Leaf Pathogen Research Articles

First line of defence: Eucalyptus leaf waxes influence infection by an aggressive fungal leaf pathogen.

Leaf epicuticular waxes provide important anatomical and chemical defences against fungi that infect leaves. In this study we analysed the leaf wax composition of Eucalyptus grandis × Eucalyptus urophylla hybrids with contrasting susceptibilities to Teratosphaeria leaf blight (TLB) caused by Teratosphaeria destructans, one of the most important foliar diseases of Eucalyptus. The Eucalyptus cuticular wax was extracted from non-inoculated and inoculated genotypes with different levels of susceptibility to TLB and analysed by gas chromatography-mass spectrometry. The results showed that a triterpenoid, cycloartenol (CAS), was abundant in a resistant genotype and that hexanedioic acid content increased in the resistant genotypes in response to T. destructans infection. In contrast, palmitic acid was significantly more abundant in the inoculated highly susceptible genotype. In-vitro and in-planta T.destructans spore germination assays with pure compounds, showed that CAS and hexanedioic acid significantly inhibited spore germination. Application of these two compounds to the leaves of a susceptible host also significantly increased resistance to infection. In contrast, palmitic acid promoted spore germination and, when applied to the leaves of a resistant genotype, increased colonization by the pathogen. This is the first study providing insights into differences in the leaf wax composition of hosts with different levels of susceptibility to T. destructans. It also showed that leaf wax compounds can modulate spore germination and, ultimately, host resistance to infection.

The impact of anthropogenic pressures on microbial diversity and river multifunctionality relationships on a global scale

Conserving biodiversity is crucial for maintaining essential ecosystem functions, as indicated by the positive relationships between biodiversity and ecosystem functioning. However, the impacts of declining biodiversity on ecosystem functions in response to mounting human pressures remain uncertain. This uncertainty arises from the complexity of trade-offs among human activities, climate change, river properties, and biodiversity, which have not been comprehensively addressed collectively. Here, we provide evidence that river biodiversity was significantly and positively associated with multifunctionality and contributed to key ecosystem functions such as microbially driven water purification, leaf litter decomposition and pathogen control. However, human pressure led to abrupt changes in microbial diversity and river multifunctionality relationships at a human pressure value of 0.5. In approximately 30 % (N = 58) of countries globally, the ratio of area above this threshold exceeded the global average (∼11 %), especially in Europe. Results show that human pressure affected ecosystem functions through direct effects and interactive effects. We provide more direct evidence that the nonadditive effects triggered by prevailing human pressure impact the multifunctionality of rivers globally. Under high levels of human stress, the beneficial effects of biodiversity on nutrient cycling, carbon storage, gross primary productivity, leaf litter decomposition, and pathogen control tend to diminish. Our findings highlight that considering interactions between human pressure and local abiotic and biotic factors is key for understanding the fate of river ecosystems under climate change and increasing human pressure.

Endophytic Fungi Inoculation Reduces Ramulosis Severity in Gossypium hirsutum Plants.

Biotic stress in cotton plants caused by the phytopathogenic fungus Colletotrichum gossypii var. cephalosporioides triggers symptoms of ramulosis, a disease characterized by necrotic spots on young leaves, followed by death of the affected branch's apical meristem, plant growth paralysis, and stimulation of lateral bud production. Severe cases of ramulosis can cause up to 85% yield losses in cotton plantations. Currently, this disease is controlled exclusively by using fungicides. However, few studies have focused on biological alternatives for mitigating the effects of contamination by C. gossypii var. cephalosporioides on cotton plants. Thus, the hypothesis raised is that endophytic fungi isolated from an Arecaceae species (Butia purpurascens), endemic to the Cerrado biome, have the potential to reduce physiological damage caused by ramulosis, decreasing its severity in these plants. This hypothesis was tested using plants grown from seeds contaminated with the pathogen and inoculated with strains of Gibberella moniliformis (BP10EF), Hamigera insecticola (BP33EF), Codinaeopsis sp. (BP328EF), G. moniliformis (BP335EF), and Aspergillus sp. (BP340EF). C. gossypii var. cephalosporioides is a leaf pathogen; thus, the evaluations were focused on leaf parameters: gas exchange, chlorophyll a fluorescence, and oxidative metabolism. The hypothesis that inoculation with endophytic strains can mitigate physiological and photochemical damage caused by ramulosis in cotton was confirmed, as the fungi improved plant growth and stomatal index and density, increased net photosynthetic rate (A) and carboxylation efficiency (A/Ci), and decreased photochemical stress (ABS/RC and DI0/RC) and oxidative stress by reducing enzyme activity (CAT, SOD, and APX) and the synthesis of malondialdehyde (MDA). Control plants developed leaves with a low adaxial stomatal index and density to reduce colonization of leaf tissues by C. gossypii var. cephalosporioides due to the absence of fungal antagonism. The Codinaeopsis sp. strain BP328EF can efficiently inhibit C. gossypii var. cephalosporioides in vitro (81.11% relative inhibition), improve gas exchange parameters, reduce photochemical stress of chlorophyll-a, and decrease lipid peroxidation in attacked leaves. Thus, BP328EF should be further evaluated for its potential effect as a biological alternative for enhancing the resistance of G. hirsutum plants and minimizing yield losses caused by C. gossypii var. cephalosporioides.

