PCR-based assays for rapid and sensitive detection of Trichoderma afroharzianum, a fungal pathogen causing new corn ear rot

PCR-based assays for rapid and sensitive detection of Trichoderma afroharzianum, a fungal pathogen causing new corn ear rot | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article PCR-based assays for rapid and sensitive detection of Trichoderma afroharzianum, a fungal pathogen causing new corn ear rot Clovis Douanla-Meli, Annette Pfordt, Andreas von Tiedemann, Gritta Schrader, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8501765/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 12 You are reading this latest preprint version Abstract The emergence of a new corn ear rot was recently described in Germany and has since received increased attention following detections in several European and Asian countries. All reports consistently identify Trichoderma afroharzianum as the causal agent. The potential impact on maize production, combined with the risk of misdiagnosis as other ear rot diseases, highlights the need for a rapid and accurate diagnostic method for the reliable detection of T. afroharzianum . In this study, we developed PCR-based diagnostic tools including an endpoint PCR and a real-time TaqMan assay targeting the TEF1α and RPB2 loci, respectively. Both assays were validated using a broad panel of Trichoderma isolates and other fungal species obtained from maize plants, additional host species from diverse geographic origins, and soils collected from German maize fields. All performance criteria required to demonstrate assay reliability were met. The assays showed high accuracy and sensitivity, with limits of detection of 1 pg for the endpoint PCR and 1 fg per reaction for the real-time PCR. The real-time assay exhibited amplification efficiencies of 105% (R² = 0.98) for pure cultures and 118% (R² = 0.998) for infected maize kernels. Furthermore, both assays reliably detected T. afroharzianum in asymptomatic maize cobs following artificial inoculation with pathogenic strains. These methods therefore enable early and reliable detection of T. afroharzianum ear rot and allow its discrimination from other maize ear rot diseases. This capability is essential for the development of targeted management strategies to address this emerging threat to maize production. Biological sciences/Biological techniques Biological sciences/Biotechnology Biological sciences/Microbiology Biological sciences/Molecular biology Biological sciences/Plant sciences Pathogenic Trichoderma Taqman PCR corn disease Trichoderma afroharzianum emerging diseases Figures Figure 1 Introduction Maize ( Zea mays ) is one of the world’s most important crops and the second most widely produced cereal, serving as an essential food source for both humans and livestock 1,2 . Beyond direct consumption, maize is a highly versatile commodity used in the production of numerous derivatives, including food products, animal feed, biofuels, and a broad range of industrial goods 3 . However, maize production is increasingly challenged by abiotic and biotic stressors that threaten yields and, more broadly, global food security 4 . Abiotic factors such as drought and extreme temperatures are largely driven by climate change 5 , biotic stressors comprise living organisms that negatively affect maize crops and can cause substantial yield losses 6 . These include a wide range of pests and pathogens—bacteria, fungi, viruses, nematodes, insects as well as weeds. Among the most prevalent maize diseases are ear rots, primarily caused by Fusarium species, but also by other fungal pathogens such as Aspergillus , Penicillium , Nigrospora , and Stenocarpella 7,8 . Previous studies have reported grain yield losses of 10–30% due to ear rot 9,10 , a figure that is likely to rise with the emergence of new ear rot types. In 2018, a previously unreported Trichoderma ear rot of maize was identified in southern Germany, and T. afroharzianum was determined as the causal agent 11 . Initial symptoms consist of white mycelium, which subsequently develops into green to grey-green growth spreading between or over the kernels. A subsequent survey revealed an extensive distribution of the disease throughout Germany 12 . Since then, Trichoderma ear rot has been reported in France, Italy, Austria, and China 12,13, 14, 15 . In all cases, T. afroharzianum was identified as the causative agent, supporting the view that this species represents a major pathogen associated with ear rot symptoms under field conditions. In addition to T. afroharzianum , pathogenic strains of T. asperellum and T. atroviride have been reported in India to cause ear rot in maize 16 . Furthermore, T. afroharzianum and T. harzianum have been associated with post-flowering stalk rot in maize 16 . Taken together, these findings suggest that an increasing number of Trichoderma species may become associated with ear rot and other maize diseases in the future. At the same time, the emergence of Trichoderma -associated diseases in maize may have been underestimated due to limited symptom-based differentiation from other ear rot diseases and the insufficient emphasis on diagnostic approaches. This interpretation is supported by the growing number of reports published within a relatively short time frame 11,13,14 . With specific regard to T. afroharzianum , this species is globally distributed and commonly occurs in soil, dead wood, and living plants as an endophyte. In addition, it is used as an active ingredient in several commercial biological control products. Its pathogenic lifestyle therefore represents a newly recognised trait of this species and underscores the need for accurate identification. This is particularly important because non-pathogenic species or strains are widely applied in biological control, and their use must be demonstrably safe. In general, early and efficient diagnosis is a critical prerequisite for the effective management of plant diseases. Accordingly, the rapid, accurate, and reliable detection and identification of T. afroharzianum is essential. Morphological characterisation alone is insufficient for the reliable diagnosis of Trichoderma species, as many taxa exhibit overlapping phenotypic traits. Consequently, the combination of multilocus phylogenetic analysis (MLPA) and DNA barcoding has become the most effective approach for species-level identification within the genus Trichoderma 17,18,19 . Phylogenetically, T. afroharzianum belongs to the Harzianum clade and can be distinguished from closely related species using MLPA. However, MLPA has inherent limitations, most notably the time required to complete the analysis, which restricts its applicability for routine diagnostic purposes 20 . In addition, the generation of pure cultures for DNA sequencing through single-spore isolation can be particularly time-consuming, especially when multiple Trichoderma species co-occur on infected maize cobs 12 . Given the global importance of maize, substantial efforts have been devoted to the development of a wide range of diagnostic technologies, from laboratory-based PCR assays to field-based remote sensing approaches for disease detection 21,22,23 . Direct methods employing non-invasive imaging sensors have been used predominantly for the diagnosis and monitoring of fungal diseases affecting maize leaves 24,25,26 . In contrast, applications addressing maize ear rot have so far been largely limited to the detection, monitoring, and quantification of mycotoxin production 27 . The implementation of molecular tools for species- and strain-specific identification, including immunological approaches, fingerprinting techniques, exogenous markers, and PCR-based methods 28,29 , can facilitate the effective detection and differentiation of Trichoderma ear rot. Among these approaches, species-specific PCR has proven to be a particularly effective tool, providing faster, more specific, and more sensitive detection and identification of pathogenic fungi 30,31 . The primary objective of this study was to develop a specific and sensitive molecular tool to facilitate the detection and identification of T. afroharzianum . Primers and hydrolysis probes were designed targeting the TEF1α and RPB2 genes and evaluated for their ability to specifically amplify T. afroharzianum from pure cultures as well as from naturally and artificially infected maize kernels. Two assays, a conventional PCR (cPCR) and a TaqMan-based real-time PCR (qPCR), were developed and validated in accordance with the EPPO standard PM 7/98 32 . Methods Isolates collection and identification The isolates of Trichoderma spp. and other fungi included in this study were primarily derived from maize samples and soil collected from maize fields. Samples of maize cobs and stalks were obtained between 2018 and 2024 from both symptomatic and asymptomatic plants. Trichoderma isolates were also recovered from soil originating from fields with and without confirmed Trichoderma ear rot infection. Sampling of soil and maize plant material was conducted throughout Germany in collaboration with farmers willing to contribute to the survey of outbreaks of Trichoderma ear rot disease. Samples stored in plastic bags were sent to the laboratory for analysis. The procedures for isolating fungi from maize and soil samples and assigning them to Trichoderma followed the methods described by 12 . Additional Trichoderma strains were obtained from fungal culture collections, research institutions, and commercial biocontrol products containing Trichoderma as active organisms. Isolation of Trichoderma spp. from biocontrol products was carried out in accordance with the methodology outlined in 12 . For each isolate, the ITS, TEFα, and RPB2 regions were sequenced, and molecular identification was conducted following the protocol proposed by 19 . All isolates had previously undergone pathogenicity testing on maize under controlled greenhouse conditions. Details pertaining to taxonomic identification and pathogenicity are available in 12 . A complete list of fungal strains and their origins used in this study is provided in Table 1. Design of primers and hydrolysis probes Sequences generated from the TEF1α and RPB2 genes for Trichoderma species in the Brevicompactum, Harzianum, Virens and Viride clades including referenced sequences retrieved from the NCBI GenBank database were composed. Alignment was performed in MAFFT v. 7 online version 33 using the iterative refinement option G-INS-i and manual optimization with MEGA v. 7. The alignments were analysed in Geneious Prime to identify regions with poly-morphisms. The design of a set of primers and hydrolysis probes was conducted for each gene alignment using the online version of Primer 3 software (http://bioinfo.ut.ee/primer3-0.4.0/). The specificity of the designed primers, as well as the absence of primer-dimer formation, was subjected to further testing using Primer-Blast (http://www.ncbi.nlm.nih.gov/tools/primer-blast). The primers and probes were custom-synthesized by Metabion (Planegg, Germany) with the probes labelled using FAM (6-carboxyfluorescein) and BHQ-1 (Black Hole Quencher®-1). PCR conditions and optimization The cPCR was performed using 25 μL PCR reactions containing 1µM (0.5 μL) of each primer TrafTef1-F and TrafTef1-R, 2x GoTaq Green Master Mix (Promega, USA) (12.5 μL), 2.5 μL of gDNA. Molecular grade water was added to the reaction up to 25 μL. PCR reactions were per-formed in a gradient block Biometra thermal cycler (AnalytiK Jena, Germany) using following cycling conditions: initial denaturation at 94°C for 3 min, followed by 35 cycles of 45 s at 94°C, 45 s annealing with a gradient temperature run from 56°C to 64°C and 1 min at 72°C and a final extension of 7 minutes at 72°C. No template control (NTC) consisting of ddH2O was included to check for the absence of contamination. PCR products (3 μL) were checked on 1% agarose gel electrophoresis stained with ROTI GelStain (Roth, Germany). A further check on the conformity of the amplicons with regard to sequence length and species identity was achieved by means of sequencing. Thereby, amplicons derived from T. afroharzianum isolates CBS 124620, AP22TRI85 and MRI349 were sequenced as described in 12 , followed by BLAST search in the NCBI GenBank. The newly generated sequences matched those with accession numbers PQ558230 (1,248 bp), PQ558225 (1,258 bp) and PQ558235 (1,250 bp), respectively, previously generated from the above isolates 12 . As just a small part of known sequences, their addition to GenBank was not considered. The qPCR was carried out in 12.5 μL final volume containing 0.25 µM (0.125 μL) of forward primer TrafRpb2-F and 0.125 µM (0.125 μL) of reverse primer TrafRpb2-R, 0.15 µM (0.19 μL) of probe TrafRpb2-P, 1x iTaq Universal Probes Supermix (6.25 μL) (Bio-Rad Laboratories, Pty, Ltd.) and 1 μL DNA template. Molecular grade water (4.81 μL) was added to obtain the final volume. The cycling conditions were an initial denaturation at 95°C for 2 min, and 40 cycles of denaturation at 95°C for 15 s following by annealing with a gradient temperature run from 60°C to 66°C for 1 min. Reactions were performed on the StepOnePlus Real-Time PCR System (Applied Biosystems). No template controls (NTC) consisted of ddH2O. The Ct values and standard deviations of all samples were recorded using the StepOnePlus™ Software v2.3. Validation of the PCR methods The performance of the PCR assays to amplify T. afroharzianum was evaluated by validating different parameters. All DNA extracts were standardized to a concentration of 1 ng µL-1. The analytical specificity was assessed using isolate composition including as non-target 37 of other Trichoderma