Effect of temperature and precipitation on the occurrence of Trichoderma ear rot in Europe

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While some species are used as effective biocontrol agents against plant pathogens, others, such as T. afroharzianum , have recently been identified as pathogenic, causing diseases like Trichoderma ear rot in maize and infections in wheat. The occurrence of Trichoderma species is strongly influenced by environmental factors, particularly temperature and precipitation. This study aims to assess the occurrence and distribution of Trichoderma ear rot in maize across Europe, and to evaluate the effect of environmental factors, especially temperature and precipitation, on their occurrence. Over a seven-year period (2018–2024), 345 samples of maize cobs, stalks, and soil samples were received from 72 locations in Europe. The obtained Trichoderma isolates (n = 130) were identified to the species level, and their pathogenicity to maize was tested under controlled conditions. Weather data, including temperature and precipitation, were gathered from each location and correlated with the occurrence of Trichoderma ear rot. Additionally, in vitro and in vivo climate chamber trials were conducted to confirm the favorable temperature conditions for disease development. Our results indicate that T. afroharzianum was the most prevalent species isolated from maize cobs and agricultural soil, exhibiting strong pathogenicity in maize cobs. Correlation analysis between the occurrence of T. afroharzianum and environmental factors revealed that high temperatures (> 19.6°C mean temperature), particularly during the summer months, strongly favored the presence of T. afroharzianum , while cooler and wetter regions (< 18.2 mean temperature) showed little to no occurrence of pathogenic Trichoderma species. In vitro and in vivo climate chamber experiments confirmed these results observed in the field. The growth rate of T. afroharzianum isolates was significantly higher compared to other Trichoderma species, with an optimal temperature range between 28 and 32°C. Trichoderma maize ear rot environmental conditions infection Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 1. Introduction Maize ( Zea mays L.) is one of the most important staple crops worldwide, playing a critical role in global agriculture. As a versatile and highly productive cereal, maize serves as a primary source of food for humans and a key ingredient in animal feed (Erenstein et al. 2022 ). For human consumption, maize is an essential food source in many developing countries, providing a substantial proportion of daily caloric intake. Maize can be processed into a variety of products, including flour for tortillas, polenta, and maize meal, which are staple foods in many regions. In addition, maize silage is a fundamental component of livestock feed, particularly for poultry, swine, and cattle due to its high energy content (Tenaillon and Charcosset 2011 ). As the global demand for both food and animal feed continues to rise, maize production faces significant challenges, including pest and disease stresses (Shiferaw et al. 2011 ). The evolving environmental conditions can significantly affect the prevalence and severity of pest and pathogen damage, with existing pathogens potentially gaining or losing significance (Anderson et al. 2004 ). Additionally, the emergence of new pathogens is anticipated to become an increasing concern in the future (King et al. 2023 ). Trichoderma species belong to the division Ascomycota and are globally distributed, inhabiting diverse ecosystems ranging from dead organic matter to forest and agricultural soils (Druzhinina et al. 2010 ; Cai and Druzhinina 2021a ). Known for their rapid growth and competitive nature, Trichoderma species produce and detoxify antimicrobial substances, contributing to their success as free-living soil-borne fungi (Kubicek and Harman 2002 ). Furthermore, Trichoderma strains from agricultural soils often form opportunistic symbiotic relationships with plants, enhancing their utility as biological control agents. Some Trichoderma species exhibit antagonistic effects against a wide variety of plant pathogens, including fungi, oomycetes, bacteria, and nematodes, through mechanisms such as rapid growth, nutrient competition, mycoparasitism, and antibiosis (Harman 2005 ). However, Trichoderma spp. has also been identified as a causative agent of ear rot in maize in the United States, typically associated with mechanical damage to the cob (Sutton 1972 ; Munkvold and White 2016 ; Wise et al. 2016 ). In 2018, a pathogenic Trichoderma species was discovered on maize cobs in Europe, identified as T. afroharzianum , which infects maize cobs and produces white mycelium that is rapidly producing dark, bearing grey-green conidia (Pfordt et al. 2020 ). Severe infections result in enzymatic decomposition of starch in the kernels, making cobs soft and watery through the secretion of alpha-amylase (Pfordt et al. 2024 ). Starch is a primary carbohydrate reserve in the grains of most cereals, and the production of enzymes like alpha-amylase by Trichoderma allows the fungus to infect not only maize but also other cereals, such as wheat and barley (Pfordt et al. 2023 ). Since the initial discovery in Southern Germany in 2018, the disease has been observed on maize in other regions of Germany, France, Italy, China, and India (Sanna et al. 2022 ; Chen et al. 2023 ; Harish et al. 2025 ), particularly following dry and hot summer periods. This pattern suggests that climatic conditions and meteorological factors determine the occurrence of Trichoderma ear rot in maize, with elevated temperatures during June, July, and August potentially enhancing natural infections during the flowering period (Pfordt et al. 2020 ). Trichoderma species are cosmopolitan, occurring in diverse habitats in temperate, tropical, and subtropical regions. However, the abundance and species composition can vary across different geographic locations (Ghorbanpour et al. 2018 ). Most Trichoderma species thrive at moderate temperatures, typically between 20–30°C. This range supports their rapid growth and efficient colonization of substrates (Begoude et al. 2007 ; Zehra et al. 2017 ). While some species, like T. harzianum and T. afroharzianum , are considered cosmopolitan and found across multiple continents, other species show more localized occurrences, which is indicative of restricted ecological adaptation. For instance, species like T. simonsii , T. atrobrunneum and T. atroviride , exhibit a preference for lower temperatures and are found in cooler regions, including temperate zones like Europe and North America. Meanwhile other species like T. asperellum and T. longibrachiatum are more commonly isolated in tropical and subtropical regions (Chaverri et al. 2015 ; Di Lelio et al. 2021 ). The aim of this study is to ascertain the existence of a correlation between local meteorological conditions and the occurrence of Trichoderma species in maize fields across Europe. In addition to field observations, in vitro and in vivo climate chamber experiments were conducted to further investigate the influence of temperature on Trichoderma species. By providing insights into the environmental drivers of Trichoderma distribution, this research seeks to contribute to better management strategies for mitigating the risks posed by pathogenic Trichoderma species, while also supporting the beneficial applications of non-pathogenic strains in agriculture. 2. Results 2.1 Occurrence of Trichoderma species obtained from maize and soil samples in Germany Over the past seven years (2018–2024), a total of 345 samples were received from symptomatic and asymptomatic maize cobs and stalks, as well as agricultural soils across 72 different locations in Europe. Among these, 130 Trichoderma isolates were cultured from 54 sampling sites (Sub. Table 1 ). Of the isolates, 48 were obtained from maize cobs and stalks, while the remaining 82 derived from soil samples. The highest incidence of visible maize cob infections and the presence of T. afroharzianum were found predominantly at sites in Southern Germany, particularly in Bavaria along the Donau river, and in Rhineland-Palatinate and along the Rhine River in the Alsace (Germany and France). Additionally, several locations in central Germany, especially in the warmer and drier regions around Magdeburg (Saxony-Anhalt) and Brandenburg along the Oder river, also exhibited maize cob infections and soil isolates of T. afroharzianum . In contrast, no T. afroharzianum isolates were detected in cooler areas of central and northern Germany. In addition to the isolates obtained in Alsace, one isolate was found in the southwest region of Aquitaine, and another in Burgundy. Three isolates were found in northern Italy and two isolates in Türkiye, in Salihli (Aegean region) and Tarsus (Belediye region). Additionally, Trichoderma ear rot disease was identified in Austria, specifically in Neustift, Heiligenkreuz, and Rudersdorf, located in the Nordburgenland region near the borders with Bratislava and Hungary (Fig. 1 ). Notably, at sites where maize ear rot was observed, T. afroharzianum was consistently isolated from both maize cobs and soil samples. This correlation suggests a close association between soil-borne populations of T. afroharzianum and the occurrence of cob infections. However, T. afroharzianum was also detected in soil samples from locations without visible cob infections, indicating that the fungus can persist in the soil even in the absence of visible disease symptoms. All T. afroharzianum isolates subjected to pathogenicity tests exhibited a high level of virulence, causing significant disease symptoms in maize plants. 2.2 Trichoderma species isolated from maize and soil samples A total of six different Trichoderma species were isolated from the maize cobs (Table 1 ). The majority of isolates (n = 27) were identified as T. afroharzianum , exhibited significant pathogenicity, causing 89.4% disease severity. In contrast, T. harzianum was also isolated, but only in a smaller number of three isolates colonizing the cobs endophytically without causing symptoms (0.8 disease severity). Additionally, isolates of T. atroviride (8 isolates) and T. gamsii (4 isolates) were recovered, both of which exhibited mild pathogenicity of 18.8% and 14.6% disease severity after artificial inoculation on the maize cobs. Five isolates of T. azevodoi and one isolate of T. tomentosum were obtained, which showed no pathogenicity. In addition to the isolates obtained from maize cobs, the study also identified several Trichoderma species from soil samples. A total of 82 isolates were recovered from the soil, depicting a diversity of thirteen species. Among these, T. afroharzianum (16 isolates) and T. harzianum (15 isolates) were the most prevalent, followed by T. peberdyi , (10 isolates), T. atroviride (8 isolates) and T. cerinum (7 isolates). All T. afroharzianum isolates from soil exhibited high pathogenicity (86.6%) while the isolates of the remaining species, including T. harzianum and T. atroviride , displayed low to moderate (0.2–37%) levels of pathogenicity. Table 1 Origin and pathogenicity (% disease severity, DS) of various Trichoderma species isolated from maize cobs and agricultural soil. Species Maize Soil Total n % DS n % DS T. afroharzianum 27 89.4 16 86.6 43 T. harzianum 3 0.8 15 2.1 18 T. atroviride 8 18.8 8 23.2 16 T. gamsii 4 14.6 1 5 T. azevodoi 5 0.7 5 T. tomentosum 1 0.0 1 T. peberdyi 10 1.1 10 T. cerinum 7 37.0 7 T. koningiopsis 5 0.9 5 T. velutinum 7 2.6 7 T. hamatum 5 1.1 5 T. atrobrunneum 2 0.2 2 T. asperellum 2 8.0 2 T. arundinaceum 2 28.6 2 T. brevicompactum 2 2.9 2 T. virens 1 1.1 1 Total 48 43.4 82 22.45 130 2.3 Effect of temperature and precipitation on the occurrence of Trichoderma ear rot in Europe The Pearson correlation coefficients (r) for temperature and precipitation from April to September (2018–2023) were assessed with the occurrence of pathogenic T. afroharzianum isolates (Sub. Table 2). Temperature in June (r = 0.36), July (r = 0.6), and August (r = 0.33), showed significant correlation with the occurrence of Trichoderma ear rot, suggesting that higher temperatures favor the occurrence of Trichoderma ear rot (Fig. 2 ) during flowering time. The correlation for precipitation shows a negative trend, especially in July (-0.4), indicating that dry conditions favour the occurrence of Trichoderma ear rot during this month. Figure 3 illustrates the correlation between the Trichoderma ear rot infection per location and weather conditions, specifically temperature and precipitation, during June, July, and August (2018–2023). In June, there is a positive correlation between temperature and Trichoderma ear rot infection (r = 0.4401), indicating that higher temperatures in this month slightly favor the disease. In contrast, June precipitation shows a weak negative correlation with Trichoderma ear rot infection (r = -0.2888), suggesting that lower rainfall levels are associated with higher disease infection. The relationship between environmental factors and Trichoderma ear rot infection becomes more pronounced in July. Here, temperature shows a strong positive correlation with the disease (r = 0.6079), highlighting that elevated temperatures during this month significantly favor Trichoderma ear rot infections. This period is particularly critical as it coincides with maize flowering, when the plants are most susceptible. July precipitation, on the other hand, displays a moderate negative correlation with Trichoderma ear rot infection (r = -0.3549), emphasizing the role of drought stress during this vulnerable growth stage. In August, the correlations between environmental factors and Trichoderma ear rot infection are decreasing. Temperature exhibits a weak positive correlation (r = 0.3315), suggesting that higher temperatures in August may still contribute to the development of Trichoderma ear rot, though to a lesser extent than in July. Precipitation shows a weak, non-significant negative correlation (r = -0.2394), indicating that the effect of rainfall becomes less pronounced in late summer. 