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Abstract: Aims: The present study was conducted to evaluate the antagonistic effect of Bacillus velezensisNRRL B‐23189 towards Penicillium roqueforti s.s. and Penicillium paneum (designated together as P. roqueforti s.l.) in silage conditions. Methods and Results: Corn silage conditions were simulated in vitro, and the impact of B. velezensis culture supernatant or cell suspension on P. roqueforti s.l. growth and roquefortine C production was evaluated. The antagonism was promising, but growth of B. velezensis in corn silage infusion was poor. Additionally, an in vivo experiment was carried out with mini‐silos containing a mixture of perennial ryegrass and white clover inoculated with P. roqueforti s.l. The applied B. velezensis cell suspension was unsuccessful in reducing P. roqueforti s.l. numbers, but did not compromise the silage acidification. Conclusions: Although the antagonism observed in vitro was promising, the applied B. velezensis cell suspension could not live up to the expectations in vivo. Significance and Impact of the Study: To our knowledge, the present study is the first one evaluating the antagonistic properties of B. velezensis towards toxigenic moulds in silage conditions, offering a good base for further research. [ABSTRACT FROM AUTHOR]

Abstract: Perennial ryegrass (Lolium perenne) is an important source of protein and energy for dairy cattle. To improve the protein/energy ratio of this forage, focus is now on improving its cell wall digestibility. The in vitro assessment of the digestible fraction of the neutral detergent fibre (NDFD) is a superior method for determining the cell wall digestibility, but requires the use of ruminal fluid, which has a highly variable composition and is often not readily available. As lignin is considered the main cell wall component that impedes NDFD, we investigated whether this “subtrait” could serve as alternative breeding selection criterion to improve NDFD. Therefore, we assessed the accuracy of two lignin quantification methods: van Soest (ADL) and Klason lignin (KL). We also considered KL estimates corrected for the solubilized lignin (total lignin or TL) and non‐solubilized protein (TL'). Although the latter is considered the truest possible lignin content, it was not always the most correlated to NDFD, due to the limited accuracy of protein quantification on lignin residue. TL is most correlated to NDFD and we therefore recommend it for use in conventional breeding if NDFD determination is not a possibility. However, NDFD is still a superior selection criterion, as it combines the effect of several subtraits and not just lignin. For marker–trait association studies, a more accurate estimate of lignin content is more important than a high correlation with NDFD, but also here, TL performs best. [ABSTRACT FROM AUTHOR]

To assess the incidence of early blight/brown spot (EB/BS) in Flanders (Belgium), potato fields in 22 locations were monitored and scored over the duration of the growing seasons 2014 and 2015. The average disease incidence was shown to be higher in 2014 than in 2015. Soil type, rainfall and temperature were also analysed in relation to disease incidence. In 2014, potato plants grown in sandy soils had more EB/BS disease than those grown in clay or loamy soils. However, the low disease incidence in 2015 meant that differences in disease levels between soil types could not be discerned for that growing season. A windowpane analysis demonstrated that rainfall and humidity accounted for the differences in disease incidence between the two growing seasons. During the course of the survey, the species composition in leaves with symptoms was assessed using real‐time PCR. Remarkably, small‐spored Alternaria species, such as A. alternata and A. arborescens, rather than the more virulent A. solani were the predominant species on potato leaves throughout the growing season. As the disease progressed, the proportion of A. solani increased. In view of these results, the virulence of a collected set of Alternaria isolates was assessed by an in vitro assay. Despite A. solani being more virulent than A. alternata or A. arborescens, the most abundant species isolated from potato leaves with symptoms was A. arborescens. [ABSTRACT FROM AUTHOR]

The objective of the present study was to calculate an optimal harvest period for both fresh and ensiled samples of forage maize and to calculate a set of harvest dates (called a harvest window), for which the variety ranking of the fresh forage corresponds with the variety ranking at the optimal harvest period calculated from the ensiled forage. Forage maize is fed almost exclusively as silage, but official variety trials with silage maize determine quality parameters in fresh (i.e. non-preserved) forage. Eight silage maize varieties were monitored at six harvest dates (from 25 to 40% dry matter content) in Merelbeke (Belgium) in 2013–15. At each harvest date, fresh samples were taken and half of the sampled material was ensiled in micro silos for 20 weeks. An optimal harvest period was calculated based on frequently measuring starch concentration and organic matter digestibility for both fresh and ensiled forage. Eventually, harvesting the silage maize at a dry matter content of 32–35% guaranteed an optimal harvest period. Based on the results of eight varieties, reporting variety ranks without going through the ensiling process continues to be a scientifically justified practice in Belgian official variety trials. Varieties with a superior fresh quality keep their leading position after ensiling, but variety differences become smaller after ensiling. [ABSTRACT FROM PUBLISHER]

