Search Results (76)

Almond decline and dieback have become significant challenges in newly established orchards, with symptoms including internal necrosis, canker, and external gummosis. This work aims to explore the potential fungal and bacterial causative agents through metabarcoding and traditional culture plate isolation across six almond cultivars. Our results emphasize the multifactorial nature of almond decline and dieback, with possible co-infections by opportunistic fungi and bacteria playing a central role. Classical isolation identified 47 fungal species or genera, including Diaporthe amygdali, Diplodia corticola, Phytophthora sp., and several Fusarium species. Almond metabarcoding revealed a more diverse microbial community, highlighting the prevalence of soilborne pathogens such as Neocosmospora rubicola, Dactylonectria estremocensis, and Plectosphaerella niemeijerarum. Soil metabarcoding suggested that these pathogens likely originate from nursery substrates or soils shared with other crops, such as olives and vineyards, that serve as a source of inoculum. ‘Soleta’ generally presented lower richness when compared to the other tested cultivars, suggesting a higher degree of biotic stress and decreased plant resilience. This study highlights the value of integrating NGS approaches to comprehensively study complex diseases and the need for further research on pathogen interactions and cultivar susceptibility for the future development of new sustainable, targeted management strategies in almond orchards. Full article

Common buckthorn (Rhamnus cathartica L.) is a small tree that forms dense stands, displacing native plant species and threatening natural forest habitats in its introduced range in North America. Removal via cutting is labor intensive and often ineffective due to vigorous resprouting. Although chemical control methods are effective, they can negatively affect sensitive ecosystems. A mycoherbicide that selectively kills buckthorn would provide an additional method for control. In the present study, fungi were collected from dying buckthorn species (Frangula alnus Mill., Rhamnus cathartica, Ventia alnifolia L’Hér) located at 19 sites across Minnesota and Wisconsin for their potential use as mycoherbicides for common buckthorn. A total of 412 fungi were isolated from samples of diseased tissue and identified via DNA extraction and sequencing. These fungi were identified as 120 unique taxa belonging to 81 genera. Of these fungi, 46 species belonging to 26 genera were considered to be canker or root-rot pathogens of woody plants, including species in Cytospora, Diaporthe, Diplodia, Dothiorella, Eutypella, Fusarium, Hymenochaete, Irpex, Phaeoacemonium, and others. A future study testing the pathogenicity of these putative pathogens of buckthorn is now needed to assess their utility as potential mycoherbicide agents for control of common buckthorn. Full article

Lemon, widely used in food, medicine, cosmetics, and other industries, has considerable value as a commodity and horticultural product. Previous research has shown that the fungus Diaporthe citri infects several citrus species, including mandarin, lemon, sweet orange, pomelo, and grapefruit, in China. Although D. citri has been reported to cause melanose disease in lemons in China, key pathological evidence, such as Koch’s postulates fulfillment on lemon fruits and detailed morphological characterization, is still lacking. In May 2018, fruits, leaves, and twigs were observed to be infected with melanose disease in lemon orchards in Chongqing municipality in China. The symptoms appeared as small black discrete spots on the surface of fruits, leaves, and twigs without obvious prominent and convex pustules. D. citri was isolated consistently from symptomatic organs and identified provisionally based on the morphological characteristics. The identification was confirmed using sequencing and multigene phylogenetic analysis of ITS, TUB, TEF, HIS, and CAL regions. Pathogenicity tests were performed using a conidium suspension, and melanose symptoms similar to those observed in the field were reproduced. To our knowledge, this study provides the first comprehensive evidence for D. citri as a causal agent of melanose disease in lemons in China, including morphological characterization and pathogenicity assays on lemon fruits. This report broadens the spectrum of hosts of D. citri in China and provides useful information for the management of melanose in lemons. Full article

Fraxinus angustifolia, the main ash species in Croatia in terms of economic and ecological importance, is affected by a severe dieback initially attributed to the fungal pathogen Hymenoscyphus fraxineus. Recently, another pathogen, Diplodia fraxini, has been shown to play a key role in ash dieback in several European countries. Therefore, because the dieback symptoms of ash trees observed in Croatia are typical of Botryosphaeriaceae attacks, the aim of this study was to define the etiology of F. angustifolia dieback. To this end, symptomatic shoots and branches and cross-sections of the main stem were sampled from 20 symptomatic trees at eight locations and analyzed for the presence of D. fraxini and other possible fungal pathogens. Diplodia fraxini was the fungus most frequently associated with branch cankers and dieback; it was isolated from 17 trees in all sites monitored, and its pathogenicity towards F. angustifolia was confirmed. The fungus was also associated with wood necrosis at the base of the trunk in two trees. Other fungi, namely H. fraxineus, Diaporthe eres, Diplodia seriata, Botryosphaeria dothidea, Armillaria gallica, and Lentinus tigrinus, were isolated sporadically. Full article