2.4 Effect of in vitro temperature on the growth rate Temperature exerts a significant effect (p < 0.0001) on the radial growth of Trichoderma. spp. in vitro . All factors, such as isolate, temperature and their interaction, are significant (p < 0.0001), whereas there was no significant effect of the replicate (Sub.Table 3). There was no fungal growth at 8°C and above 40°C while growth was observed in the range from 12 to 36°C for all isolates (Table 3). T. afroharzianum isolates, including AP18TRI2 and APP18TRI1, demonstrated the highest growth rates (Sub. Figure 1 ), especially at the optimal temperature range of 28°C to 32°C. At 32°C, AP18TRI2 exhibited the fastest growth (40.6 mm), followed by APP18TRI1 (37.4 mm). In contrast, T. asperellum (T34) and T. harzianum (T39) showed comparatively slower growth, with T. asperellum reaching 29.1 mm at 32°C. All isolates experienced reduced growth at extreme temperatures, particularly at 36°C and 12°C, though T. afroharzianum isolates maintained relatively higher growth rates compared to non-pathogenic strains like CBS 124.620 at low temperatures. For instance, at 12°C, T. afroharzianum isolates grew better (10.9 mm for AP18TRI2) than CBS 124.620 (7.9 mm). 2.5 Effect of temperature on disease severity in vivo The results indicate that temperature and isolate as well as the interaction has a significant (p < 0.0001) impact on the disease severity of Trichoderma spp. on maize cobs (Sub. Table 3). The impact of temperature on disease severity caused by four Trichoderma isolates (CBS124620, AP18TRI2, T34, TS1) was assessed across a temperature range from 10°C to 35°C (Fig. 4 ). The results indicate that disease severity increases with rising temperature for all isolates, with significant differences in pathogenicity observed between the isolates. The reference strain CBS124620 exhibited low disease severity at temperatures below 20°C, with minimal infection (below 10%) at 10°C and 15°C. However, at 30°C and 35°C, disease severity increased significantly, reaching nearly 50% at 35°C, suggesting that even mild-pathogenic isolates can exhibit higher disease severity under favorable conditions. The pathogenic T. afroharzianum isolate AP18TRI2 demonstrated the highest disease severity across the temperature range. At 20°C, AP18TRI2 caused severe infection, with disease severity peaking at nearly 80%. Disease severity decreased at higher temperatures (30°C and 35°C), with values of 60% and 50%, respectively. The T. afroharzianum isolate TS1 followed a similar trend as AP18TRI2, showing severe infection at 20°C (over 70%) and slightly decreasing disease severity (62%) at 35°C. Surprisingly, the biocontrol T. asperellum isolate T34 also showed severe disease symptoms on maize cobs with disease severity up to 58% at both 30°C and 35°C. 3. Discussion A total of 109 Trichoderma isolates from sixteen different species were obtained from agricultural soils and maize tissue. The most prevalent species in both, agricultural soil samples and maize tissue was T. afroharzianum , showing high pathogenicity on maize cobs. The highest variety of Trichoderma species was found in agricultural soil, with a greater number of different species like T. peberdyi , T. cerinum, T. koningiopsis , and T. hamatum , which were not found in maize tissue. In contrast, maize tissue yielded fewer species, predominantly consisting of T. afroharzianum , T. harzianum , T. atroviride , and T. gamsii . However, T. afroharzianum was the only species exhibiting high pathogenicity on the cob, other species like T. harzianum appeared to colonize maize cobs endophytically without causing symptoms. This finding aligns with previous observations that T. afroharzianum is a prominent species with the potential to cause Trichoderma ear rot in maize. It also underscores the capacity of this species to persist in soil while actively infecting maize under favorable conditions (Pfordt et al. 2020 ; Pfordt et al. 2024 ). The study highlights distinct distribution patterns of pathogenic Trichoderma species across Europe, with T. afroharzianum being prevalent in warmer and dry regions, and absent in cooler, northern regions. This suggests that temperature and precipitation are key factors influencing the occurrence of pathogenic Trichoderma species (Zehra et al. 2017 ; Qiu et al. 2017 ). Southern Germany, France and Italy typically experience higher temperatures throughout the year when compared to the northern part of the Germany. The southern regions of Germany, especially in areas like Bavaria and Baden-Württemberg, have a more continental climate, characterized by hotter summers and colder winters with average monthly temperatures ranging from 18°C and 25°C (Flohn 1984 ; Kaspar et al. 2016 ). In contrast, Northern Germany, including regions like Lower Saxony and Schleswig-Holstein, generally experiences cooler summer temperatures, often ranging from 15°C to 20°C with a more temperate maritime climate due to its proximity to the North Sea and the Baltic Sea (Maier et al. 2003; Müller-Westermeier and Rocznik 2006 ; Schönwiese 2020 ). This temperature gradient is crucial for the growth and prevalence of pathogenic Trichoderma species, as warmer conditions in the south favor the proliferation of these strains, explaining their higher occurrence in Southern Germany (Wong et al. 2002 ). In addition, Southern Germany and regions in Saxony-Anhalt are more prone to longer and more intense drought periods, particularly during the summer months, creating conditions that stress plants more severely (Müller-Westermeier and Rocznik 2006 ; Kaspar et al. 2013 ). Similarly, the Nordburgenland region in Austria, near the borders with Bratislava and Hungary, features a Pannonian climate with warm to hot summers (25°C to 30°C) and mild winters (0°C to 5°C). Annual precipitation is relatively low, ranging from 500 to 600 mm, relatively dry compared to other parts of Austria (Hammerl 2001 ; GeoSphere Austria). In Italy, pathogenic T. afroharzianum isolates have so far only been identified in the northern part, particularly in Piemont, Lombardy, and Venetia, where maize is extensively cultivated due to the favorable climate and fertile soils of the Po Valley (Bocchiola et al. 2013 ; Borrelli et al. 2014 ). The climate here is characterized by warm summers with moderate precipitation, creating conditions conducive to both maize growth and the proliferation of T. afroharzianum . In contrast, Southern Italy experiences a Mediterranean climate, with milder winters and hotter, dry summers. While maize is cultivated in some southern regions, it is less dominant compared to the north (Rigutti 2007 ). So far, Trichoderma ear rot has only been reported in Europe from France, Germany, Austria, and Italy. However, we hypothesize that it may also occur in neighboring countries with similar climatic conditions. In particular, regions with warm and dry conditions in summer are likely to provide an environment conducive to the growth and proliferation of T. afroharzianum , facilitating infection of maize cobs. Trichoderma ear rot and stalk rot have also been reported in the United States (Iowa, Kentucky, Ohio), India, and China where similar climatic and agricultural characteristics are prevalent (Munkvold and White 2016 ; Chen et al. 2023 ; Harish et al. 2025 ). Additionally, the widespread distribution of Trichoderma spp. in agricultural soils across Europe further suggests that Trichoderma ear rot may also occur in other countries. Given these factors, a broader geographic range may be expected and considered, particularly in countries with similar agricultural practices and environmental conditions. This observations are confirmed by the Pearson correlation indicating a significant correlation between temperature and precipitation with the occurrence of T. afroharzianum . It reveals that higher temperatures favour the occurrence of T. afroharzianium , while precipitation correlates negatively with Trichoderma spp. presence, thereby suggesting that drought conditions may promote the occourence of the disease. The correlation of temperature and precipitation increases until flowering in July and decreases thereafter, suggesting that specific conditions during flowering are crucial for infection. During flowering, maize reproductive organs (silk and tassels) are exposed, providing potential entry points for pathogens (Reid et al. 1992 ). Fungal pathogens can take advantage of this window when the plant tissue is soft and moist, making it easier for the fungus to colonize the cob (Thompson and Raizada 2018 ). After July, as maize cobs develop and the kernels begin to harden, the plant becomes more resistant to fungal infection. The silks dry out and fall off, reducing the pathogen access points (Duncan and Howard 2010 ). The kernels also develop tougher outer layers, making it harder for pathogens to penetrate (Sampietro et al. 2009 ). The monitoring results align closely with the findings from in vitro and in vivo experiments, reinforcing the role of temperature in influencing the growth and pathogenicity of T. afroharzianum . In both settings, pathogenic isolates of T. afroharzianum exhibited significantly higher growth rates compared to non-pathogenic strains, with an optimal temperature range between 28°C and 32°C. Pathogenic T. afroharzianum isolates, in particular, exhibit superior growth across a broader range of temperatures, especially at cooler conditions (12°C) where they outperform non-pathogenic strains. All isolates exhibited no growth at 8°C or above 40°C on PDA, consistent with typical growth patterns observed for most Trichoderma species (Danielson & Davey, 1973). Growth rates increased with rising temperatures for all isolates, reaching an optimum at 28°C or 32°C. This aligns with previous findings for T. afroharzianum , T. harzianum , and T. asperellum (Begoude et al. 2007 ; Singh A. L. et al. 2013; Zehra et al. 2017 ). The results of this study demonstrate a clear influence of temperature on the disease severity on maize cobs in climate chamber experiments. Disease severity increased with rising temperature, with only light infection observed at 10°C and strong severity at 35°C. Trichoderma afroharzianum displayed the highest pathogenicity, suggesting that temperature influences both pathogen virulence and host plant susceptibility. Environmental conditions play a critical role in influencing both the pathogen and the host plant, shaping the dynamics of disease development. On the one hand, it can be assumed that drought stress weakens the plant, making it more susceptible to fungal infection by accelerating tissue senescence and affecting the production of defense-related compounds (Ristaino and Duniway J. M. 1989; Mayek-Perez et al. 2002 ; Achuo et al. 2006 ; Ramegowda and Senthil-Kumar 2015 ). At elevated temperatures, plant metabolism is altered, which can hinder its ability to mount an effective defense response (Dikilitas et al. 2016 ; Haddoudi et al. 2021 ). In particular, stress-induced hormonal changes (e.g., increased abscisic acid and reduced salicylic acid) can limit plant defense against fungal pathogens (Thaler and Bostock 2004 ). On the other hand, temperature directly affects growth and virulence of the pathogen . At higher temperatures, Trichoderma spp. can grow more rapidly and colonize maize tissue more effectively (Qiu et al. 2017 ). With increasing temperatures and prolonged drought conditions during summer months, current climatic trends suggest that Trichoderma ear rot on maize is likely to become more frequent in the future. This prediction is supported by the findings of Juroszek and von Tiedemann ( 2013 ), who proposed that the incidence of ear rots caused by fungal pathogens, particularly Fusarium and Aspergillus , will rise under such conditions (Pfordt et al. 2020 ). 