Defense Pathways of Wheat Plants Inoculated with Zymoseptoria tritici under NaCl Stress Conditions: An Overview

Due to being sessile, plants develop a broad range of defense pathways when they face abiotic or biotic stress factors. Although plants are subjected to more than one type of stress at a time in nature, the combined effects of either multiple stresses of one kind (abiotic or biotic) or more kinds (abiotic and biotic) have now been realized in agricultural lands due to increases in global warming and environmental pollution, along with population increases. Soil-borne pathogens, or pathogens infecting aerial parts, can have devastating effects on plants when combined with other stressors. Obtaining yields or crops from sensitive or moderately resistant plants could be impossible, and it could be very difficult from resistant plants. The mechanisms of combined stress in many plants have previously been studied and elucidated. Recent studies proposed new defense pathways and mechanisms through signaling cascades. In light of these mechanisms, it is now time to develop appropriate strategies for crop protection under multiple stress conditions. This may involve using disease-resistant or stress-tolerant plant varieties, implementing proper irrigation and drainage practices, and improving soil quality. However, generation of both stress-tolerant and disease-resistant crop plants is of crucial importance. The establishment of a database and understanding of the defense mechanisms under combined stress conditions would be meaningful for the development of resistant and tolerant plants. It is clear that leaf pathogens show great tolerance to salinity stress and result in pathogenicity in crop plants. We noticed that regulation of the stomata through biochemical applications and some effort with the upregulation of the minor gene expressions indirectly involved with the defense mechanisms could be a great way to increase the defense metabolites without interfering with quality parameters. In this review, we selected wheat as a model plant and Zymoseptoria tritici as a model leaf pathogen to evaluate the defense mechanisms under saline conditions through physiological, biochemical, and molecular pathways and suggested various ways to generate tolerant and resistant cereal plants.

First records of Microstroma album (Desmazières) Saccardo, 1878 (Basidiomycota: Microstromataceae) and Phylloxera glabra (von Heyden, 1837) (Hemiptera: Phylloxeridae) in oak stands in Bulgaria

The current study reports the first records of an aphid-like pest and leaf pathogen on oak species in Bulgaria. In 2020-2023, leaf samples presenting characteristic yellow spots and lesions were collected from mature trees of Quercus petraea, Q. cerris, Q. frainetto and Q. robur in different localities of the country. Damage caused by oak obligate leaf pathogen Microstroma album, the leaf phylloxera, Phylloxera glabra, and the oak lace bug, Corythucha arcuata were identified on the lower surface of oak leaves. M. album was identified on all studied Quercus species. Angular white patches, the fruiting stage of the fungus, appeared on the lower surfaces of the leaves. P. glabra caused yellowish mosaic discoloration on the leaf surface of Quercus cerris, Q. frainetto and Q. pertaea. Sucking damage displayed a stippled pattern of yellow, brown, or whitish dots. In places where M. album develops, the invasive oak lace bug was often found as the only sucking species on oak leaves, which is most likely also a vector of the disease. Severe damage caused by established both obligate pathogen (M. album) and sucking insect pests (P. glabra, C. arcuata) could result in a weakening of the oak trees. On sufficiently weakened trees, secondary fungal pathogens or boring insects, commonly infested oak trees, can cause their death.

