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Plant Pathology

Wiley

Preprints posted in the last 30 days, ranked by how well they match Plant Pathology's content profile, based on 16 papers previously published here. The average preprint has a 0.01% match score for this journal, so anything above that is already an above-average fit.

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Danish profile of soft rot Pectobacteriaceae; A three-year field sampling study proving several clonal clades of soft rot isolates across diverse locations implicating a common origin

Pedersen, J. S.; Junco, L. M. F.; Streubel, A.; Jensen, B.; Kot, W.; Roy, C.; Carstens, A. B.; Hansen, L. H.; Hille, F.; Franz, C. M. A. P.; Rothgardt, M. M.; Nielsen, T. K.

2026-05-15 microbiology 10.64898/2026.05.11.724364 medRxiv
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Soft rot Pectobacteriaceae (SRP) are among the most economically important plant pathogenic bacteria and are especially known to be problematic in potato production. The epidemiology of disease transmission has been investigated for almost a century, and several aspects have been highlighted as plausible infection routes. However, it is generally accepted that the major source of disease is the latently infected mother tuber, but several parameters are still influencing disease prevalence including contaminated equipment, soil water status as well as temperature. Management of the disease is limited to hygiene practices, dry storage and seed certification systems but several studies have also proven biocontrol agents such as bacteriophages (phages) as promising tools. Despite the severity of SRP on potato production, little is known about the genetic diversity of SRPs in Denmark, and since only few isolates are available, the possibility to design a broadly effective phage cocktail is limited. Here we describe a three-year field study utilizing an agri-citizen science approach where Danish farmers provided symptomatic potato plants or tubers, together with metadata such as date, location, potato variety and origin. By using whole genome sequencing (Illumina and Nanopore) together with metadata we were able to investigate and monitor the epidemiological disease spread across the country using 103 complete genomes, sampled across all three years. In this study we provide epidemiological evidence of disease origins and a suite of phages that could be used as a biocontrol tool for early disease intervention. Our results revealed several clonal clades across diverse locations (SNPs < 20) which strongly indicate common origin. A total of 17 Pectobacterium phages were tested and did target > 80% of clonal clades. Based on the clonality across the soft rot isolates we propose the possibility to set in early on using phages targeting strains relevant for soft rot development, with the possibility of a surveillance program together with customizing the phage preference.

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Morphology of petiole bending, senescence, epinasty, along with necrotic scarring in tomato leaves infiltrated with virulent Ralstonia pseudosolanacearum

Jain, M.; Kalita, S.; Daimari, P. R.; Rabha, Z.; Begum, S.; Dutta, L.; Giri, S. J.; Bhuyan, S.; Kushwah, S.; Kumar, A.; Ray, S. K.

2026-05-23 pathology 10.64898/2026.05.21.711296 medRxiv
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Ralstonia pseudosolanacearum (Rps) belongs to the Ralstonia solanacearum species complex (RSSC). It is a vascular pathogen that causes lethal bacterial wilt disease in many plants, including tomato and eggplant. In this study, we infiltrated tomato leaves with the phytopathogenic bacterium at 109 CFU/mL and observed the development of necrotic scars in the infiltrated area at 48 hours post-infiltration. Interestingly, this response was followed by petiole bending toward the ground of the compound leaf. This was followed by the gradual senescence of the infiltrated leaflet only. In addition, the terminal leaflet infiltrated with the pathogen exhibited epinasty. None of the above symptoms were observed in leaves infiltrated with the known virulent deficient hrpB::{Omega} mutant. Surprisingly, all of the above symptoms were observed in leaves infiltrated with another well-known virulence-deficient mutant phcA::{Omega}. It indicated that the necrotic lesion caused in tomato leaves was hrp-dependent. Infiltration in eggplant leaves caused necrotic scarring and leaf senescence, which were relatively delayed. Necrotic scarring without petiole bending or senescence in tomato leaves was also observed due to infiltration of Pseudomonas aeruginosa SPT08, a tomato endophyte having plant growth promotion activity. The patho-phenotypes such as petiole bending, epinasty, and senescence observed in the case of tomato in this study were not reported earlier. We believe these phenotypes produced in tomato after leaf infiltration may be useful to study the virulence of this pathogen.

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Revisiting the diversity of secondary endosymbionts in the major pest oat aphid, Rhopalosiphum padi

Yang, Q.; Zhu, B.; Yu, W.; Zhao, Z.; Gill, A.; Kaur, J.; Jonge, N. d.; Luan, J.-B.; Kristensen, T.; Liang, P.; Hoffmann, A. A.