4. Materials and Methods 4.1 Isolation of Trichoderma spp. from maize plants and soil samples Five to ten maize cobs and stalks were randomly collected from symptomatic and non-symptomatic plants of silage and grain maize fields in Germany and France from 2018 to 2023. Twenty randomly chosen kernels from each cob were surface sterilized for 10 min with 0.25% silver nitrate and placed on potato dextrose agar (PDA) with 400 µg/ml streptomycin (Duchefa Biochemie, Haarlem, Netherlands) and 30 µg/ml rifampicin (AppliChem, Darmstadt, Germany). Plates were stored at 22°C under 12h/12h light-dark cycle in climate chambers. After two days, outgrown Trichoderma -like mycelium was transferred to PDA plates. Individual cultures were grown for two weeks at 22°C under 12h/12h light-dark cycle in growth chambers (Mytron, Heiligenstadt, Germany) and determined morphologically under a light microscope at genus level. Single conidia cultures were produced and isolates were stored on synthetic low nutrition agar (SNA) plates at 4°C. Soil samples were obtained in 2021, 2022 and 2023 either from agricultural fields with previous Trichoderma ear rot infection or non-infected fields. From each field site, four soil samples from the upper 15 cm of the soil profile were collected after removal of surface plant material. The samples were stored in a cooling chamber at 5°C until use. Soil samples were passed through a 5 mm gauze to remove coarse debris and plant material. Ten g of soil were dissolved in 100 ml water and shaken for 20 min. After sedimentation, soil suspension was diluted with sterile water to 10 − 2 and 1 ml was added to sterile PDA plates containing Bengal Rose (50 ppm) and antibiotics (200 ppm streptomycin, 200 ppm rifampicin). Plates were incubated at 25°C with 12 h light cycle for four days. After incubation, individual colonies with Trichoderma -like appearance were picked with a sterile loop and transferred to PDA. Individual cultures were grown for two weeks at 22°C under 12h/12h light-dark cycle in growth chambers (Mytron, Heiligenstadt, Germany), and determined morphologically as above. Isolate identification to species level involved the sequence analysis. 4.2 Molecular identification DNA extraction, PCR amplification and sequencing The isolates selected for this study were incubated on PDA at 25°C for seven days. Approximately 50 mg of fresh mycelium was scraped from the agar surface and subjected to genomic DNA extraction using the Qiagen Plant Mini Kit (QIAGEN GmbH, Hilden, Germany) according to the manufacturer’s protocol. The nuclear rDNA ITS region was amplified using the primer pairs ITS1F and ITS4 (Gardes and Bruns 1993 ). The fragments of translation elongation factor 1-α (tef1-α) was amplified with EF1-728F and tef1LLErev (Carbone and Kohn 1999 ) (Jaklitsch et al. 2005 ), while the fragments of the RNA polymerase II second largest subunit B (rbp2) was obtained using RBP2-5F and RPB2-7R (Liu et al. 1999 ). PCR amplifications were performed in a total reaction volume of 25 µl, containing 2.5 µl Buffer, 5 µl MgCl2 (5 mM), 1 µl dNTPs (0.2 mM), 0.4 µl of each primer (1 µM), 0.125 µl Taq polymerase (1 U), 2 µl DNA template, and 13.575 µl sterile deionized water. Amplification was conducted with a Biometra Thermal Cycler (Analytik Jena, Germany) following the cycling conditions outlined by Gu et al. 2017 . The PCR products were visualized by ROTI®GelStain (Roth, Germany) staining on 1% agarose gel electrophoresis. After purification with the QIAquick PCR Purification Kit (QIAGEN GmbH, Hilden, Germany), amplicons were sequenced bidirectionally using the respective PCR primers by Macrogen Europe (Netherlands) and the raw nucleotide sequences were edited in MEGA v. 7 (Kumar et al. 2016 ). Barcoding species delimitation Identification followed the protocol outlined by Cai and Druzhinina (2021), applying the formula Trichoderma [ITS76] ~ sp∃!(RPB299 ≅ TEF197) (Cai and Druzhinina 2021b ). The obtained ITS sequences were incorporated into the ITS56 dataset (available at www.trichokey.com ), which represents the intrageneric polymorphism of the Trichoderma genus (Cai and Druzhinina 2021b ). The resulting alignment was used to calculate pairwise similarity in BioEdit (Hall 1999 ). Isolates with ITS sequences sharing ≥ 76% similarity with at least one known Trichoderma species were selected for further identification. These isolates were then assigned to Trichoderma species based on pairwise similarity of their RPB2 and TEF1-α sequences with reference sequences. Using TrichoBLAST ( www.trichokey.com ), Trichoderma species exhibiting high sequence similarity to the query were identified, and their corresponding reference sequences were retrieved from databases. The sequences were aligned and trimmed to the length of a phylogenetic marker (Kopchinskiy et al. 2005 ; Cai and Druzhinina 2021b ) and pairwise similarity calculation conducted as mentioned above. A similarity threshold of ≥ 99% for unambiguous identification was expected, with ≥ 97% considered acceptable. If the condition ∃!(RPB299 ≅ TEF197) was not fulfilled, phylogenetic analysis was conducted on both single-locus and concatenated RPB2 and TEF1-α sequences. 4.3 Plant cultivation and pathogenicity assessment in the greenhouse Maize seeds were sown in 18 cm diameter pots filled with a mixture of potting soil, sand and compost (3/1/3). Pots were placed in the greenhouse at 25°C temperature and 14h/10h day-/night cycle. Plants were watered as needed. Spores from single spore cultures were transferred to PDA plates containing antibiotics as above, and incubated at 23°C in a growth chamber. After two weeks, sterile water was added to plates and conidia were scraped off with a microscope slide. Spore concentration was measured with a Thoma haemocytometer (Merck, Darmstadt, Germany) and adjusted to 1x10 6 conidia per ml. The primary cobs of maize plants were inoculated seven days after silk channel emergence (BBCH 65). A needle pin was dipped in conidia suspension and engraved in the cob through the husk, hurting the kernels. Ten plants in two repetitions were inoculated and twenty plants were inoculated with sterile water, which served as control. Four weeks (28 dpi) after inoculation, husk leaves of inoculated and control cobs were removed and percent disease severity was visually estimated (0-100%) according to EPPO Guidelines PP1/285 (EPPO Bulletin 2015 ). 4.4 Cultures and strains used for temperature trials in vivo and in vitro Fungal isolates utilized in this study were obtained from different sources and are currently kept in Trichoderma strain collection at the University of Göttingen's Department of Plant Pathology and Plant Protection. TS1 was provided by the Department of Agricultural, Forestry and Food Sciences (DISAFA-Plant Pathology Unit), AGROINNOVA - Interdepartmental Centre of the University of Torino (Sanna et al. 2022 ). The holotype of T. afroharzianum , CBS 124.620, along with four strains recently isolated from maize cobs in Germany, France, and Italy, were included in the investigation. Additionally, two Trichoderma strains used as active ingredients in biopesticide products, T34 ( T. asperellum ) and T39 ( T. harzianum ), were included. To ensure the purity of the strains, single spore colonies were isolated and cultured for subsequent experiments. Table 1 Isolates used for in vivo and in vitro climate chamber trials Isolate Species Origin Year CBS124620 T. afroharzianum Westerdijk Fungalbiodiversity Inst. - AP18TRI1 T. afroharzianum Maize cob, Croix des Pardis, France 2018 AP18TRI2 T. afroharzianum Maize cob, Kuenzing, Germany 2018 TS1 T. afroharzianum Maize cob, DISAFA-Plant Pathology Unit, Carmagnola, Italy 2020 AP18TRI7 T. harzianum Maize cob, Grucking, Germany 2018 T39 T. harzianum Trichodex, Makhteshim Agan Industries T34 T. asperellum Xilon®, Kwizda Agro GmbH 4.5 In vitro temperature trial The effect of temperature was assessed on PDA. The medium was inoculated with a 6 mm PDA plug obtained from a seven-day-old culture. The plates were placed in darkness and incubated at temperatures ranging from 8 to 44°C, with temperature increments of 4°C, using temperature chambers (Rumed Rubarth Apparate GmbH). The position of the plates within the chamber was randomized. A total of four plates were inoculated per isolate for each temperature and medium. Radial growth was assessed by measuring using a ruler along two perpendicular lines marked on the bottom of each Petri dish at twelve-hour intervals up to seven days post-inoculation (dpi) or until the plates were completely colonized. The radial growth at a given time was obtained by dividing the sum of the two radial measures in millimetres (mm) by four. 4.6 Plant cultivation and inoculation procedure for climate chamber experiments The inoculum for all experiments was obtained from single spore cultures grown on PDA, incubated for seven days in total darkness at 25°C in a growth chamber (Mytron Bio und Solartechnik GmbH). The spore suspension was adjusted to 1x10 6 colony-forming units (CFU) per milliliter using a Thoma haemocytometer. The maize variety LIKEit (K 180, Deutsche Saatveredelung AG) was sown into 16x16x13 cm pots with a mixture of 3:3:1 of potting soil, compost, and sand. The plants were cultivated under greenhouse conditions at a temperature of approximately 25°C, in accordance with the natural day-night rhythm and irrigated as necessary. The maize cobs were inoculated at BBCH 65 after the first silks had begun to dry-off. Silk channel inoculation was conducted by injecting 1 ml spore suspension, with a concentration of 1x10 6 spores per milliliter, into the tip of the first developed cob using a needle and syringe. Six plants per isolate were inoculated (BBCH 65), in two repetitions and placed afterwards in the climate chamber (RUMED). A total of 60 plants were placed in 5 climate chambers. The relative humidity was maintained at a constant level of 70%, with temperature variations ranging from 10°C, 15°C, 20°C, 30°C, to 35°C and with the day- /night light cycle set to 12/10 hours. Disease severity was assessed after four weeks as described above. 4.7 Statistical analysis Statistical analysis was conducted using STATISTICA version 13 (Statistica GmbH, Germany). All experiments were fully randomized. To assess treatment effects and differences among the means of experimental units, two- or three-factorial analysis of variance (ANOVA) was performed. Analysis of variance (ANOVA) was carried out by Tukey-HSD-test at 5% probability. Pearson’s correlation coefficients were used to examine the relationship between temperature and precipitation in June, July, August, and September and infection of T. afroharzianum on maize cobs and soil. In addition, a multiple regression was performed to determine the relationship of Trichoderma ear rot infection with temperature and precipitation in June, July and August for each sample location. Trichoderma ear rot infection was calculated by the following equation: $$\:Trichoderma\:ear\:rot\:infection\:\left[\%\right]=\frac{Number\:of\:T.\:afroharzianum\:isolates\:}{Total\:number\:of\:samples\:}\:÷\:Location$$ The impact of weather conditions on the occurrence of T. afroharzianum was analyzed by variance components derived from the overall variance estimated with the restricted maximum likelihood model. Declarations Supplementary Materials: The following supporting information can be downloaded at: www.mdpi.com/xxx/s1, Figure S1: title; Table S1: title; Video S1: title. Ethics approval and consent to participate: Not applicable Consent for publication: Written informed consent has been obtained from the authors to publish this paper Availability of data and material: The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. Competing interest: The authors declare no conflict of interest Funding: This research was funded by Federal Ministry of Agriculture, Food and Regional Identity, grant number FKZ2221NR014 Author Contributions: Conceptualization, A.P.; methodology, W.F.V.; investigation, A.P, and W.F.V.; writing—original draft preparation, A.P. and C.D-M.; writing—review, C.D-M., G.S, B.C.S and A.v.T.; visualization, A.P.; supervision, B.C.S and A.v.T.; project administration, A.P.; funding acquisition, A.P, C.D-M., G.S, B.C.S and A.v.T. All authors have read and agreed to the published version of the manuscript.” Acknowledgments: We would like to thank the breeding companies, universities and federal research institutions that provided us with maize and soil samples. We also thank Manuela Mücke and Tobias Wille for technical support of the experiments and for performing DNA sequencing, respectively. References Achuo EA, Prinsen E, Höfte M. