Plant Pathology

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.

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.

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 [≥] 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.

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

Microbial biological control agents (mBCAs) offer a promising alternative to conventional disease management, but identifying effective strains requires screening large microbial collections against many pathogens. Here, we developed a scalable screening approach to characterize the antagonistic activity of potato-associated bacteria against multiple pathogens and investigated how bacterial origin and phylogeny relate to antagonistic phenotypes. A collection of 600 bacterial strains isolated from two potato cultivars, three plant compartments using four cultivation media was screened against six major potato pathogens: the bacteria Dickeya solani and Pectobacterium carotovorum, the fungi Alternaria solani and Rhizoctonia solani, and the oomycetes Phytophthora infestans and Pythium ultimum. A scalable confrontation method was developed for non-filamentous bacteria, while complementary lower-throughput assays were used for filamentous strains. This approach enabled successful assessment of 92% of non-filamentous and 97% of filamentous strain-pathogen combinations. Isolation cultivar, plant compartment and cultivation medium had only limited effects on the proportion of antagonistic strains. In contrast, pathogen sensitivity varied markedly: P. infestans was the most susceptible pathogen, whereas D. solani and P. carotovorum were generally resistant to bacterial antagonism. Antagonistic activity clustered phylogenetically, with Bacillus and Streptomyces containing many broad-spectrum inhibitors. In addition, several taxa showed preferential activity against the oomycetes, including P. infestans-specific antagonists from the Pseudomonas genus and P. ultimum-specific antagonists from the Frigoribacterium, Curtobacterium, Pedobacter and Phyllobacterium genera. Overall, multi-pathogen screening revealed distinct generalist and specialist antagonistic profiles and identified candidate strains and taxa for further evaluation as mBCAs.

Red crown rot (RCR), caused by Calonectria ilicicola, is an emerging soybean disease in the U.S. Midwest for which scalable approaches to characterize within-field disease distribution are lacking. This study evaluated high-resolution PlanetScope satellite imagery for mapping RCR-affected soybean canopies across 15 commercial fields in Illinois surveyed during the 2024 and 2025 growing seasons. A total of 2,921 georeferenced canopy plots were classified as asymptomatic or RCR-affected and paired with six multispectral bands and seven vegetation indices. Spectral differences between classes were evaluated using linear mixed-effects models, and seven machine-learning classifiers representing linear, tree-based, neural-network, kernel, and probabilistic approaches were compared using spatially independent leave-one-field-out cross-validation. RCR-affected canopies exhibited increased reflectance in the visible and red-edge regions, reduced near-infrared reflectance, and lower vegetation-index values relative to asymptomatic canopies. All classifiers showed strong discrimination, with ROC-AUC values ranging from 0.963 to 0.982. Regularized logistic regression achieved the highest overall performance, with an accuracy of 0.945, balanced accuracy of 0.945, F1-score of 0.948, and ROC-AUC of 0.982 at the optimized decision threshold. Permutation analysis identified EVI, NDVI, and red reflectance as the most influential predictors across representative model architectures. Satellite-derived probability and classification maps generally corresponded with symptomatic canopy patterns observed in high-resolution UAV imagery, although mixed pixels reduced precision near disease-patch boundaries. These results demonstrate the potential of high-resolution satellite imagery for within-field mapping of RCR-associated canopy symptoms across independent commercial soybean fields.