species from maize plants, soil from maize fields and other sources and 14 of non- Trichoderma species from maize plants (Table 1). The inclusivity was assessed with a panel of 37 T. afroharzianum isolates from different sources (predominantly from maize cobs and soil from maize fields, mycoparasite, biostimulant products) and geographical origin (Germany, France, Italy, Peru). All assays were conducted with three replicates of each isolate. The pDNA of T. afroharzianum strain CBS 124620 served as positive control and the no template control consisting of ddH2O was included. The analytical sensitivity was evaluated through a ten-fold serial dilution of pDNA in TE buffer and of pDNA pooled with maize kernel DNA at 1:1 ratio from 1 ng µL -1 concentration with ten replicates being conducted for each template. The standard curve was generated using the obtained cycle threshold (Ct) values and the efficiency of the assay calculated using the equation E = -1 +10 (-1/slope) 34 . The limit of detection (LOD) was defined as the lowest concentration that could still be amplified in all replicates. The sensitivity of the assays was further evaluated by testing on asymptomatic artificially infected maize cobs obtained from a greenhouse pathogenicity experiment (as described in 12 ). The collection of these asymptomatic infected cobs was carried out prior to the manifestation of first symptoms. Further criteria were evaluated using templates consisting of the pDNA of T. afroharzianum (strain CBS 124620) set at 10×LOD and 100×LOD, 0.1 ng µL-1 of T. afroharzianum (strain CBS124620) and T. afroharzianum (from sample 159 as natural infected maize kernels), 1 ng µL-1 of T. asperellum (AP22TRI100) and T. brevicompactum (AP22TRI140). Repeatability and reproducibility were assessed by performing each assay (cPCR and qPCR) with these templates in ten and three replicates, respectively. Reproducibility was also assessed by two different operators repeating the assays on two different qPCR platforms: the StepOnePlus Real-Time PCR System (Applied Biosystems) and qTower3 thermocycler (Analytik Jena, Germany). The robustness of the qPCR assays was assessed in ten replicates using reaction volumes of ±10% (11,25 µL and 13,75 µL) at ±2°C modified hybridisation temperatures (60°C, 62°C and 64°C). For the transferability, the qPCR was performed in ten replicates on two qPCR platforms above, and with four different master mixes: iTaq Universal Probes Supermix (Bio-Rad), Maxima Probe/ROX qPCR Master Mix (Fischer scientific), Applied Biosystems™ TaqMan™ Universaler PCR-Mastermix (Fischer scientific) and TaKaRa Premix Ex TaqTM (Probe qPCR); TaKaRa, Kusatsu, Japan). Data analysis For the qPCR assay, the StepOnePlus software automatically generated the baseline range, threshold cycle (Ct) values and standard curves. The results were checked to ensure they aligned with the provider's analysis guidelines. The mean threshold cycle (Ct) values were calculated along with their standard deviations (SD). Precision of the assay was evaluated by computing the coefficient of variation (CV). All statistical calculations were performed using Microsoft Excel. Results Selection of T. afroharzianum specific primers and PCR conditions The design of primers and probes for both cPCR and qPCR was initially based on TEFα and RPB2 sequences. The positions of the selected primers and probe are illustrated in Figure S1 and the sequences are presented in Table 2. The few qPCR primer combinations obtained on TEF1α predominantly showed cross-reactivity with many non-target Trichoderma species (data not shown), despite optimization efforts. By contrast, several qPCR primer combinations were identified based on the RPB2 region. Initial screening using 3 annealing temperatures (60, 62, 64°C) suggested selecting the TrafRpb2q-F/ TrafRpb2q-R pair and the corresponding TrafRpb2q-P probe, which successfully amplified all target templates at an annealing temperature of 62°C with no cross-reactivity. The PCR products obtained at annealing temperature of 62°C show a single band of the expected size of 138 bp in agarose electrophoresis. For the specificity, the selected cPCR primers yielded 100% analytical specificity leading to the amplification of a single 217 bp fragment, for all and only the T. afroharzianum isolates. In order to ascertain and extend the inclusivity of the cPCR primers, a series of tests were carried out on a panel of samples consisting of kernels from naturally infected and greenhouse-inoculated maize plants. These tests yielded satisfactory results. Amplicon sequencing further confirmed the suitability of the primers to amplify the TEF1α region of T. afroharzianum . Comparatively, the qPCR assay demonstrated 100% inclusivity by amplifying gDNA from all the T. afroharzianum isolates (mean Ct = 19.25-25.38, SD = 0.2-1.47; n=10) (Table 1), regardless of the host and geographical origin. In general, the qPCR assay did not amplify DNA from non- Trichoderma species, with the exception of the T. gamsii isolate AP24TRI434. Despite implementing all possible options to optimize the primer specificity (e.g. varying the annealing temperature), this issue persistently occurred. Reliability of the PCR assays to detect T. afroharzianum Analytical sensitivity differed between the species-specific cPCR and qPCR assays. In a tenfold serial dilution of plasmid DNA ranging from 1 ng to 10 -8 ng µL⁻¹, the cPCR consistently produced amplicons of the expected size (217 bp) in 100% of replicates (n = 10) down to a concentration of 1 pg µL⁻¹ (= 10 -3 ng µL⁻¹) using DNA from pure cultures. In contrast, the qPCR consistently amplified target DNA in all replicates (n = 10) down to a concentration of 1 fg µL⁻¹ (= 10 -6 ng µL⁻¹) (Fig. 1A). Linear calibration curves generated from mean Ct values plotted against the logarithm of template concentration are shown in Fig. 1B–C. Fully repeatable positive results (100%) were obtained down to the determined limit of detection (LOD) of 1 fg µL⁻¹ using plasmid DNA (pDNA) diluted in TE buffer (mean Ct = 30.4) and pDNA diluted in 1 ng µL⁻¹ genomic DNA extracted from healthy maize kernels (mean Ct = 30.8). For pDNA diluted in TE buffer, the standard curve showed a correlation coefficient (R²) of 0.98, a slope of −3.20, and an amplification efficiency of 105%. When pDNA was diluted in genomic DNA from healthy maize kernels, the corresponding values were R² = 0.996, a slope of −2.99, and an efficiency of 118%. Both cPCR and qPCR assays successfully detected T. afroharzianum in all samples derived from maize kernels collected from asymptomatic infected cobs. The qPCR assay yielded Ct values ranging from 27 to 29, indicating its suitability for detecting T. afroharzianum in asymptomatic samples at concentrations close to the determined LOD. Both assays demonstrated satisfactory repeatability, reproducibility, robustness, and transferability when evaluated using a panel comprising plasmid target DNA and genomic DNA from pure cultures as well as naturally infected samples. Both the cPCR and qPCR assays amplified exclusively templates containing target DNA, whereas no amplification was observed for non-target DNA in any replicate, resulting in 100% repeatable and reproducible outcomes (Supplementary Table S1). The coefficients of variation (CVs) ranged from 0.44% to 1.05% for repeatability and from 0.48% to 0.72% for reproducibility. The qPCR assay exhibited high robustness with respect to variations in experimental parameters. Alterations in reaction volume (12.5 µL) or hybridisation temperature (60 °C) had little effect on assay performance. However, a slight decrease in Ct values was observed at an annealing temperature of 64 °C (Supplementary Table S2). Furthermore, analysis of the template panel by two independent operators yielded the expected results for both target and non-target templates. These results were consistent across operators and showed very low variation in Ct values, confirming the repeatability and reproducibility of the assay (data not shown). To evaluate the transferability of the qPCR assay, additional analyses were performed using three different commercial master mixes and an alternative PCR platform. Changes in the master mix or PCR platform did not affect assay specificity, and repeatability and reproducibility were maintained across all tested conditions (Supplementary Table S3). The observed differences were primarily related to assay sensitivity. When using the StepOnePlus system, the TaKaRa Premix Ex Taq Probe qPCR produced Ct values comparable to those obtained with the iTaq Universal Probes Supermix (Bio-Rad). The Maxima Probe/ROX qPCR Master Mix increased assay sensitivity, resulting in Ct values that were 0.67–2.04 cycles lower than those obtained with the iTaq Universal Probes Supermix. In contrast, use of the iTaq Universal Probes Supermix on the qTower3 thermocycler (Analytik, Jena) resulted in reduced robustness, with coefficients of variation ranging from 2.90% to 3.32%, and Ct values that were 1.25–3.24 cycles higher than those obtained using the StepOnePlus system. Discussion Trichoderma-associated diseases in maize production are an increasing concern. The threat to this key cereal crop is likely to intensify due to the widespread occurrence and persistence of Trichoderma species in agricultural soils worldwide 19 , 35 , 36 , 37 . Moreover, Trichoderma species are widely used as biological control agents, and the pathogenic potential of certain species or strains therefore represents a significant challenge for the safe application, regulation, and acceptance of Trichoderma -based biocontrol products. Accordingly, the development of molecular tools for the accurate identification of pathogenic Trichoderma species is essential for improving the understanding and management of these diseases. Here, we provide a PCR-based tool for the rapid and sensitive detection of T. afroharzianum . Current knowledge indicates that multiple Trichoderma species are associated with diseases in maize 12 , 15 , 16 , therefore, the implementation of multiplex PCR assays enabling the simultaneous detection of multiple pathogens represents a rational and efficient diagnostic approach 38 . However, epidemiological data from Europe indicate that T. afroharzianum has consistently been identified as the predominant etiological agent of Trichoderma ear rot 11 , 13 . Under these circumstances, species-specific detection of this pathogen using simplex PCR remains justified. The most significant achievement of this study was the development of a diagnostic tool based on complementary cPCR and qPCR assays. Despite the longer processing time associated with cPCR, it remains a cost-effective option compared with the more expensive qPCR approach 39 , which in turn provides the advantage of higher sensitivity 40 . The selection of target genes for primer design was guided by established molecular identification practices for Trichoderma species, which rely primarily on the ITS, TEF1α, and RPB2 regions for phylogenetic analysis and DNA barcoding 19 . For PCR-based detection, ITS markers are generally effective at the genus level 41 , 42 , 43 , whereas TEF1α and RPB2 are more suitable for species-specific discrimination 19 , 28 , 44 , 45 , 46 . Both TEF1α and RPB2 have previously been demonstrated to reliably target T. afroharzianum 37 , 45 , 46 . In the present study, TEF1α was identified as the most suitable gene for cPCR, whereas RPB2 was found to be more appropriate for the qPCR assay. A previous study developed a qPCR assay for the detection and quantification of T. afroharzianum causing maize ear rot in Italy 46 . That assay targeted the TEF1α gene, based on the assumption that conserved single-nucleotide polymorphisms within this region allow discrimination of T. afroharzianum from other species within the Harzianum clade. However, the qPCR primers designed in the same TEF1α region as in study 46 exhibited unexpected cross-reactivity with certain T. asperellum and T. atrobrunneum isolates. Full specificity for T. afroharzianum was only achieved when primers targeting the RPB2 gene were employed. This single-copy and highly conserved RPB2 gene plays a central role in resolving the phylogeny of Trichoderma species 12 , 19 and has increasingly been recognised as a suitable and reliable marker for PCR-based detection of Trichoderma species (Prabhakaran et al. 2014; Gerin et al. 2018). Regarding real-time PCR chemistry, SYBR Green used by 46 has been widely applied for the detection and quantification of Trichoderma species, particularly in studies addressing their biocontrol potential 47 , 48 . The TaqMan PCR system is likewise well established for diagnostic and quantitative applications 45 , 49 , 50 and was selected in the present study for two main reasons: first, to provide a user-friendly diagnostic tool, and second, because of its superior accuracy and specificity 51 . A rigorous validation process demonstrated that the proposed assays largely fulfil the requirements for new diagnostic methods as defined by 32 . Both assays exhibited high specificity against all tested Trichoderma species and other fungi associated with maize cobs and a range of additional host plants. Although cross-reactivity of the qPCR assay was observed with the T. gamsii isolate AP14TRI434, this isolate originated from soil and was shown to be non-pathogenic to maize. Consequently, it can be inferred that neither the cPCR nor the qPCR assay amplifies Trichoderma isolates capable of causing field or greenhouse symptoms comparable to those induced by T. afroharzianum 12 , 15 . Both assays achieved 100% inclusivity for all T. afroharzianum isolates tested, encompassing a broad range of geographic