Diaporthe species are critical plant pathogens that contribute to a disease complex responsible for substantial yield losses in soybean production worldwide. However, reports on the primary Diaporthe species causing soybean stem blight and their sensitivity to various fungicides are scarce in China. In this study, a total of 46 D. longicolla strains were isolated and identified from diseased soybean stems and rots collected from 14 regions of Heilongjiang province in 2021 and 2022. Among the eight fungicides examined, fludioxonil, mefentrifluconazole, tebuconazole, and azoxystrobin demonstrated effective inhibition for D. longicolla, with EC₅₀ values < 0.3 µg/mL. Interestingly, the EC₅₀ values of D. longicolla to two succinate dehydrogenase inhibitors (SDHIs), pydiflumetofen and fluopyram, were 5.47 µg/mL and over 100 µg/mL, respectively. In molecular dynamics simulations, pydiflumetofen exhibited a smaller RMSD, while fluopyram had a higher binding free energy with Sdh proteins compared to pydiflumetofen. This difference may contribute to the higher activity of pydiflumetofen in D. longicolla. Further analysis of the electrostatic potential and structural conformations of the binding pocket revealed that pydiflumetofen formed more hydrophobic interactions with SdhC and SdhD and was positioned closer to the SdhD subunit. A mixture of fludioxonil and mefentrifluconazole at a ratio of 1:5, as well as fludioxonil and pydiflumetofen at a ratio of 1:5, exhibited synergistic effects. These findings demonstrated that several fungicides could be utilized to control Diaporthe stem blight, and the difference in binding affinity to the Sdh subunit impacts sensitivity to fluopyram and pydiflumetofen. Full article

Endophytic fungi are valuable sources of bioactive secondary metabolites, with potential applications in pharmaceutical and agricultural fields. This study investigates the metabolic potential of Diaporthe hongkongensis, an endophytic fungus isolated from Minquartia guianensis. To date, no secondary metabolites have been identified from this species, highlighting the novelty of this research and its contribution to understanding the chemical diversity of endophytic fungi. The fungus was cultivated on parboiled rice under static and dark conditions for 28 days, leading to the isolation of the following three compounds: 5,6-dihydro-5,6-epoxymultiplolide A (1), cytosporone C (2), and uridine (3). Structural identification was carried out using nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry. The results revealed the metabolic versatility of D. hongkongensis, as demonstrated by its ability to produce structurally diverse substances with biological relevance. Hence, it describes the first isolation of secondary metabolites from the endophytic fungus D. hongkongensis, marking a significant step in understanding its chemical profile. The identification of a known antifungal compound and a lactone derivative underscores the biosynthetic potential of this endophytic fungus, while the isolation of a nucleoside expands the chemical repertoire of fungal metabolites, suggesting possible roles in cellular metabolism and stress adaptation. These findings highlight the role of endophytic fungi as prolific sources of structurally diverse and potentially bioactive natural products, supporting further exploration of their biotechnological applications. Full article

The rapid expansion of pomegranate (Punica granatum L.) cultivation in central and southern Italy has revealed emerging phytosanitary challenges, including “pomegranate wood decay syndrome”, characterised by cortical cankers, wood browning, and progressive plant decline. This study investigates the fungal microbiota associated with symptomatic pomegranate wood using a combined approach of traditional fungal isolation and ITS2 metabarcoding analysis. Samples from two orchards in Lazio were examined, revealing a complex fungal community with a high prevalence of Neofusicoccum parvum (putative) and species belonging to the genus Diaporthe. Pathogenicity tests confirmed the role of N. parvum in causing significant wood browning, while other isolates showed variable virulence. Statistical analyses validated the pathogenicity of select isolates, with the putative Diaporthe eres (Nitschke) consistently demonstrating potential pathogenic activity across all trials. Metabarcoding identified 289 taxa, highlighting a richer fungal diversity in the symptomatic wood compared to the asymptomatic sections. Notably, Coniella granati, previously implicated in pomegranate decline, was absent in the studied orchards. The findings reveal that pomegranate wood decay is a complex syndrome driven by fungal pathogens and environmental stressors, such as low temperatures. This study highlights the value of integrative approaches for understanding and managing fungal-associated wood diseases in pomegranate orchards. Full article