Fusarium oxysporum f. sp. lactucae (Fola), the cause of lettuce fusarium wilt, poses an increasing threat to global production due to the rapid emergence of resistance breaking isolates. However, the evolutionary origins and mechanisms underpinning this diversification remain poorly understood. We characterised 103 Fola isolates using molecular profiling, selecting ten key isolates for long-read whole-genome sequencing and performed in planta virulence assays for selected isolates. This included the first assemblies for Fola2 and Fola3, recent resistance-breaking variants and a highly divergent isolate from France designated as a putative novel race, FolaX. Phylogenomic analysis revealed that Fola is polyphyletic, with lettuce pathogenicity acquired independently across divergent F. oxysporum clades. Despite this, all lineages share a conserved "PanFola" effector set with near-identical sequences, indicating extensive horizontal gene transfer. Resistance-breaking Fola variants within established races were linked to copy number variations and targeted deletions, such as the loss of the SIX8/PSE1 locus in Fola4+. Conversely, FolaX possessed a divergent core genome and a chimeric accessory genome derived from multiple Fola lineages via segmental chromosome transfer and transposon-mediated reshuffling. Our findings highlight the remarkable evolutionary plasticity of Fola and the need for breeding strategies that address the dynamic nature of effector repertoires.

Hybridization between invasive and native species can generate novel genetic variation with ecological and evolutionary consequences, yet introgression often remains cryptic. We investigated natural hybridization between the invasive pinewood nematode Bursaphelenchus xylophilus, the causal agent of pine wilt disease, and native Bursaphelenchus mucronatus in Japan. Using multiplexed inter-simple sequence repeat genotyping by sequencing, we genotyped 342 field-collected nematodes and then conducted pedigree-calibrated analysis using HIest v2.0, based on 48 highly informative diagnostic single-nucleotide polymorphisms (SNPs), followed by ADMIXTURE, complementary NeighborNet and principal component analyses of a broader 2,756-SNP dataset. The results identified nine high-confidence hybrids (2.6%) across two consecutive years (2023-2024), including relatively balanced F1-like individuals and strongly parentally biased advanced backcross-like individuals, demonstrating recurrent, multigenerational introgression. Four hybrids (44.4%) were undetectable using a single-locus diagnostic marker, showing that advanced introgression can escape conventional surveillance. Chemotaxis assays detected no volatile-mediated pre-mating discrimination, whereas post-mating barriers were incomplete and asymmetric. Crosses with B. xylophilus as the maternal parent showed reduced hatchability, particularly with B. mucronatus mucronatus, while reciprocal crosses generally retained higher hatchability, although some showed reduced fecundity. These results provide a mechanistic basis for natural gene flow in a recent secondary contact zone. Because B. mucronatus kolymensis is widespread in Europe and produces viable hybrid offspring with B. xylophilus in both directions, pathogen establishment could create new opportunities for introgression. Our findings reveal cryptic, multigenerational introgression between an invasive forest pathogen and its native congener, highlighting the importance of genomic surveillance for forest disease management and invasion-risk assessment under climate change.

The bean thrips, Caliothrips phaseoli, is an emergent pest affecting peanut production. Thus, gaining a deeper understanding of the naturally occurring viral populations associated with this insect is of major importance, as such knowledge could support the design of novel pest management strategies that are both ecologically responsible and sustainable over the long term. In this study, we performed the first RNA high-throughput sequencing (HTS) analysis of C. phaseoli to unravel its virome, discovering a wide number of novel RNA viruses. Our HTS analysis revealed the presence of 31 viral segments, corresponding to 18 viruses, including double-stranded RNA viruses as well as positive- and negative-sense single-stranded RNA viruses. All these are novel viruses and cover multiple viral lineages belonging to diverse families. Overall, these results indicates that C. phaseoli hides a complex virome. The findings presented here provide an important framework for subsequent studies aimed at understanding the ecological significance, evolutionary patterns, and possible use of these viruses as components of biologically based management programs for this emerging peanut pest.