Studies on variability in mode of infection by the isolates of leaf pathogens isolated from diseased leaves of Musa paradisiaca from Uttar Dinajpur District

Banana is (Musa sp) one of the most exoteric and materialistic fruit crop of many tropical and subtropical areas in India. Banana leaf diseases, caused by several fungal pathogens are one of the major diseases. Among the fungi causing banana leaf diseases, Curvularia sp/C. lunata is one of the main causal organisms causing leaf spot disease in banana plant. Two fungal isolates- MUSLF-2, MUSLF-3 and one fungal isolate- MUSLF-1 were isolated from the infected leaf from Ramganj and Gotgaon locations. Based on light microscopic and scanning electron microscopic studies, MUSLF-2 and MUSLF-3 were identified as Curvularia sp. After Koch’s postulate, three fungal isolates- MUSLF-2, MUSLF-3 [Curvularia sp] and MUSLF-1 were applied in banana field along with foliar spray of spore suspensions. The symptoms were similar to those observed in the in vitro detached leaf inoculation technique. The present research work depicts about the severity of leaf spot diseases of Musa paradisiaca and the nature of mode of infections exhibited by the fungal pathogens.

The interplay between defaunation and phylogenetic diversity affects leaf damage by natural enemies in tropical plants

Abstract Natural enemies play an important role in controlling plant population growth and vegetation dynamics. Tropical rainforests host the greatest diversity of herbivores, from large mammalian ungulates to microscopic pathogens, generating and maintaining plant diversity. By feeding on the same resources, large mammalian herbivores may interfere with plant consumption and leaf damage by important enemy guilds such as invertebrate herbivores and pathogens, triggering indirect trophic cascades. However, the impact of local extinctions of large herbivores on plant–enemy interactions is relatively unknown. We experimentally tested the effects of defaunation of large mammalian herbivores (e.g. peccaries, tapirs and brocket deer; hereafter, large herbivores) on the leaf damage of 3350 understorey plants in tropical rainforests of Brazil. We examined leaf damage in 10,050 leaves from 333 morphospecies by assigning the area consumed or damaged by five guilds of insect herbivores and leaf pathogens within 86 paired open‐closed plots and investigating the joint effects of defaunation and plant phylogenetic diversity. Plants released from large herbivores had 9% less leaf damage; this difference was due to the lower leaf pathogens incidence (29%) rather than insect herbivory. Evolutionary distinctness was positively correlated with leaf damage in a similar way in all treatments, suggesting additive effects of defaunation and phylogenetic diversity. Total and pathogenic leaf damage (but not insect damage) decreased with plant richness across treatments, and large herbivores exclusion resulted in increased plant species richness. This suggests that large herbivores exclusion leads to a dilution of total and pathogens' leaf damage by increasing plant species richness. Our results suggest that indirect effects of large herbivores decrease the dilution potential of plant communities against pathogens and rather reinforce their top‐down impact on vegetation, demonstrating a previously overlooked cascading effect of large herbivore extinction on forest ecosystems. Synthesis: The extinction of large mammalian herbivores can lead to a decrease in pathogen‐driven leaf damage, a previously unknown indirect effect in forest ecosystems, which might have consequences for plant fitness and ultimately for plant diversity. Large herbivores and plant pathogens might have synergistic effects in regulating the diversity of plant communities in some of the most diverse ecosystems on Earth.

Studies on variability in mode of infection by the isolates of leaf pathogens isolated from diseased leaves of <i>Musa paradisiaca</i> from Uttar Dinajpur District, West Bengal

Studies on variability in mode of infection by the isolates of leaf pathogens isolated from diseased leaves of <i>Musa paradisiaca</i> from Uttar Dinajpur District, West Bengal

Neofusicoccum parvum: A NOVEL PATHOGEN SPECIES CAUSING WILTED LEAF AND DIEBACK PETIOLES ON LOTUS (Nelumbo nucifera) IN THUA THIEN HUE, VIETNAM