Silage maize variety testing systems usually evaluate new varieties within properly defined maturity groups; within a maturity group, all varieties are harvested on a single harvest date. In a small country like Belgium where growing conditions are similar across the maize growing areas, all tested silage maize varieties, comprising different maturity groups, are harvested on a single date. Under this testing system, only few varieties are harvested at the physiological stage where they theoretically show their optimal performance. In the current research, eight silage maize varieties were monitored from 25 to 40% whole-crop dry matter (DM) content at three locations in Belgium during the period 2007–2009. The optimal harvest date was calculated as the date where whole-crop DM yield, ear DM yield and organic matter digestibility were maximal. The variety rank at the optimal harvest date was compared with the variety rank at any studied single harvest date. Harvest dates where the variety rank was not statistically different to the rank at the optimal harvest date were pooled in a ‘harvest window’. Based on the current data, the harvest window comprised a flexible harvest period of about 14 days. It was therefore concluded that applying a single harvest date is scientifically justified for the ranking of silage maize varieties in Belgium, when the whole-crop DM content is between 28·1 and 40·6% with a maximum difference of 7·2% between all compared varieties. [ABSTRACT FROM PUBLISHER]

The relevance of particular architectural characteristics for a good regrowth has not been elucidated yet in red clover. Here, we present the shoot architecture and regrowth of six red clover genotypes analysed under controlled and field conditions, with and without competition from perennial ryegrass. Among the genotypes investigated, Lemmon and Crossway displayed the highest number of nodes and branches, the highest increase in the number of nodes in the basal 7 cm and the highest outgrowth capacity of the nodes in this zone resulting in the highest dry-matter yields. Our results indicate that selecting genotypes with a high number of first-order branches, short internodes and the ability to resume growth from the nodes that remain in the uncut zone will allow the development of cultivars with good regrowth. [ABSTRACT FROM AUTHOR]

In monocots, lignin content has a strong impact on the digestibility of the cell wall fraction. Engineering lignin biosynthesis requires a profound knowledge of the role of paralogues in the multigene families that constitute the monolignol biosynthesis pathway. We applied a bioinformatics approach for genome-wide identification of candidate genes in Lolium perenne that are likely to be involved in the biosynthesis of monolignols. More specifically, we performed functional subtyping of phylogenetic clades in four multigene families: 4CL, COMT, CAD and CCR. Essential residues were considered for functional clade delineation within these families. This classification was complemented with previously published experimental evidence on gene expression, gene function and enzymatic activity in closely related crops and model species. This allowed us to assign functions to novel identified L. perenne genes, and to assess functional redundancy among paralogues. We found that two 4CL paralogues, two COMT paralogues, three CCR paralogues and one CAD gene are prime targets for genetic studies to engineer developmentally regulated lignin in this species. Based on the delineation of sequence conservation between paralogues and a first analysis of allelic diversity, we discuss possibilities to further study the roles of these paralogues in lignin biosynthesis, including expression analysis, reverse genetics and forward genetics, such as association mapping. We propose criteria to prioritise paralogues within multigene families and certain SNPs within these genes for developing genotyping assays or increasing power in association mapping studies. Although L. perenne was the target of the analyses presented here, this functional subtyping of phylogenetic clades represents a valuable tool for studies investigating monolignol biosynthesis genes in other monocot species. [ABSTRACT FROM AUTHOR]