2026-05-21 microbiology 10.64898/2026.05.19.726398 medRxiv
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There is disagreement on whether secondary endosymbionts are found in the major cereal pest aphid, Rhopalosiphum padi. Some papers report a diversity of secondary bacterial endosymbionts while others have failed to find evidence of these bacteria in this species. Here we revisit this issue by summarizing the relevant literature and through additional sampling of the species in Australia, China and Denmark using a combination of molecular approaches. We find a general absence of secondary endosymbionts beyond the obligate endosymbiont Hamiltonella defensa in R. padi. While the inconsistency in survey results may reflect rapid changes in endosymbiont turnover in populations and/or the impact of ecological factors such as host plant type on endosymbiont diversity, we are concerned that technical issues may be at least partly responsible for inconsistencies in the literature. This leads us to emphasize the importance of multiple sources of evidence required to establish and characterize endosymbiont infections, including PCR and qPCR assays, DNA Sanger sequencing and 16SrRNA gene metabarcoding. We note that several major aphid pests show a low incidence of secondary endosymbionts which raises issues about the importance of these endosymbionts in aphids that constitute pests, even though endosymbionts can in some cases increase host fitness and therefore pest impact.

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Compatibility and Multi-Season Field Evaluation of Trichoderma koningiopsis Integrated with Fungicides for Soybean Charcoal Rot Management

Bleckwedel, J.; Nieva, R. E.; Gonzalez, V.; Ploper, L. D.; Reznikov, S.

2026-05-13 plant biology 10.64898/2026.05.11.724353 medRxiv
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Soybean (Glycine max [L.] Merr.) productivity is frequently compromised by soil-borne pathogens. Macrophomina phaseolina (Mp), the causal agent of charcoal rot, can produce important soybean yield losses especially when hot and dry weather prevails. Integrating biological control agents with chemical seed treatments represents a promising strategy for improving disease management. This study aimed to (i) assess the in vitro compatibility of Trichoderma koningiopsis with commercial fungicide seed treatments, and (ii) evaluate the field performance of T. koningiopsis, alone or combined with compatible fungicides, across three soybean growing seasons. Compatibility assays revealed fungicide-specific effects, with Acronis(R) classified as non-fungitoxic and Topseed Extra as moderately fungitoxic. Across field seasons, Mp inoculation reduced seedling emergence, while several seed treatments improved emergence compared to the inoculated control, however, treatment effects varied markedly among years. Disease severity did not differ significantly among treatments in any season, and yield responses were strongly modified by environmental conditions rather than treatment effects. Temperature-response assays showed that T. koningiopsis exhibited optimal growth between 28 to 30{degrees}C and complete inhibition above 40{degrees}C, indicating high thermal sensitivity. The results demonstrate that T. koningiopsis can be integrated with compatible fungicides and may enhance early stand establishment under favorable conditions, but its field performance is strongly limited by high temperatures. These findings highlight the importance of environmental conditions when biological seed treatments are used.

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A decade of disease survey data in a progeny-provenance trial: Dothistroma needle blight in Scots pine

Perry, A.; Moore, B.; Jones, S.; Kaur, S.; Crampton, B.; Gurung, A.; Stockan, J. A.; Cottrell, J. E.; Beaton, J. K.; Cavers, S.

2026-05-14 ecology 10.64898/2026.05.12.724484 medRxiv
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Longitudinal data on disease susceptibility in forest trees are rare but essential for understanding host-pathogen dynamics and genetic variation in susceptibility traits. We present a long-term multisite common garden dataset quantifying susceptibility of Scots pine (Pinus sylvestris) to Dothistroma needle blight. The dataset comprises annual disease assessments collected from the same trees across 11 years, spanning 168 families and 21 Scottish provenances. This design enables partitioning of genetic and environmental sources of variation, evaluation of temporal stability in host response, and estimation of variance components and narrow-sense heritability of susceptibility. The data support analyses of phenotypic plasticity, provenance-level responses, and interactions between disease susceptibility and other adaptive traits. This resource will facilitate predictive modelling of host susceptibility under current and future environmental conditions.

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Characterization of a Novel Pseudomonad with Biocontrol Activity Against Aphanomyces euteiches

Kirk, A.; Workman, S. D.; Tiefenbach, A. M.; Hemmingsen, S. M.; Yost, C. K.