origins, host associations, and pathogenicity profiles 12 . It should be noted, however, that not all T. afroharzianum strains are pathogenic to maize, and the determinants underlying pathogenic versus non-pathogenic lifestyle, as well as reliable molecular markers to discriminate between these isolates, remain unknown. Importantly, neither the phylogenetic analyses 12 nor the PCR assays developed in the present study are able to distinguish between pathogenic and non-pathogenic T. afroharzianum isolates. This limitation has direct implications for the interpretation of diagnostic results. In particular, pathogenicity testing remains necessary to substantiate a positive diagnostic result, especially when latent infections are tested, in order to confirm that the detected T. afroharzianum isolate is capable of causing disease, in accordance with Koch’s postulates. Conversely, maize samples exhibiting characteristic Trichoderma ear rot symptoms but yielding negative results in these assays should be further evaluated for the presence of other Trichoderma species that may also be pathogenic to maize. The high sensitivity of the qPCR assay is consistent with previous assays targeting Trichoderma species 44 , 52 . The LOD of 1 fg µL⁻¹ determined for the TaqMan assay represents a substantial improvement over the LOD of 50 fg µL⁻¹ reported for a SYBR Green based assay 46 . This pronounced difference in sensitivity is unlikely to be attributable solely to the detection chemistry and is more likely related to differences in sample preparation or reaction composition 53 . The presence of host plant material had no detectable effect on analytical sensitivity, as indicated by comparable results obtained from pure cultures, naturally infected samples, and artificially inoculated material. Although minor inhibitory effects caused by co-extracted DNA cannot be entirely excluded 52 , 54 , such effects are unlikely to compromise assay performance when testing maize kernels. Both assays successfully detected T. afroharzianum in asymptomatic material at early infection stages (7–10 days post inoculation), underscoring their suitability for early diagnosis. As expected, both assays demonstrated high repeatability, reproducibility, and robustness. While the cPCR assay was readily transferable across platforms, the qPCR assay required careful verification prior to diagnostic implementation, particularly when using platforms other than the StepOnePlus system. For example, testing on the qTower qPCR device resulted in delayed amplification signals, which could lead to false-negative results in samples containing low levels of target DNA, a limitation that is especially relevant for latent infections. However, the risk of false negatives in symptomatic samples is considered low, as maize tissues affected by Trichoderma ear rot typically exhibit extensive fungal colonization 12 , 13 , 15 , providing sufficiently high concentrations of target DNA. Conclusion The aim of this study was to facilitate the diagnosis of the newly emerging Trichoderma ear rot by providing a rapid and efficient molecular tool. The two PCR assays developed and validated here are well suited to achieve this objective. A key advantage of this approach is the use of two complementary PCR systems endpoint and TaqMan real-time PCR targeting different genetic loci. This diagnostic framework enables the rapid detection of T. afroharzianum directly from infected maize tissue, thereby eliminating the need for culture-based isolation. Furthermore, the high sensitivity of the qPCR assay allows the detection of latent infections, which is essential for early diagnosis. The ability to identify T. afroharzianum ear rot at early stages is critical for the timely implementation of management strategies aimed at minimizing yield losses and preventing further spread of the disease. Declarations Competing interests The authors declare no competing interests. Funding Declaration This study was supported by the Federal Ministry of Food and Agriculture – BMEL (actually Federal Ministry of Agriculture, Food and Regional Identity – BMLEH), grant number FKZ2221NR014, subproject 2221NR014B. The funder had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript. Author Contribution CDM, AP, GS, BCS, and AvT designed the project and secured the funding. CDM designed and conducted the experiments, and analysed the data. CDM wrote the manuscript. AP, GS, BCS, and AvT reviewed it. All authors read and approved the final version of the manuscript. Acknowledgement We would like to express our gratitude to the companies, universities and research institutions that provided us with Trichoderma isolates. Additionally, we thank Julia Krebs and Tobias Wille for their technical support during the method development process. Data Availability All the data is available in the main manuscript References Rouf Shah, T., Prasad, K. & Kumar, P. Maize- A potential source of human nutrition and health: A review. Cogent. Food Agric. 2 , 1–9. doi: 10.1080/23311932.2016.1166995 (2016). Chen, C., Chaudhary, A. & Mathys A. Nutrient adequacy of global food production. Front. Nutr. 8 , 739755. doi: 10.3389/fnut.2021.739755 (2021) Lütke, E. N., Schwarz, F. J. & Heilmann H. Handbuch Mais: Grundlagen Anbau Verwertung Ökonomie (DLG-verlag, Frankfurt 2013). Daszak, P., Cunningham, A. A. & Hyatt AD. Anthropogenic environmental change and the emergence of infectious diseases in wildlife. Acta Trop. 78 (2), 103–116 (2001) Juroszek, P. & von Tiedemann, A. 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Species Isolate Year Source/Host Location Assay (target) cPCR qPCR T. afroharzianum P AP18TRI1 2018 Symptomatic corncobs France + 21,70 ± 0,54 T. afroharzianum P AP18TRI2 2018 Symptomatic corncobs Germany + 20,45 ± 0,78 T. afroharzianum P AP18TRI3 2018 Symptomatic corncobs Germany + 20,26 ± 0,24 T. afroharzianum P AP19TRI5 2019 Symptomatic corncobs Germany + 22,6 ± 0,57 T. afroharzianum NP CBS 124620 - Theobroma cacao Peru + 21,58 ± 0,59 T. afroharzianum P AP20TRI15 2020 Symptomatic corncobs Germany + 19,25 ± 0,89 T. afroharzianum P AP20TRI16 2020 Symptomatic corncobs Germany + 20,14 ± 0,35 T. afroharzianum NP KG10 - Pleurotus ostreatus substrate - + 20,97 ± 0,71 T. afroharzianum NP KG13 - P. ostreatus substrate - + 25,83 ± 0,31 T. afroharzianum NP MRI349 - Biostimulant - + 21,53 ± 0,28 T. afroharzianum NP T42 - Marchantia polymorpha - + 21,87 ± 0,41 T. afroharzianum NP T138 - M. polymorpha - + 21,23 ± 0,62 T. afroharzianum P DISAFATS-1 - Symptomatic corncobs Italy + 19,93 ± 1,08 T. afroharzianum P AP22TRI81 2022 Symptomatic corncobs Germany + 21,29 ± 0,57 T. afroharzianum P AP22TRI19 2022 Symptomatic corncobs Germany + 22,87 ± 0,51 T. afroharzianum P AP22TRI99 2022 Soil maize field Germany + 22,87 ± 0,51 T. afroharzianum P AP22TRI108 2022 Soil maize field Germany + 20,82 ± 1,07 T. afroharzianum P AP22TRI118 2022 Soil maize field Germany + 19,73 ± 0,82 T. afroharzianum P AP22TRI121 2022 Soil maize field Germany + 20,66 ± 0,76 T. afroharzianum P AP22TRI129 2022 Soil maize field Germany + 21,11±1,47 T. afroharzianum P AP22TRI130 2022 Soil maize field Germany + 19,70 ± 0,71 T. afroharzianum P AP22TRI131 2022 Soil maize field Germany + 23,27 ± 1,76 T. afroharzianum P AP22TRI134 2022 Soil maize field Germany + 24,94 ± 0,77 T. afroharzianum P AP23TRI136 2022 Soil maize field Germany + 23,36 ± 0,67 T. afroharzianum P AP23TRI221 2023 Soil maize field Germany + 24,17 ± 1,75 T. afroharzianum P AP22TRI225 2023 Soil maize field Germany + 23,97 ± 1,56 T. afroharzianum P AP24TRI401 2024 Symptomatic corncobs Germany + 24,32 ± 1,14 T. afroharzianum P AP24TRI402 2024 Symptomatic corncobs Germany + 23,64 ± 1,24 T. afroharzianum P AP24TRI403 2024 Symptomatic corncobs Germany + 23,37 ± 1,05 T. afroharzianum P AP24TRI404 2024 Symptomatic corncobs Germany + 24,24 ± 0,70 T. afroharzianum P AP14TRI409 2024 Symptomatic corncobs Germany + 24,08 ± 0,76 T. afroharzianum P AP14TRI414 2024 Symptomatic corncobs Germany + 23,84 ± 1,10 T. afroharzianum P AP14TRI426 2024 Soil maize field Germany + 24,19 ± 0,85 T. afroharzianum P AP14TRI430 2024 Soil maize field Germany + 22,67 ± 0,08 T. afroharzianum P AP14TRI450 2024 Corncobs Germany + 23,34 ± 0,16 T. afroharzianum P AP14TRI458 2024 Corncobs Germany + 23,58 ±1,34 T. afroharzianum P AP14TRI460 2024 Corncobs Germany + 24,48 ±1,23 T. afroharzianum P AP14TRI461 2024 Corncobs Germany + 21,70 ± 0,54 T. afroharzianum P AP14TRI462 2024 Corncobs Germany + 20,45 ± 0,78 T. arundinaceum NP TR1 2016 Tomato root Serbia - >40 T. asperelloides NP HOHTR22 - University Hohenheim - >40 T. asperellum P ٭ TR4 Apricot, fruit Serbia - >40 T. asperellum P XILONT34 Biostimulant Kwizda Agro GmbH - >40 T. asperellum P ABITEP02 ABiTEP GmbH Berlin Biostimulant - >40 T. asperellum P ٭ AP22TRI100 Soil maize field Germany - >40 T. atrobrunneum NP AP22TRI118 2022 Soil maize field Germany - >40 T. atrobrunneum NP T54 A. bisporus fruitbodies Serbia - >40 T. atroviride NP IPP0316 1976 Infant food - - >40 T. atroviride NP T33 A. bisporus fruitbodies Serbia - >40 T. atroviride P ٭ T60 2008 A. bisporus fruitbodies Serbia - >40 T. atroviride NP TR10 2019 Tomato, rhizosphere Serbia - >40 T. atroviride P ٭ VINTEC_SCI 2016 Vintec (crop protection) Biostimulant - >40 T. atroviride NP AP22TRI103 2022 Soil maize field Germany - >40 T. azevedoi NP IPP0320 1992 Soil Thailand - >40 T. azevedoi NP AP19TRI6 2019 Corncobs KWS - >40 T. azevedoi NP AP19TRI7 2019 Corncobs Germany - >40 T. brevicompactum NP AP22TRI129 2022 Soil maize field Germany - >40 T. cerinum NP AP22TRI112 2022 Soil maize field Germany - >40 T. gamsii P ٭ AP23TRI278 2023 Maize stalks Germany - >40 T. gamsii NP AP23TRI283 2023 Corncobs Germany - >40 T. gamsii P ٭ AP23TRI285 2023 Corncobs Germany - >40 T. gamsii P ٭ AP23TRI286 2023 Corncobs Germany - >40 T. gamsii NP AP24TRI434 2024 Soil maize field Germany + 21,16±0,33 T. hamatum NP AP22TRI127 2022 Soil maize field Germany - >40 T. harzianum AP19TRI12 2019 Corncobs Germany - >40 T. harzianum NP AP19TRI14 2019 Maize stalks Germany - >40 T. koningii NP IPP1657 Spore suspension Trichodex - >40 T. koningii NP T39 A. bisporus fruitbodies Serbia - >40 T. koningiopsis NP AP22TRI98 2022 Soil Germany - >40 T. paratroviride P ٭ VINTEC_SC1 Biostimulant Vintec - >40 T. paraviridescens NP HOHUHBot Collection Uni. Hohenheim - - >40 T. peberdyi NP AP22TRI96 2022 Soil maize field Germany - >40 T. peberdyi NP AP22TRI101 2022 Soil maize field Germany - >40 T. simmonsii NP BIOHEALTH_T50 Biostimulant Biohealth GmbH - >40 T. velutinum NP AP22TRI95 2022 Soil maize field Germany - >40 T. velutinum NP AP22TRI108 2022 Soil maize field Germany - >40 T. virens NP AP22TRI141 2022 Soil maize field Germany - >40 Fusarium proliferatum IPP1663 Spore suspension Collection Karlovsky, - >40 F. proliferatum 90 Symptomatic corncobs Germany - >40 F. graminearum 69 Symptomatic corncobs Germany - >40 F. graminearum F. G Symptomatic corncobs Germany - >40 F. graminearum IFA66 corncobs Germany - >40 F. temperatum 22.4. corncobs Germany - >40 F. temperatum 81.1 Symptomatic corncobs Germany - >40 F. temperatum F. V corncobs Germany - >40 F. subglutinans 209.4 Symptomatic corncobs Germany - >40 Penicillium paneum AP24TRI412 Symptomatic corncobs Germany - >40 P. brevicompactum AP18TRI4 corncobs Germany - >40 Pseudogymnoascus sp. 75 corncobs Germany - >40 Pseudogymnoascus sp. 80 corncobs Germany - >40 P. pannorum 64 corncobs Germany - >40 P: Pathogenic; P٭: Moderately pathogenic; NP: Non-pathogenic on maize as tested in the greenhouse Table 2. Details on the primers and probes designed and used in this study for conventional and real-time PCR to target Trichoderma afroharzianum . Primer name Sequence (5‘ to 3‘) Locus Product size (bp) TrafTef1c-F TTCAGCGACGCTAACCACTT TEF1α 217 TrafTef1c-R TGTTAGCACTGGTCCGCAAT TEF1α TrafRpb2q-F GAGGAGACGGCCATGATCTG RPB2 138 TrafRpb2q-R GTGAGTTGTCGGGTTCGTCT RPB2 TrafRpb2q-P Fam -CGTCTTCAGAAGGCCGGTAT- BHQ1 RPB2 Additional Declarations No competing interests reported. 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Plants, Institute for National and International Plant Health","correspondingAuthor":false,"prefix":"","firstName":"Gritta","middleName":"","lastName":"Schrader","suffix":""},{"id":579121332,"identity":"779d5f33-8875-4d7c-a5b4-348b539909c6","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":""}],"badges":[],"createdAt":"2026-01-02 15:08:31","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8501765/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8501765/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":100971480,"identity":"a03ba781-23ad-4574-ade0-b0ff2a889d5d","added_by":"auto","created_at":"2026-01-23 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10:13:19","extension":"html","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":225359,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-8501765/v1/7c1a41885ec2a5bfd0295d0c.html"},{"id":100971468,"identity":"a93db405-86fa-4bec-8c40-52d2f2cb0894","added_by":"auto","created_at":"2026-01-23 10:13:19","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":320218,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eReal-time assay. \u003c/strong\u003eA. Amplification curve of the RPB2 gene using a 10-fold serial dilution of pDNA for sensitivity determination. The amplification curves of 9 out of 10 replicates for each dilution were hidden to ensure visibility. B-C. Standard curve assessed with a 10-fold serial dilution of the T. afroharzianum (strain CBS 124620) plasmid DNA (pDNA) positive control. Dilution started from 1 ng µL\u003csup\u003e-1\u003c/sup\u003e concentration. B. pDNA in TE buffer; C. pDNA in 1 ng µL\u003csup\u003e-1\u003c/sup\u003e of DNA from healthy maize kernels. The mean Ct values were calculated with ten replicates.