Lotus (Nelumbo nucifera) is an essential species in many countries. In Vietnam, the lotus is a plant with cultural and spiritual significance, representing purity, spiritual growth, and enlightenment. However, petiole dieback and wilted dry leaves are severe diseases that weaken the host and decrease leaf photosynthesis, reducing lotus production. In this study, a new dieback and wilted leaf pathogen were identified via its morphology, phylogeny, and pathogenicity. Four representative Botryosphaeriaceae isolates from lotus fields in Hue, Phong Dien, and Phu Loc were selected for identification and pathogenicity testing. Based on morphological and phylogenetic analyses and by using the ribosomal internal transcribed spacer region (ITS) and β-tubulin (tub-2) gene sequences, we identified four isolates as Neofusicoccum parvum. In the pathogenicity test, typical symptoms appear on the inoculated lotus petioles, including dieback, curving, and wilted leaves. These symptoms are consistent with those observed in the field. In addition to identifying the pathogen species responsible for lotus disease, this study provides valuable insights into the taxonomy and phylogenetic relationships of new fungal pathogens that affect lotus fields. These findings can contribute to effective management strategies to control these diseases and improve lotus production. To the best of our knowledge, this is the first report of characterization and phylogenetic analysis of N. fusicoccum as the causal agent of wilted leaves and dieback disease in Thua Thien Hue, Vietnam.

A GH81-type β-glucan-binding protein enhances colonization by mutualistic fungi in barley

Cell walls are important interfaces of plant-fungal interactions, acting as robust physical and chemical barriers against invaders. Upon fungal colonization, plants deposit phenolics and callose at the sites of fungal penetration to prevent further fungal progression. Alterations in the composition of plant cell walls significantly impact host susceptibility. Furthermore, plants and fungi secrete glycan hydrolases acting on each other's cell walls. These enzymes release various sugar oligomers into the apoplast, some of which activate host immunity via surface receptors. Recent characterization of cell walls from plant-colonizing fungi has emphasized the abundance of β-glucans in different cell wall layers, which makes them suitable targets for recognition. To characterize host components involved in immunity against fungi, we performed a protein pull-down with the biotinylated β-glucan laminarin. Thereby, we identified a plant glycoside hydrolase family 81-type glucan-binding protein (GBP) as a β-glucan interactor. Mutation of GBP1 and its only paralog, GBP2, in barley led to decreased colonization by the beneficial root endophytes Serendipita indica and S.vermifera, as well as the arbuscular mycorrhizal fungus Rhizophagus irregularis. The reduction of colonization was accompanied by enhanced responses at the host cell wall, including an extension of callose-containing cell wall appositions. Moreover, GBP mutation in barley also reduced fungal biomass in roots by the hemibiotrophic pathogen Bipolaris sorokiniana and inhibited the penetration success of the obligate biotrophic leaf pathogen Blumeria hordei. These results indicate that GBP1 is involved in the establishment of symbiotic associations with beneficial fungi-a role that has potentially been appropriated by barley-adapted pathogens.

Ironwood/hophornbeam leaf rust, an emergent disease across the southeastern United States affiliated to Melampsoridium asiaticum

In the late fall of 2018, foliar rust (referred to as ironwood/hophornbeam leaf rust [IHLR]) was discovered in several counties in Florida, United States, on ironwood (Carpinus caroliniana) and hophornbeam (Ostrya virginiana), both members of the Betulaceae. Uredinia were observed on leaves and, in some cases, samaras of both species at numerous locations on trees of all age classes. Similar reports across the southeastern United States (Georgia, the Carolinas, Tennessee, and Texas) were detected the following year, with European hornbeam (Carpinus betulus) being reported as an additional host, while Ostrya virginiana var. guatemalensis in El Salvador was also discovered showing signs of uredinial infection. Field observations and analyses of morphological data obtained with light and scanning electron microscopy on IHLR and related Melampsoridium herbarium samples and combined molecular data from the ITS and LSU loci indicate that (i) IHLR across samples from the southeastern United States and El Salvador belong to the same taxon; (ii) IHLR is closely affiliated to M. asiaticum; and (iii) some taxonomic modifications might be necessary at the genus level. No alternative (gametophyte) host has been identified, and the rust is likely overwintering in the uredinial stage. This disease represents a novel leaf pathogen on these hosts in North America, and efforts are needed to monitor future disease epidemiology and impacts on these native and cultivated tree species.

First Report of Alternaria alternata Causing Leaf Spot on Broccoli in Ecuador.

