Plant Pathology

Cultivar susceptibility strongly influences the epidemiology of vector-borne plant diseases, and understanding cultivar-specific variation can inform management strategies. This is particularly relevant for flavescence doree, an incurable grapevine disease associated with a phytoplasma and transmitted by the leafhopper Scaphoideus titanus. In this study, we investigated the susceptibility of the main Swiss varieties, by combining controlled insect-mediated inoculation experiments with complementary field analyses conducted at progressively finer spatial scales. Together, these approaches allowed us to compare both infection probability and phytoplasma relative titre under standardised transmission conditions with disease incidence and relative titre under natural epidemiological conditions. For most cultivars, laboratory results were broadly consistent with field observations. However, a marked discrepancy emerged in the relative infection pattern between the two main grapevine cultivars grown in Switzerland: Chasselas and Pinot Noir. Under controlled conditions, they did not differ significantly in either their probability of infection or the phytoplasma relative titre, indicating no detectable difference in susceptibility to phytoplasma infection. In contrast, Pinot Noir consistently showed higher disease incidence than Chasselas under natural conditions. This pattern was observed across all spatial scales examined, from regional surveys to neighbouring vineyard plots, and was mirrored by higher phytoplasma relative titres. Importantly, under controlled conditions, S. titanus mortality during the one-week inoculation period was significantly higher on Chasselas than on Pinot Noir, indicating that Chasselas may provide a less favourable host for S. titanus. Together, these findings support the hypothesis that differences in field disease incidence between these cultivars may arise from differences in vector performance rather than intrinsic susceptibility to phytoplasma infection. This highlights the importance of considering plant-vector interactions, alongside susceptibility to infection, when assessing cultivar-specific vulnerability to vector-borne plant diseases.

To reduce reliance on pesticides in potato production, alternative partial control methods are needed, yet their efficacy and reliability are seldom quantified over multiple years. Canopy architecture is a potential lever against late blight, caused by Phytophthora infestans, a pathogen whose infection and development are highly driven by humidity and temperature. However, its practical value under field conditions in pure crop stands remains poorly documented. We compared two commercial potato cultivars with contrasting canopy architectures but similar leaf tissue susceptibility to P. infestans, in replicated field trials conducted over six years (2011 2016) under irrigation, and over three of these years also without irrigation. Disease and canopy development (height, closure, leaf area, stem number) were monitored throughout the growing seasons and analysed using non linear growth models. Monalisa, an erect and ramose cultivar, consistently slowed epidemic progress relative to Bintje, a semi-erect and leafy cultivar. This effect was significant in every non irrigated trial but in only half of the irrigated ones. No single canopy trait explained this suppression across all years and conditions. However, canopy closure at the time of inoculation was the most consistent correlate, supporting the hypothesis of a microclimate mediated effect. Disease reduction seldom translated into a yield benefit, except under irrigation, where Monalisa occasionally out yielded Bintje. Our results confirm that canopy architecture can complement other levers within integrated late blight management. Nevertheless, its significant year to year variability and dependence on environmental conditions must be accounted for if it is to be deployed as a reliable control method.

L-phenylalanine (Phe) is converted via the phenylpropanoid pathway into phenolic compounds, and elevated phenolic/flavonoid levels are generally associated with enhanced plant resistance. We previously showed that exogenous Phe suppresses the necrotrophic fungus Botrytis cinerea in petunia, chrysanthemum, determinant tomato, and several postharvest diseases. Here, we expanded evaluation to a broad range of pathosystems, including an indeterminate greenhouse tomato cultivar, multiple dicot species, and the monocot wheat. Phe, applied as spray or drench across concentrations, was consistently effective at [≥]4 mM. It reduced disease severity in diverse systems: Sclerotinia sclerotiorum (tomato, sweet basil, cucumber, lettuce), Leveillula taurica and Oidium neolycopersici (tomato), Podosphaera xanthii (cucumber), wheat foliar pathogens (Blumeria graminis f. sp. tritici, Zymoseptoria tritici, Puccinia triticina, P. striiformis f. sp. tritici), the oomycetes Pseudoperonospora cubensis (cucumber leaves) and Pythium aphanidermatum (roots), bacterial pathogens (Pseudomonas syringae pv. tomato and Clavibacter michiganensis subsp. michiganensis) and Tomato brown rugose fruit (ToBRFV) that belongs to the Tobamovirus genus. Phe was effective on both young and mature leaves and often performed comparably to chemical fungicides; combinations rarely improved control, except for enhanced activity with pyrimethanil against B. cinerea in tomato. Synergistic effects were observed when Phe was combined with an adjuvant against tomato powdery mildews. A formulated product (NaturaFend 550 SP) was more effective than non-formulated Phe in B. cinerea (tomato) and P. xanthii (cucumber). Application timing also influenced efficacy, with treatment 5 days before infection providing superior control compared with 0, 3, or 7 days. Overall, Phe effectively controlled biotrophic and necrotrophic fungi, oomycetes, bacterial pathogens and a virus across diverse crops under experimental greenhouses and commercial like conditions.