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6871003","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":473665432,"identity":"66e288d0-a1d5-4b0c-8942-70fdc9b825a4","order_by":0,"name":"Annette Pfordt","email":"data:image/png;base64,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","orcid":"","institution":"University of Göttingen","correspondingAuthor":true,"prefix":"","firstName":"Annette","middleName":"","lastName":"Pfordt","suffix":""},{"id":473665433,"identity":"43345d21-6124-4150-be22-cf78743241c1","order_by":1,"name":"Clovis Douanla-Meli","email":"","orcid":"","institution":"Julius Kühn-Institut (JKI) - Federal Research Centre for Cultivated Plants, Institute for National and International Plant Health","correspondingAuthor":false,"prefix":"","firstName":"Clovis","middleName":"","lastName":"Douanla-Meli","suffix":""},{"id":473665434,"identity":"b9ea23a3-e592-4d46-89b8-f9b3185d56a3","order_by":2,"name":"Felix Voll","email":"","orcid":"","institution":"University of Göttingen","correspondingAuthor":false,"prefix":"","firstName":"Felix","middleName":"","lastName":"Voll","suffix":""},{"id":473665435,"identity":"0f0d5073-e020-4146-bcf7-c11a14fe079b","order_by":3,"name":"Gritta Schrada","email":"","orcid":"","institution":"Julius Kühn-Institut (JKI) - Federal Research Centre for Cultivated Plants, Institute for National and International Plant Health","correspondingAuthor":false,"prefix":"","firstName":"Gritta","middleName":"","lastName":"Schrada","suffix":""},{"id":473665436,"identity":"e5798f88-6c05-4064-ba78-59ad3facf7bd","order_by":4,"name":"Bernhard Carl Schäfer","email":"","orcid":"","institution":"Julius Kühn-Institut (JKI) - Federal Research Centre for Cultivated Plants, Institute for National and International Plant Health","correspondingAuthor":false,"prefix":"","firstName":"Bernhard","middleName":"Carl","lastName":"Schäfer","suffix":""},{"id":473665437,"identity":"941bc094-842d-41c5-ae5f-6274577ddc4e","order_by":5,"name":"Andreas von Tiedemann","email":"","orcid":"","institution":"University of Göttingen","correspondingAuthor":false,"prefix":"","firstName":"Andreas","middleName":"","lastName":"von Tiedemann","suffix":""}],"badges":[],"createdAt":"2025-06-11 10:53:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6871003/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6871003/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s42483-025-00396-4","type":"published","date":"2026-02-10T15:58:18+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":85068696,"identity":"34338b7b-5183-4ec2-8296-cb0100934caa","added_by":"auto","created_at":"2025-06-20 15:24:45","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":264202,"visible":true,"origin":"","legend":"\u003cp\u003eGeographic distribution of received pathogenic \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates from maize cobs and agricultural soil samples in Europe\u003c/p\u003e","description":"","filename":"Picture1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6871003/v1/a0d5e88454cf7d32004620cb.jpg"},{"id":85068691,"identity":"0b177598-efed-481a-8272-08a8d2a8b1b0","added_by":"auto","created_at":"2025-06-20 15:24:45","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":149069,"visible":true,"origin":"","legend":"\u003cp\u003ePearson correlation coefficients (r) between mean monthly temperature and precipitation with the occurrence of T. afroharzianum infected cobs per location from April to September in 2018-2023 (n= 59). The white bars represent the correlation of temperature, while the grey bars indicate the correlation of precipitation. Asterisks represent significant (p≤ 0.05) correlation.\u003c/p\u003e","description":"","filename":"Picture2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6871003/v1/a2e8a665fd35b587d6ccc091.jpg"},{"id":85069848,"identity":"90e61220-65b5-4f2d-94a5-594f4a4cb250","added_by":"auto","created_at":"2025-06-20 15:32:45","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":233214,"visible":true,"origin":"","legend":"\u003cp\u003eCorrelation between Trichoderma ear rot infection per location (%) and weather conditions (temperature, left and precipitation, right) during June, July, and August in 2018-2023.\u003c/p\u003e","description":"","filename":"Picture3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6871003/v1/45198512818ca328659b812c.jpg"},{"id":85069847,"identity":"7eed7c7f-cf9e-453e-89d2-df1d2620df35","added_by":"auto","created_at":"2025-06-20 15:32:45","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":46604,"visible":true,"origin":"","legend":"\u003cp\u003eMean radial growth rate [mm] at different temperatures (12 to 44°C) 44 hours after inoculation with T. afroharzianum (TS1, AP18Tri1, AP18Tri2 and CBS124620), T. harzianum (T39, AP19Tri7) and T. asperellum (T34). Error bars representing the standard error.\u003c/p\u003e","description":"","filename":"Picture4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6871003/v1/93b27606e502394cded03dda.jpg"},{"id":85071558,"identity":"4aca2aaa-8b6a-49ea-844a-f239df872230","added_by":"auto","created_at":"2025-06-20 15:40:45","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":58449,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of different temperature levels (10°C, 15°C, 20°C, 30°C, and 35°C) on disease severity caused by \u003cem\u003eT. afroharzianum\u003c/em\u003eisolates (CBS124620, AP18TRI2, TS1) and \u003cem\u003eT. asperellum\u003c/em\u003e (T34) on maize cobs. Bars show the mean disease severity, with error bars representing the standard error (SE). Letters show significant differences between temperatures (Tukey-test, p ≤ 0.05).\u003c/p\u003e","description":"","filename":"Picture5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6871003/v1/1b539edae062758b59136de4.jpg"},{"id":102785346,"identity":"15075140-b1b6-4fc4-be37-65b3633834fd","added_by":"auto","created_at":"2026-02-16 16:05:24","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2179299,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6871003/v1/8b5beeb8-9120-4712-81f4-e498695a4a1c.pdf"},{"id":85068690,"identity":"6a1f3b17-0b4b-41f3-8170-5589cc6978b1","added_by":"auto","created_at":"2025-06-20 15:24:45","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":78735,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementalMaterial.docx","url":"https://assets-eu.researchsquare.com/files/rs-6871003/v1/384d9759e3072787397dd17a.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Effect of temperature and precipitation on the occurrence of Trichoderma ear rot in Europe","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eMaize (\u003cem\u003eZea mays\u003c/em\u003e L.) is one of the most important staple crops worldwide, playing a critical role in global agriculture. As a versatile and highly productive cereal, maize serves as a primary source of food for humans and a key ingredient in animal feed (Erenstein et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). For human consumption, maize is an essential food source in many developing countries, providing a substantial proportion of daily caloric intake. Maize can be processed into a variety of products, including flour for tortillas, polenta, and maize meal, which are staple foods in many regions. In addition, maize silage is a fundamental component of livestock feed, particularly for poultry, swine, and cattle due to its high energy content (Tenaillon and Charcosset \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). As the global demand for both food and animal feed continues to rise, maize production faces significant challenges, including pest and disease stresses (Shiferaw et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The evolving environmental conditions can significantly affect the prevalence and severity of pest and pathogen damage, with existing pathogens potentially gaining or losing significance (Anderson et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Additionally, the emergence of new pathogens is anticipated to become an increasing concern in the future (King et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). \u003cem\u003eTrichoderma\u003c/em\u003e species belong to the division Ascomycota and are globally distributed, inhabiting diverse ecosystems ranging from dead organic matter to forest and agricultural soils (Druzhinina et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Cai and Druzhinina \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2021a\u003c/span\u003e). Known for their rapid growth and competitive nature, \u003cem\u003eTrichoderma\u003c/em\u003e species produce and detoxify antimicrobial substances, contributing to their success as free-living soil-borne fungi (Kubicek and Harman \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). Furthermore, \u003cem\u003eTrichoderma\u003c/em\u003e strains from agricultural soils often form opportunistic symbiotic relationships with plants, enhancing their utility as biological control agents. Some \u003cem\u003eTrichoderma\u003c/em\u003e species exhibit antagonistic effects against a wide variety of plant pathogens, including fungi, oomycetes, bacteria, and nematodes, through mechanisms such as rapid growth, nutrient competition, mycoparasitism, and antibiosis (Harman \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). However, \u003cem\u003eTrichoderma\u003c/em\u003e spp. has also been identified as a causative agent of ear rot in maize in the United States, typically associated with mechanical damage to the cob (Sutton \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e1972\u003c/span\u003e; Munkvold and White \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Wise et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). In 2018, a pathogenic \u003cem\u003eTrichoderma\u003c/em\u003e species was discovered on maize cobs in Europe, identified as \u003cem\u003eT. afroharzianum\u003c/em\u003e, which infects maize cobs and produces white mycelium that is rapidly producing dark, bearing grey-green conidia (Pfordt et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Severe infections result in enzymatic decomposition of starch in the kernels, making cobs soft and watery through the secretion of alpha-amylase (Pfordt et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Starch is a primary carbohydrate reserve in the grains of most cereals, and the production of enzymes like alpha-amylase by \u003cem\u003eTrichoderma\u003c/em\u003e allows the fungus to infect not only maize but also other cereals, such as wheat and barley (Pfordt et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSince the initial discovery in Southern Germany in 2018, the disease has been observed on maize in other regions of Germany, France, Italy, China, and India (Sanna et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Chen et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Harish et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), particularly following dry and hot summer periods. This pattern suggests that climatic conditions and meteorological factors determine the occurrence of Trichoderma ear rot in maize, with elevated temperatures during June, July, and August potentially enhancing natural infections during the flowering period (Pfordt et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eTrichoderma\u003c/em\u003e species are cosmopolitan, occurring in diverse habitats in temperate, tropical, and subtropical regions. However, the abundance and species composition can vary across different geographic locations (Ghorbanpour et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Most \u003cem\u003eTrichoderma\u003c/em\u003e species thrive at moderate temperatures, typically between 20\u0026ndash;30\u0026deg;C. This range supports their rapid growth and efficient colonization of substrates (Begoude et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Zehra et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). While some species, like \u003cem\u003eT. harzianum\u003c/em\u003e and \u003cem\u003eT. afroharzianum\u003c/em\u003e, are considered cosmopolitan and found across multiple continents, other species show more localized occurrences, which is indicative of restricted ecological adaptation. For instance, species like \u003cem\u003eT. simonsii\u003c/em\u003e, \u003cem\u003eT. atrobrunneum\u003c/em\u003e and \u003cem\u003eT. atroviride\u003c/em\u003e, exhibit a preference for lower temperatures and are found in cooler regions, including temperate zones like Europe and North America. Meanwhile other species like \u003cem\u003eT. asperellum\u003c/em\u003e and \u003cem\u003eT. longibrachiatum\u003c/em\u003e are more commonly isolated in tropical and subtropical regions (Chaverri et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Di Lelio et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe aim of this study is to ascertain the existence of a correlation between local meteorological conditions and the occurrence of \u003cem\u003eTrichoderma\u003c/em\u003e species in maize fields across Europe. In addition to field observations, \u003cem\u003ein vitro\u003c/em\u003e and \u003cem\u003ein vivo\u003c/em\u003e climate chamber experiments were conducted to further investigate the influence of temperature on \u003cem\u003eTrichoderma\u003c/em\u003e species. By providing insights into the environmental drivers of \u003cem\u003eTrichoderma\u003c/em\u003e distribution, this research seeks to contribute to better management strategies for mitigating the risks posed by pathogenic \u003cem\u003eTrichoderma\u003c/em\u003e species, while also supporting the beneficial applications of non-pathogenic strains in agriculture.