . Root and shoot length of grown tomato plants was also affected by treatments while differences between NPs and bulk counterparts varied. A marked oxidative stress response was recorded in all cases of NPs/bulk counterparts as indicated by increased MDA and H 2 O 2 levels of treated plants. Treated plants had significantly reduced chlorophyl-a and carotenoid levels compared to the untreated control. NPs and counterparts did not affect FsK colonization of roots indicating a possible shielding effect of tomato plants once the endophyte was established inside the roots. Vice versa, a possible alleviation of CuO-NPs, ZnO-NPs, and ZnSO 4 toxicity was observed in the presence of FsK inside tomato roots in terms of plant dry weight. The results suggest that phytotoxicity of NPs in tomato treated plants should be considered before application and while both FsK and tomato are sensitive to NPs, their reciprocal benefits may extent to resistance towards these toxic agents., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Fusarium solani FsK is patented (20070100563/1006119, issued by the Industrial Property Organization to NK and KKP)., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The potential of Ag-NPs to suppress Monilia fructicola isolates and to broaden the effectiveness of fungicides to overcome resistance was tested in vitro and in vivo. Twenty-three M. fructicola isolates were subjected to fungitoxicity screening with a number of fungicides in vitro, which resulted in the detection of 18 isolates resistant to benzimidazoles (BEN-R) thiophanare methyl (TM) and carbendazim (CARB). DNA sequencing revealed the E198A resistance mutation in the β-tubulin gene, target site of the benzimidazole fungicides in all resistant isolates. Ag-NPs effectively suppressed mycelial growth in both sensitive (BENS) and resistant isolates. The combination of Ag-NPs with TM led to a significantly enhanced fungitoxic effect compared to the individual treatments regardless resistant phenotype (BEN-R/S) both in vitro and when applied on apple fruit. The above observed additive/synergistic action is probably associated with an enhanced Ag-NPs activity/availability as indicated by the positive correlation between Ag-NPs and TM + Ag-NPs treatments. No correlation was found between AgNO 3 and Ag-NPs suggesting that difference(s) exist in the fungitoxic mechanism of action between nanoparticles and their ionic counterparts. Synergy observed between Ag-NPs and the oxidative phosphorylation-uncoupler fluazinam (FM) against both resistance phenotypes indicates a possible role of energy (ATP) metabolism in the mode of action of Ag-NPs. Additionally, the role of released silver ions on the fungitoxic action of Ag-NPs against M. fructicola was found to be limited because the combination with NaCl revealed a synergistic rather than the antagonistic effect that would be expected from silver ion binding with chlorine ions. The results of this study suggested that Ag-NPs can be effectively used against M. fructicola and when used in combination with conventional fungicides they could provide the means for countering benzimidazole resistance and at the same time reduce the environmental impact of synthetic fungicides by reducing doses needed for the control of the pathogen., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Combating drug-resistance is a daunting task, especially due to the shortage of available drug alternatives with multisite modes of action. In this study, the potential of copper nanoparticles (Cu-NPs) to suppress 15 Botrytis cinerea isolates, which are sensitive or resistant to fungicides, alone or in combination with conventional fungicides, was tested in vitro and in vivo. Sensitivity screening in vitro revealed two fungicide resistance phenotypes, resulting from target site mutations. DNA sequencing revealed three B. cinerea isolates highly resistant to benzimidazoles (BEN-R), thiophanare methyl (TM), and carbendazim, bearing the E198A resistance mutation in the β-tubulin gene, and four isolates highly resistant to the QoI pyraclostrobin (PYR-R) with a G143A mutation in the cytb gene. Cu-NPs were equally effective against sensitive and resistant isolates. An additive/synergistic effect was observed between Cu-NPs and TM in the case of BEN-S isolates both in vitro and when applied in apple fruit. A positive correlation was observed between TM and TM + Cu-NPs treatments, suggesting that an increased TM availability in the target site could be related with the observed additive/synergistic