Pectobacterium, a member of the soft rot Pectobacteriaceae (SRP), can cause highly destructive soft rot and blackleg diseases on many important crop and other host plants, particularly potato (Solanum tuberosum). Pectobacterium and its sister genus Dickeya have common virulence strategies, yet a highly diverse "open" pangenome containing many unique and hypothetical proteins. Here, we used randomly-barcoded transposon insertion-site sequencing (RB-TnSeq) on six Pectobacterium species to identify genes involved in bacterial survival and growth in potato tubers, as well as in minimal media conditions containing different single carbon sources. The strains evaluated varied broadly in genomic content as well as host plant and year of isolation. We focused on identifying conditionally essential genes across all strains tested, as well as genes that were important for growth in one or few of the tested strains. These results highlight the importance of many common genes involved in metabolism and maintaining cellular homeostasis, as well as shared and rare virulence traits including polygalacturonate catabolism, genes involved in polysaccharide biosynthesis, immunity proteins, and (for P. brasiliense) self-tolerance of the antibiotic carbapenem. This large dataset of conditionally essential genes, generated across Pectobacterium species during soft rot infection of the agriculturally important plant host potato, can serve as a rich dataset for further hypothesis generation and identification of important and/or novel in planta fitness factors.

Sudden Death Syndrome (SDS), caused by Fusarium virguliforme, can significantly impact soybean yields. To advance the understanding of SDS resistance in plant introductions, we conducted genome-wide association studies (GWAS) on a set of 351 accessions from the USDA Soybean Germplasm Collection. The analysis revealed seven single nucleotide polymorphisms (SNPs) significantly associated with foliar SDS resistance; three significant QTL (on chromosomes 3, 9 and 11) were detected with Fixed and Random Model Circulating Probability Unification (FarmCPU) and four (on chromosomes 5, 6 and two on chromosome 7) with Bayesian-information and Linkage disequilibrium Iterative Nested Keyway (BLINK). Localized linkage disequilibrium blocks were estimated for these regions. Notably, all of them overlapped with regions where QTL associated with SDS resistance had been previously reported and where within, or close to, sequences annotated as implicated in plant defense mechanisms, such as toxin transporters and transcription factors. The architecture of SDS resistance appears quantitative and polygenic, with most loci explaining modest phenotypic variance. Six plant introductions in this study--PI 175188, PI 307889A, PI 323559, PI 578307C, PI 445683, and PI 323558--harbor favorable alleles at all seven loci, providing high-value resources for resistance introgression. This study underscores the value of leveraging the USDA Soybean Germplasm Collection for the discovery of resistance loci and provides a foundation for marker-assisted breeding to enhance SDS resistance in elite soybean lines.

Stink bugs (Hemiptera Pentatomidae) are major agricultural pests worldwide, including both native and invasive species. In France, for instance, Halyomorpha halys has rapidly expanded its range, affecting key crops such as hazelnuts, apples, and vegetables. Meanwhile, several native, polyphagous species - Nezara viridula, Palomena prasina and, to a lesser extent, Graphosoma italicum - are re-emerging and impacting diverse crops because of changes in agricultural practices and climatic conditions. Egg parasitoids represent promising biological control agents for these pests, be it through conservation, augmentation or introduction biological control. However, their actual potential is not precisely documented due to a lack of ambitious surveys and possible taxonomic issues. This study aims to address this gap by compiling results obtained from various of egg parasitism surveys of Pentatomidae in France and implementing a large-scale DNA Barcoding approach (2000 individuals) for the emerged parasitoids. Our results indicate a high diversity of egg parasitoids, and detected some taxonomic issues and molecular inconsistencies that warrant further study. Among the three families of egg parasitoids recovered (Encyrtidae, Eupelmide, and Scelionidae), Scelionidae was the most abundant and diverse with nine species of Trissolcus (including the two adventive species, Tr. japonicus and Tr. mitsukurii) and four Telenomus species (including two unidentified to species). Scelionidae dominated the parasitoid assemblages associated with most of the Pentatomidae host species, except for H. halys for which the eupelmid Anastatus bifasciatus was the most common species.