\u003c/p\u003e"},{"header":"2. Results","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Occurrence of \u003cem\u003eTrichoderma\u003c/em\u003e species obtained from maize and soil samples in Germany\u003c/h2\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eOver the past seven years (2018\u0026ndash;2024), a total of 345 samples were received from symptomatic and asymptomatic maize cobs and stalks, as well as agricultural soils across 72 different locations in Europe. Among these, 130 \u003cem\u003eTrichoderma\u003c/em\u003e isolates were cultured from 54 sampling sites (Sub. Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Of the isolates, 48 were obtained from maize cobs and stalks, while the remaining 82 derived from soil samples.\u003c/p\u003e \u003cp\u003eThe highest incidence of visible maize cob infections and the presence of \u003cem\u003eT. afroharzianum\u003c/em\u003e were found predominantly at sites in Southern Germany, particularly in Bavaria along the Donau river, and in Rhineland-Palatinate and along the Rhine River in the Alsace (Germany and France). Additionally, several locations in central Germany, especially in the warmer and drier regions around Magdeburg (Saxony-Anhalt) and Brandenburg along the Oder river, also exhibited maize cob infections and soil isolates of \u003cem\u003eT. afroharzianum\u003c/em\u003e. In contrast, no \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates were detected in cooler areas of central and northern Germany. In addition to the isolates obtained in Alsace, one isolate was found in the southwest region of Aquitaine, and another in Burgundy. Three isolates were found in northern Italy and two isolates in T\u0026uuml;rkiye, in Salihli (Aegean region) and Tarsus (Belediye region). Additionally, Trichoderma ear rot disease was identified in Austria, specifically in Neustift, Heiligenkreuz, and Rudersdorf, located in the Nordburgenland region near the borders with Bratislava and Hungary (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eNotably, at sites where maize ear rot was observed, \u003cem\u003eT. afroharzianum\u003c/em\u003e was consistently isolated from both maize cobs and soil samples. This correlation suggests a close association between soil-borne populations of \u003cem\u003eT. afroharzianum\u003c/em\u003e and the occurrence of cob infections. However, \u003cem\u003eT. afroharzianum\u003c/em\u003e was also detected in soil samples from locations without visible cob infections, indicating that the fungus can persist in the soil even in the absence of visible disease symptoms.\u003c/p\u003e \u003cp\u003eAll \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates subjected to pathogenicity tests exhibited a high level of virulence, causing significant disease symptoms in maize plants.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 \u003cem\u003eTrichoderma\u003c/em\u003e species isolated from maize and soil samples\u003c/h2\u003e \u003cp\u003eA total of six different \u003cem\u003eTrichoderma\u003c/em\u003e species were isolated from the maize cobs (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The majority of isolates (n\u0026thinsp;=\u0026thinsp;27) were identified as \u003cem\u003eT. afroharzianum\u003c/em\u003e, exhibited significant pathogenicity, causing 89.4% disease severity. In contrast, \u003cem\u003eT. harzianum\u003c/em\u003e was also isolated, but only in a smaller number of three isolates colonizing the cobs endophytically without causing symptoms (0.8 disease severity). Additionally, isolates of \u003cem\u003eT. atroviride\u003c/em\u003e (8 isolates) and \u003cem\u003eT. gamsii\u003c/em\u003e (4 isolates) were recovered, both of which exhibited mild pathogenicity of 18.8% and 14.6% disease severity after artificial inoculation on the maize cobs. Five isolates of \u003cem\u003eT. azevodoi\u003c/em\u003e and one isolate of \u003cem\u003eT. tomentosum\u003c/em\u003e were obtained, which showed no pathogenicity.\u003c/p\u003e \u003cp\u003eIn addition to the isolates obtained from maize cobs, the study also identified several \u003cem\u003eTrichoderma\u003c/em\u003e species from soil samples. A total of 82 isolates were recovered from the soil, depicting a diversity of thirteen species. Among these, \u003cem\u003eT. afroharzianum\u003c/em\u003e (16 isolates) and \u003cem\u003eT. harzianum\u003c/em\u003e (15 isolates) were the most prevalent, followed by \u003cem\u003eT. peberdyi\u003c/em\u003e, (10 isolates), \u003cem\u003eT. atroviride\u003c/em\u003e (8 isolates) and \u003cem\u003eT. cerinum\u003c/em\u003e (7 isolates). All \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates from soil exhibited high pathogenicity (86.6%) while the isolates of the remaining species, including \u003cem\u003eT. harzianum\u003c/em\u003e and \u003cem\u003eT. atroviride\u003c/em\u003e, displayed low to moderate (0.2\u0026ndash;37%) levels of pathogenicity.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eOrigin and pathogenicity (% disease severity, DS) of various \u003cem\u003eTrichoderma\u003c/em\u003e species isolated from maize cobs and agricultural soil.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSpecies\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eMaize\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eSoil\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTotal\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003en\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e% DS\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003en\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e% DS\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. afroharzianum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e89.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e86.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e43\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. harzianum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. atroviride\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e18.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e23.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. gamsii\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e14.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. azevodoi\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. tomentosum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. peberdyi\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. cerinum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e37.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. koningiopsis\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. velutinum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. hamatum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. atrobrunneum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. asperellum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. arundinaceum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e28.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. brevicompactum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT. virens\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e48\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e43.4\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e82\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e22.45\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e130\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Effect of temperature and precipitation on the occurrence of Trichoderma ear rot in Europe\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eThe Pearson correlation coefficients (r) for temperature and precipitation from April to September (2018\u0026ndash;2023) were assessed with the occurrence of pathogenic \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates (Sub. Table\u0026nbsp;2). Temperature in June (r\u0026thinsp;=\u0026thinsp;0.36), July (r\u0026thinsp;=\u0026thinsp;0.6), and August (r\u0026thinsp;=\u0026thinsp;0.33), showed significant correlation with the occurrence of Trichoderma ear rot, suggesting that higher temperatures favor the occurrence of Trichoderma ear rot (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) during flowering time. The correlation for precipitation shows a negative trend, especially in July (-0.4), indicating that dry conditions favour the occurrence of Trichoderma ear rot during this month.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e illustrates the correlation between the Trichoderma ear rot infection per location and weather conditions, specifically temperature and precipitation, during June, July, and August (2018\u0026ndash;2023). In June, there is a positive correlation between temperature and Trichoderma ear rot infection (r\u0026thinsp;=\u0026thinsp;0.4401), indicating that higher temperatures in this month slightly favor the disease.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn contrast, June precipitation shows a weak negative correlation with Trichoderma ear rot infection (r = -0.2888), suggesting that lower rainfall levels are associated with higher disease infection. The relationship between environmental factors and Trichoderma ear rot infection becomes more pronounced in July. Here, temperature shows a strong positive correlation with the disease (r\u0026thinsp;=\u0026thinsp;0.6079), highlighting that elevated temperatures during this month significantly favor Trichoderma ear rot infections. This period is particularly critical as it coincides with maize flowering, when the plants are most susceptible. July precipitation, on the other hand, displays a moderate negative correlation with Trichoderma ear rot infection (r = -0.3549), emphasizing the role of drought stress during this vulnerable growth stage. In August, the correlations between environmental factors and Trichoderma ear rot infection are decreasing. Temperature exhibits a weak positive correlation (r\u0026thinsp;=\u0026thinsp;0.3315), suggesting that higher temperatures in August may still contribute to the development of Trichoderma ear rot, though to a lesser extent than in July. Precipitation shows a weak, non-significant negative correlation (r = -0.2394), indicating that the effect of rainfall becomes less pronounced in late summer.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Effect of \u003cem\u003ein vitro\u003c/em\u003e temperature on the growth rate\u003c/h2\u003e \u003cp\u003eTemperature exerts a significant effect (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) on the radial growth of \u003cem\u003eTrichoderma.