\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8501765/v1/4e19e089d22afcc7442e222f.png"},{"id":102298563,"identity":"5e8f37bb-9fed-457c-98c6-0504745eff82","added_by":"auto","created_at":"2026-02-10 10:47:00","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1490564,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8501765/v1/26352149-9de7-4fe6-aefa-2dfeb96ba368.pdf"},{"id":100971469,"identity":"c8e2ce87-fdd6-4a48-89e4-602ba569f1e5","added_by":"auto","created_at":"2026-01-23 10:13:19","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1547924,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementaryinformation.docx","url":"https://assets-eu.researchsquare.com/files/rs-8501765/v1/3a494d1a20a9efc6e090019c.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"PCR-based assays for rapid and sensitive detection of Trichoderma afroharzianum, a fungal pathogen causing new corn ear rot","fulltext":[{"header":"Introduction","content":"\u003cp\u003eMaize (\u003cem\u003eZea mays\u003c/em\u003e) is one of the world\u0026rsquo;s most important crops and the second most widely produced cereal, serving as an essential food source for both humans and livestock\u003csup\u003e1,2\u003c/sup\u003e. Beyond direct consumption, maize is a highly versatile commodity used in the production of numerous derivatives, including food products, animal feed, biofuels, and a broad range of industrial goods\u003csup\u003e3\u003c/sup\u003e. However, maize production is increasingly challenged by abiotic and biotic stressors that threaten yields and, more broadly, global food security\u003csup\u003e4\u003c/sup\u003e. Abiotic factors such as drought and extreme temperatures are largely driven by climate change\u003csup\u003e5\u003c/sup\u003e, biotic stressors comprise living organisms that negatively affect maize crops and can cause substantial yield losses\u003csup\u003e6\u003c/sup\u003e. These include a wide range of pests and pathogens\u0026mdash;bacteria, fungi, viruses, nematodes, insects as well as weeds. Among the most prevalent maize diseases are ear rots, primarily caused by \u003cem\u003eFusarium\u003c/em\u003e species, but also by other fungal pathogens such as \u003cem\u003eAspergillus\u003c/em\u003e, \u003cem\u003ePenicillium\u003c/em\u003e, \u003cem\u003eNigrospora\u003c/em\u003e, and \u003cem\u003eStenocarpella\u003c/em\u003e\u003csup\u003e7,8\u003c/sup\u003e. Previous studies have reported grain yield losses of 10\u0026ndash;30% due to ear rot\u003csup\u003e9,10\u003c/sup\u003e, a figure that is likely to rise with the emergence of new ear rot types.\u003c/p\u003e\n\u003cp\u003eIn 2018, a previously unreported Trichoderma ear rot of maize was identified in southern Germany, and \u003cem\u003eT. afroharzianum\u003c/em\u003e was determined as the causal agent\u003csup\u003e11\u003c/sup\u003e. Initial symptoms consist of white mycelium, which subsequently develops into green to grey-green growth spreading between or over the kernels. A subsequent survey revealed an extensive distribution of the disease throughout Germany\u003csup\u003e12\u003c/sup\u003e. Since then, Trichoderma ear rot has been reported in France, Italy, Austria, and China\u003csup\u003e12,13, 14, 15\u003c/sup\u003e. In all cases, \u003cem\u003eT. afroharzianum\u003c/em\u003e was identified as the causative agent, supporting the view that this species represents a major pathogen associated with ear rot symptoms under field conditions. In addition to \u003cem\u003eT. afroharzianum\u003c/em\u003e, pathogenic strains of \u003cem\u003eT. asperellum\u003c/em\u003e and \u003cem\u003eT. atroviride\u003c/em\u003e have been reported in India to cause ear rot in maize\u003csup\u003e16\u003c/sup\u003e. Furthermore, \u003cem\u003eT. afroharzianum\u003c/em\u003e and \u003cem\u003eT. harzianum\u003c/em\u003e have been associated with post-flowering stalk rot in maize\u003csup\u003e16\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eTaken together, these findings suggest that an increasing number of \u003cem\u003eTrichoderma\u003c/em\u003e species may become associated with ear rot and other maize diseases in the future. At the same time, the emergence of \u003cem\u003eTrichoderma\u003c/em\u003e-associated diseases in maize may have been underestimated due to limited symptom-based differentiation from other ear rot diseases and the insufficient emphasis on diagnostic approaches. This interpretation is supported by the growing number of reports published within a relatively short time frame\u003csup\u003e11,13,14\u003c/sup\u003e. With specific regard to \u003cem\u003eT. afroharzianum\u003c/em\u003e, this species is globally distributed and commonly occurs in soil, dead wood, and living plants as an endophyte. In addition, it is used as an active ingredient in several commercial biological control products. Its pathogenic lifestyle therefore represents a newly recognised trait of this species and underscores the need for accurate identification. This is particularly important because non-pathogenic species or strains are widely applied in biological control, and their use must be demonstrably safe.\u003c/p\u003e\n\u003cp\u003eIn general, early and efficient diagnosis is a critical prerequisite for the effective management of plant diseases. Accordingly, the rapid, accurate, and reliable detection and identification of \u003cem\u003eT.\u003c/em\u003e \u003cem\u003eafroharzianum\u003c/em\u003e is essential. Morphological characterisation alone is insufficient for the reliable diagnosis of \u003cem\u003eTrichoderma\u003c/em\u003e species, as many taxa exhibit overlapping phenotypic traits. Consequently, the combination of multilocus phylogenetic analysis (MLPA) and DNA barcoding has become the most effective approach for species-level identification within the genus \u003cem\u003eTrichoderma\u003c/em\u003e\u003csup\u003e17,18,19\u003c/sup\u003e. Phylogenetically, \u003cem\u003eT. afroharzianum\u003c/em\u003e belongs to the Harzianum clade and can be distinguished from closely related species using MLPA. However, MLPA has inherent limitations, most notably the time required to complete the analysis, which restricts its applicability for routine diagnostic purposes\u003csup\u003e20\u003c/sup\u003e. In addition, the generation of pure cultures for DNA sequencing through single-spore isolation can be particularly time-consuming, especially when multiple \u003cem\u003eTrichoderma\u003c/em\u003e species co-occur on infected maize cobs\u003csup\u003e12\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eGiven the global importance of maize, substantial efforts have been devoted to the development of a wide range of diagnostic technologies, from laboratory-based PCR assays to field-based remote sensing approaches for disease detection\u003csup\u003e21,22,23\u003c/sup\u003e. Direct methods employing non-invasive imaging sensors have been used predominantly for the diagnosis and monitoring of fungal diseases affecting maize leaves\u003csup\u003e24,25,26\u003c/sup\u003e. In contrast, applications addressing maize ear rot have so far been largely limited to the detection, monitoring, and quantification of mycotoxin production\u003csup\u003e27\u003c/sup\u003e. The implementation of molecular tools for species- and strain-specific identification, including immunological approaches, fingerprinting techniques, exogenous markers, and PCR-based methods\u003csup\u003e28,29\u003c/sup\u003e, can facilitate the effective detection and differentiation of Trichoderma ear rot. Among these approaches, species-specific PCR has proven to be a particularly effective tool, providing faster, more specific, and more sensitive detection and identification of pathogenic fungi\u003csup\u003e30,31\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eThe primary objective of this study was to develop a specific and sensitive molecular tool to facilitate the detection and identification of \u003cem\u003eT. afroharzianum\u003c/em\u003e. Primers and hydrolysis probes were designed targeting the TEF1\u0026alpha; and RPB2 genes and evaluated for their ability to specifically amplify \u003cem\u003eT. afroharzianum\u003c/em\u003e from pure cultures as well as from naturally and artificially infected maize kernels. Two assays, a conventional PCR (cPCR) and a TaqMan-based real-time PCR (qPCR), were developed and validated in accordance with the EPPO standard PM 7/98\u003csup\u003e32\u003c/sup\u003e.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003e\u003cstrong\u003eIsolates collection and identification\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe isolates of \u003cem\u003eTrichoderma\u003c/em\u003e spp. and other fungi included in this study were primarily derived from maize samples and soil collected from maize fields. Samples of maize cobs and stalks were obtained between 2018 and 2024 from both symptomatic and asymptomatic plants. \u003cem\u003eTrichoderma\u003c/em\u003e isolates were also recovered from soil originating from fields with and without confirmed Trichoderma ear rot infection. Sampling of soil and maize plant material was conducted throughout Germany in collaboration with farmers willing to contribute to the survey of outbreaks of Trichoderma ear rot disease. Samples stored in plastic bags were sent to the laboratory for analysis. The procedures for isolating fungi from maize and soil samples and assigning them to \u003cem\u003eTrichoderma\u003c/em\u003e followed the methods described by\u003csup\u003e12\u003c/sup\u003e. Additional \u003cem\u003eTrichoderma\u003c/em\u003e strains were obtained from fungal culture collections, research institutions, and commercial biocontrol products containing \u003cem\u003eTrichoderma\u003c/em\u003e as active organisms. Isolation of \u003cem\u003eTrichoderma\u003c/em\u003e spp. from biocontrol products was carried out in accordance with the methodology outlined in\u003csup\u003e12\u003c/sup\u003e. For each isolate, the ITS, TEF\u0026alpha;, and RPB2 regions were sequenced, and molecular identification was conducted following the protocol proposed by\u003csup\u003e19\u003c/sup\u003e. All isolates had previously undergone pathogenicity testing on maize under controlled greenhouse conditions. Details pertaining to taxonomic identification and pathogenicity are available in\u003csup\u003e12\u003c/sup\u003e. A complete list of fungal strains and their origins used in this study is provided in Table 1.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDesign of primers and hydrolysis probes\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSequences generated from the TEF1\u0026alpha; and RPB2 genes for \u003cem\u003eTrichoderma\u003c/em\u003e species in the Brevicompactum, Harzianum, Virens and Viride clades including referenced sequences retrieved from the NCBI GenBank database were composed. Alignment was performed in MAFFT v. 7 online version\u003csup\u003e33\u003c/sup\u003e using the iterative refinement option G-INS-i and manual optimization with MEGA v. 7. The alignments were analysed in Geneious Prime to identify regions with poly-morphisms. The design of a set of primers and hydrolysis probes was conducted for each gene alignment using the online version of Primer 3 software (http://bioinfo.ut.ee/primer3-0.4.0/). The specificity of the designed primers, as well as the absence of primer-dimer formation, was subjected to further testing using Primer-Blast (http://www.ncbi.nlm.nih.gov/tools/primer-blast). The primers and probes were custom-synthesized by Metabion (Planegg, Germany) with the probes labelled using FAM (6-carboxyfluorescein) and BHQ-1 (Black Hole Quencher\u0026reg;-1).\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003ePCR conditions and optimization\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eThe cPCR was performed using 25 \u0026mu;L PCR reactions containing 1\u0026micro;M (0.5 \u0026mu;L) of each primer TrafTef1-F and TrafTef1-R, 2x GoTaq Green Master Mix (Promega, USA) (12.5 \u0026mu;L), 2.5 \u0026mu;L of gDNA. Molecular grade water was added to the reaction up to 25 \u0026mu;L. PCR reactions were per-formed in a gradient block Biometra thermal cycler (AnalytiK Jena, Germany) using following cycling conditions: initial denaturation at 94\u0026deg;C for 3 min, followed by 35 cycles of 45 s at 94\u0026deg;C, 45 s annealing with a gradient temperature run from 56\u0026deg;C to 64\u0026deg;C and 1 min at 72\u0026deg;C and a final extension of 7 minutes at 72\u0026deg;C. No template control (NTC) consisting of ddH2O was included to check for the absence of contamination. PCR products (3 \u0026mu;L) were checked on 1% agarose gel electrophoresis stained with ROTI GelStain (Roth, Germany). A further check on the conformity of the amplicons with regard to sequence length and species identity was achieved by means of sequencing. Thereby, amplicons derived from \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates CBS 124620, AP22TRI85 and MRI349 were sequenced as described in\u003csup\u003e12\u003c/sup\u003e, followed by BLAST search in the NCBI GenBank. The newly generated sequences matched those with accession numbers PQ558230 (1,248 bp), PQ558225 (1,258 bp) and PQ558235 (1,250 bp), respectively, previously generated from the above isolates\u003csup\u003e12\u003c/sup\u003e. As just a small part of known sequences, their addition to GenBank was not considered.