In Ecuador, broccoli (Brassica oleracea var. italica) production is located in the Andean region, specifically Cotopaxi-Ecuador (INEC, 2019). A leaf pathogen has been constantly observed in this area, showing brown circular necrosis surrounded by yellowish halo-like spots causing leaf death (Fig. 1a). This pathogen was believed to be Alternaria sp.; however, the species was not determined either using classical or molecular tools. In 2021, ten leaves showing similar symptoms were collected in Cotopaxi and sent for pathogen identification. Here, leaf explants (0.25 cm2) showing disease symptoms were surface sterilized with 2% sodium hypochlorite (NaClO) and 70% ethanol (C2H6O), rinsed with sterile water, and transferred to Potato Dextrose Agar (PDA) media. Petri dishes were incubated in darkness at 25°C for five days. The single hyphal tip method was used to purify the cultures on PDA. Fifteen pure isolates were obtained after incubating for 14 days. Isolates were incubated under blacklight for two days to induce fungal sporulation. All isolates presented early white cotton-like mycelium that later became dark green (Fig 1b). Under the microscope, we observed straight primary conidia in simple or branched chains. Also, the conidia were obclavate, long ellipsoids, moderate in size (19.5-43.9 μm in length, 7.1-17.2 μm in width), and septate with few longitudinal septa. Lastly, the conidium body can narrow itself into a secondary conidia (Fig 1c) (Woudenberg et al., 2013). According to colony and conidia morphology, isolates were identified as Alternaria sp. (Woudenberg et al., 2013). Five isolates were randomly selected for DNA extraction and sequencing of ITS (internal transcribed spacer; Chou, H.H. and Wu, W.S. 2002), TEF (translation elongation factor; O'Donnell et al., 1998), and RPB2 (RNA polymerase II second largest subunit; Liu et al., 1999) gene regions. DNA sequences obtained from each marker were identical for all isolates. Consensus sequences and alignment were built using ClustalX in MEGA X (Kumar et al., 2018). Consensus sequences were deposited in GenBank with the following accession numbers: ITS, ON982232; TEF, ON983964; RPB2, ON983963. A multilocus Bayesian inference phylogenetic tree was constructed in Beast software (version 1.8.4) using the concatenated sequences (Drummond et al., 2012; Maharachchikumbura et al., 2014). The isolates in our study clustered with isolates of Alternaria alternata, confirming their identity (Figure 2). For Koch's postulates, healthy broccoli plants were grown in sterile soil for six weeks. The fungal conidia were suspended in sterile distilled water (1×106 conidia/ml), and the leaves were inoculated by spraying the spore solution. The control treatment was sprayed with sterile distilled water alone. Plants were maintained at 28°C and had more than 85% relative humidity (Sigillo et al., 2020). Seven days after inoculation, plants showed chlorosis and necrosis. Ten days later, 100% of the treated leaves presented brown circular necrosis (Fig. 1d). Control plants showed no disease symptoms. Re-isolation of the pathogen from the diseased leaf tissue was performed as previously described. The isolates presented the exact morphology of pure cultures obtained from field-diseased leaves. The pathogenicity test was performed twice. To our knowledge, this is the first report on A. alternata being the causal agent of leaf spot on broccoli in Ecuador. Disease diagnosis contributes to providing strategies against this pathogen. Further investigations are needed to find biological/chemical techniques or cultivar resistance to control this pathogen in broccoli.

Effects of tree species diversity and conspecific seedling density on insect herbivory and pathogen infection on big‐leaf mahogany seedlings

The Janzen–Connell hypothesis (JCH) predicts that attack by specialist enemies on seedlings increases with conspecific seedling density, but studies have rarely experimentally tested for the contingency of such effects on the tree community context (e.g. diversity, composition) ormeasured responses by different enemies (e.g. herbivores, pathogens). We conducted a field study in a large‐scale system evaluating tree species diversity and conspecific density effects on leaf damage on mahogany Swietenia macrophylla seedlings. We established quadrats of eight levels of seedling density across mahogany tree monocultures and tree species polycultures including mahogany, and recorded percent leaf damage by insects and percent leaf necrosis by a pathogenic fungus on mahogany seedlings. We found contrasting effects of tree species diversity on insects and pathogens. Whereas tree diversity did not affect leaf damage by insects, it had a significant negative effect on leaf necrosis by pathogens whereby, on average, percent leaf necrosis on mahogany seedlings in polyculture was half of that observed in monoculture. We discuss the potential influences of changes in mahogany tree density versus frequency (relative to other tree species) driving this diversity effect. Unexpectedly, we found no effect of seedling conspecific density on either leaf insect or pathogen damage (i.e. density‐independent attack). Likewise, we found no significant tree diversity by seedling density interaction, indicating that plant enemies were consistently unresponsive to variation in seedling density acrosslevels of tree diversity. Overall, this study provides a unique test of the JCH by experimentally evaluating seedling density and tree diversity effects on contrasting plant enemies and, in turn, the controls they exert over plant recruitment.