\u003c/em\u003e spp. \u003cem\u003ein vitro\u003c/em\u003e. All factors, such as isolate, temperature and their interaction, are significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001), whereas there was no significant effect of the replicate (Sub.Table\u0026nbsp;3). There was no fungal growth at 8\u0026deg;C and above 40\u0026deg;C while growth was observed in the range from 12 to 36\u0026deg;C for all isolates (Table\u0026nbsp;3). \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates, including AP18TRI2 and APP18TRI1, demonstrated the highest growth rates (Sub. Figure\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), especially at the optimal temperature range of 28\u0026deg;C to 32\u0026deg;C. At 32\u0026deg;C, AP18TRI2 exhibited the fastest growth (40.6 mm), followed by APP18TRI1 (37.4 mm). In contrast, \u003cem\u003eT. asperellum\u003c/em\u003e (T34) and \u003cem\u003eT. harzianum\u003c/em\u003e (T39) showed comparatively slower growth, with \u003cem\u003eT. asperellum\u003c/em\u003e reaching 29.1 mm at 32\u0026deg;C. All isolates experienced reduced growth at extreme temperatures, particularly at 36\u0026deg;C and 12\u0026deg;C, though \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates maintained relatively higher growth rates compared to non-pathogenic strains like CBS 124.620 at low temperatures. For instance, at 12\u0026deg;C, \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates grew better (10.9 mm for AP18TRI2) than CBS 124.620 (7.9 mm).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Effect of temperature on disease severity \u003cem\u003ein vivo\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eThe results indicate that temperature and isolate as well as the interaction has a significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) impact on the disease severity of \u003cem\u003eTrichoderma\u003c/em\u003e spp. on maize cobs (Sub. Table\u0026nbsp;3).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe impact of temperature on disease severity caused by four \u003cem\u003eTrichoderma\u003c/em\u003e isolates (CBS124620, AP18TRI2, T34, TS1) was assessed across a temperature range from 10\u0026deg;C to 35\u0026deg;C (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The results indicate that disease severity increases with rising temperature for all isolates, with significant differences in pathogenicity observed between the isolates. The reference strain CBS124620 exhibited low disease severity at temperatures below 20\u0026deg;C, with minimal infection (below 10%) at 10\u0026deg;C and 15\u0026deg;C. However, at 30\u0026deg;C and 35\u0026deg;C, disease severity increased significantly, reaching nearly 50% at 35\u0026deg;C, suggesting that even mild-pathogenic isolates can exhibit higher disease severity under favorable conditions. The pathogenic \u003cem\u003eT. afroharzianum\u003c/em\u003e isolate AP18TRI2 demonstrated the highest disease severity across the temperature range. At 20\u0026deg;C, AP18TRI2 caused severe infection, with disease severity peaking at nearly 80%. Disease severity decreased at higher temperatures (30\u0026deg;C and 35\u0026deg;C), with values of 60% and 50%, respectively. The \u003cem\u003eT. afroharzianum\u003c/em\u003e isolate TS1 followed a similar trend as AP18TRI2, showing severe infection at 20\u0026deg;C (over 70%) and slightly decreasing disease severity (62%) at 35\u0026deg;C. Surprisingly, the biocontrol \u003cem\u003eT. asperellum\u003c/em\u003e isolate T34 also showed severe disease symptoms on maize cobs with disease severity up to 58% at both 30\u0026deg;C and 35\u0026deg;C.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Discussion","content":"\u003cp\u003e \u003cb\u003eA total of 109 Trichoderma isolates from sixteen different species were obtained from agricultural soils and maize tissue.\u003c/b\u003e The most prevalent species in both, agricultural soil samples and maize tissue was \u003cem\u003eT. afroharzianum\u003c/em\u003e, showing high pathogenicity on maize cobs. The highest variety of \u003cem\u003eTrichoderma\u003c/em\u003e species was found in agricultural soil, with a greater number of different species like \u003cem\u003eT. peberdyi\u003c/em\u003e, \u003cem\u003eT. cerinum, T. koningiopsis\u003c/em\u003e, and \u003cem\u003eT. hamatum\u003c/em\u003e, which were not found in maize tissue. In contrast, maize tissue yielded fewer species, predominantly consisting of \u003cem\u003eT. afroharzianum\u003c/em\u003e, \u003cem\u003eT. harzianum\u003c/em\u003e, \u003cem\u003eT. atroviride\u003c/em\u003e, and \u003cem\u003eT. gamsii\u003c/em\u003e. However, \u003cem\u003eT. afroharzianum\u003c/em\u003e was the only species exhibiting high pathogenicity on the cob, other species like \u003cem\u003eT. harzianum\u003c/em\u003e appeared to colonize maize cobs endophytically without causing symptoms.\u003c/p\u003e \u003cp\u003eThis finding aligns with previous observations that \u003cem\u003eT. afroharzianum\u003c/em\u003e is a prominent species with the potential to cause Trichoderma ear rot in maize. It also underscores the capacity of this species to persist in soil while actively infecting maize under favorable conditions (Pfordt et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Pfordt et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cb\u003eThe study highlights distinct distribution patterns of pathogenic\u003c/b\u003e \u003cb\u003eTrichoderma\u003c/b\u003e \u003cb\u003especies across Europe, with\u003c/b\u003e \u003cb\u003eT. afroharzianum\u003c/b\u003e \u003cb\u003ebeing prevalent in warmer and dry regions, and absent in cooler, northern regions.\u003c/b\u003e This suggests that temperature and precipitation are key factors influencing the occurrence of pathogenic \u003cem\u003eTrichoderma\u003c/em\u003e species (Zehra et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Qiu et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Southern Germany, France and Italy typically experience higher temperatures throughout the year when compared to the northern part of the Germany. The southern regions of Germany, especially in areas like Bavaria and Baden-W\u0026uuml;rttemberg, have a more continental climate, characterized by hotter summers and colder winters with average monthly temperatures ranging from 18\u0026deg;C and 25\u0026deg;C (Flohn \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e1984\u003c/span\u003e; Kaspar et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). In contrast, Northern Germany, including regions like Lower Saxony and Schleswig-Holstein, generally experiences cooler summer temperatures, often ranging from 15\u0026deg;C to 20\u0026deg;C with a more temperate maritime climate due to its proximity to the North Sea and the Baltic Sea (Maier et al. 2003; M\u0026uuml;ller-Westermeier and Rocznik \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Sch\u0026ouml;nwiese \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). This temperature gradient is crucial for the growth and prevalence of pathogenic \u003cem\u003eTrichoderma\u003c/em\u003e species, as warmer conditions in the south favor the proliferation of these strains, explaining their higher occurrence in Southern Germany (Wong et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). In addition, Southern Germany and regions in Saxony-Anhalt are more prone to longer and more intense drought periods, particularly during the summer months, creating conditions that stress plants more severely (M\u0026uuml;ller-Westermeier and Rocznik \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Kaspar et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2013\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSimilarly, the Nordburgenland region in Austria, near the borders with Bratislava and Hungary, features a Pannonian climate with warm to hot summers (25\u0026deg;C to 30\u0026deg;C) and mild winters (0\u0026deg;C to 5\u0026deg;C). Annual precipitation is relatively low, ranging from 500 to 600 mm, relatively dry compared to other parts of Austria (Hammerl \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; GeoSphere Austria).\u003c/p\u003e \u003cp\u003eIn Italy, pathogenic \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates have so far only been identified in the northern part, particularly in Piemont, Lombardy, and Venetia, where maize is extensively cultivated due to the favorable climate and fertile soils of the Po Valley (Bocchiola et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Borrelli et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). The climate here is characterized by warm summers with moderate precipitation, creating conditions conducive to both maize growth and the proliferation of \u003cem\u003eT. afroharzianum\u003c/em\u003e. In contrast, Southern Italy experiences a Mediterranean climate, with milder winters and hotter, dry summers. While maize is cultivated in some southern regions, it is less dominant compared to the north (Rigutti \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2007\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSo far, Trichoderma ear rot has only been reported in Europe from France, Germany, Austria, and Italy. However, we hypothesize that it may also occur in neighboring countries with similar climatic conditions. In particular, regions with warm and dry conditions in summer are likely to provide an environment conducive to the growth and proliferation of \u003cem\u003eT. afroharzianum\u003c/em\u003e, facilitating infection of maize cobs. Trichoderma ear rot and stalk rot have also been reported in the United States (Iowa, Kentucky, Ohio), India, and China where similar climatic and agricultural characteristics are prevalent (Munkvold and White \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Chen et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Harish et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Additionally, the widespread distribution of \u003cem\u003eTrichoderma\u003c/em\u003e spp. in agricultural soils across Europe further suggests that Trichoderma ear rot may also occur in other countries. Given these factors, a broader geographic range may be expected and considered, particularly in countries with similar agricultural practices and environmental conditions.\u003c/p\u003e \u003cp\u003e \u003cb\u003eThis observations are confirmed by the Pearson correlation indicating a significant correlation between temperature and precipitation with the occurrence of\u003c/b\u003e \u003cb\u003eT. afroharzianum\u003c/b\u003e. It reveals that higher temperatures favour the occurrence of \u003cem\u003eT. afroharzianium\u003c/em\u003e, while precipitation correlates negatively with \u003cem\u003eTrichoderma\u003c/em\u003e spp. presence, thereby suggesting that drought conditions may promote the occourence of the disease. The correlation of temperature and precipitation increases until flowering in July and decreases thereafter, suggesting that specific conditions during flowering are crucial for infection.\u003c/p\u003e \u003cp\u003eDuring flowering, maize reproductive organs (silk and tassels) are exposed, providing potential entry points for pathogens (Reid et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e1992\u003c/span\u003e). Fungal pathogens can take advantage of this window when the plant tissue is soft and moist, making it easier for the fungus to colonize the cob (Thompson and Raizada \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). After July, as maize cobs develop and the kernels begin to harden, the plant becomes more resistant to fungal infection. The silks dry out and fall off, reducing the pathogen access points (Duncan and Howard \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). The kernels also develop tougher outer layers, making it harder for pathogens to penetrate (Sampietro et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003e \u003cb\u003eThe monitoring results align closely with the findings from\u003c/b\u003e \u003cb\u003ein vitro\u003c/b\u003e \u003cb\u003eand\u003c/b\u003e \u003cb\u003ein vivo\u003c/b\u003e \u003cb\u003eexperiments, reinforcing the role of temperature in influencing the growth and pathogenicity of\u003c/b\u003e \u003cb\u003eT. afroharzianum\u003c/b\u003e. In both settings, pathogenic isolates of \u003cem\u003eT. afroharzianum\u003c/em\u003e exhibited significantly higher growth rates compared to non-pathogenic strains, with an optimal temperature range between 28\u0026deg;C and 32\u0026deg;C. Pathogenic \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates, in particular, exhibit superior growth across a broader range of temperatures, especially at cooler conditions (12\u0026deg;C) where they outperform non-pathogenic strains. All isolates exhibited no growth at 8\u0026deg;C or above 40\u0026deg;C on PDA, consistent with typical growth patterns observed for most \u003cem\u003eTrichoderma\u003c/em\u003e species (Danielson \u0026amp; Davey, 1973). Growth rates increased with rising temperatures for all isolates, reaching an optimum at 28\u0026deg;C or 32\u0026deg;C. This aligns with previous findings for \u003cem\u003eT. afroharzianum\u003c/em\u003e, \u003cem\u003eT. harzianum\u003c/em\u003e, and \u003cem\u003eT. asperellum\u003c/em\u003e (Begoude et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Singh A. L. et al. 2013; Zehra et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe results of this study demonstrate a clear influence of temperature on the disease severity on maize cobs in climate chamber experiments. Disease severity increased with rising temperature, with only light infection observed at 10\u0026deg;C and strong severity at 35\u0026deg;C. \u003cem\u003eTrichoderma afroharzianum\u003c/em\u003e displayed the highest pathogenicity, suggesting that temperature influences both pathogen virulence and host plant susceptibility.