An LC-MS/MS method was developed and validated for the simultaneous determination of deoxynivalenol, zearalenone, T-2-toxin, HT-2-toxin and metabolites, including 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, deoxynivalenol-3-glucoside, α-zearalenol, β-zearalenol, zearalenone-4-glucoside, α-zearalenol-4-glucoside, β-zearalenol-4-glucoside and zearalenone-4-sulfate in maize, wheat, oats, cornflakes and bread. Extraction was performed with acetonitrile/water/acetic acid (79/20/1, v/v/v) followed by a hexane defatting step. After filtration, the extract was evaporated and the residue was redissolved in mobile phase for injection. The mobile phase, which consisted of a mixture of methanol and water with 10 mM ammonium acetate, was adjusted to pH 3 with glacial acetic acid. A sample clean-up procedure was not included because of the low recoveries of free and masked mycotoxins and their differences in polarity. The method allowed the simultaneous determination of 13 Fusarium mycotoxins in a one-step chromatographic run using a Waters Acquity UPLC system coupled to a Quattro Premier XE mass spectrometer. The method was validated for several parameters such as linearity, apparent recovery, limit of detection, limit of quantification, precision, expanded measurement uncertainty and specificity. The limits of detection varied from 5 to 13 ng g−1; those for the limit of quantification from 10 to 26 ng g−1. The results of the performance characteristics of the developed LC-MS/MS method were in good agreement with the criteria mentioned in Commission Regulation (EC) No. 401/2006. Thirty samples of a variety of food and feed matrices were sampled and analysed between July 2010 and January 2011. [ABSTRACT FROM PUBLISHER]

The effect of humic substances on the nutrient uptake, herbage production and nutritive value of herbage from sown grass pastures was studied in six field experiments. Commercial humic substances were applied in combination with mineral fertilizer or slurry, either as a solution (HF liquid; 8·3 kg humic substances ha−1) or incorporated into the mineral fertilizer (HF incorporated; 3·6 to 6·4 kg humic substances ha−1). A series of cuts, ranging from two to five cuts, was taken during the growing season. The general response in herbage production to application of humic substances was an increase in herbage mass of dry matter (DM) at the first cut although this was only significant in two experiments for the HF incorporated treatment. Total herbage production of DM over the growing season, however, was similar for treatments with or without application of humic substances. The overall effect of HF incorporated and HF liquid on the herbage mass of DM at the first cut across the experiments was calculated using a meta-analysis technique and it was shown that there was a significant proportional increase of 0·14 ( P < 0·05) with the HF incorporated treatment and a non-significant increase of 0·08 with the HF liquid treatment compared to the control treatment. The nutritive value of the herbage at the first cut was similar across all treatments. In general nitrogen, phosphorus and potassium uptake at the first grass cut was higher after application of humic substances but only in one experiment was this increase statistically significant. [ABSTRACT FROM AUTHOR]

The effect of humic substances on yield and nutrient uptake of grass, maize, potato and spinach was investigated in six field experiments and two pot experiments in a high input cropping system. The humic substances originating from leonardite formations in Canada were applied as liquid solution to the soil (Humifirst liquid) or as a solid incorporated in mineral fertilizers (Humifirst incorporated). Formal meta-analysis of the results of all executed experiments showed that the application of humic substances had an overall positive effect on dry matter yield of the crops and this effect was statistically significant for Humifirst incorporated. In the case of permanent grassland, humic substances promoted mainly the production of the first grass cut, which has the highest grass quality among all cuts during the growing season. Tuber production on the potato field trial showed a high response on the application of humic substances. Total potato yield increased with 13 and 17% for Humifirst liquid and Humifirst incorporated, respectively. The effect of humic substances on maize yield was limited, probably due to the rather high nutrient status of both soils. Finally, the formal meta-analysis showed a consequent increase in nitrogen and phosphorus uptake of all studied crops as well. The effect on potassium and magnesium uptake was also mainly positive, while sodium and calcium uptake were not affected in most of the experiments. [ABSTRACT FROM AUTHOR]

Biotransformation of mycotoxins in animals comprises phase I and phase II metabolisation reactions. For the trichothecene deoxynivalenol (DON), several phase II biotransformation reactions have been described resulting in DON-glutathiones, DON-glucuronides and DON-sulfates made by glutathione-S-transferases, uridine-diphosphoglucuronyl transferases and sulfotransferases, respectively. These metabolites can be easily excreted and are less toxic than their free compounds. Here, we demonstrate for the first time in the animal kingdom the conversion of DON to DON-3-glucoside (DON-3G) via a model system with plant pathogenic aphids. This phase II biotransformation mechanism has only been reported in plants. As the DON-3G metabolite was less toxic for aphids than DON, this conversion is considered a detoxification reaction. Remarkably, English grain aphids (Sitobion avenae) which co-occur with the DON producer Fusarium graminearum on wheat during the development of fusarium symptoms, tolerate DON much better and convert DON to DON-3G more efficiently than pea aphids (Acyrthosiphon pisum), the latter being known to feed on legumes which are no host for F. graminearum. Using a non-targeted high resolution mass spectrometric approach, we detected DON-diglucosides in aphids probably as a result of sequential glucosylation reactions. Data are discussed in the light of an eventual co-evolutionary adaptation of S. avenae to DON. [ABSTRACT FROM AUTHOR]