2026-05-19 microbiology 10.64898/2026.05.18.726007 medRxiv
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Aphanomyces euteiches, the causative agent of Aphanomyces root rot (ARR), is of major concern for pea and other legume crops globally. This oomycete pathogen causes substantial decreases in crop yields, is unaffected by most fungicides, and persists in the soil for many years via its resilient oospores. Given the significance of pea crops in sustainable agriculture, namely the ability to fix nitrogen and act as a sustainable protein source, solutions to ARR are of high importance. We used RNA-seq in a novel strain of Pseudomonas donghuensis to identify two biosynthetic gene clusters under GacA/S control that are involved in producing bioactive molecules capable of inhibiting A. euteiches. Based on similarity to other reported clusters in Pseudomonas, the first is predicted to encode for a pseudoiodinine compound, while the second is predicted to produce the siderophore 7-hydroxytropolone. Individual knockouts of each cluster showed loss of inhibitory action of P. donghuensis NRC29 against A, euteiches in vivo. This is the first report highlighting the potential of P. donghuensis and the products of the two identified biosynthetic pathways as biocontrol agents for A. euteiches. Further investigations into the efficacy of P. donghuensis NRC29 and its metabolites in inhibiting A. euteiches in field trials will be of high value in developing sustainable strategies for ARR mitigation. ImportanceModern fungicidal treatments for control of root rot in pulse crops are ineffective for control of A. euteiches, leaving limited strategies for management of A. euteiches infected fields. We describe a novel P. donghuensis strain with potential for biocontrol against this persistent pathogen. Given the economic value of peas and other pulses globally, further work into harnessing the bioactive metabolites produced by this strain into a practical in-field treatment will be valuable.

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The impact of Cronartium ribicola inoculum density on quantitative disease resistance in whitebark pine.

Johnson, J. S.; Wilhite, B.; Kegley, A.; Danchok, R.; Sniezko, R. A.

2026-05-06 genetics 10.64898/2026.05.02.722345 medRxiv
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Whitebark pine (Pinus albicaulis), a wide-ranging high-elevation conifer in western North America, is listed as threatened in the U.S. and as endangered in Canada. A major threat to whitebark pine is the non-native, invasive white pine blister rust disease, caused by the fungal pathogen Cronartium ribicola. In many pathosystems (including white pine blister rust), seedling inoculation trials are used to identify parent trees with genetic resistance. However, many of these trials use only one spore density for inoculation, and little information exists on the effectiveness of quantitative disease resistance (QDR) under varying spore densities and the corresponding implications for field performance. In this study, we examine the levels of infection and survival present within six whitebark pine seedling families previously rated for QDR (three susceptible and three resistant families) under six widely varying inoculum densities. The susceptible families showed very high infection and mortality at all inoculum densities, while performance of the resistant families varied with spore density treatment. The information gathered from the study will be useful in updating the projections of the future of whitebark pine populations under field conditions in areas of different rust hazard. The results also serve as a caution to those working in other pathosystems where seedling inoculation trials based on one spore density level are used to rate the resistance level of parent trees and their associated progeny.

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Evaluating β-glucanases as cell wall-permeabilising agents against Phytophthora agathidicida oospores

Pierson, E.; Mainwaring, J. C.; Patrick, W. M.; Gerth, M. L.

2026-05-12 biochemistry 10.64898/2026.05.08.723360 medRxiv
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The persistence of specialised survival spores produced by microbial pathogens represents a primary bottleneck in the management of plant diseases. In oomycetes, these spores (known as oospores) are largely impervious to chemical control, allowing them to persist in soil and initiate new infection cycles over many years. A prominent example is the soil-borne pathogen Phytophthora agathidicida, the causal agent of kauri dieback disease, where long-lived oospores hinder conservation efforts in native forests. The resilience of oospores is attributed to their thick wall composed of complex {beta}-glucan layers that render the oospores impermeable to most conventional biocides. Here we have investigated an enzyme-based approach for weakening the oospore cell wall. We searched enzyme databases to select {beta}-glucanases targeting a variety of linkages found in Phytophthora oospore walls. Eight of these {beta}-glucanases were successfully purified and tested for their digestive activity against intact oospores in vitro using a phenol-sulfuric acid assay. We showed that combining these enzymes was crucial to achieve significant digestion through synergies and additive effects. The optimal combination, comprising 1,3-, 1,6-, and 1,3(4)-{beta}-glucanases, was evaluated for its ability to permeabilise oospores to five biocides typically effective only on other, more sensitive lifecycle stages of the pathogen. Using a live/dead fluorescence assay, we observed that the effects of the membrane-targeting biocides were potentiated in oospores that were pre-treated with the {beta}-glucanase mixture. Our results highlight enzymatic cell wall permeabilisation as a promising strategy toward improved management of oospore persistence in kauri forest soils and against broader oomycete threats. KeypointsO_LIOur phenol-sulfuric acid assay can be used to screen for oospore-degrading enzymes. C_LIO_LISynergistic enzyme combinations are essential for effective oospore wall digestion. C_LIO_LIEnzyme pre-treatment sensitises oospores to membrane-targeting biocides. C_LI

9
Method for maintaining translocated wild roses under laboratory conditions for controlled gall induction by Diplolepis rosae and D. mayri

Laszlo, Z.; Denes, A. L.; Witiak, S. M.; Peterfi, E.; Podar, D.