Global change has a major impact on rivers and their associated terrestrial compartment, the riparian forest. The black poplar (Populus nigra L.), the dominant species of the European riparian forest, is considered an indicator of the biodiversity dynamics of these ecosystems, but is threatened with extinction in several parts of its range. The species is dioecious and shows wide variability in many traits, enabling it to adapt to a broad range of environmental conditions. A detailed characterization of this natural variability is therefore a key issue both for the conservation of natural populations and for biological engineering aimed at restoring riverbanks. Phenology is both an indicator of climate change and an adaptation lever for populations, particularly in perennial species such as trees. In this study, we characterized the spatial and temporal variability in flowering phenology by studying 13 natural black poplar populations along the Loire River, France. Flowering date and duration showed substantial variability both within and between populations, and males generally flowered earlier than females. Despite this variability, the ranking of individual trees' flowering dates was highly consistent across years. Most male-female pairs shared several days of overlapping flowering each year, and this overlap was moderately stable across years. Flowering dates advanced by about five to six days per degree Celsius during early spring, with no detectable difference in sensitivity between sexes. This work contributes to the characterization of flowering variability in an economically important tree species and may inform both the conservation of genetic resources and breeding.

A native entomopathogenic nematode isolate, Heterorhabditis indica Che4.1, was recovered from soils in Valencia (Spain), and its symbiotic bacterium, Photorhabdus akhurstii Che4.1, was isolated and characterized. Nematode identification was confirmed by ITS and 28S rDNA sequencing, while genome-based analyses supported the taxonomic assignment of the bacterial symbiont. The nematode-bacterium complex exhibited insecticidal activity against Spodoptera exigua and Spodoptera littoralis. H. indica Che4.1 showed an LC of 53.7 infective juveniles (IJs)/mL against S. exigua, comparable to that of the commercial reference Larvanem (54.3 IJs/mL). Against S. littoralis, Che4.1 exhibited an approximately twofold lower LC than Larvanem (77.3 versus 155.9 IJs/mL), with non-overlapping 95% confidence intervals. The complete genome of P. akhurstii Che4.1 consists of a single circular chromosome of 5.66 Mb encoding a diverse repertoire of putative virulence factors, including toxin complex (Tc) components, YenTc-like proteins, Mcf toxins, App-family proteins, Pra/Prb proteins, and chitinases, together with several predicted secondary metabolite biosynthetic gene clusters. LC-MS/MS analyses of bacterial culture supernatants and infected insect cadavers detected multiple predicted virulence-associated proteins, including Tc-related, YenTc-like, App-family, and Pra/Prb proteins. In addition, bacterial cell-free supernatants caused 68.8-81.3% mortality in S. exigua neonates under laboratory conditions. Together, these findings reveal a diverse repertoire of potential virulence determinants in P. akhurstii Che4.1 and demonstrate the insecticidal potential of its native nematodebacterium complex. The performance of H. indica Che4.1 relative to a commercial reference supports further evaluation of this isolate for biological control of Spodoptera spp., particularly under greenhouse and field conditions.

The potential of alternative agricultural systems, such as organic or conservation agriculture, for biodiversity restoration has been widely investigated. However, variability of practices within each of these two categories of agricultural system has rarely been accounted for. Furthermore, these systems have most often been studied separately, isolating either organic or conservation agriculture (itself often reduced to a soil tillage reduction), and rarely analyzing these systems side by side. This study attempts to disentangle these multiple interrelationships by sampling carabid beetles in a network of 48 commercial Belgian farms including organic agriculture (OA), conservation agriculture (CA), farms combining the two approaches (OCA) and farms applying conventional practices (CVA). In each farm, we sampled carabids using pitfall and emergence traps in one winter cereal field and collected a detailed description of crop management practices. We found that community composition was primarily structured by seasonality early in the season (March, April) followed by convergence (May, June) driven by the aggregation of the most dominant species. Based on agroecosystem classification, organic certification (i.e., the absence of synthetic pesticide use) exerted a twofold stronger effect than application of conservation agriculture principles (i.e., reduced or no soil tillage) on carabid beetle community composition. Notably, OCA supported carabid assemblages that were structurally distinct from those of CA and CVA. Furthermore, carabid communities associated with OA differed significantly in composition from those of CVA. Observed and estimated species richness were also significantly higher in OCA than in CA and CVA, while OA differed from CVA only for observed richness. Beyond differences among agricultural systems, community responses were associated with specific management practices, which explained nearly a quarter of the compositional variation with significant influence of the duration since organic conversion and the frequency of synthetic pesticide application. Carabid abundances were negatively affected by synthetic pesticides and moldboard ploughing, whereas richness was more strongly related to the time since conversion to OA, cover crops, and organic amendments. Overall, these results position synthetic inputs and time since conversion as the main filters over carabid communities. Our study highlights the importance of jointly considering system level analyses and detailed management practices to better understand the complex relationships between agricultural practices and carabid communities.