Abstract Background Powdery mildew, caused by the obligate biotrophic fungus Blumeria graminis, is a major problem in cereal production as it can reduce quality and yield. B. graminis has evolved eight distinct formae speciales (f.sp.) which display strict host specialization. In the last decade, powdery mildew has emerged on triticale, the artificial intergeneric hybrid between wheat and rye. This emergence is probably triggered by a host range expansion of the wheat powdery mildew B. graminis f.sp. tritici. To gain more precise information about the evolutionary processes that led to this host range expansion, we pursued a combined pathological and genetic approach. Results B. graminis isolates were sampled from triticale, wheat and rye from different breeding regions in Europe. Pathogenicity tests showed that isolates collected from triticale are highly pathogenic on most of the tested triticale cultivars. Moreover, these isolates were also able to infect several wheat cultivars (their previous hosts), although a lower aggressiveness was observed compared to isolates collected from wheat. Phylogenetic analysis of nuclear gene regions identified two statistically significant clades, which to a certain extent correlated with pathogenicity. No differences in virulence profiles were found among the sampled regions, but the distribution of genetic variation demonstrated to be geography dependent. A multilocus haplotype network showed that haplotypes pathogenic on triticale are distributed at different sites in the network, but always clustered at or near the tips of the network. Conclusions This study reveals a genetic structure in B. graminis with population differentiation according to geography and host specificity. In addition, evidence is brought forward demonstrating that the host range expansion of wheat isolates to the new host triticale occurred recently and multiple times at different locations in Europe.

Abstract Background Fusarium head blight is a very important disease of small grain cereals with F. graminearum as one of the most important causal agents. It not only causes reduction in yield and quality but from a human and animal healthcare point of view, it produces mycotoxins such as deoxynivalenol (DON) which can accumulate to toxic levels. Little is known about external triggers influencing DON production. Results In the present work, a combined in vivo/in vitro approach was used to test the effect of sub lethal fungicide treatments on DON production. Using a dilution series of prothioconazole, azoxystrobin and prothioconazole + fluoxastrobin, we demonstrated that sub lethal doses of prothioconazole coincide with an increase in DON production 48 h after fungicide treatment. In an artificial infection trial using wheat plants, the in vitro results of increased DON levels upon sub lethal prothioconazole application were confirmed illustrating the significance of these results from a practical point of view. In addition, further in vitro experiments revealed a timely hyperinduction of H2O2 production as fast as 4 h after amending cultures with prothioconazole. When applying H2O2 directly to germinating conidia, a similar induction of DON-production by F. graminearum was observed. The effect of sub lethal prothioconazole concentrations on DON production completely disappeared when applying catalase together with the fungicide. Conclusions These cumulative results suggest that H2O2 induced by sub lethal doses of the triazole fungicide prothioconazole acts as a trigger of DON biosynthesis. In a broader framework, this work clearly shows that DON production by the plant pathogen F. graminearum is the result of the interaction of fungal genomics and external environmental triggers.

An abstract of the article "Impact of Fungicide Timing on the Composition of the Fusarium Head Blight Disease Complex and the Presence of Deoxynivalenol (Don) in Wheat" by K. Audenaert and colleagues is presented.