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eThe qPCR was carried out in 12.5 \u0026mu;L final volume containing 0.25 \u0026micro;M (0.125 \u0026mu;L) of forward primer TrafRpb2-F and 0.125 \u0026micro;M (0.125 \u0026mu;L) of reverse primer TrafRpb2-R, 0.15 \u0026micro;M (0.19 \u0026mu;L) of probe TrafRpb2-P, 1x iTaq Universal Probes Supermix (6.25 \u0026mu;L) (Bio-Rad Laboratories, Pty, Ltd.) and 1 \u0026mu;L DNA template. Molecular grade water (4.81 \u0026mu;L) was added to obtain the final volume. The cycling conditions were an initial denaturation at 95\u0026deg;C for 2 min, and 40 cycles of denaturation at 95\u0026deg;C for 15 s following by annealing with a gradient temperature run from 60\u0026deg;C to 66\u0026deg;C for 1 min. Reactions were performed on the StepOnePlus Real-Time PCR System (Applied Biosystems). No template controls (NTC) consisted of ddH2O. The Ct values and standard deviations of all samples were recorded using the StepOnePlus\u0026trade; Software v2.3.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eValidation of the PCR methods\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe performance of the PCR assays to amplify \u003cem\u003eT. afroharzianum\u003c/em\u003e was evaluated by validating different parameters. All DNA extracts were standardized to a concentration of 1 ng \u0026micro;L-1. The analytical specificity was assessed using isolate composition including as non-target 37 of other \u003cem\u003eTrichoderma\u003c/em\u003e species from maize plants, soil from maize fields and other sources and 14 of non-\u003cem\u003eTrichoderma\u003c/em\u003e species from maize plants (Table 1). The inclusivity was assessed with a panel of 37 \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates from different sources (predominantly from maize cobs and soil from maize fields, mycoparasite, biostimulant products) and geographical origin (Germany, France, Italy, Peru). All assays were conducted with three replicates of each isolate. The pDNA of \u003cem\u003eT. afroharzianum\u003c/em\u003e strain CBS 124620 served as positive control and the no template control consisting of ddH2O was included.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe analytical sensitivity was evaluated through a ten-fold serial dilution of pDNA in TE buffer and of pDNA pooled with maize kernel DNA at 1:1 ratio from 1 ng \u0026micro;L\u003csup\u003e-1\u003c/sup\u003e concentration with ten replicates being conducted for each template. The standard curve was generated using the obtained cycle threshold (Ct) values and the efficiency of the assay calculated using the equation E = -1 +10\u003csup\u003e(-1/slope) 34\u003c/sup\u003e. The limit of detection (LOD) was defined as the lowest concentration that could still be amplified in all replicates. The sensitivity of the assays was further evaluated by testing on asymptomatic artificially infected maize cobs obtained from a greenhouse pathogenicity experiment (as described in\u003csup\u003e12\u003c/sup\u003e). The collection of these asymptomatic infected cobs was carried out prior to the manifestation of first symptoms.\u003c/p\u003e\n\u003cp\u003eFurther criteria were evaluated using templates consisting of the pDNA of \u003cem\u003eT. afroharzianum\u003c/em\u003e (strain CBS 124620) set at 10\u0026times;LOD and 100\u0026times;LOD, 0.1 ng \u0026micro;L-1 of \u003cem\u003eT. afroharzianum\u003c/em\u003e (strain CBS124620) and \u003cem\u003eT. afroharzianum\u003c/em\u003e (from sample 159 as natural infected maize kernels), 1 ng \u0026micro;L-1 of \u003cem\u003eT. asperellum\u003c/em\u003e (AP22TRI100) and \u003cem\u003eT. brevicompactum\u003c/em\u003e (AP22TRI140). Repeatability and reproducibility were assessed by performing each assay (cPCR and qPCR) with these templates in ten and three replicates, respectively. Reproducibility was also assessed by two different operators repeating the assays on two different qPCR platforms: the StepOnePlus Real-Time PCR System (Applied Biosystems) and qTower3 thermocycler (Analytik Jena, Germany). The robustness of the qPCR assays was assessed in ten replicates using reaction volumes of \u0026plusmn;10% (11,25 \u0026micro;L and 13,75 \u0026micro;L) at \u0026plusmn;2\u0026deg;C modified hybridisation temperatures (60\u0026deg;C, 62\u0026deg;C and 64\u0026deg;C). For the transferability, the qPCR was performed in ten replicates on two qPCR platforms above, and with four different master mixes: iTaq Universal Probes Supermix (Bio-Rad), Maxima Probe/ROX qPCR Master Mix (Fischer scientific), Applied Biosystems\u0026trade; TaqMan\u0026trade; Universaler PCR-Mastermix (Fischer scientific) and TaKaRa Premix Ex TaqTM (Probe qPCR); TaKaRa, Kusatsu, Japan).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData analysis\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFor the qPCR assay, the StepOnePlus software automatically generated the baseline range, threshold cycle (Ct) values and standard curves. The results were checked to ensure they aligned with the provider\u0026apos;s analysis guidelines. The mean threshold cycle (Ct) values were calculated along with their standard deviations (SD). Precision of the assay was evaluated by computing the coefficient of variation (CV). All statistical calculations were performed using Microsoft Excel.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eSelection of \u003cem\u003eT. afroharzianum\u003c/em\u003e specific primers and PCR conditions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe design of primers and probes for both cPCR and qPCR was initially based on TEF\u0026alpha; and RPB2 sequences. The positions of the selected primers and probe are illustrated in Figure S1 and the sequences are presented in Table 2. The few qPCR primer combinations obtained on TEF1\u0026alpha; predominantly showed cross-reactivity with many non-target \u003cem\u003eTrichoderma\u003c/em\u003e species (data not shown), despite optimization efforts. By contrast, several qPCR primer combinations were identified based on the RPB2 region. Initial screening using 3 annealing temperatures (60, 62, 64\u0026deg;C) suggested selecting the TrafRpb2q-F/ TrafRpb2q-R pair and the corresponding TrafRpb2q-P probe, which successfully amplified all target templates at an annealing temperature of 62\u0026deg;C with no cross-reactivity. The PCR products obtained at annealing temperature of 62\u0026deg;C show a single band of the expected size of 138 bp in agarose electrophoresis.\u003c/p\u003e\n\u003cp\u003eFor the specificity, the selected cPCR primers yielded 100% analytical specificity leading to the amplification of a single 217 bp fragment, for all and only the \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates. In order to ascertain and extend the inclusivity of the cPCR primers, a series of tests were carried out on a panel of samples consisting of kernels from naturally infected and greenhouse-inoculated maize plants. These tests yielded satisfactory results. Amplicon sequencing further confirmed the suitability of the primers to amplify the TEF1\u0026alpha; region of \u003cem\u003eT. afroharzianum\u003c/em\u003e. Comparatively, the qPCR assay demonstrated 100% inclusivity by amplifying gDNA from all the \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates (mean Ct = 19.25-25.38, SD = 0.2-1.47; n=10) (Table 1), regardless of the host and geographical origin. In general, the qPCR assay did not amplify DNA from non-\u003cem\u003eTrichoderma\u003c/em\u003e species, with the exception of the \u003cem\u003eT. gamsii\u003c/em\u003e isolate AP24TRI434. Despite implementing all possible options to optimize the primer specificity (e.g. varying the annealing temperature), this issue persistently occurred.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eReliability of the PCR assays to detect \u003cem\u003eT. afroharzianum\u003c/em\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAnalytical sensitivity differed between the species-specific cPCR and qPCR assays. In a tenfold serial dilution of plasmid DNA ranging from 1 ng to 10\u003csup\u003e-8\u003c/sup\u003e ng \u0026micro;L⁻\u0026sup1;, the cPCR consistently produced amplicons of the expected size (217 bp) in 100% of replicates (n = 10) down to a concentration of 1 pg \u0026micro;L⁻\u0026sup1; (= 10\u003csup\u003e-3\u003c/sup\u003e ng \u0026micro;L⁻\u0026sup1;) using DNA from pure cultures. In contrast, the qPCR consistently amplified target DNA in all replicates (n = 10) down to a concentration of 1 fg \u0026micro;L⁻\u0026sup1; (= 10\u003csup\u003e-6\u003c/sup\u003e ng \u0026micro;L⁻\u0026sup1;) (Fig. 1A). Linear calibration curves generated from mean Ct values plotted against the logarithm of template concentration are shown in Fig. 1B\u0026ndash;C. Fully repeatable positive results (100%) were obtained down to the determined limit of detection (LOD) of 1 fg \u0026micro;L⁻\u0026sup1; using plasmid DNA (pDNA) diluted in TE buffer (mean Ct = 30.4) and pDNA diluted in 1 ng \u0026micro;L⁻\u0026sup1; genomic DNA extracted from healthy maize kernels (mean Ct = 30.8). For pDNA diluted in TE buffer, the standard curve showed a correlation coefficient (R\u0026sup2;) of 0.98, a slope of \u0026minus;3.20, and an amplification efficiency of 105%. When pDNA was diluted in genomic DNA from healthy maize kernels, the corresponding values were R\u0026sup2; = 0.996, a slope of \u0026minus;2.99, and an efficiency of 118%. Both cPCR and qPCR assays successfully detected \u003cem\u003eT. afroharzianum\u003c/em\u003e in all samples derived from maize kernels collected from asymptomatic infected cobs. The qPCR assay yielded Ct values ranging from 27 to 29, indicating its suitability for detecting \u003cem\u003eT. afroharzianum\u003c/em\u003e in asymptomatic samples at concentrations close to the determined LOD.\u003c/p\u003e\n\u003cp\u003eBoth assays demonstrated satisfactory repeatability, reproducibility, robustness, and transferability when evaluated using a panel comprising plasmid target DNA and genomic DNA from pure cultures as well as naturally infected samples. Both the cPCR and qPCR assays amplified exclusively templates containing target DNA, whereas no amplification was observed for non-target DNA in any replicate, resulting in 100% repeatable and reproducible outcomes (Supplementary Table S1). The coefficients of variation (CVs) ranged from 0.44% to 1.05% for repeatability and from 0.48% to 0.72% for reproducibility. The qPCR assay exhibited high robustness with respect to variations in experimental parameters. Alterations in reaction volume (12.5 \u0026micro;L) or hybridisation temperature (60 \u0026deg;C) had little effect on assay performance. However, a slight decrease in Ct values was observed at an annealing temperature of 64 \u0026deg;C (Supplementary Table S2). Furthermore, analysis of the template panel by two independent operators yielded the expected results for both target and non-target templates. These results were consistent across operators and showed very low variation in Ct values, confirming the repeatability and reproducibility of the assay (data not shown).\u003c/p\u003e\n\u003cp\u003eTo evaluate the transferability of the qPCR assay, additional analyses were performed using three different commercial master mixes and an alternative PCR platform. Changes in the master mix or PCR platform did not affect assay specificity, and repeatability and reproducibility were maintained across all tested conditions (Supplementary Table S3). The observed differences were primarily related to assay sensitivity. When using the StepOnePlus system, the TaKaRa Premix Ex Taq Probe qPCR produced Ct values comparable to those obtained with the iTaq Universal Probes Supermix (Bio-Rad). The Maxima Probe/ROX qPCR Master Mix increased assay sensitivity, resulting in Ct values that were 0.67\u0026ndash;2.04 cycles lower than those obtained with the iTaq Universal Probes Supermix. In contrast, use of the iTaq Universal Probes Supermix on the qTower3 thermocycler (Analytik, Jena) resulted in reduced robustness, with coefficients of variation ranging from 2.90% to 3.32%, and Ct values that were 1.25\u0026ndash;3.24 cycles higher than those obtained using the StepOnePlus system.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eTrichoderma-associated diseases in maize production are an increasing concern. The threat to this key cereal crop is likely to intensify due to the widespread occurrence and persistence of \u003cem\u003eTrichoderma\u003c/em\u003e species in agricultural soils worldwide\u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e,\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e,\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e,\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u003c/sup\u003e. Moreover, \u003cem\u003eTrichoderma\u003c/em\u003e species are widely used as biological control agents, and the pathogenic potential of certain species or strains therefore represents a significant challenge for the safe application, regulation, and acceptance of \u003cem\u003eTrichoderma\u003c/em\u003e-based biocontrol products. Accordingly, the development of molecular tools for the accurate identification of pathogenic \u003cem\u003eTrichoderma\u003c/em\u003e species is essential for improving the understanding and management of these diseases. Here, we provide a PCR-based tool for the rapid and sensitive detection of \u003cem\u003eT. afroharzianum\u003c/em\u003e. Current knowledge indicates that multiple \u003cem\u003eTrichoderma\u003c/em\u003e species are associated with diseases in maize\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e,\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e,\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e, therefore, the implementation of multiplex PCR assays enabling the simultaneous detection of multiple pathogens represents a rational and efficient diagnostic approach\u003csup\u003e\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/sup\u003e. However, epidemiological data from Europe indicate that \u003cem\u003eT. afroharzianum\u003c/em\u003e has consistently been identified as the predominant etiological agent of Trichoderma ear rot\u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e,\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. Under these circumstances, species-specific detection of this pathogen using simplex PCR remains justified.