Tomato worldwide production is increasingly challenged by complex disease outbreaks involving multiple interacting pathogens. In Israel, recent years have seen a marked rise in vascular collapse symptoms in greenhouse-grown tomatoes, coinciding with the widespread emergence of tomato brown rugose fruit virus (ToBRFV) and pepino mosaic virus (PepMV). Here, we investigated the bacterial and viral agents associated with these outbreaks and examined how viral infection influences the development and severity of bacterial diseases that cause vascular collapse. Surveys conducted between 2021 and 2026 revealed that tomato pith necrosis outbreaks were associated with a diverse bacterial community dominated by members of the Pseudomonadales and Enterobacterales, while bacterial canker outbreaks were exclusively linked to Clavibacter michiganensis. Multilocus sequence analysis showed that pith necrosis-associated Pseudomonas isolates clustered primarily within the P. syringae, and P. corrugata phylogroups. Pathogenicity assays demonstrated that only a subset of pith necrosis-associated bacteria, P. mediterranea, P. capsici, P. viridiflava, and Xanthomonas euvesicatoria pv. perforans, induced pith necrosis under controlled conditions, with high variability in symptom severity. Co-inoculation experiments showed that ToBRFV- and PepMV-infected plants exhibited a 40-100% increase in lesion size following inoculation with pith necrosis-associated bacteria, without a corresponding increase in bacterial colonization, whereas the same viral infections attenuated wilt symptoms caused by C. michiganensis. Together, our findings demonstrate that endemic viral infections differentially modulate bacterial disease outcomes, either exacerbating or attenuating symptoms depending on the pathogen. These results highlight the importance of multi-pathogen interactions in disease severity and have important implications for tomato disease management.

Peanut smut, caused by Thecaphora frezzii, is an important constraint to peanut production in Argentina, but quantitative estimates of yield losses across environments remain limited. We quantified the relationship between disease incidence and kernel yield using 922 observations from 26 field studies conducted in Cordoba, Argentina, between 2021 and 2025. Study-specific incidence-yield relationships were analyzed using linear regression, random-effects meta-analysis, and linear mixed-effects models. Peanut smut incidence was consistently associated with yield reduction across studies. The estimated damage coefficient ranged from 24.2 to 28.7 kg ha-{superscript 1} per 1% increase in disease incidence, corresponding to a relative yield reduction of 0.74-0.87% of attainable yield. In contrast, attainable yield varied markedly among studies, ranging from 1,370 to 5,409 kg ha-{superscript 1}. Although an exploratory segmented analysis suggested a breakpoint near 12% incidence, subsequent moderator analyses, study- specific regressions, and normalized response curves provided no evidence of a biologically meaningful change in the damage coefficient across incidence or yield classes. These results indicate that differences among environments were primarily associated with attainable yield rather than with changes in the magnitude of disease-associated yield loss. The resulting damage function provides a quantitative basis for yield-loss assessment and disease management in peanut.

Cucurbit yellow vine disease (CYVD) is a phloem disease caused by a group of Serratia ureilytica bacteria within the S. marcescens complex. The CYVD pathogen is distinct from other phloem pathogens in two fundamental ways: i) unlike other bacterial phloem pathogens, the CYVD pathogen readily grows on standard laboratory media; and ii) whereas most bacterial phloem diseases are transmitted solely by hemipteran insects, CYVD can be transmitted by both hemipteran (squash bug) and non-hemipteran (cucumber beetle) vectors. The phloem of cucurbits is also distinct, with cucurbits having a unique secondary phloem system, the extrafascicular phloem (EFP), for which a role in CYVD has not been investigated. The distinctive features of this pathosystem prompted a re-evaluation of pathogen localization in planta using a green fluorescent protein-tagged CYVD strain during infection of squash (Cucurbita pepo). In epifluorescence and confocal microscopy images of transverse and longitudinal sections of stem and petiole tissues, the fluorescence patterns of the CYVD pathogen were consistent with pathogen colonization of all four known EFP tissues, namely the peripheral, entocyclic, ectocyclic, and commissural sieve tubes. The CYVD pathogen may enter the EFP during herbivory by pathogen-infected vectors that cause mechanical damage, as the EFP exhibits much slower blockage following injury than the conventional fascicular (bundle) phloem. Moreover, although EFP has been proposed to have an antimicrobial function, EFP exudate collected from the stem and petioles of C. pepo supported rapid pathogen growth. These results provide evidence for a unique tissue tropism of the newly emerged plant pathogen S. ureilytica.