2026-05-21 ecology 10.64898/2026.05.19.726150 medRxiv
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Plant-gall wasp systems provide unique models for studying multitrophic interactions and unique developmental trajectories, yet standardized laboratory protocols for maintaining wild rose hosts (Rosa spp.) and sustaining gall inducers (Diplolepis spp.) are lacking. We developed and tested a method for growing and maintaining translocated individuals of Rosa canina, R. rubiginosa, R. spinosissima, R. gallica, R. tomentosa, and R. pendulina under laboratory conditions over three consecutive years (2023-2026). The goal was to have a constant supply of plant host material for reliably producing galls of D. rosae and D. mayri for experimental use. The protocol integrates soil and substrate composition, photoperiod and humidity regimes, pruning, dormancy management, and controlled exposure to gall-inducing wasps. More than 75% of rose individuals survived the full 3-year period, with consistent annual gall induction across some of the species. This work represents the first reproducible laboratory method for long-term maintenance of wild rose hosts and controlled gall induction by Diplolepis species, while also providing a transferable framework for maintaining perennial woody hosts and experimentally manipulating specialized plant-insect interactions under laboratory conditions, thereby providing a platform for ecological, physiological, and evolutionary studies on these interactions.

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Unlocking the <Alphitobius diaperinus> (Coleoptera: Tenebrionidae)-Combatting Capabilities of <Bacillus thuringiensis> INTA Mo4-4 through genomic and phenotypic characterization

Perez, M. P.; Palma, L.; Berretta, M. F.; Benintende, G. B.; Sauka, D. H.

2026-05-13 microbiology 10.64898/2026.05.13.724709 medRxiv
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Bacillus thuringiensis INTA Mo4-4 was characterized phenotypically, genomically, and for insecticidal activity against Alphitobius diaperinus. Microscopy revealed rare flat rectangular parasporal crystals, and SDS-PAGE identified a ca. 67 kDa protein, similar to B. thuringiensis serovar morrisoni strain tenebrionis DSM-2803, which was proteolytically processed to a ca. 55 kDa fragment. Genomic analysis showed a 5.99 Mb genome with 99.43% completeness, clustering phylogenetically with B. cereus and B. thuringiensis. High genomic similarity was observed with B. thuringiensis svar. morrisoni BGSC 4AA1, confirmed by MLST analysis assigning it to ST-23. The genome encodes an interesting arsenal of pesticidal proteins showing significant similarity to Cry3Aa, Mpp23Aa, Xpp37Aa, Mpp5Ab, Vpb1Ad, Vpb1Ae, Vpa2Ab, Vpa2Ba, Vpa2Bb and Spp1Aa, with demonstrated toxicity against coleopteran pests. Biosynthetic gene clusters for toyoncin, fengycin, and bacillibactin were identified. Dose-response bioassays showed that INTA Mo4-4 was nearly four times more toxic to A. diaperinus larvae (LC50 136.9 {micro}g/ml) than DSM-2803 (LC50 540.5 {micro}g/ml), with the difference being statistically significant. No teratological effects were observed on Musca domestica. These findings suggest that INTA Mo4-4 is a promising candidate for the biological control of A. diaperinus.

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Chromosomal variability in a clonal crop: Somaclonal change follows the emergence of triploid saffron crocus

El-nagish, A.; Dhar, M. K.; Mann, L.; An, R.; Houben, A.; Blattner, F.; Harpke, D.; Heitkam, T.