Colletotrichum fructicola Causal agent of Shot-Hole Symptoms on Leavesof Prunus sibirica in China.

Prunus sibiricaL. (Siberian apricot) is a member of the Rosaceae family and an ecologically important tree species in China (Buer et al., 2022). Shot hole symptoms on the leaves were observed in five Siberian apricot groves in Chengdu (103.81 E, 30.97 N), Sichuan province in July 2020. The symptoms first appeared as small purplish-brown spots with yellow rings around them. As the disease progressed, the damaged area (diameter 1.5-3.0 cm) became necrotic and fell off. The disease incidence was about 60% and the disease index was 28.6 of leaves in the grove. in most severe cases. Fifteen symptomatic leaves were collected from 5 different trees in an orchard. Pathogen isolation was performed from symptomatic leaf tissue (5 × 5 mm) though surface disinfection (in 70% ethanol and 2% NaClO) and incubation on Potato Dextrose Agar (PDA) at 28℃ for 3 days. Overall 10 isolates with similar colony morphology wereobtained from the 15 infected tissue pieces, and three representative isolates (XCK 2-4) were selected for further study. Colonies of the isolates on PDA were initially cottony, pale white to grayish-green with abundant aerial hyphae and produced conidial masses after 7 days. Conidiogenous cells were clavate and aggregated in acervuli. Conidia were smooth-walled, single-celled, straight, and slightly obtusely rounded at both ends, 12.8 to 18.7 × 4.3 to 5.7 μm in size (Fig. 1). The morphological characteristics of the three isolates were consistent with the description of species in the Colletotrichum gloeosporioides complex. DNA was amplified using the following primers pairs for the internal transcribed spacer (ITS) region of rDNA and partial sequences of beta-tubulin (TUB2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), chitin synthase (CHS-1), and translation elongation factor (TEF-1), respectively: ITS1/ITS4, T1/Bt2b, GDF/GDR, CHS-F/CHS-R, and EF-F/EF-R (Vieira et al., 2014). Accession numbers (MW228049, MW284974, MW284976, MW284975 and MW284977, respectively) were obtained afterepositing all the resulting sequences in GenBank. Nucleotide blast showed 99 to 100% identities with Colletotrichum fructicola (GenBank accessions nos. MZ961683, MW284974, MN525881, MN525860, MF627961). Phylogenetic analysis of combined ITS-TUB-GAPDH genes using the Mrbayes inference method showed that the three isolates clustered with three reference isolates of C. fructicola as a distinct clade(Fig. 2). To verify Koch's postulates, ten 3-year-old healthy potted plants ofP.sibirica were inoculated by spraying a conidial suspension (6 × 105 conidia/mL) of isolate XCK2 on both sides of leaves, and the control leaves were sprayed with sterile water. Then, all treatments were placed in a moist environment (25±2°C, 80% relative humidity, natural light). The inoculated plants showed typical symptoms of plants with natural infections, while the controls remained asymptomatic after 14 days. The pathogen C. fructicola was re-isolated from all inoculated plants, and the culture and fungus characteristics were the same as those of the original isolate. Colletotrichum fructicola was not isolated from the control plants. The results indicated that C. fructicola is the causal agent of the disease. Colletotrichum fructicola was reported as a leaf pathogen on Camellia chrysanthain China (Zhao et al., 2021). This is the first report of C. fructicola causing P. sibirica leaf shot-hole in the world. The identification of C. fructicola could provide relevant information for applying management strategies and research on the Siberian apricot disease.

Increased soil cadmium concentrations enhanced poplar defence against a leaf pathogenic fungus infection

AbstractSince heavy metals are toxic to herbivores and pathogens, the Elemental defence hypothesis suggested that metals accumulated in plants might serve as defence against herbivores and pathogen attacks. However, several studies have tested this hypothesis with regard to pathogen resistance are limited. Most existing studies used herbaceous species as study model while studies on tree species are still lacking. In this study, we used poplar as a study model, to investigate whether soil treatment with five different cadmium (Cd) concentrations can protect plants from artificial infection of a leaf pathogenic fungus. The results showed that the Cd concentrations accumulated in plant leaves were positively correlated with increasing Cd concentrations amended in the soil while negatively correlated with the disease severity index (DSI) of plants after pathogen infection. Additional partial correlation analysis indicated that the plant DSI was only significantly correlated with Cd concentration accumulated in plant leaves while plant growth parameters and other leaf substances showed no clear correlations. The in vitro toxicity test of fungal mycelium grown on a series of Cd concentrations amended in the artificial medium further showed that the mycelium growth decreased with increasing Cd concentrations contained in the medium, which confirmed the biological toxicity of Cd on this fungus independently of other organic defensive substances present in poplar leaves. In conclusion, our study supported the Elemental defence hypothesis and proved that even a lower accumulation of Cd in poplar plants can provide sufficient protection against leaf pathogen infection.