action. No correlation between Cu(OH) 2 and Cu-NPs sensitivity was found, indicating that different mechanisms govern the fungitoxic activity between nano and bulk counterparts. A synergistic profile was observed between Cu-NPs and fluazinam (FM) - an oxidative phosphorylation inhibitor - in all isolates regardless of resistance phenotype, suggesting that ATP metabolism could be involved in the mode of action of Cu-NPs. Furthermore, the observed cross sensitivity and antagonistic action between Cu-NPs and NaCl also provided evidence for copper ions contribution to the fungitoxic action of Cu-NPs. The results suggested that Cu-NPs in combination with conventional fungicides can provide the means for an environmentally safe, sustainable resistance management strategy by reducing fungicide use and combating resistance against B. cinerea., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Nano-fungicides are expected to play an important role in future plant disease management as eco-friendly alternatives of conventional synthetic fungicides. In the present study, the sensitivity of seven fungal species, known to cause foliar and soil-borne diseases, to nanoparticles (NPs) containing copper (Cu-NPs, CuO-NPs), silver (Ag-NPs) and zinc (ZnO-NPs) was assessed in vitro. Mycelial growth assays revealed that Cu-NPs with mean inhibition rates, EC 50 , ranging between 162 and 310 μg/mL were most effective among the NPs tested in inhibiting fungal growth, followed by ZnO-NPs with EC 50 ranging between 235 and 848 μg/mL. All fungal species were practically insensitive to CuO-NPs and Ag-NPs except for B. cinerea, which was equally sensitive to Ag-NPs and Cu-NPs (EC 50  = 307 μg/mL). Cu-NPs were more fungitoxic in terms of mycelial growth, to almost all species tested, than a protective fungicide containing Cu(OH) 2 , which was used as a reference. Fungitoxicity experiments with the NPs tested and bulk size reagents containing the respective metals revealed that ZnO-NPs were more toxic to all fungal species tested than ZnSO 4 , whereas Cu-NPs were more fungitoxic than CuSO 4 in all cases, except for B. cinerea, A. alternata and M. fructicola. The existence of a positive correlation between Cu-NPs and CuO-NPs toxicity and, at the same time, the absence of any correlation between NPs tested and their respective bulk metal counterparts indicated potential differences in the mode of action between bulk and nanosized antifungal ingredients. Although there was considerable variation between fungal species, all NPs were generally 10 to 100 fold more fungitoxic to spores than hyphae and in the majority of cases more effective than Cu(OH) 2 , as revealed by colony formation bioassays. NPs significantly suppressed grey mold symptoms on plum fruit, especially Ag-NPs, which completely inhibited disease development. Consequently, tested NPs have the potential to be used as protective antifungal agents., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Nanofungicides are expected to play an important role in future plant disease management as eco-friendly alternatives of conventional synthetic fungicides. The suppressive properties of metal nanoparticles containing copper (Cu-NPs, CuO-NPs), silver (Ag-NPs) and zinc (ZnO-NPs) were assessed in vitro against seven causal agents of economically important foliar and soil-borne plant diseases. Cu-NPs were proven to be most successful in inhibiting mycelial growth (EC50 values ranging between 162 and 310 μg/mL) followed by ZnO-NPs exhibiting mean inhibition rates ranging between 235 and 848 μg/mL. All fungal species were practically insensitive to CuO-NPs while Ag-NPs exhibited significant inhibitory effect only against B. cinerea.. Comparative mycelial growth fungitoxicity experiments between the nanoparticles tested and their bulk sized counterparts revealed that ZnO-NPs were more toxic to all fungal species tested than ZnSO4 whereas CuNPs were more fungitoxic to CuSO4 in all cases except for B. cinerea, A. alternata and M. fructicola. A positive correlation between Cu-NPs and CuO-NPs toxicity and the absence of any correlation between NPs and their respective bulk metal counterparts is indicative of potential differences in the mode of action between bulk and nanosized metals. Although there was considerable variation between fungal species, fungitoxicity of all nanoparticles dramatically increased when applied to spores rather than fungal hyphae indicating a great potential of these compounds to be used as protective antifungal agents. [ABSTRACT FROM AUTHOR]