BACKGROUNDBlackleg, caused by Leptosphaeria maculans, is a major disease limiting oilseed rape production worldwide, and its management increasingly requires sustainable alternatives to chemical fungicides. In this study, we evaluated the antagonistic activity and plant growth-promoting potential of three endophytic bacteria, Bacillus velezensis Bro5, Bacillus subtilis Bro11, and Pantoea agglomerans Bru13, against a geographically diverse collection of 139 L. maculans isolates from five oilseed rape-producing regions of Argentina. RESULTSDual culture assays revealed strong inhibitory activity by Bro5 and Bro11, with mean inhibition rates of [~]80% across isolates, while Bru13 showed variable inhibition (<75% for most isolates). Greenhouse and growth chamber assays confirmed the protective potential of these strains. At the cotyledon stage, Bro11 and Bro5 reduced lesion size by 47% and 28%, respectively, while their combination achieved a 51% reduction. In greenhouse trials, combined application of Bro5 and Bro11 reduced stem base necrosis by 45% and increased the proportion of plants with [&le;]50% damage to 98%, compared to only 70% in controls. Key disease metrics, including disease index, incidence, and severity, decreased by 60%, 23%, and 26%, respectively. Beyond pathogen suppression, inoculation with the Bro5-Bro11 consortium enhanced plant growth, increasing shoot biomass by 89% at early stages, and improving stem dry weight and diameter by 10% and 35%, respectively, at maturity. CONCLUSIONThese findings highlight the robustness of Bacillus endophytes as biocontrol agents, their capacity to suppress diverse pathogen isolates, and their dual role in plant growth promotion, supporting their potential integration into sustainable blackleg management programs.

Black pod disease is a major threat to global cacao (Theobroma cacao L.) production. It is caused by multiple Phytophthora species with geographic ranges from regionally restricted to pantropical, and developing resistant cultivars requires screening against the diversity of these pathogens. This study evaluated the response of 25 cacao clones to 13 isolates of P. palmivora (eight), P. megakarya (two), and P. theobromicola (three) by nonwounding inoculation of detached pods. Lesion size and lesion expansion status were evaluated 4 days after inoculation. The linear mixed model included 178 combinations among 17 clones and 12 isolates, each represented by at least two biological replicate pods. Lesion size differed among clones (F(16, 449.2) = 25.16) and among isolates (F(11, 416.0) = 32.90), and the clone by isolate interaction was significant (F(150, 465.4) = 1.72; all P < 0.001). PENTAGONA-17 had the smallest estimated marginal lesion size (9.3 mm) and fitted values of 3.8 to 16.9 mm across 11 isolates representing all three species. ARF-31, ICS-47, POUND-7, and PMCT-73 had the next four smallest estimates, whereas NA-246 had the largest (36.7 mm). Isolate estimates varied more than fourfold, from 9.6 to 42.0 mm. Despite a marginal estimate of 12.9 mm, POUND-7 had a fitted estimate of 49.2 mm with P. megakarya GH34. The probability of lesion expansion also differed among clones and isolates (both P < 0.001). Together, these findings identify candidates for further resistance evaluation and show that multi-isolate screening provides a stronger basis for selecting materials for black pod resistance breeding.