Species of the genus Diaporthe have a mating-type system with the two mating types MAT1-1 and MAT1-2, like other ascomycetes. They can either be heterothallic, which means that any isolate only possesses one of the two mating types and needs a mating partner for sexual reproduction, or homothallic, which means that they possess both mating types and are self-fertile. For several Diaporthe species, no sexual reproduction has been observed so far. Using PCR with primers specific to the defining genes MAT1-1-1 and MAT1-2-1, we determined the mating types of 33 isolates of Diaporthe caulivora, D. eres, D. longicolla, and D. novem from central Europe. In addition, we partially sequenced the mating-type genes of 25 isolates. We found that different D. longicolla isolates either possess MAT1-1-1 or MAT1-2-1, making the species heterothallic, which is in contrast to previous studies and the general assumption that D. longicolla only reproduces asexually. D. eres and D. novem were also found to be heterothallic. Using genomic sequence information and re-sequencing of DNA and RNA, we identified the MAT1-1-1 gene in D. caulivora and present here the full sequence of the mating-type locus of this homothallic species. Finally, we used sequence information from MAT1-1-1 and MAT1-2-1, respectively, for improved phylogenetic resolution of our isolates. Full article

Blueberry has been a burgeoning fruit in China in recent years, but its perishable nature places a constant strain on industrial development. To determine the pathogens infecting blueberry fruits, diseased samples were collected from Guizhou and Fujian Provinces. Isolates from the samples were identified by morphological characterization and phylogenetic analyses. Pathogenicity assays were conducted on fresh blueberry fruits using spore suspensions. Sixteen isolates were identified as seven species, namely, Botryosphaeria dothidea, Botrytis cinerea, Cladosporium guizhouense, Colletotrichum fioriniae, Diaporthe anacardii, Fusarium annulatum, and Neopestalotiopsis surinamensis, and their pathogenicity on blueberry fruits were confirmed following Koch’s postulates. The current study reported Cladosporium guizhouense, Fusarium annulatum, and Neopestalotiopsis surinamensis for the first time on blueberry. The study (1) demonstrated that fruit rot disease results from a mixed infection of multiple pathogens; and (2) expanded the understanding of causal agents of blueberry fruit rot during the growth stage, highlighting their potential as latent pathogens that contribute to post-harvest losses. Relevant results provide a reference for the etiological research and disease management in blueberry fruit diseases. Full article

Humulus lupulus L. (hop) is a crucial crop within the brewing industry and a rich source of bioactive compounds. Traditionally concentrated in northeast regions of Europe, hop cultivation has expanded towards southern territories such as Italy over recent decades. Managing phytosanitary threats in Mediterranean climates poses challenges due to limited knowledge and registered agrochemicals. In pursuit of eco-friendly alternatives for disease management, we isolated 262 endophytic fungal strains from wild hop roots, stems, leaves, and flowers. Through phylogenetic analyses, we identified 51 operational taxonomic units. Dominant species such as Ilyonectria macrodidyma, Penicillium sp., Diaporthe columnaris, Plectosphaerella cucumerina, and Fusarium oxysporum were exclusive to roots. In contrast, Alternaria spp. and Epicoccum spp. were prevalent in other tissues, and Botrytis cinerea was exclusively detected in female flowers. We tested seven isolates—Epicoccum sp., Aureobasidium pullulans, Plectosphaerella cucumerina, Stemphylium vesicarium, Periconia byssoides, Talaromyces wortmannii, and Nigrospora sphaerica—against the four phytopathogenic fungi Alternaria sp., Fusarium oxysporum, Botrytis cinerea, and Sclerotinia sclerotiorum. All endophytes exhibited antagonistic effects against at least one pathogen, with Plectosphaerella cucumerina showing the strongest inhibition against Alternaria sp. This study marks the first exploration of endophytic fungi from various hop tissues. All isolated strains were ex situ conserved for future bioactivity assessments and biotechnological applications. Original data with a key relevance for the environmentally friendly management of plant diseases are provided. Full article

Considering that fungi display a great morphological, ecological, metabolic, and phylogenetic diversity, their taxonomic identification is extremely important because it helps us establish important information about each species and its possible biochemical and ecological roles. Traditionally, the identification of fungi at the species level has been carried out with molecular tools such as DNA sequencing, but it still represents a huge challenge today due to the heterogeneity of the fungal kingdom, making the task of identification a complex and difficult process. Biotyping, a type of chemotaxonomy, has been developed in the field of the identification/differentiation and classification of micro-fungi through tools such as mass spectrometry (MS). Here, two endophytic strains isolated from two different hosts were cultivated and studied regarding their morphology and molecular biology. Morphology analysis determined the strains as Diaporthe, and the molecular analysis results grouped them as D. melongenae. We sought a faster and less complex way of differentiating these fungal strains of interest through an MS chemical profile and MS/MS data using a low-resolution mass spectrometer. Additionally, we linked this information with the structure of compounds previously isolated in the genus Diaporthe. Studies conducted using this technique allowed us to propose the structure of distinctive molecules that are unique to each strain and share compounds common to this genus (13 compounds in total). In addition, this is the first report of secondary metabolites in D. melongenae. The dataset demonstrates that the two strains under investigation can be distinguished via mass spectrometry, suggesting host affinity; both exhibits pronounced differences in their chemical profiles across all culture media and incubation periods with the parameters described herein. Full article