2026-05-07 plant biology 10.64898/2026.05.04.722608 medRxiv
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(1) BackgroundSaffron crocus (Crocus sativus) is the source of saffron, the most expensive spice in the world. It evolved about 3000 years ago as a sterile triploid clone in Greece. Since then, saffron has spread across the globe, where regionally distinct practices of saffron cultivation have developed. Despite differences in morpho-physiological traits, genetic variability is low, if present at all. Here, we aim to resolve chromosomal and sequence-associated variability across saffron crocus cultivars from the crops main cultivation areas in Africa, Asia and Europe. (2) MethodsWe used genome-wide DNA polymorphisms obtained through genotyping-by-sequencing (GBS) of 33 saffron and 14 closely related Crocus accessions, which we place into a phylogenetic context. For karyotyping, we compare nine saffron accessions by multi-color fluorescent in situ hybridisation (FISH) with repetitive DNA probes. (3) Key resultsPhylogenetic analyses confirmed the single origin and clonal nature of all saffron accessions. We detected slight DNA differences among saffron crocus genotypes, which were minor compared with those in wild C. cartwrightianus populations. Still, the Iranian saffron accessions form a genetically very narrow group that differs from the other proveniences in population genetic analyses. However, chromosomes of some saffron accessions display variable FISH signals, likely resulting from gains and losses of tandemly repeated DNA. (4) Main conclusionsBased on the high genetic identity and small karyotypic differences, we confirm the clonal origin of the saffron accessions. Nevertheless, as we detected small and regional chromosomal variability, we conclude that at least four somaclonal saffron lineages emerged after saffrons origin. Societal Impact StatementFor millennia, many cultures developed cultivation practices and regional crop varieties. A notable case is saffron, the worlds most expensive spice that is harvested from stigmas of saffron crocus. This flower crop arose 3000 years ago in a singular genome triplication event and since then spread clonally across the globe. By identifying genetic and chromosomal variability in clonal saffron accessions, we highlight regional diversity, support the preservation of traditional knowledge, and underscore the risk of relying on only one clonal lineage. This informs strategies for saffron cultivation, linking cultural heritage with modern genomics to address biodiversity, evolution, and food security.

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Insect pests survey of Prosopis juliflora, Afar rangeland, Ethiopia

Mekonnen, B. B.; Ali, S. E.; Lemma, E. G.

2026-05-05 ecology 10.64898/2026.05.04.722396 medRxiv
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Prosopis juliflora is an invasive alien plant species and a problematic weed that poses significant ecological and socio-economic challenges in Ethiopia, particularly in the Afar rangelands. The study explored the diversity and effects of insect herbivores communities feeding on the flowers and pods of P. juliflora to determine their role in limiting reproductive success across three selected ecological sites: Amibara, Gewanne, and Aysayita. A total of 118 adult insect specimens were collected between January and November 2021 using a sweep net and hand collection methods. Community structure, analysis via the Shannon Wiener diversity index, strongly influenced damage pattern. Amibara exhibited the highest insect diversity resulting in significant reproductive damage, including 5.98% of flower loss and 10.39% pods tunneling, primarily caused by Chrysomelidae and Pyralidae. Conversely, Gewanne was showed lower diversity, but higher sap-sucking (13.39 % shriveled pods; 5.11 % flower curling) were caused by Aphididae. Overall, 18.41 % of the pods, and 11.59 % of the flowers were exhibited insect related injury. These finding confirm that more internal seed predation and nutrient depletion were revealed significantly reduce viable seed production. The result was suggested that natural insect communities currently function as partial biological control agents. This indicates strong potential for developing integrated biological control strategies to manage P. juliflora invasion in Ethiopia rangelands.

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Easy to use and low cost leaf disease quantification workflow using Ilastik

Prouvost, A.; Connesson, L.; Le Gourrierec, T.; Freville, H.; David, J.; Plessis, C.; Magnier, B.

2026-05-16 plant biology 10.64898/2026.05.14.719059 medRxiv
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Accurate and reproducible assessment of foliar disease severity is essential for evaluating the performance of heterogeneous plant communities and understanding host-pathogen interactions. However, traditional visual scoring methods remain subjective, with limited precision, and difficult to scale in large phenotyping experiments. Here, we present a semi-automated image analysis workflow designed to quantify multiple foliar disease symptoms simultaneously on wheat flag leaves sampled from varietal mixtures. The workflow combines three methodological components: (i) a standardized protocol for leaf sampling and imaging, (ii) supervised machine learning segmentation using Random Forest implemented in Ilastik to classify multiple symptoms (powdery mildew and yellow rust), and (iii) a graphical user interface facilitating pipeline deployment by non-specialist operators. To evaluate the influence of image representation on classification performance, four color spaces (RGB, HSV, HLS, LAB) were systematically compared. The approach was validated using images of durum wheat flag leaves collected from a field experiment assessing eight-way varietal mixtures under natural fungal pressure. Cross-validation against manually annotated images demonstrated high segmentation accuracy across all symptom. Comparison among color spaces revealed only minor differences in performance. Overall, this workflow offers a cost-effective, annotation-efficient and reproducible alternative to deep learning approaches, leveraging open-source and actively maintained tools while requiring limited training data and enabling objective, reproducible and scalable disease phenotyping.