Vein clearing of zucchini (VCZ) and bacterial leaf spot (BLS) are caused by various strains of the Pseudomonas syringae species complex that infect zucchini (Cucurbita pepo) seeds. VCZ strains have a narrow host range of cucurbits and affect only seedlings, whereas BLS strains have a broader host range and cause symptoms on adult plants. A multiplex qPCR test showed that VCZ strains predominated in infected seed lots produced in different countries. We surveyed hybrid seed crops grown in parallel in two French regions to address inoculum sources. According to DNA-based approach, parental seed lots were positive to BLS strains, although no culturable bacteria were recovered. Hybrid seed lots produced in the Rhone Valley (southeastern France) showed higher infection rates than those produced in Limagne (central France), and VCZ strains were recovered only from the Rhone Valley. Two representative strains of VCZ and BLS colonized seeds through the vascular and floral pathways, whereas only the BLS strain was transmitted through the pericarp. These findings suggest that floral transmission, potentially mediated by pollinators, could explain the predominance of VCZ strains under favorable regional conditions, and that pericarp transmission in BLS strains could be linked to their capacity to cause disease on adult plants. Furthermore, some infections undetected in seeds became apparent after germination, indicating that testing germinated seeds rather than seeds could help seed industry to take in account only bacterial infections transmitted to the seedling. Together, these results provide valuable insights into the epidemiology of P. syringae transmission to zucchini seeds.

Xylella fastidiosa is one of the most important plant pathogens worldwide, infecting more than 700 plant species and causing major economic losses in affected agricultural regions. In the absence effective curative treatments under field conditions, experimental research is essential. However, research efforts are severely constrained by strict phytosanitary regulations and the pathogen's restricted geographic distribution, as well as its demanding culture requirements. To the best of our knowledge, no studies have compared different host infection models under controlled growth-chamber conditions. To address these limitations, we developed and compared a microinjection-based inoculation protocol across herbaceous and woody hosts maintained in biosafety growth chambers. Stems of Vitis vinifera, Nicotiana tabacum, N. benthamiana, and Medicago sativa were inoculated with X. fastidiosa by microinjection and maintained under controlled growth-chamber conditions. Pathogen establishment was assessed by qPCR analysis of petiole tissue, together with disease progression, plant growth, and leaf scorch development. Approximately one month after inoculation, bacterial detection rates reached 100% in V. vinifera and 80% in N. tabacum. Both species subsequently exhibited measurable disease development, whereas bacterial establishment and phenotypic responses were less consistent in N. benthamiana and M. sativa.

Background: Plant defensins are small cysteine-rich antimicrobial peptides. In Arabidopsis thaliana, the inducible Class I gene AtPDF1.2a is widely used as a transcriptional marker of jasmonate/ethylene immunity, whereas the constitutively expressed Class II members have received less functional attention. AtPDF2.2 was compared with AtPDF1.2a during infection of Arabidopsis by the vascular fungal pathogen Verticillium longisporum, the necrotrophic fungus Sclerotinia sclerotiorum and the bacterium Pseudomonas syringae pv. tomato DC3000. Results: Profiling of the AtPDF family showed that AtPDF2.2, together with the Class II genes AtPDF2.3 and AtPDF2.5, was significantly reduced during V. longisporum infection, whereas AtPDF1.2a was induced. A promoter-reporter line showed a corresponding decrease in AtPDF2.2 promoter activity during V. longisporum infection, strongest in infected roots. KD-pdf2.2 plants contained more V. longisporum DNA, whereas OE-PDF2.2 plants contained less. The same reciprocal disease pattern was observed during S. sclerotiorum and Pst DC3000 infection. Histochemical Prom::AtPDF2.2::GUS reporter signal decreased during S. sclerotiorum infection but increased following Pst DC3000 infection. Basal AtPDF2.2 expression was lower in JA and ET-signaling mutants and higher in the SA-deficient sid2 background, while the analyzed promoter lacked the GCC-box that mediates inducible AtPDF1.2a regulation. AtPDF1.2a overexpression reduced symptoms across the three pathosystems, but loss of AtPDF1.2a increased disease only during V. longisporum infection under the conditions tested and did not alter Pst DC3000 DNA abundance. AtPDF2.2 overexpression was not associated with reduced vegetative growth under the conditions tested, whereas AtPDF1.2a overexpression reduced rosette size and leaf number. Conclusions: AtPDF2.2 is a constitutively expressed Class II defensin whose expression level is associated with disease outcome across two fungal pathogens and a bacterium. Its promoter shows tissue-dependent regulation during V. longisporum infection, while histochemical reporter signal decreases during S. sclerotiorum infection and increases during Pst DC3000 infection. This regulatory behavior differs from that of the inducible reference gene AtPDF1.2a. The reciprocal disease phenotypes of the AtPDF2.2 transgenic backgrounds support a protective association that may involve direct antimicrobial activity, modulation of host responses or both. The absence of reduced vegetative growth in OE-PDF2.2 under the conditions tested supports further evaluation of PDF2-type genes in Brassica.