\u003c/p\u003e\u003cp\u003e \u003cb\u003eEnvironmental conditions play a critical role in influencing both the pathogen and the host plant, shaping the dynamics of disease development.\u003c/b\u003e On the one hand, it can be assumed that drought stress weakens the plant, making it more susceptible to fungal infection by accelerating tissue senescence and affecting the production of defense-related compounds (Ristaino and Duniway J. M. 1989; Mayek-Perez et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Achuo et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Ramegowda and Senthil-Kumar \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). At elevated temperatures, plant metabolism is altered, which can hinder its ability to mount an effective defense response (Dikilitas et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Haddoudi et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In particular, stress-induced hormonal changes (e.g., increased abscisic acid and reduced salicylic acid) can limit plant defense against fungal pathogens (Thaler and Bostock \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). On the other hand, temperature directly affects growth and virulence of \u003cem\u003ethe pathogen\u003c/em\u003e. At higher temperatures, \u003cem\u003eTrichoderma\u003c/em\u003e spp. can grow more rapidly and colonize maize tissue more effectively (Qiu et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eWith increasing temperatures and prolonged drought conditions during summer months, current climatic trends suggest that Trichoderma ear rot on maize is likely to become more frequent in the future. This prediction is supported by the findings of Juroszek and von Tiedemann (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2013\u003c/span\u003e), who proposed that the incidence of ear rots caused by fungal pathogens, particularly \u003cem\u003eFusarium\u003c/em\u003e and \u003cem\u003eAspergillus\u003c/em\u003e, will rise under such conditions (Pfordt et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e"},{"header":"4. Materials and Methods","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\n \u003ch2\u003e4.1 Isolation of \u003cem\u003eTrichoderma\u003c/em\u003e spp. from maize plants and soil samples\u003c/h2\u003e\n \u003cdiv class=\"BlockQuote\"\u003e\n \u003cp\u003eFive to ten maize cobs and stalks were randomly collected from symptomatic and non-symptomatic plants of silage and grain maize fields in Germany and France from 2018 to 2023. Twenty randomly chosen kernels from each cob were surface sterilized for 10 min with 0.25% silver nitrate and placed on potato dextrose agar (PDA) with 400 \u0026micro;g/ml streptomycin (Duchefa Biochemie, Haarlem, Netherlands) and 30 \u0026micro;g/ml rifampicin (AppliChem, Darmstadt, Germany). Plates were stored at 22\u0026deg;C under 12h/12h light-dark cycle in climate chambers. After two days, outgrown \u003cem\u003eTrichoderma\u003c/em\u003e-like mycelium was transferred to PDA plates. Individual cultures were grown for two weeks at 22\u0026deg;C under 12h/12h light-dark cycle in growth chambers (Mytron, Heiligenstadt, Germany) and determined morphologically under a light microscope at genus level. Single conidia cultures were produced and isolates were stored on synthetic low nutrition agar (SNA) plates at 4\u0026deg;C.\u003c/p\u003e\n \u003cp\u003eSoil samples were obtained in 2021, 2022 and 2023 either from agricultural fields with previous \u003cem\u003eTrichoderma\u003c/em\u003e ear rot infection or non-infected fields. From each field site, four soil samples from the upper 15 cm of the soil profile were collected after removal of surface plant material. The samples were stored in a cooling chamber at 5\u0026deg;C until use. Soil samples were passed through a 5 mm gauze to remove coarse debris and plant material. Ten g of soil were dissolved in 100 ml water and shaken for 20 min. After sedimentation, soil suspension was diluted with sterile water to 10\u003csup\u003e\u0026minus;\u0026thinsp;2\u003c/sup\u003e and 1 ml was added to sterile PDA plates containing Bengal Rose (50 ppm) and antibiotics (200 ppm streptomycin, 200 ppm rifampicin). Plates were incubated at 25\u0026deg;C with 12 h light cycle for four days. After incubation, individual colonies with \u003cem\u003eTrichoderma\u003c/em\u003e-like appearance were picked with a sterile loop and transferred to PDA. Individual cultures were grown for two weeks at 22\u0026deg;C under 12h/12h light-dark cycle in growth chambers (Mytron, Heiligenstadt, Germany), and determined morphologically as above. Isolate identification to species level involved the sequence analysis.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003ch2\u003e4.2 Molecular identification\u003c/h2\u003e\n \u003cp\u003eDNA extraction, PCR amplification and sequencing\u003c/p\u003e\n \u003cp\u003eThe isolates selected for this study were incubated on PDA at 25\u0026deg;C for seven days. Approximately 50 mg of fresh mycelium was scraped from the agar surface and subjected to genomic DNA extraction using the Qiagen Plant Mini Kit (QIAGEN GmbH, Hilden, Germany) according to the manufacturer\u0026rsquo;s protocol. The nuclear rDNA ITS region was amplified using the primer pairs ITS1F and ITS4 (Gardes and Bruns \u003cspan class=\"CitationRef\"\u003e1993\u003c/span\u003e). The fragments of translation elongation factor 1-\u0026alpha; (tef1-\u0026alpha;) was amplified with EF1-728F and tef1LLErev (Carbone and Kohn \u003cspan class=\"CitationRef\"\u003e1999\u003c/span\u003e) (Jaklitsch et al. \u003cspan class=\"CitationRef\"\u003e2005\u003c/span\u003e), while the fragments of the \u003cem\u003eRNA polymerase II second largest subunit B\u003c/em\u003e (rbp2) was obtained using RBP2-5F and RPB2-7R (Liu et al. \u003cspan class=\"CitationRef\"\u003e1999\u003c/span\u003e). PCR amplifications were performed in a total reaction volume of 25 \u0026micro;l, containing 2.5 \u0026micro;l Buffer, 5 \u0026micro;l MgCl2 (5 mM), 1 \u0026micro;l dNTPs (0.2 mM), 0.4 \u0026micro;l of each primer (1 \u0026micro;M), 0.125 \u0026micro;l Taq polymerase (1 U), 2 \u0026micro;l DNA template, and 13.575 \u0026micro;l sterile deionized water. Amplification was conducted with a Biometra Thermal Cycler (Analytik Jena, Germany) following the cycling conditions outlined by Gu et al. \u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e. The PCR products were visualized by ROTI\u0026reg;GelStain (Roth, Germany) staining on 1% agarose gel electrophoresis. After purification with the QIAquick PCR Purification Kit (QIAGEN GmbH, Hilden, Germany), amplicons were sequenced bidirectionally using the respective PCR primers by Macrogen Europe (Netherlands) and the raw nucleotide sequences were edited in MEGA v. 7 (Kumar et al. \u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eBarcoding species delimitation\u003c/p\u003e\n \u003cp\u003eIdentification followed the protocol outlined by Cai and Druzhinina (2021), applying the formula \u003cem\u003eTrichoderma\u003c/em\u003e [ITS76]\u0026thinsp;~\u0026thinsp;sp\u0026exist;!(RPB299\u0026thinsp;\u0026cong;\u0026thinsp;TEF197) (Cai and Druzhinina \u003cspan class=\"CitationRef\"\u003e2021b\u003c/span\u003e). The obtained ITS sequences were incorporated into the ITS56 dataset (available at \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ewww.trichokey.com\u003c/span\u003e\u003c/span\u003e), which represents the intrageneric polymorphism of the \u003cem\u003eTrichoderma\u003c/em\u003e genus (Cai and Druzhinina \u003cspan class=\"CitationRef\"\u003e2021b\u003c/span\u003e). The resulting alignment was used to calculate pairwise similarity in BioEdit (Hall \u003cspan class=\"CitationRef\"\u003e1999\u003c/span\u003e). Isolates with ITS sequences sharing\u0026thinsp;\u0026ge;\u0026thinsp;76% similarity with at least one known \u003cem\u003eTrichoderma\u003c/em\u003e species were selected for further identification. These isolates were then assigned to \u003cem\u003eTrichoderma\u003c/em\u003e species based on pairwise similarity of their RPB2 and TEF1-\u0026alpha; sequences with reference sequences. Using TrichoBLAST (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ewww.trichokey.com\u003c/span\u003e\u003c/span\u003e), \u003cem\u003eTrichoderma\u003c/em\u003e species exhibiting high sequence similarity to the query were identified, and their corresponding reference sequences were retrieved from databases. The sequences were aligned and trimmed to the length of a phylogenetic marker (Kopchinskiy et al. \u003cspan class=\"CitationRef\"\u003e2005\u003c/span\u003e; Cai and Druzhinina \u003cspan class=\"CitationRef\"\u003e2021b\u003c/span\u003e) and pairwise similarity calculation conducted as mentioned above. A similarity threshold of \u0026ge;\u0026thinsp;99% for unambiguous identification was expected, with \u0026ge;\u0026thinsp;97% considered acceptable. If the condition \u0026exist;!(RPB299\u0026thinsp;\u0026cong;\u0026thinsp;TEF197) was not fulfilled, phylogenetic analysis was conducted on both single-locus and concatenated RPB2 and TEF1-\u0026alpha; sequences.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003e4.3 Plant cultivation and pathogenicity assessment in the greenhouse\u003c/h2\u003e\n \u003cdiv class=\"BlockQuote\"\u003e\n \u003cp\u003eMaize seeds were sown in 18 cm diameter pots filled with a mixture of potting soil, sand and compost (3/1/3). Pots were placed in the greenhouse at 25\u0026deg;C temperature and 14h/10h day-/night cycle. Plants were watered as needed. Spores from single spore cultures were transferred to PDA plates containing antibiotics as above, and incubated at 23\u0026deg;C in a growth chamber. After two weeks, sterile water was added to plates and conidia were scraped off with a microscope slide. Spore concentration was measured with a Thoma haemocytometer (Merck, Darmstadt, Germany) and adjusted to 1x10\u003csup\u003e6\u003c/sup\u003e conidia per ml. The primary cobs of maize plants were inoculated seven days after silk channel emergence (BBCH 65). A needle pin was dipped in conidia suspension and engraved in the cob through the husk, hurting the kernels. Ten plants in two repetitions were inoculated and twenty plants were inoculated with sterile water, which served as control. Four weeks (28 dpi) after inoculation, husk leaves of inoculated and control cobs were removed and percent disease severity was visually estimated (0-100%) according to EPPO Guidelines PP1/285 (EPPO Bulletin \u003cspan class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003ch2\u003e4.4 Cultures and strains used for temperature trials \u003cem\u003ein vivo\u003c/em\u003e and \u003cem\u003ein vitro\u003c/em\u003e\u003c/h2\u003e\n \u003cp\u003eFungal isolates utilized in this study were obtained from different sources and are currently kept in \u003cem\u003eTrichoderma\u003c/em\u003e strain collection at the University of G\u0026ouml;ttingen\u0026apos;s Department of Plant Pathology and Plant Protection. TS1 was provided by the Department of Agricultural, Forestry and Food Sciences (DISAFA-Plant Pathology Unit), AGROINNOVA - Interdepartmental Centre of the University of Torino (Sanna et al. \u003cspan class=\"CitationRef\"\u003e2022\u003c/span\u003e). The holotype of \u003cem\u003eT. afroharzianum\u003c/em\u003e, CBS 124.620, along with four strains recently isolated from maize cobs in Germany, France, and Italy, were included in the investigation. Additionally, two \u003cem\u003eTrichoderma\u003c/em\u003e strains used as active ingredients in biopesticide products, T34 (\u003cem\u003eT. asperellum\u003c/em\u003e) and T39 (\u003cem\u003eT. harzianum\u003c/em\u003e), were included. To ensure the purity of the strains, single spore colonies were isolated and cultured for subsequent experiments.