\u003c/p\u003e \u003cp\u003eThe most significant achievement of this study was the development of a diagnostic tool based on complementary cPCR and qPCR assays. Despite the longer processing time associated with cPCR, it remains a cost-effective option compared with the more expensive qPCR approach\u003csup\u003e\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e, which in turn provides the advantage of higher sensitivity\u003csup\u003e\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u003c/sup\u003e. The selection of target genes for primer design was guided by established molecular identification practices for \u003cem\u003eTrichoderma\u003c/em\u003e species, which rely primarily on the ITS, TEF1α, and RPB2 regions for phylogenetic analysis and DNA barcoding\u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e. For PCR-based detection, ITS markers are generally effective at the genus level\u003csup\u003e\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e,\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e,\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e\u003c/sup\u003e, whereas TEF1α and RPB2 are more suitable for species-specific discrimination\u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e,\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e,\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e,\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e,\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e. Both TEF1α and RPB2 have previously been demonstrated to reliably target \u003cem\u003eT. afroharzianum\u003c/em\u003e \u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e, \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e. In the present study, TEF1α was identified as the most suitable gene for cPCR, whereas RPB2 was found to be more appropriate for the qPCR assay.\u003c/p\u003e \u003cp\u003eA previous study developed a qPCR assay for the detection and quantification of \u003cem\u003eT. afroharzianum\u003c/em\u003e causing maize ear rot in Italy\u003csup\u003e\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e. That assay targeted the TEF1α gene, based on the assumption that conserved single-nucleotide polymorphisms within this region allow discrimination of \u003cem\u003eT. afroharzianum\u003c/em\u003e from other species within the Harzianum clade. However, the qPCR primers designed in the same TEF1α region as in study\u003csup\u003e\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e exhibited unexpected cross-reactivity with certain \u003cem\u003eT. asperellum\u003c/em\u003e and \u003cem\u003eT. atrobrunneum\u003c/em\u003e isolates. Full specificity for \u003cem\u003eT. afroharzianum\u003c/em\u003e was only achieved when primers targeting the RPB2 gene were employed. This single-copy and highly conserved RPB2 gene plays a central role in resolving the phylogeny of \u003cem\u003eTrichoderma\u003c/em\u003e species\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e,\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e and has increasingly been recognised as a suitable and reliable marker for PCR-based detection of \u003cem\u003eTrichoderma\u003c/em\u003e species (Prabhakaran et al. 2014; Gerin et al. 2018). Regarding real-time PCR chemistry, SYBR Green used by\u003csup\u003e46\u003c/sup\u003e has been widely applied for the detection and quantification of \u003cem\u003eTrichoderma\u003c/em\u003e species, particularly in studies addressing their biocontrol potential\u003csup\u003e\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e,\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e\u003c/sup\u003e. The TaqMan PCR system is likewise well established for diagnostic and quantitative applications\u003csup\u003e\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e,\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e,\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e\u003c/sup\u003e and was selected in the present study for two main reasons: first, to provide a user-friendly diagnostic tool, and second, because of its superior accuracy and specificity\u003csup\u003e\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eA rigorous validation process demonstrated that the proposed assays largely fulfil the requirements for new diagnostic methods as defined by\u003csup\u003e32\u003c/sup\u003e. Both assays exhibited high specificity against all tested \u003cem\u003eTrichoderma\u003c/em\u003e species and other fungi associated with maize cobs and a range of additional host plants. Although cross-reactivity of the qPCR assay was observed with the \u003cem\u003eT. gamsii\u003c/em\u003e isolate AP14TRI434, this isolate originated from soil and was shown to be non-pathogenic to maize. Consequently, it can be inferred that neither the cPCR nor the qPCR assay amplifies \u003cem\u003eTrichoderma\u003c/em\u003e isolates capable of causing field or greenhouse symptoms comparable to those induced by \u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e,\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e. Both assays achieved 100% inclusivity for all \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates tested, encompassing a broad range of geographic origins, host associations, and pathogenicity profiles\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. It should be noted, however, that not all \u003cem\u003eT. afroharzianum\u003c/em\u003e strains are pathogenic to maize, and the determinants underlying pathogenic versus non-pathogenic lifestyle, as well as reliable molecular markers to discriminate between these isolates, remain unknown. Importantly, neither the phylogenetic analyses\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e nor the PCR assays developed in the present study are able to distinguish between pathogenic and non-pathogenic \u003cem\u003eT. afroharzianum\u003c/em\u003e isolates. This limitation has direct implications for the interpretation of diagnostic results. In particular, pathogenicity testing remains necessary to substantiate a positive diagnostic result, especially when latent infections are tested, in order to confirm that the detected \u003cem\u003eT. afroharzianum\u003c/em\u003e isolate is capable of causing disease, in accordance with Koch\u0026rsquo;s postulates. Conversely, maize samples exhibiting characteristic Trichoderma ear rot symptoms but yielding negative results in these assays should be further evaluated for the presence of other \u003cem\u003eTrichoderma\u003c/em\u003e species that may also be pathogenic to maize.\u003c/p\u003e \u003cp\u003eThe high sensitivity of the qPCR assay is consistent with previous assays targeting \u003cem\u003eTrichoderma\u003c/em\u003e species\u003csup\u003e\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e,\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e\u003c/sup\u003e. The LOD of 1 fg \u0026micro;L⁻\u0026sup1; determined for the TaqMan assay represents a substantial improvement over the LOD of 50 fg \u0026micro;L⁻\u0026sup1; reported for a SYBR Green based assay\u003csup\u003e\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e. This pronounced difference in sensitivity is unlikely to be attributable solely to the detection chemistry and is more likely related to differences in sample preparation or reaction composition\u003csup\u003e\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e\u003c/sup\u003e. The presence of host plant material had no detectable effect on analytical sensitivity, as indicated by comparable results obtained from pure cultures, naturally infected samples, and artificially inoculated material. Although minor inhibitory effects caused by co-extracted DNA cannot be entirely excluded\u003csup\u003e\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e,\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e\u003c/sup\u003e, such effects are unlikely to compromise assay performance when testing maize kernels. Both assays successfully detected \u003cem\u003eT. afroharzianum\u003c/em\u003e in asymptomatic material at early infection stages (7\u0026ndash;10 days post inoculation), underscoring their suitability for early diagnosis. As expected, both assays demonstrated high repeatability, reproducibility, and robustness. While the cPCR assay was readily transferable across platforms, the qPCR assay required careful verification prior to diagnostic implementation, particularly when using platforms other than the StepOnePlus system. For example, testing on the qTower qPCR device resulted in delayed amplification signals, which could lead to false-negative results in samples containing low levels of target DNA, a limitation that is especially relevant for latent infections. However, the risk of false negatives in symptomatic samples is considered low, as maize tissues affected by Trichoderma ear rot typically exhibit extensive fungal colonization\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e,\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e,\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e, providing sufficiently high concentrations of target DNA.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe aim of this study was to facilitate the diagnosis of the newly emerging Trichoderma ear rot by providing a rapid and efficient molecular tool. The two PCR assays developed and validated here are well suited to achieve this objective. A key advantage of this approach is the use of two complementary PCR systems endpoint and TaqMan real-time PCR targeting different genetic loci. This diagnostic framework enables the rapid detection of \u003cem\u003eT. afroharzianum\u003c/em\u003e directly from infected maize tissue, thereby eliminating the need for culture-based isolation. Furthermore, the high sensitivity of the qPCR assay allows the detection of latent infections, which is essential for early diagnosis. The ability to identify \u003cem\u003eT. afroharzianum\u003c/em\u003e ear rot at early stages is critical for the timely implementation of management strategies aimed at minimizing yield losses and preventing further spread of the disease.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eCompeting interests\u003c/h2\u003e \u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eDeclaration\u003c/p\u003e \u003cp\u003eThis study was supported by the Federal Ministry of Food and Agriculture \u0026ndash; BMEL (actually Federal Ministry of Agriculture, Food and Regional Identity \u0026ndash; BMLEH), grant number FKZ2221NR014, subproject 2221NR014B. The funder had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eCDM, AP, GS, BCS, and AvT designed the project and secured the funding. CDM designed and conducted the experiments, and analysed the data. CDM wrote the manuscript. AP, GS, BCS, and AvT reviewed it. All authors read and approved the final version of the manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eWe would like to express our gratitude to the companies, universities and research institutions that provided us with Trichoderma isolates. Additionally, we thank Julia Krebs and Tobias Wille for their technical support during the method development process.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eAll the data is available in the main manuscript\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eRouf Shah, T., Prasad, K. \u0026amp; Kumar, P. Maize- A potential source of human nutrition and health: A review. \u003cem\u003eCogent. Food Agric.\u003c/em\u003e\u003cstrong\u003e2\u003c/strong\u003e, 1\u0026ndash;9. doi: 10.1080/23311932.2016.1166995 (2016).\u003c/li\u003e\n\u003cli\u003eChen, C., Chaudhary, A. \u0026amp; Mathys A. Nutrient adequacy of global food production. \u003cem\u003eFront. Nutr.\u003c/em\u003e\u003cstrong\u003e8\u003c/strong\u003e, 739755. doi: 10.3389/fnut.2021.739755 (2021)\u003c/li\u003e\n\u003cli\u003eL\u0026uuml;tke, E. N., Schwarz, F. 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Technol.\u003c/em\u003e\u003cstrong\u003e102\u003c/strong\u003e, 2793\u0026ndash;2798. doi: 10.1016/j.biortech.2010.09.120 (2011).\u003c/li\u003e\n\u003cli\u003eL\u0026oacute;pez-Mond\u0026eacute;jar, R., Ant\u0026oacute;n, A., Raidl, S., Ros, M. \u0026amp; Pascual, J. A. Quantification of the biocontrol agent \u003cem\u003eTrichoderma harzianum\u003c/em\u003e with real-time TaqMan PCR and its potential extrapolation to the hyphal biomass. \u003cem\u003eBioresour. Technol.\u003c/em\u003e\u003cstrong\u003e101\u003c/strong\u003e, 2888\u0026ndash;2891. doi: 10.1016/j.biortech.2009.10.019 (2010).\u003c/li\u003e\n\u003cli\u003eAlvarez, M.L. \u0026amp; Don\u0026eacute;, S.C. SYBR\u0026reg; Green and TaqMan\u0026reg; quantitative PCR arrays: expression profile of genes relevant to a pathway or a disease state. Methods Mol. Biol. 1182,321-359. doi: 10.1007/978-1-4939-1062-5_27. PMID: 25055922 (2014).\u003c/li\u003e\n\u003cli\u003eDodd, L. S., Hill, R. A. \u0026amp; Stewart, A. A duplex-PCR bioassay to detect a \u003cem\u003eTrichoderma virens\u003c/em\u003e biocontrol isolate in non-sterile soil. \u003cem\u003eSoil Biol. Biochem. \u003c/em\u003e\u003cstrong\u003e36\u003c/strong\u003e, 1955\u0026ndash;1965. doi: 10.1016/j.soilbio.2004.03.012 (2004).\u003c/li\u003e\n\u003cli\u003eMao, F., Leung, W. Y. \u0026amp; Xin, X. Characterization of EvaGreen and the implication of its physicochemical proper-ties for qPCR applications. \u003cem\u003eBMC Biotechnol.