Nucleoredoxin gene SINRX1 negatively regulates tomato immunity by activating SA signaling pathway

The tomato (Solanum lycopersicum) is widely consumed globally and renowned for its health benefits, including the reduction of cardiovascular disease and prostate cancer risk. However, tomato production faces significant challenges, particularly due to various biotic stresses such as fungi, bacteria, and viruses. To address this challenges, we employed the CRISPR/Cas9 system to modify the tomato NUCLEOREDOXIN (SlNRX) genes (SlNRX1 and SlNRX2) belonging to the nucleocytoplasmic THIOREDOXIN subfamily. CRISPR/Cas9-mediated mutations in SlNRX1 (slnrx1) plants exhibited resistance against bacterial leaf pathogen Pseudomonas syringae pv. maculicola (Psm) ES4326, as well as the fungal pathogen Alternaria brassicicola. However, the slnrx2 plants did not display resistance. Notably, the slnrx1 demonstrated elevated levels of endogenous salicylic acid (SA) and reduced levels of jasmonic acid after Psm infection, in comparison to both wild-type (WT) and slnrx2 plants. Furthermore, transcriptional analysis revealed that genes involved in SA biosynthesis, such as ISOCHORISMATE SYNTHASE 1 (SlICS1) and ENHANCED DISEASE SUSCEPTIBILITY 5 (SlEDS5), were upregulated in slnrx1 compared to WT plants. In addition, a key regulator of systemic acquired resistance, PATHOGENESIS-RELATED 1 (PR1), exhibited increased expression in slnrx1 compared to WT. These findings suggest that SlNRX1 acts as a negative regulator of plant immunity, facilitating infection by the Psm pathogen through interference with the phytohormone SA signaling pathway. Thus, targeted mutagenesis of SlNRX1 is a promising genetic means to enhance biotic stress resistance in crop breeding.

Variation in the Outcome of Plant-Mediated Pathogen Interactions in Potato: Effects of Initial Infections on Conspecific vs. Heterospecific Subsequent Infections

Plants are often attacked sequentially by multiple enemies. Pathogen sequential co-infections can lead to indirect interactions mediated by plant induced responses whose outcome is contingent on differences in the magnitude and type of plant induced defences elicited by different species or guilds. To date, however, most studies have tested unidirectional effects of one pathogen on another, not discerning between conspecific vs. heterospecific infections, and often not measuring plant induced responses underlying such outcomes. To address this, we conducted a greenhouse experiment testing for the impact of initial infection by two leaf pathogens (Alternaria solani and Phytophthora infestans) on subsequent infection by each of these pathogens on potato (Solanum tuberosum) plants, and also measured induced plant defences (phenolic compounds) to inform on interaction outcomes. We found contrasting results depending on the identity of the initially infecting pathogen. Specifically, initial infection by A. solani drove induced resistance (lower necrosis) by subsequently infecting A. solani (conspecific induced resistance) but had no effect on subsequent infection by P. infestans. In contrast, initial infection by P. infestans drove induced resistance to subsequent infection by both conspecifics and A. solani. Patterns of plant induced defences correlated with (and potentially explained) induced resistance to conspecific but not heterospecific (e.g., in the case of P. infestans) subsequent infection. Overall, these results further our understanding of plant-mediated pathogen interactions by showing that plant-mediated interactions between pathogen species can be asymmetrical and in some cases not reciprocal, that pathogen species can vary in the importance of conspecific vs. heterospecific effects, and shed mechanistic insight into the role of plant induced responses driving such interactions.

Synchrospora gen. nov., a New Peronosporaceae Genus with Aerial Lifestyle from a Natural Cloud Forest in Panama.