Fungal contamination of metalworking fluids (MWF) is a dual problem in automated processing plants because resulting fungal biofilms obstruct cutting, drilling, and polishing machines. Moreover, some fungal species of MWF comprise pathogens such as Fusarium solani. Therefore, the development of an accurate analytical tool to evaluate conidial viability in MWF is important. We developed a flow cytometric method to measure fungal viability in MWF using F. solani as the model organism. To validate this method, viable and dead conidia were mixed in several proportions and flow was cytometrically analyzed. Subsequently, we assessed the fungicidal activity of two commercial MWF using flow cytometry (FCM) and compared it with microscopic analyses and plating experiments. We evaluated the fungal growth in both MWF after 7 days using quantitative PCR (qPCR) to assess the predictive value of FCM. Our results showed that FCM distinguishes live from dead conidia as early as 5 h after exposure to MWF, whereas the microscopic germination approach detected conidial viability much later and less accurately. At 24 h, microscopic analyses of germinating conidia and live/dead analyses by FCM correlated well, although the former consistently underestimated the proportion of viable conidia. In addition, the reproducibility and sensitivity of the flow cytometric method were high and allowed assessment of the fungicidal properties of two commercial MWF. Importantly, the obtained flow cytometric results on viability of F. solani conidia at both early time points (5 h and 24 h) correlated well with fungal biomass measurements assessed via a qPCR methodology 7 days after the start of the experiment. IMPORTANCE This result shows the predictive power of flow cytometry (FCM) in assessing the fungicidal capacity of MWF formulations. It also implies that FCM can be implemented as a rapid detection tool to estimate the viable fungal load in an industrial processing matrix (MWF). [ABSTRACT FROM AUTHOR]

Triticale (xTriticosecale) is the intergeneric hybrid between the female parent wheat and the male parent rye. With the expansion of the triti-cale growing area, powdery mildew emerged on this new host and has become a significant disease on triticale. Recent research demonstrated that this "new" powdery mildew on triticale has emerged through a host range expansion of powdery mildew of wheat. Moreover, this expansion occurred recently and multiple times at different locations in Europe. An effective and environmentally sensitive approach to con-trolling powdery mildew involves breeding crop plants for resistance. The main goal of this study was to identify the presence of powdery mildew resistance in commercial triticale cultivars. First, the avirulence (AVR) genes and gene complexity carried by this new powdery mildew population on triticale were characterized. Virulence was identified for all the resistance genes evaluated in the present study, and virulence frequencies higher than 50% were recorded on the genes Pm3f, Pm5b, Pm6, Pm7, Pm8, and Pm17. Using molecular markers, the presence of resistance genes Pm3f and Pm17 was identified in certain triticale cultivars. The triticale cultivars were also evaluated for the presence of quantitative resistance at adult plant growth stages in a 2-year field experiment. Despite the high disease pressure, cultivars highly resistant at the adult-plant growth stages were identified. Because 'Grenado' also showed effective race-specific resistance, this cultivar could be of high value for breeding for durable resistance to powdery mildew. Altogether, this study reveals valuable information on the presence of powdery mildew resistance in commercial triticale cultivars, which can be used in breeding programs in triticale. Additionally, this study underscores the need to broaden the base of powdery mildew re-sistance in triticale through introgression and deployment of new sources of mildew resistance, including quantitative resistance. [ABSTRACT FROM AUTHOR]

Despite great efforts to forecast plant diseases, many of the existing systems often fall short in providing farmers with accurate predictions. One of the main problems arises from the existence of year and location effects, so that more advanced procedures are required for evaluating existing systems in an unbiased manner. This paper illustrates the case of Fusarium head blight of winter wheat in Belgium. We present a new cross-validation strategy that enables the evaluation of the predictive performance of a forecasting system for years and locations that are different from the years and locations on which the forecast was developed. Four different cross-validation strategies and five regression techniques are used. The results demonstrated that traditional evaluation strategies are too optimistic in their predictions, whereas the crossyear cross-location validation strategy yielded more realistic outcomes. Using this procedure, the mean squared error increased and the coefficient of determination decreased in predicting disease severity and deoxynivalenol content, suggesting that existing evaluation strategies may generate a substantial optimistic bias. The strongest discrepancies between the cross-validation strategies were observed for multiple linear regression models. [ABSTRACT FROM AUTHOR]

Buckwheat starch has attracted worldwide attention in the food industry as a valuable raw material or food additive. Nitrogen (N) and sulfur (S) are two nutrients essential to ensure grain quality. This study investigated the combined application of N fertilizer (0, 45 and 90 kg N ha -1 ) and S fertilizer (0 and 45 kg SO 3 ha -1 ) on the chemical composition, structure and physicochemical properties of buckwheat starch. The results showed that increasing the fertilizer application decreased amylose content and starch granule size but increased light transmittance, water solubility and swelling power. The stability of the absorption peak positions and the decrease in short-range order degree suggested that fertilization influenced the molecular structure of buckwheat starch. In addition, increases in viscosity and gelatinization enthalpy as well as decreases in gelatinization temperatures and dynamic rheological properties indicated changes in the processing characteristics and product quality of buckwheat-based foods., 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 © 2024 Elsevier Ltd. All rights reserved.)