\u003c/em\u003e\u003cstrong\u003e7\u003c/strong\u003e, 76. doi: 10.1186/1472-6750-7-76 (2007).\u003c/li\u003e\n\u003cli\u003eCordier, C., Edel-Hermann, V., Martin-Laurent, F., Blal, B., Steinberg, C. \u0026amp; Alabouvette C. SCAR-based real time PCR to identify a biocontrol strain (T1) of \u003cem\u003eTrichoderma atroviride\u003c/em\u003e and study its population dynamics in soils. \u003cem\u003eJ. Microbiol. Methods\u003c/em\u003e\u003cstrong\u003e68\u003c/strong\u003e, 60\u0026ndash;68. doi: 10.1016/j.mimet.2006.06.006 (2007).\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTable 1 Fungal isolates used in this study and results of the conventional and real-time PCR assays carried out on their DNA extracts. For each isolate following details are provided: taxonomy, name, year of collection, source or host, location and the results of testing with endpoint and real-time PCR. The pathogenicity of all \u003cem\u003eTrichoderma\u003c/em\u003e isolates on maize was tested in previous work\u003csup\u003e12\u003c/sup\u003e in a greenhouse.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" align=\"\" width=\"624\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpecies\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eIsolate\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eYear\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSource/Host\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLocation\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAssay (target)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ecPCR\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eqPCR\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP18TRI1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2018\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003eFrance\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e21,70 \u0026plusmn; 0,54\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP18TRI2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2018\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e20,45\u0026nbsp;\u0026plusmn; 0,78\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP18TRI3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2018\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e20,26\u0026nbsp;\u0026plusmn;\u0026nbsp;0,24\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP19TRI5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2019\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e22,6\u0026nbsp;\u0026plusmn; 0,57\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eCBS 124620\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cem\u003eTheobroma cacao\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003ePeru\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e21,58\u0026nbsp;\u0026plusmn; 0,59\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP20TRI15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2020\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e19,25\u0026nbsp;\u0026plusmn;\u0026nbsp;0,89\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP20TRI16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2020\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e20,14\u0026nbsp;\u0026plusmn; 0,35\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eKG10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cem\u003ePleurotus ostreatus\u003c/em\u003e substrate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e20,97\u0026nbsp;\u0026plusmn; 0,71\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eKG13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cem\u003eP. ostreatus\u003c/em\u003e substrate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e25,83 \u0026plusmn; 0,31\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eMRI349\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eBiostimulant\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e21,53 \u0026plusmn; 0,28\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eT42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cem\u003eMarchantia polymorpha\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e21,87\u0026nbsp;\u0026plusmn; 0,41\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eT138\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cem\u003eM. polymorpha\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e21,23\u0026nbsp;\u0026plusmn; 0,62\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eDISAFATS-1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eItaly\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e19,93\u0026nbsp;\u0026plusmn; 1,08\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e21,29 \u0026plusmn; 0,57\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e22,87\u0026nbsp;\u0026plusmn; 0,51\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e22,87\u0026nbsp;\u0026plusmn; 0,51\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI108\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e20,82\u0026nbsp;\u0026plusmn; 1,07\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI118\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e19,73\u0026nbsp;\u0026plusmn;\u0026nbsp;0,82\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI121\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e20,66 \u0026plusmn; 0,76\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI129\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e21,11\u0026plusmn;1,47\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT.\u0026nbsp;\u003c/em\u003e\u003cem\u003eafroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI130\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e19,70\u0026nbsp;\u0026plusmn; 0,71\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI131\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e23,27\u0026nbsp;\u0026plusmn; 1,76\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI134\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e24,94 \u0026plusmn; 0,77\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP23TRI136\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e23,36\u0026nbsp;\u0026plusmn; 0,67\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP23TRI221\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2023\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e24,17\u0026nbsp;\u0026plusmn; 1,75\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI225\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2023\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e23,97\u0026nbsp;\u0026plusmn; 1,56\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP24TRI401\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e24,32\u0026nbsp;\u0026plusmn; 1,14\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP24TRI402\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e23,64\u0026nbsp;\u0026plusmn; 1,24\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP24TRI403\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e23,37\u0026nbsp;\u0026plusmn; 1,05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP24TRI404\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e24,24\u0026nbsp;\u0026plusmn; 0,70\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP14TRI409\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e24,08\u0026nbsp;\u0026plusmn; 0,76\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP14TRI414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e23,84\u0026nbsp;\u0026plusmn; 1,10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP14TRI426\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e24,19\u0026nbsp;\u0026plusmn; 0,85\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP14TRI430\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e22,67\u0026nbsp;\u0026plusmn; 0,08\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP14TRI450\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eCorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e23,34\u0026nbsp;\u0026plusmn;\u0026nbsp;0,16\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP14TRI458\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eCorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" align=\"left\" width=\"605\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 100%;\"\u003e\n \u003cp\u003e23,58\u0026nbsp;\u0026plusmn;1,34\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP14TRI460\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eCorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e24,48 \u0026plusmn;1,23\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP14TRI461\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eCorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e21,70 \u0026plusmn; 0,54\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. afroharzianum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP14TRI462\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eCorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e20,45\u0026nbsp;\u0026plusmn; 0,78\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. arundinaceum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eTR1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2016\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eTomato root\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eSerbia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. asperelloides\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eHOHTR22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eUniversity Hohenheim\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. asperellum\u003c/em\u003e\u003csup\u003e\u0026nbsp;P\u003c/sup\u003e\u003csup\u003e\u003cspan dir=\"RTL\"\u003e٭\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eTR4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eApricot, fruit\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eSerbia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. asperellum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eXILONT34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eBiostimulant\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eKwizda Agro GmbH\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. asperellum\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eABITEP02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eABiTEP GmbH Berlin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eBiostimulant\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. asperellum\u003c/em\u003e\u003csup\u003e\u0026nbsp;P\u003c/sup\u003e\u003csup\u003e\u003cspan dir=\"RTL\"\u003e٭\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. atrobrunneum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI118\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. atrobrunneum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eT54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cem\u003eA. bisporus\u003c/em\u003e fruitbodies\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eSerbia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. atroviride\u003csup\u003eNP\u003c/sup\u003e\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eIPP0316\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e1976\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eInfant food\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. atroviride\u003csup\u003eNP\u003c/sup\u003e\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eT33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cem\u003eA. bisporus\u003c/em\u003e fruitbodies\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eSerbia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. atroviride\u003c/em\u003e\u003csup\u003eP\u003c/sup\u003e\u003csup\u003e\u003cspan dir=\"RTL\"\u003e٭\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eT60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2008\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cem\u003eA. bisporus\u003c/em\u003e fruitbodies\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eSerbia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. atroviride\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eTR10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2019\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eTomato, rhizosphere\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eSerbia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. atroviride\u003c/em\u003e\u003csup\u003e\u0026nbsp;P\u003c/sup\u003e\u003csup\u003e\u003cspan dir=\"RTL\"\u003e٭\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eVINTEC_SCI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2016\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eVintec (crop protection)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eBiostimulant\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. atroviride\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI103\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. azevedoi\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eIPP0320\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e1992\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eThailand\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. azevedoi\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP19TRI6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2019\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eCorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eKWS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. azevedoi\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP19TRI7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2019\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eCorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. brevicompactum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI129\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. cerinum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI112\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. gamsii\u003c/em\u003e\u003csup\u003e\u0026nbsp;P\u003c/sup\u003e\u003csup\u003e\u003cspan