Bulk phloem flow is driven by pressure gradients generated by water drawn from the xylem, coupling source-sink transport with water status. Huanglongbing (HLB), a phloem-affecting bacterial disease of citrus, disrupts phloem transport, and causes canopy water deficit. Although improved water management can enhance HLB-affected tree health, the effects of HLB on water-driven source-sink relations are unknown. Fruits function as both carbon sink and water reservoir, but the role of fruit in regulating whole-tree water status remain poorly understood. We examined leaf-fruit water relations in HLB-affected citrus and evaluated how HLB exposure duration, fruit developmental stage, and source-sink manipulation shape these dynamics. Source-sink relations were manipulated in branches of symptomatic trees with long-term (four years) and recent (six months) HLB exposure. We girdled branches with a single fruit as the sink and defoliated at (0%, 50%, or 75%), and an ungirdled, non-defoliated branch served as control. We measured stomatal conductance, leaf hydraulic traits, leaf and fruit soluble sugars, diurnal fruit shrinkage (to estimate xylem backflow) and fruit water loss. Source-sink manipulation did not affect shoot total soluble sugar. HLB-exposure resulted in less negative osmotic potential, more negative turgor potential and reduced modulus of elasticity and relative water content at turgor loss point. HLB exposure also increased diurnal fruit shrinkage, with shrinkage responses to source-sink manipulation varying across fruit development stages. Collectively, these findings suggest that long-term HLB exposure promotes leaf acclimation to water deficits, however, high sink demand may diminish the leaf water status regulation at Stage II, when fruit sink carbon demand is highest.

Ralstonia pseudosolanacearum causes a lethal bacterial wilt disease in many plants. The pathogen enters through roots and systemically colonizes the xylem tissue, resulting in wilting and eventual death of the host. Using transposon mutagenesis followed by screening in tomato seedlings, we identified a mutant exhibiting virulence deficiency by root inoculation, but not by leaf inoculation. The mutation was recreated in the wild-type genome via natural transformation to prove the linkage of virulence deficiency with the transposon insertion. Whole genome sequencing revealed that the transposon insertion was in a cyp450 homologue, encoding cytochrome P450 monooxygenase. Two independent targeted mutations were generated in the cyp450 gene, and both the mutants recapitulated the root-specific virulence deficiency. The mutants rarely cause disease in mature plants following soil drenching but remained fully virulent upon petiole pricking inoculation. Complementation with the full length cyp450 gene amplified from the wild-type strain F1C1 restored virulence in the mutant. Compared to the wild-type F1C1, the cyp450 mutants exhibited greater sensitivity to tomato root extracts. HR-LCMS of tomato root extracts revealed the occurrence of several antimicrobial compounds. Cytochrome P450 enzymes are known to mediate the degradation or biosynthesis of diverse metabolites in bacteria. Considering the virulence difference due to the mode of inoculation, our findings indicate that cyp450 contributes to tissue-specific adaptation of the pathogen during infection has opened an avenue for integrating tissue metabolomics with bacterial wilt susceptibility research.