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First occurrence of Corynespora cassiicola infecting chia plant in Bangladesh and its sensitivity to selected fungicides

Badhon, A. K.; Gupta, D. R.; Paul, S. K.; Ali, J.; Rahman, M. M.; Islam, T.

2026-05-06 molecular biology 10.64898/2026.05.01.722373 medRxiv
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Chia (Salvia hispanica L.) is an emerging crop in Bangladesh valued for its medicinal properties and economic significance. In March 2024, target spot-like symptoms were observed in an experimental chia field (24.75{degrees} N, 90.50{degrees} E) at Bangladesh Agricultural University in Mymensingh, Bangladesh with disease incidence ranging from 23% to 47% across approximately 0.25 ha. Initially appearing as brick-red spots, these symptoms developed into target-shaped concentric rings, affecting leaves, stems, and inflorescences. A total of 24 fungal isolates were recovered from infected tissue; two representative isolates (BGECh-3 and BGECh-4) were randomly selected for details characterization. Pathogen identity was established through morphological traits, multilocus phylogenetic analysis of internal transcribed spacer (ITS) and elongation factor 1-alpha (EF-1) genes sequence, and pathogenicity confirmation through Kochs postulates, collectively identifying the causal agent as Corynespora cassiicola. The isolates demonstrated a broad host range, successfully infecting brinjal, chili, bottle gourd, country bean, tomato, and soybean. In vitro fungicide sensitivity assays with seven commercial fungicides showed that both isolates were highly sensitive to Goldzim (50% carbendazim), which completely inhibited mycelial growth at 10 {micro}g mL-{superscript 1}. Conza (10% Hexaconazole) and Amister top (18.2% azoxystrobin + 11.4% difenoconazole) reduced growth by up to 85% and 67%, respectively at equal concentration. Other fungicides showed comparatively lower efficacy even at higher concentrations. This study represents the first report of target spot disease of chia caused by C. cassiicola in Bangladesh and provides insights for effective disease management strategies.

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Genetic diversity of tomato brown rugose fruit virus in Morocco

Maachi, A.; Donaire, L.; Aranda, M. A.

2026-05-12 microbiology 10.64898/2026.05.11.724243 medRxiv
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Tomato brown rugose fruit virus (Tobamovirus fructirugosum) is an emerging virus that affects tomatoes, capsicum, and chili. Since its first detection in Jordan in 2015, the virus was reported in more than 40 countries across all the continents. In Morocco, the virus was reported for the first time in October 2021. However, its genetic diversity remains unexplored. In this work, we used a collection of tomato fruits from local markets to investigate the variability of the virus in the country. We explored the different pressures acting on the N-terminus of the RNA-dependent RNA polymerase, the movement protein, and the coat protein genes. Then, we used haplotype network analyses to reveal the population structure within the Moroccan isolates and studied their relationships with the ones from the world. We found that genetic diversity is low, which is consistent with the global situation. No signatures of diversifying selection were detected across the analyzed genes. However, the virus sequences from Morocco showed a clear geographic structure, suggesting that geographic factors probably combined with agricultural practices may contribute to shaping the population structure of ToBRFV in Morocco.

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Effects of leaf removal on photosynthetic activity, fruit yield, and quality of micro-dwarf tomatoes

Usenko, D.; Giladi, C.; Ziv, C.; Helman, D.

2026-05-13 plant biology 10.64898/2026.05.10.724098 medRxiv
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Micro-dwarf tomato cultivars are increasingly considered for urban and controlled-environment agriculture due to their compact architecture and suitability for high-density planting. However, optimal canopy management strategies for these cultivars remain poorly defined. In this study, we evaluated the effects of different leaf removal intensities on leaf-level physiological performance, fruit yield, and fruit quality in three micro-dwarf tomato cultivars (Mohammed, Hahms Gelbe Topftomate, and Red Robin) grown under contrasting seasonal light conditions. Plants were subjected to low (15%), moderate (30%), or severe (90%) leaf removal, and leaf-level gas exchange was measured across canopy layers, along with yield and fruit quality assessments. Severe leaf removal (90%) increased carbon assimilation, transpiration, and stomatal conductance in middle and lower canopy leaves by up to approximately twofold compared with control plants, indicating improved light availability at the leaf level. However, these physiological enhancements did not consistently translate into higher yield, reflecting reduced whole-plant source capacity under excessive leaf removal. Low to moderate leaf removal (15-30%) generally increased or maintained yield and fruit number, whereas severe leaf removal reduced yield in Hahms Gelbe and Red Robin, particularly under low seasonal radiation. In contrast, Mohammed exhibited yield increases of up to 220% under low leaf removal and maintained increased yield even under severe leaf removal under high-light conditions. Fruit quality was largely unaffected by leaf removal, except for total soluble solids, which declined by approximately 12% under severe leaf removal across cultivars, consistent with sugar dilution under source limitation. Overall, these results demonstrate that optimal leaf removal in micro-dwarf tomatoes requires balancing improved canopy light distribution with maintenance of sufficient leaf area for carbon assimilation. Leaf removal thresholds are strongly cultivar- and light-dependent, emphasizing the need for cultivar-specific canopy management strategies in compact tomato systems and controlled-environment agriculture.