Rubber vines (Cryptostegia madagascariensis and C. grandiflora) are highly invasive weeds in the semi-arid landscapes of many tropical regions. The rust fungus Maravalia cryptostegiae has provided a spetatular and sustainable control of invasive C. grandiflora populations in Australia, after its scientific and quarantine-authorized release in the context of a classical biological control program. The Caatinga biome of Northeastern Brazil is under a devastating invasion by C. madagascariensis. This species is also susceptible to M. cryptostegia but its epidemic potential towards C. madagascariensis in Brazil remains unknown. We developed a weather-driven epidemiological model integrating experimentally derived relationships for infection as a function of temperature and wetness duration and for temperature-dependent latent development. Three infection-response models were fitted to 20 experimental temperature x wetness combinations from the literature and compared using information criteria, grouped cross-validation and a parametric bootstrap. The selected model, in which thermal breadth increases with wetness duration, was best supported (R2 = 0.931; {Delta}AICc [&ge;] 3.3 relative to the alternatives), and prolonged wetness significantly broadened the thermal range favorable for infection ({gamma} = 0.30 oC h-1; bootstrap P = 0.002). The infection and latent-development models were coupled with 25 years (2001-2025) of hourly NASA POWER data for 65 grid cells representing rubber-vine-infested locations in Northeastern Brazil, with wetness defined as hours with relative humidity equal or greater than 90%. Infection potential was concentrated in March-June (73% of the annual total, peaking in April), coinciding with the regional rainy season, and was highest in the interior of northern and central Ceara. Predicted potential varied more than a hundredfold among grid cells and was lowest along parts of the Ceara coast and the lower Jaguaribe valley, which include some of the largest infestations; the magnitude of these contrasts depended on the humidity threshold used to define wetness. Applied with identical rules to 13 historical release sites in Queensland, Australia, the model reproduced the documented wet-season timing of rust activity, and predicted potential was on average higher at sites with reported high rust impact, although the difference was not statistically significant. These results indicate strong seasonal and geographic structuring of weather-driven epidemic potential and provide testable hypotheses for selecting release periods and locations for M. cryptostegiae in Brazil.

Gray leaf spot can cause severe turfgrass damage, particularly in intensively managed sports fields, while disease management remains challenging due to the rapid development of symptoms under favorable environmental conditions. Early detection of Pyricularia oryzae could therefore provide valuable information for anticipating disease development and supporting management decisions. This study aimed to develop a digital PCR (dPCR) molecular tool to support the monitoring and management of gray leaf spot disease in stadium turfgrass. The assay showed high sensitivity, consistently detecting P. oryzae when infected tissue represented only 0.05% of a mixture with healthy turfgrass, and enabled pathogen detection before symptom appearance under both controlled and field conditions. Different matrices were also investigated across 15 stadiums to identify the most relevant material for disease monitoring. Roots showed the highest prevalence of P. oryzae detection, including during asymptomatic periods, highlighting their potential as a relevant matrix for pathogen surveillance. In a stadium monitored several months after a gray leaf spot outbreak, P. oryzae DNA was still detected in roots from 27 of the 32 sampled zones despite the absence of visible symptoms. Overall, these results demonstrate the potential of dPCR for the early detection and monitoring of P. oryzae in stadium turfgrass, particularly through root sampling during periods when environmental conditions are not favorable for gray leaf spot development, and a spatio-temporal surveillance at stadium scale.

Red crown rot of soybean (RCR), caused by Calonectria ilicicola, is an emerging soilborne disease whose quantification is challenging due to its complex symptom development across root and foliage levels. This study developed and evaluated a multi-scale framework to improve the assessment of RCR severity from controlled environments to field conditions using root imaging and standardized visual scales. Under controlled conditions, a standard area diagram (SAD) for root necrosis was developed and validated, and SAD-assisted evaluations significantly improved accuracy, precision, and inter-rater agreement compared with unaided assessments. In field conditions, a diagrammatic symptom scale (DSS) was developed using consensus-rated images from experts and showed high reliability, repeatability, and reproducibility across 18 raters, with strong intra- and inter-rater agreement. This study developed and evaluated complementary methods to improve the assessment of RCR severity from controlled environments to field conditions using root imaging and standardized visual scales.