Maize lethal necrosis (MLN) is a maize disease caused by the maize chlorotic mottle virus (MCMV), a potyvirus which causes yield losses of 30-100%. The present study aimed to isolate nanobodies against the MCMV coat protein (CP) for the diagnosis of MLN. MCMV CP expressed in Escherichia coli was used for llama immunization. VHH (i.e. variable heavy domain of heavy chain) gene fragments were prepared from blood drawn from the immunized llama and used to generate a library in E. coli TG1 cells. MCMV specific nanobodies were selected by three rounds of phage display and panning against MCMV CP. The selected nanobodies were finally expressed in E. coli WK6 cells and purified. Eleven MCMV-specific nanobodies were identified and shown to detect MCMV in infected maize plants. Thus, our results show that nanobodies isolated from llama immunized with MCMV CP can distinguish infected and healthy maize plants, potentially enabling development of affordable MCMV detection protocols., (© 2024 The Author(s). FEBS Open Bio published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

The present study investigated the effect of nitrogen fertilization (NF) at the levels of 0, 45, and 90 kg·ha -1 combined with selected sulfur complex fertilization (SCF) levels of 0 and 45 kg·ha -1 on the nutritional and technological characteristics of buckwheat flour from five varieties. The results showed that the genotype was a critical factor affecting the chemical composition and physicochemical properties of buckwheat flour. NF significantly increased protein, total starch, and amylose content as well as mineral composition but decreased particle size, color value, and water hydration properties. However, SCF enhanced the ash content and decreased the protein content but had no significant effect on the pasting temperature. In addition, the combination of NF and SCF significantly reduced granule size, water solubility, viscosity, and rheological properties with increasing fertilization levels. This study can guide the cultivation of buckwheat with the desired physicochemical properties and provide information for buckwheat-based products in the food industry.

Fusarium wilt, caused by Fusarium oxysporum f. sp. spinaciae , causes a significant challenge on vegetative spinach and seed production. Addressing this issue necessitates continuous research focused on innovative treatments and protocols through comprehensive bioassays. Recent studies have highlighted the potential of plant-based compounds in controlling fungal diseases. The present work aims to conduct a series of experiments, encompassing both in vitro and in planta assessments, to investigate the biocontrol capabilities of different essential oils (EOs) at various application rates, with the ultimate goal of reducing the incidence of Fusarium wilt in spinach. The inhibitory effect of four plant EOs (marjoram, thyme, oregano, and tea tree) was initially assessed on the spore germination of five unknown Fusarium strains. The outcomes revealed diverse sensitivities of Fusarium strains to EOs, with thyme exhibiting the broadest inhibition, followed by oregano at the highest concentration (6.66 μL/mL) in most strains. The tested compounds displayed a diverse range of median effective dose (ED50) values (0.69 to 7.53 µL/mL), with thyme and oregano consistently showing lower ED50 values. The direct and indirect inhibitory impact of these compounds on Fusarium mycelial growth ranged from ~14% to ~100%, wherein thyme and oregano consistently exhibiting the highest effectiveness. Following the results of five distinct inoculation approaches and molecular identification, the highly pathogenic strain F-17536 ( F. oxysporum f.sp. spinaciae ) was chosen for Fusarium wilt assessment in spinach seedlings, employing two promising EO candidates through seed and soil treatments. Our findings indicate that colonized grain (CG) proved to be a convenient and optimal inoculation method for consistent Fusarium wilt assessment under greenhouse conditions. Seed treatments with thyme and oregano EOs consistently resulted in significantly better disease reduction rates, approximately 54% and 36% respectively, compared to soil treatments (P > 0.05). Notably, thyme, applied at 6.66 µL/mL, exhibited a favorable emergence rate (ERI), exceeding seven, in both treatments, emphasizing its potential for effective disease control in spinach seedlings without inducing phytotoxic effects. This study successfully transitions from in vitro to in planta experiments, highlighting the potential incorporation of EOs into integrated disease management for Fusarium wilt in spinach production., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Mirmajlessi, Najdabbasi, Sigillo and Haesaert.)