dir=\"RTL\"\u003e٭\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP23TRI278\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2023\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eMaize stalks\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. gamsii\u003c/em\u003e\u003csup\u003e\u0026nbsp;NP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP23TRI283\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2023\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eCorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. gamsii\u003c/em\u003e\u003csup\u003e\u0026nbsp;P\u003c/sup\u003e\u003csup\u003e\u003cspan dir=\"RTL\"\u003e٭\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP23TRI285\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2023\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eCorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. gamsii\u003c/em\u003e\u003csup\u003e\u0026nbsp;P\u003c/sup\u003e\u003csup\u003e\u003cspan dir=\"RTL\"\u003e٭\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP23TRI286\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2023\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eCorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. gamsii\u003c/em\u003e\u003csup\u003e\u0026nbsp;NP\u003c/sup\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP24TRI434\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e21,16\u0026plusmn;0,33\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. hamatum\u003csup\u003eNP\u003c/sup\u003e\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI127\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. harzianum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP19TRI12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2019\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eCorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. harzianum\u003c/em\u003e\u003cem\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP19TRI14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2019\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eMaize stalks\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. koningii\u003csup\u003eNP\u003c/sup\u003e\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eIPP1657\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSpore suspension\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eTrichodex\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. koningii\u003csup\u003eNP\u003c/sup\u003e\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eT39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e\u003cem\u003eA. bisporus\u003c/em\u003e fruitbodies\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eSerbia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. koningiopsis\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. paratroviride\u003c/em\u003e\u003csup\u003e\u0026nbsp;P\u003c/sup\u003e\u003csup\u003e\u003cspan dir=\"RTL\"\u003e٭\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eVINTEC_SC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eBiostimulant\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eVintec\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. paraviridescens\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eHOHUHBot\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eCollection Uni. Hohenheim\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. peberdyi\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. peberdyi\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI101\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. simmonsii\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eBIOHEALTH_T50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eBiostimulant\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eBiohealth GmbH\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. velutinum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. velutinum\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI108\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eT. virens\u003c/em\u003e\u003csup\u003eNP\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP22TRI141\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSoil maize field\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eFusarium proliferatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eIPP1663\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSpore suspension\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eCollection Karlovsky,\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e\u003cem\u003e-\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eF. proliferatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eF. graminearum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eF. graminearum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eF. G\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eF. graminearum\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eIFA66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003ecorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eF. temperatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e22.4.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003ecorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eF. temperatum\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e81.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eF. temperatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eF. V\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003ecorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eF. subglutinans \u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e209.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003ePenicillium paneum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP24TRI412\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eSymptomatic corncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eP. brevicompactum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003eAP18TRI4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003ecorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003ePseudogymnoascus\u0026nbsp;\u003c/em\u003esp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003ecorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003ePseudogymnoascus\u0026nbsp;\u003c/em\u003esp.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003ecorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003e\u003cem\u003eP. pannorum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 95px;\"\u003e\n \u003cp\u003e64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 38px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003ecorncobs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 113px;\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u0026gt;40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;P: Pathogenic; P٭: Moderately pathogenic; NP: Non-pathogenic on maize as tested in the greenhouse\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 2. Details on the primers and probes designed and used in this study for conventional and real-time PCR to target \u003cem\u003eTrichoderma afroharzianum\u003c/em\u003e.\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" align=\"\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePrimer name\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 283px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSequence (5\u0026lsquo; to 3\u0026lsquo;)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLocus\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 123px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eProduct size (bp)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 104px;\"\u003e\n \u003cp\u003eTrafTef1c-F\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 283px;\"\u003e\n \u003cp\u003eTTCAGCGACGCTAACCACTT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eTEF1\u0026alpha;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" style=\"width: 123px;\"\u003e\n \u003cp\u003e217\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 104px;\"\u003e\n \u003cp\u003eTrafTef1c-R\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 283px;\"\u003e\n \u003cp\u003eTGTTAGCACTGGTCCGCAAT \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eTEF1\u0026alpha;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 104px;\"\u003e\n \u003cp\u003eTrafRpb2q-F\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 283px;\"\u003e\n \u003cp\u003eGAGGAGACGGCCATGATCTG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eRPB2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"3\" style=\"width: 123px;\"\u003e\n \u003cp\u003e138\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 104px;\"\u003e\n \u003cp\u003eTrafRpb2q-R\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 283px;\"\u003e\n \u003cp\u003eGTGAGTTGTCGGGTTCGTCT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eRPB2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 104px;\"\u003e\n \u003cp\u003eTrafRpb2q-P\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 283px;\"\u003e\n \u003cp\u003eFam -CGTCTTCAGAAGGCCGGTAT- BHQ1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003eRPB2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"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":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Pathogenic Trichoderma, Taqman PCR, corn disease, Trichoderma afroharzianum, emerging diseases","lastPublishedDoi":"10.21203/rs.3.rs-8501765/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8501765/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe emergence of a new corn ear rot was recently described in Germany and has since received increased attention following detections in several European and Asian countries. All reports consistently identify \u003cem\u003eTrichoderma afroharzianum\u003c/em\u003e as the causal agent. The potential impact on maize production, combined with the risk of misdiagnosis as other ear rot diseases, highlights the need for a rapid and accurate diagnostic method for the reliable detection of \u003cem\u003eT. afroharzianum\u003c/em\u003e. In this study, we developed PCR-based diagnostic tools including an endpoint PCR and a real-time TaqMan assay targeting the TEF1α and RPB2 loci, respectively. Both assays were validated using a broad panel of \u003cem\u003eTrichoderma\u003c/em\u003e isolates and other fungal species obtained from maize plants, additional host species from diverse geographic origins, and soils collected from German maize fields. All performance criteria required to demonstrate assay reliability were met. The assays showed high accuracy and sensitivity, with limits of detection of 1 pg for the endpoint PCR and 1 fg per reaction for the real-time PCR. The real-time assay exhibited amplification efficiencies of 105% (R\u0026sup2; = 0.98) for pure cultures and 118% (R\u0026sup2; = 0.998) for infected maize kernels. Furthermore, both assays reliably detected \u003cem\u003eT. afroharzianum\u003c/em\u003e in asymptomatic maize cobs following artificial inoculation with pathogenic strains. These methods therefore enable early and reliable detection of \u003cem\u003eT. afroharzianum\u003c/em\u003e ear rot and allow its discrimination from other maize ear rot diseases. This capability is essential for the development of targeted management strategies to address this emerging threat to maize production.\u003c/p\u003e","manuscriptTitle":"PCR-based assays for rapid and sensitive detection of Trichoderma afroharzianum, a fungal pathogen causing new corn ear rot","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-23 10:11:52","doi":"10.21203/rs.3.rs-8501765/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-02-16T06:13:12+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-10T06:21:34+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"413173169938901304677884883777977582","date":"2026-02-01T07:34:56+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"210660053489765102268040550153869084988","date":"2026-02-01T04:45:37+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-29T10:51:03+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"214466632624584478016027312087178885613","date":"2026-01-27T02:16:37+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"293910547250010010673069745958120252613","date":"2026-01-22T06:39:04+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-01-22T06:34:32+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-01-21T00:50:46+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2026-01-09T10:18:40+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-01-07T15:36:56+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2026-01-07T15:30:11+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"48fc9c5f-fbc7-4c78-ae7e-36696cf91ecf","owner":[],"postedDate":"January 23rd, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":61619100,"name":"Biological sciences/Biological techniques"},{"id":61619101,"name":"Biological sciences/Biotechnology"},{"id":61619102,"name":"Biological sciences/Microbiology"},{"id":61619103,"name":"Biological sciences/Molecular biology"},{"id":61619104,"name":"Biological sciences/Plant sciences"}],"tags":[],"updatedAt":"2026-04-27T04:53:28+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-23 10:11:52","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8501765","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8501765","identity":"rs-8501765","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}