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Isolation of rhizobia from Ontario soils that are effective at fixing nitrogen with common bean (Phaseolus vulgaris)

Harrison, T. L.; Pandher, U. S.; Dixon, A.; Esme, O.; Gagnon, E. M.; Naranjo-Robayo, N.; Doyle, R. T.; Oresnik, I. J.; diCenzo, G. C.

2026-05-04 microbiology 10.64898/2026.05.01.722220 medRxiv
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Common bean (Phaseolus vulgaris) is an important crop in Canada and globally. Like other legumes, common bean (Phaseolus vulgaris) establishes symbiotic interactions with nitrogen fixing bacteria called rhizobia. However, nitrogen fixation by rhizobia in association with common bean is often suboptimal, constraining its productivity and necessitating the application of nitrogen fertilizer. To support the development of high-performing, locally adapted rhizobial inoculants for Ontario common bean growers, we isolated 216 common bean-nodulating rhizobia from southern Ontario soils using a nodule trapping approach with four common bean cultivars. Whole genome sequencing followed by phylogenomic analyses of the 216 rhizobial isolates revealed substantial diversity, assigning them to 11 Rhizobium species, including two novel species. Nearly all isolates belong to the symbiovar phaseoli, spanning the nodC {gamma}-a, {gamma}-b, and alleles, with four isolates belonging to the symbiovar gallica. Soil origin had a significant impact on the species-level community composition recovered during the nodule trapping experiments, indicative of biogeographical structuring of common bean-nodulating rhizobia across southern Ontario. In contrast, host trapping cultivar had only a minor influence of the recovered Rhizobium population diversity. Greenhouse assays demonstrated that one of the novel Rhizobium species exhibited the highest average symbiotic effectiveness, although high-quality isolates were found across multiple species. Together, these results revealed a diverse and genomically variable Rhizobium community capable of forming effective symbioses with common bean in southern Ontario soils. Importantly, our genome-sequenced Rhizobium collection will serve as a valuable resource for identifying competitive and high-quality strains for the development of inoculants tailored to Ontario common bean production. IMPORTANCECommon bean is a globally important food crop, yet its productivity is often limited by suboptimal nitrogen fixation, forcing growers to rely on synthetic fertilizers. Consequently, identifying high-performing, locally adapted inoculant strains is essential for reducing dependence on synthetic nitrogen fertilizers and improving the sustainability of temperate agroecosystems. Our study provides a genome-sequenced collection of common bean-nodulating Rhizobium from southern Ontario, revealing substantial species and genomic diversity across sampling locations. Greenhouse studies allowed us to identify multiple isolates, including isolates from a novel Rhizobium species, that consistently fix nitrogen with, and enhance the growth of, common bean plants. Our findings highlight strong biogeographical structuring of rhizobial communities and demonstrate that Ontario soils already harbour strains with high symbiotic potential. In addition, our Rhizobium collection represents a foundational resource to support future inoculant development and enables future work on the ecology, evolution, and applied optimization of legume-rhizobium symbioses.

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Beauty at risk: A taxonomic synopsis of Belemia (Nyctaginaceae), an endangered and endemic genus of vines in Brazil

Cunha-Neto, I. L.; Rossetto, E. F. S.; Goncalves, D. V.; Nogueira, M. G. C.; Antar, G. M.; Rodrigues, V. R. C.; Silva, A. O.; Angyalossy, V.; Sa, C. F. C.

2026-05-13 plant biology 10.64898/2026.05.12.724086 medRxiv
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Belemia belongs to Nyctaginaceae and comprises two species of delicate vines. Both species are endemic to Brazil. Belemia fucsioides, the type species, described in 1981, occurs in a restricted area of the Atlantic Forest in southeastern Brazil. Belemia cordata, described in 2020, is known from only two records from the same area in the Cerrado of northern Brazil. Here, we describe the taxonomic history of Belemia and provide the first synopsis for the genus. We include species description, distribution map, identification key, and anatomical data. We used field observations over the past decade and modeled anthropogenic changes in the species range to conduct a conservation assessment in accordance with the IUCN Red List criteria. Conservation assessments indicate significant concerns for Belemia, classified as either endangered (B. fucsioides) or critically endangered (B. cordata). The species are threatened primarily by habitat loss to land used for agriculture, forestry, and livestock production. This study contributes to ongoing initiatives exploring plant diversity in the Neotropics and supports efforts to identify threats to biodiversity.