Fungal biota represents important constituents of phyllosphere microorganisms. It is taxonomically highly diverse and influences plant physiology, metabolism and health. Members of the order Diaporthales are distributed worldwide and include devastating plant pathogens as well as endophytes and saprophytes. However, many phyllosphere Diaporthales species remain uncharacterized, with studies examining their diversity needed. Here, we report on the identification of several diaporthalean taxa samples collected from diseased leaves of Cinnamomum camphora (Lauraceae), Castanopsis fordii (Fagaceae) and Schima superba (Theaceae) in Fujian province, China. Based on morphological features coupled to multigene phylogenetic analyses of the internal transcribed spacer (ITS) region, the large subunit of nuclear ribosomal RNA (LSU), the partial beta-tubulin (tub2), histone H3 (his3), DNA-directed RNA polymerase II subunit (rpb2), translation elongation factor 1-α (tef1) and calmodulin (cal) genes, three new species of Diaporthales are introduced, namely, Diaporthe wuyishanensis, Gnomoniopsis wuyishanensis and Paratubakia schimae. This study contributes to our understanding on the biodiversity of diaporthalean fungi that are inhabitants of the phyllosphere of trees native to Asia. Full article

Pathogen-induced fruit decay is a significant threat to the kiwifruit industry, leading to considerable economic losses annually. The cell-wall-degrading enzymes (CWDEs) secreted by these pathogens are crucial for penetrating the cell wall and accessing nutrients. Among them, Diaporthe species are recognized as major causal agents of soft rot in kiwifruit, yet their pathogenic mechanisms are not well understood. In this study, we explored the production of various CWDEs secreted by Diaporthe Z1-1N, including polygalacturonase (PG), polymethylgalacturonase (PMG), polygalacturonic acid transeliminase (PGTE), pectin methyltranseliminase (PMTE), endoglucanase (Cx), and β-glucosidase (β-glu), both in liquid cultures and within infected kiwifruit tissues. Our findings revealed significant activities of two pectinases (PG and PMG) and cellulases (Cx and β-glu) in the infected tissues. In contrast, very low levels of PMTE and PGTE activities were observed under the same conditions. When orange pectin served as the carbon source, PG and PMG showed notable activities, while PMTE and PGTE remained inactive. Moreover, the activities of Cx and β-glu significantly decreased by more than 63 times in the liquid medium with carboxymethyl cellulose (CMC) as the carbon source compared to their levels in infected kiwifruit. A further analysis indicated that the necrotic lesions produced by pectinase extracts were larger than those produced by cellulase extracts. Notably, four enzymes—PG, PMG, Cx, and β-glu—exhibited high activities on the third or fourth day post-infection with Diaporthe Z1-1N. These results suggest that Diaporthe Z1-1N secretes a range of CWDEs that contribute to kiwifruit decay by enhancing the activities of PG, PMG, Cx, and β-glu. This study sheds light on the pathogenicity of Diaporthe in kiwifruit and highlights the importance of these enzymes in the decay process. Full article

Diaporthe (anamorph: Phomopsis) species are endophytes or fungal pathogens for many different plant species. Soybean (Glycine max) can be infected by many different Diaporthe species; among them, D. caulivora and D. longicolla are responsible for the most significant damages. Diaporthe goulteri is a species that was only recently described and has so far been found on sunflower (Helianthus annuus) in Australia and an unknown host in Thailand. Here, we report isolation of D. goulteri from soybean in southern Germany, molecular species identification, and additional morphological description. We also show that D. goulteri can infect soybean and describe the symptoms we observed, both on the plant where the isolate came from and following artificial inoculation. Full article

Peach (Prunus persica L.) is one of the most important and oldest stone fruits grown in China. Though Diaporthe species have more commonly been reported as plant pathogens, endophytes and saprophytes with a wide range of plant hosts, little is known about the Diaporthe species associated with peach trunk diseases in China. In the present study, forty-four Diaporthe isolates were obtained from trees with peach branch canker, shoot blight and gummosis symptoms in four provinces in China. Based on a combination of morphology and multi-locus sequence analysis of the rDNA internal transcribed spacer region (ITS), calmodulin (cal), translation elongation factor 1-α (tef1) and β-tubulin (tub2), these Diaporthe isolates were assigned to four species. Detailed descriptions and illustrations of all of the species, D. arecae, D. caulivora, D. discoidispora and D. eres, are provided. This study further reports the first host association of D. caulivora and D. discoidispora on peaches worldwide. The pathogenicity experiment results revealed that D. arecae was the most aggressive species, whereas D. discoidispora was the least aggressive on detached peach shoots. This study provides new insights into the fungi associated with peach trunk diseases in China, and the results of this study may help to facilitate routine diagnosis and planning of suitable plant disease management strategies. Full article