Leptosphaeria maculans is a plant-pathogenic fungus that infects Brassica species, including Brassica napus (oilseed rape). Breeding oilseed rape varieties with genetic resistance is an efficient way to control the disease; however, L. maculans can adapt and overcome these resistances. Understanding the mechanisms that enable L. maculans to adapt is crucial for managing the emergence of better-adapted isolates. Brassica carinata, the Ethiopian Mustard, although closely related to B. napus, is considered a nonhost species of L. maculans because this fungus cannot infect it. Despite the extreme resistance of B. carinata, one natural L. maculans isolate has been identified as unable to infect B. napus, causing moderate and atypical symptoms on this species. We performed a cross between this isolate and an isolate adapted to B. napus, followed by a QTL analysis, which identified seven QTL, each encompassing candidate genes involved in L. maculans adaptation to B. carinata or B. napus. Additionally, we observed transgression in the progeny, wherein a few strains caused significantly more or less aggressive symptoms on both species of Brassica. We found that epistasis within the L. maculans genome contributes to the observed transgression. These initial findings provide further opportunities to study the adaptive capacities of L. maculans, as well as data to initiate analysis of the extreme resistance of B. carinata to L. maculans. HighlightsO_LISeven pathogenicity QTL identified, carrying several interesting candidate genes C_LIO_LITransgression of some progeny isolates on B. napus and B. carinata was reported C_LIO_LIEpistasis plays a significant role in the adaptation of L. maculans toward host and nonhost Brassica species C_LI

Background: Alternaria leaf spot (ALS), caused by fungi in the genus Alternaria (family Pleosporaceae), is a significant fungal disease that affects cotton production in the Lake Zone of Tanzania, the country's main cotton-producing region. The disease reduces photosynthetic capacity, boll retention, and fiber quality, leading to substantial yield losses in affected fields. Despite its increasing prevalence, systematic baseline epidemiological data remain scarce. Objective: This study aimed to determine the occurrence, incidence, and severity of Alternaria leaf spot across the major cotton-growing districts of the Lake Zone, Tanzania. Methods: Field surveys were conducted during the 2024-2025 growing season in the districts of Bariadi, Bunda, Chato, and Magu; 20 representative cotton fields were systematically assessed using a diagonal sampling approach. Disease incidence, prevalence, and severity were recorded for 1,000 cotton plants. Fungal isolates recovered from symptomatic leaves were sent to the laboratory for isolation and morphological identification. Results: Field prevalence was 100%, and the overall mean incidence was 64.4%, with significant inter-district variation: Bariadi (77.6%), Bunda (68.8%), Chato (60.0%), and Magu (52.4%). Mean DSI ranged from 44.0% (Magu) to 70.1% (Bariadi), indicating moderate-to-severe infection. Disease incidence and Disease severity (DSI) were strongly correlated when uninfected plants were included (r = 0.942, P < 0.001). However, this relationship weakened substantially and became non-significant when restricted to infected plants only (r = 0.396, P = 0.084). This suggests that higher incidence reflected more plants becoming infected rather than greater severity among already infected plants. Morphological characterization identified all 40 isolates as belonging to the genus Alternaria. Conclusion: These findings provide the first systematic epidemiological baseline for Alternaria leaf spot across the major cotton-growing districts of Tanzania's Lake Zone. Disease incidence and severity varied significantly across districts, underscoring the need for tailored disease management strategies. This study recommends ongoing surveillance and integrated disease management to safeguard cotton productivity among smallholder farmers in the region.

Cowpea, a crop of great importance in several regions in West Africa, faces many biotic and abiotic challenges. Several species of the genus Colletotrichum cause cowpea anthracnose disease (CAD), a major constraint to cowpea production severely affecting foliage and yield. Three Colletotrichum species: C. cliviicola, C. truncatum, and C. chrysophilum recently reported as causal agents of CAD in Nigeria were investigated in their infective processes and pathogenicity on cowpea leaves using histopathological and microscopic examination of stained leaf tissues. Inoculated leaves were examined at intervals to monitor fungal establishment, colonization, and reproductive development within host tissues. The three species completed the infection cycle, progressing from conidial germination and host penetration to tissue colonization, and sporulation. However, differences in both infective ability and pathogenicity were observed in representative isolates of each species. C. truncatum isolate was the most virulent, had extensive branching hyphae, produced severe tissue necrosis, abundant acervuli, and prominent seta formation. C. cliviicola also had substantial colonization and sporulation ability with dense conidial masses and mature acervuli. In contrast, C. chrysophilum had moderate infectivity with less extensive reproductive structures and tissue degradation. Here we provide the first histopathological report of infection and pathogenic development of these three species recently reported as causal agents of CAD