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003cdiv align=\"left\" class=\"colspec\"\u003e\u003cbr\u003e\u003c/div\u003e\u0026nbsp;\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eIsolates used for \u003cem\u003ein vivo\u003c/em\u003e and \u003cem\u003ein vitro\u003c/em\u003e climate chamber trials\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"4\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIsolate\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSpecies\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eOrigin\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eYear\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCBS124620\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWesterdijk Fungalbiodiversity Inst.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAP18TRI1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMaize cob, Croix des Pardis, France\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2018\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAP18TRI2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMaize cob, Kuenzing, Germany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2018\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTS1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMaize cob, DISAFA-Plant Pathology Unit, Carmagnola, Italy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2020\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAP18TRI7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eT. harzianum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMaize cob, Grucking, Germany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2018\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eT. harzianum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTrichodex, Makhteshim Agan Industries\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eT34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eT. asperellum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eXilon\u0026reg;, Kwizda Agro GmbH\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003ch2\u003e4.5 \u003cem\u003eIn vitro\u003c/em\u003e temperature trial\u003c/h2\u003e\n \u003cp\u003eThe effect of temperature was assessed on PDA. The medium was inoculated with a 6 mm PDA plug obtained from a seven-day-old culture. The plates were placed in darkness and incubated at temperatures ranging from 8 to 44\u0026deg;C, with temperature increments of 4\u0026deg;C, using temperature chambers (Rumed Rubarth Apparate GmbH). The position of the plates within the chamber was randomized. A total of four plates were inoculated per isolate for each temperature and medium. Radial growth was assessed by measuring using a ruler along two perpendicular lines marked on the bottom of each Petri dish at twelve-hour intervals up to seven days post-inoculation (dpi) or until the plates were completely colonized. The radial growth at a given time was obtained by dividing the sum of the two radial measures in millimetres (mm) by four.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n \u003ch2\u003e4.6 Plant cultivation and inoculation procedure for climate chamber experiments\u003c/h2\u003e\n \u003cp\u003eThe inoculum for all experiments was obtained from single spore cultures grown on PDA, incubated for seven days in total darkness at 25\u0026deg;C in a growth chamber (Mytron Bio und Solartechnik GmbH). The spore suspension was adjusted to 1x10\u003csup\u003e6\u003c/sup\u003e colony-forming units (CFU) per milliliter using a Thoma haemocytometer.\u003c/p\u003e\n \u003cp\u003eThe maize variety LIKEit (K 180, Deutsche Saatveredelung AG) was sown into 16x16x13 cm pots with a mixture of 3:3:1 of potting soil, compost, and sand. The plants were cultivated under greenhouse conditions at a temperature of approximately 25\u0026deg;C, in accordance with the natural day-night rhythm and irrigated as necessary. The maize cobs were inoculated at BBCH 65 after the first silks had begun to dry-off. Silk channel inoculation was conducted by injecting 1 ml spore suspension, with a concentration of 1x10\u003csup\u003e6\u003c/sup\u003e spores per milliliter, into the tip of the first developed cob using a needle and syringe. Six plants per isolate were inoculated (BBCH 65), in two repetitions and placed afterwards in the climate chamber (RUMED). A total of 60 plants were placed in 5 climate chambers. The relative humidity was maintained at a constant level of 70%, with temperature variations ranging from 10\u0026deg;C, 15\u0026deg;C, 20\u0026deg;C, 30\u0026deg;C, to 35\u0026deg;C and with the day- /night light cycle set to 12/10 hours. Disease severity was assessed after four weeks as described above.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n \u003ch2\u003e4.7 Statistical analysis\u003c/h2\u003e\n \u003cdiv class=\"BlockQuote\"\u003e\n \u003cp\u003eStatistical analysis was conducted using STATISTICA version 13 (Statistica GmbH, Germany). All experiments were fully randomized. To assess treatment effects and differences among the means of experimental units, two- or three-factorial analysis of variance (ANOVA) was performed. Analysis of variance (ANOVA) was carried out by Tukey-HSD-test at 5% probability. Pearson\u0026rsquo;s correlation coefficients were used to examine the relationship between temperature and precipitation in June, July, August, and September and infection of \u003cem\u003eT. afroharzianum\u003c/em\u003e on maize cobs and soil. In addition, a multiple regression was performed to determine the relationship of Trichoderma ear rot infection with temperature and precipitation in June, July and August for each sample location.\u003c/p\u003e\n \u003cp\u003eTrichoderma ear rot infection was calculated by the following equation:\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Equa\" class=\"Equation\"\u003e\n \u003cdiv class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e$$\\:Trichoderma\\:ear\\:rot\\:infection\\:\\left[\\%\\right]=\\frac{Number\\:of\\:T.\\:afroharzianum\\:isolates\\:}{Total\\:number\\:of\\:samples\\:}\\:\u0026divide;\\:Location$$\u003c/div\u003e\n \u003c/div\u003e\n \u003cdiv class=\"BlockQuote\"\u003e\n \u003cp\u003eThe impact of weather conditions on the occurrence of \u003cem\u003eT. afroharzianum\u003c/em\u003e was analyzed by variance components derived from the overall variance estimated with the restricted maximum likelihood model.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eSupplementary Materials:\u0026nbsp;\u003c/strong\u003eThe following supporting information can be downloaded at: www.mdpi.com/xxx/s1, Figure S1: title; Table S1: title; Video S1: title.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate:\u0026nbsp;\u003c/strong\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication:\u003c/strong\u003e Written informed consent has been obtained from the authors to publish this paper\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and material:\u0026nbsp;\u003c/strong\u003eThe datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interest:\u003c/strong\u003e The authors declare no conflict of interest\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e This research was funded by Federal Ministry of Agriculture, Food and Regional Identity, grant number FKZ2221NR014\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions:\u003c/strong\u003e Conceptualization, A.P.; methodology, W.F.V.; investigation, A.P, and W.F.V.; writing\u0026mdash;original draft preparation, A.P. and C.D-M.; writing\u0026mdash;review, C.D-M., G.S, B.C.S and A.v.T.; visualization, A.P.; supervision, B.C.S and A.v.T.; project administration, A.P.; funding acquisition, A.P, C.D-M., G.S, B.C.S and A.v.T. All authors have read and agreed to the published version of the manuscript.\u0026rdquo;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments:\u003c/strong\u003e We would like to thank the breeding companies, universities and federal research institutions that provided us with maize and soil samples. We also thank Manuela M\u0026uuml;cke and Tobias Wille for technical support of the experiments and for performing DNA sequencing, respectively.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAchuo EA, Prinsen E, H\u0026ouml;fte M. Influence of drought, salt stress and abscisic acid on the resistance of tomato to \u003cem\u003eBotrytis cinerea\u003c/em\u003e and \u003cem\u003eOidium neolycopersici\u003c/em\u003e. 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JEB. 2017;38(2):197\u0026ndash;203. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.22438/jeb/38/2/ms-251\u003c/span\u003e\u003cspan address=\"10.22438/jeb/38/2/ms-251\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Trichoderma, maize, ear rot, environmental conditions, infection","lastPublishedDoi":"10.21203/rs.3.rs-6871003/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6871003/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e \u003cem\u003eTrichoderma\u003c/em\u003e species are widespread soil-living fungi known for their roles in both agriculture and natural ecosystems. While some species are used as effective biocontrol agents against plant pathogens, others, such as \u003cem\u003eT. afroharzianum\u003c/em\u003e, have recently been identified as pathogenic, causing diseases like Trichoderma ear rot in maize and infections in wheat. The occurrence of \u003cem\u003eTrichoderma\u003c/em\u003e species is strongly influenced by environmental factors, particularly temperature and precipitation. This study aims to assess the occurrence and distribution of Trichoderma ear rot in maize across Europe, and to evaluate the effect of environmental factors, especially temperature and precipitation, on their occurrence. Over a seven-year period (2018\u0026ndash;2024), 345 samples of maize cobs, stalks, and soil samples were received from 72 locations in Europe. The obtained \u003cem\u003eTrichoderma\u003c/em\u003e isolates (n\u0026thinsp;=\u0026thinsp;130) were identified to the species level, and their pathogenicity to maize was tested under controlled conditions. Weather data, including temperature and precipitation, were gathered from each location and correlated with the occurrence of Trichoderma ear rot. Additionally, \u003cem\u003ein vitro\u003c/em\u003e and \u003cem\u003ein vivo\u003c/em\u003e climate chamber trials were conducted to confirm the favorable temperature conditions for disease development. Our results indicate that \u003cem\u003eT. afroharzianum\u003c/em\u003e was the most prevalent species isolated from maize cobs and agricultural soil, exhibiting strong pathogenicity in maize cobs. Correlation analysis between the occurrence of \u003cem\u003eT. afroharzianum\u003c/em\u003e and environmental factors revealed that high temperatures (\u0026gt;\u0026thinsp;19.6\u0026deg;C mean temperature), particularly during the summer months, strongly favored the presence of \u003cem\u003eT. afroharzianum\u003c/em\u003e, while cooler and wetter regions (\u0026lt;\u0026thinsp;18.2 mean temperature) showed little to no occurrence of pathogenic \u003cem\u003eTrichoderma\u003c/em\u003e species. \u003cem\u003eIn vitro\u003c/em\u003e and \u003cem\u003ein vivo\u003c/em\u003e climate chamber experiments confirmed these results observed in the field. The growth rate of \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates was significantly higher compared to other \u003cem\u003eTrichoderma\u003c/em\u003e species, with an optimal temperature range between 28 and 32\u0026deg;C.\u003c/p\u003e","manuscriptTitle":"Effect of temperature and precipitation on the occurrence of Trichoderma ear rot in Europe","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-20 15:24:40","doi":"10.21203/rs.3.rs-6871003/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"12e56fff-14b1-45e9-9198-c48f76b95250","owner":[],"postedDate":"June 20th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-02-16T16:02:25+00:00","versionOfRecord":{"articleIdentity":"rs-6871003","link":"https://doi.org/10.1186/s42483-025-00396-4","journal":{"identity":"phytopathology-research","isVorOnly":false,"title":"Phytopathology Research"},"publishedOn":"2026-02-10 15:58:18","publishedOnDateReadable":"February 10th, 2026"},"versionCreatedAt":"2025-06-20 15:24:40","video":"","vorDoi":"10.1186/s42483-025-00396-4","vorDoiUrl":"https://doi.org/10.1186/s42483-025-00396-4","workflowStages":[]},"version":"v1","identity":"rs-6871003","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6871003","identity":"rs-6871003","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}