During a survey of Phytophthora diversity in Panama, fast-growing oomycete isolates were obtained from naturally fallen leaves of an unidentified tree species in a tropical cloud forest. Phylogenetic analyses of sequences from the nuclear ITS, LSU and ßtub loci and the mitochondrial cox1 and cox2 genes revealed that they belong to a new species of a new genus, officially described here as Synchrospora gen. nov., which resided as a basal genus within the Peronosporaceae. The type species S. medusiformis has unique morphological characteristics. The sporangiophores show determinate growth, multifurcating at the end, forming a stunted, candelabra-like apex from which multiple (8 to >100) long, curved pedicels are growing simultaneously in a medusa-like way. The caducous papillate sporangia mature and are shed synchronously. The breeding system is homothallic, hence more inbreeding than outcrossing, with smooth-walled oogonia, plerotic oospores and paragynous antheridia. Optimum and maximum temperatures for growth are 22.5 and 25-27.5 °C, consistent with its natural cloud forest habitat. It is concluded that S. medusiformis as adapted to a lifestyle as a canopy-dwelling leaf pathogen in tropical cloud forests. More oomycete explorations in the canopies of tropical rainforests and cloud forests are needed to elucidate the diversity, host associations and ecological roles of oomycetes and, in particular, S. medusiformis and possibly other Synchrospora taxa in this as yet under-explored habitat.

Arbuscular mycorrhizal fungus changes alfalfa (Medicago sativa) metabolites in response to leaf spot (Phoma medicaginis) infection, with subsequent effects on pea aphid (Acyrthosiphon pisum) behavior.

Plant disease occurs simultaneously with insect attack. Arbuscular mycorrhizal fungi (AMF) modify plant biotic stress response. Arbuscular mycorrhizal fungi and pathogens may modify plant volatile organic compound (VOC) production and insect behavior. Nevertheless, such effects are rarely studied, particularly for mesocosms where component organisms interact with each other. Plant-mediated effects of leaf pathogen (Phoma medicaginis) infection on aphid (Acyrthosiphon pisum) infestation, and role of AMF (Rhizophagus intraradices) in modifying these interactions were elucidated in a glasshouse experiment. We evaluated alfalfa disease occurrence, photosynthesis, phytohormones, trypsin inhibitor (TI) and total phenol response to pathogen and aphid attack, with or without AMF, and aphid behavior towards VOCs from AMF inoculated and non-mycorrhizal alfalfa, with or without pathogen infection. AM fungus enhanced alfalfa resistance to pathogen and aphid infestation. Plant biomass, root : shoot ratio, net photosynthetic rate, transpiration rate, stomatal conductance, salicylic acid, and TI were significantly increased in AM-inoculated alfalfa. Arbuscular mycorrhizal fungi and pathogen significantly changed alfalfa VOCs. Aphids preferred VOCs of AM-inoculated and nonpathogen-infected to nonmycorrhizal and pathogen-infected alfalfa. We propose that AMF alter plant response to multiple biotic stresses in ways both beneficial and harmful to the plant host, providing a basis for strategies to manage pathogens and herbivore pests.

Leaf functional traits and pathogens: Linking coffee leaf rust with intraspecific trait variation in diversified agroecosystems.

Research has demonstrated that intraspecific functional trait variation underpins plant responses to environmental variability. However, few studies have evaluated how trait variation shifts in response to plant pathogens, even though pathogens are a major driver of plant demography and diversity, and despite evidence of plants expressing distinct strategies in response to pathogen pressures. Understanding trait-pathogen relationships can provide a more realistic understanding of global patterns of functional trait variation. We examined leaf intraspecific trait variability (ITV) in response to foliar disease severity, using Coffea arabica cv. Caturra as a model species. We quantified coffee leaf rust (CLR) severity-a fungal disease prominent in coffee systems-and measured key coffee leaf functional traits under contrasting, but widespread, management conditions in an agroforestry system. We found that coffee plants express significant ITV, which is largely related to shade tree treatment and leaf position within coffee canopy strata. Yet within a single plant canopy stratum, CLR severity increased with increasing resource conserving trait values. However, coffee leaves with visible signs of disease expressed overall greater resource acquiring trait values, as compared to plants without visible signs of disease. We provide among the first evidence that leaf traits are correlated with foliar disease severity in coffee, and that functional trait relationships and syndromes shift in response to increased disease prevalence in this plant-pathogen system. In doing so, we address a vital gap in our understanding of global patterns of functional trait variation and highlight the need to further explore the potential role of pathogens within established global trait relationships and spectra.