Calonectria ilicicola, an emerging and economically important soilborne pathogen in the U.S. Midwest, causes red crown rot (RCR) disease of soybean. It is essential to have an early and accurate detection method for effective management of RCR, since disease symptoms are nearly identical to those of other soybean diseases. In this study, we developed and validated a high resolution melting (HRM) assay targeting the translation elongation factor 1&alpha (TEF-1&alpha) gene for the specific detection of C. ilicicola. The assay was evaluated using a specificity panel of 78 DNA samples, including 67 fungal and oomycete DNA samples and 11 soybean host-DNA samples, as well as DNA from 56 field-collected symptomatic and asymptomatic soybean tissues. The HRM assay generated distinct melting profiles and, where applicable, absence or delayed amplification that reliably differentiated C. ilicicola from non target fungi and oomycetes. Additionally, the marker distinguished three haplotypes among C. ilicicola isolates, consistent with nucleotide polymorphisms in the TEF-1&alpha target region. The assay demonstrated strong quantitative performance, exhibiting high linearity (R^2= 0.9992) and acceptable amplification efficiency (94.05%) across a broad DNA concentration range. In field samples, HRM based detection aligned with disease status, accurately identifying C. ilicicola in symptomatic plants and no detection in asymptomatic tissues. Overall, this HRM assay provides a rapid, sensitive, and reliable diagnostic tool for routine molecular detection of C. ilicicola.

Carob (Ceratonia siliqua) is an important yet often underestimated Mediterranean tree species. Historically used as a sweetener by ancient Greeks and Arabs, it also supported populations during periods of famine. Beyond its cultural value, carob exhibits notable agronomic traits, including drought and temperature tolerance, soil erosion prevention, and potential contributions to rural development. Although a Cretan carob genome assembly has recently become available, little is known about pathogens associated with this species, particularly viruses. Here, we characterized the virome of carob trees in Crete using high throughput sequencing (HTS) and targeted molecular validation. We collected 134 leaf samples from 34 growers across the island four prefectures. HTS analysis revealed novel isolates of two Secoviridae members (Artichoke yellow ringspot virus - AYRSV) and Strawberry latent ringspot virus - SLRSV), as well as Soybean ilarvirus 1 (SolV1), a putative and incompletely characterized jivivirus, and a previously undescribed coguvirus of the family Phenuiviridae, all confirmed by PCR screening. Phylogenetic and sequence analyses support the classification of the coguvirus as a distinct viral species, for which we propose the name Carob coguvirus 1 (CaCoV1). To our knowledge, this study provides the first comprehensive characterization of the carob virome. These findings expand current knowledge of viral diversity in carob and establish a foundation for future epidemiological studies, certification of propagation material, and the sustainable expansion of carob cultivation in Crete and other Mediterranean regions.

Background: Rvi6, introgressed from Malus floribunda 821, is the most widely deployed apple scab resistance gene, but Venturia inaequalis races overcoming it are widespread. Transcriptomic differences between Rvi6-effective and Rvi6-breaking interactions in one host genotype remain unresolved. Aims: To compare these interactions in cv. Ametyst and nominate genes for subsequent validation. Methods: Own-rooted Ametyst plants were mock-inoculated or inoculated with an Rvi6-avirulent isolate (Rin; incompatible) or an Rvi6-virulent isolate (Rola; compatible) and harvested at 4 days post-inoculation (21 mRNA libraries). Expression was quantified using GDDH13 reference-based and de novo Trinity strategies, followed by DESeq2, WGCNA and GSEA. Results: Rin vs Rola yielded 1,339 candidate genes (more than two-fold change, FDR < 0.01) in the reference-based analysis. Curation of Rin-induced defense candidates retained 151 genes, 92 of which concentrated in a single co-expression module. Thirty-one also passed the Rin-vs-Rola thresholded test and were prioritized for genetic validation. Conclusions: The incompatible interaction was associated with a coherent defense program and the compatible interaction with a distinct transcriptional state. With one isolate per class, isolate identity and Rvi6 virulence cannot be separated.