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Heat tolerance and its seasonal acclimation in Fagus sylvatica compared to Fagus orientalis and Pseudotsuga menziesii

Hauck, M.; Csapek, G.; Kraemer, K.; Schmidt, O.; Lucas, Y.; Popp, L.; Szafranek, L.; Dulamsuren, C.

2026-05-18 ecology 10.64898/2026.05.17.725742 medRxiv
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Heat tolerance determines the vitality of tree species under climate change independently of drought tolerance, but has been much less studied than tree water relations. We studied species-specific differences and the capacity for seasonal heat acclimation in Central Europes naturally most important tree species, Fagus sylvatica, in comparison with two exotic tree species (Fagus orientalis, Pseudotsuga menziesii) that are considered for silvicultural climate change adaptation in managed forests. Foliage of mature trees was incubated at temperatures from 35-50 {degrees}C for up to 4 h to simulate daily heat maxima during heat waves. The maximum quantum yield (Fv/Fm) of photosystem II (PS II) of dark-adapted leaves was measured, because the PS II is particularly sensitive to heat and its functionality can decide on plant survival under heat. Fagus sylvatica was much more tolerant to heat than Pseudotsuga menziesii, but weakly (albeit significantly) less tolerant than Fagus orientalis. Within its limits, Pseudotsuga menziesii showed high seasonal heat acclimation with constantly increasing tolerance during the growing season. Fagus orientalis, but practically not Fagus sylvatica, also acclimated to heat. This makes Fagus orientalis slightly superior over Fagus sylvatica in terms of heat tolerance, whereas the suitability of Pseudotsuga menziesii for silvicultural climate change adaptation is questionable. Strong heat acclimation, but also overall low heat tolerance, in Pseudotsuga menziesii might be the result of evergreenness, which requires the generation of both cold and heat tolerance during the year.

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Identification of septoria nodorum blotch susceptibility genes in hard winter wheat

Ara, A. M.; Holmes, D. J.; Friesen, T. L.; Carver, B. F.; Bai, G.; St. Amand, P.; Bernado, A.; Sharma, R.; Aoun, M.

2026-05-15 genetics 10.64898/2026.05.13.724689 medRxiv
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Key message Characterized and unknown septoria nodorum blotch susceptibility/resistance genes were identified in contemporary U.S. hard winter wheat. The necrotrophic fungus Parastagonospora nodorum is the causal agent of septoria nodorum blotch (SNB) of wheat. To determine the prevalence of SNB sensitivity genes in a contemporary U.S. hard winter wheat (HWW), we evaluated a panel of 619 breeding lines and cultivars against five P. nodorum isolates and five necrotrophic effectors (NEs), SnToxA, SnTox1, SnTox3, SnTox267 and SnTox5, and genotyped the panel using genotyping-by-sequencing (GBS) markers and diagnostic Kompetetive-allele specific PCR (KASP) markers for the sensitivity genes Tsn1-B1, Snn1-B1, and Snn3-B1/B2. GBS analysis identified 34,357 GBS-single nucleotide polymorphism (SNP) markers. Evaluations against P. nodorum isolates showed that 40-67% of the genotypes were susceptible in the panel. Toxin infiltration assays showed that 54%, 2%, 37%, 13%, and 15% of the genotypes were sensitive to SnToxA, SnTox1, SnTox3, SnTox267, and SnTox5, respectively. Diagnostic KASP markers for Tsn1-B1, Snn1-B1, and Snn3-B1/B2 showed prediction accuracies of 98%, 75%, and 92% for the corresponding effectors SnToxA, SnTox1, and SnTox3, respectively. Genome-wide association studies (GWAS) not only confirmed the presence of the previously characterized sensitivity genes Tsn1-B1, Snn1-B1, Snn2, Snn3-B1/B2, and Snn5-B1, but also identified new loci to be associated with responses to P. nodorum isolates and NEs. Of which, Qsnb.osu-2AS on chromosome 2AS was associated with responses to all five isolates. We developed KASP markers KASP_S4B_643615365, KASP_ S2D_16184991, and KASP_S2A_9833162 linked to Snn5-B1, Snn2, and Qsnb.osu-2AS, respectively. These findings should guide breeding for SNB resistance in hard winter wheat.