Search Results (14)

Grapevine trunk diseases, particularly Botryosphaeria dieback, pose a major threat to vineyard sustainability, a risk that is further intensified by climate variability and increasing environmental stress. This study evaluated pathogenicity and bacterial biocontrol efficacy against Neofusicoccum parvum and Diplodia seriata under vineyard conditions, analyzing the combined effects of cultivar (Vitis vinifera L. cv. Cabernet Sauvignon and Sauvignon Blanc), tissue type (young shoots and lignified arms), and phenological season (autumn/winter and spring/summer). Pathogenicity assays revealed clear tissue-age specialization: N. parvum was more aggressive in young shoots, whereas D. seriata caused the most severe vascular lesions in lignified wood. Seasonality further modulated disease expression, with higher lesion development during spring/summer, particularly for N. parvum in young shoots, while D. seriata maintained high aggressiveness in lignified tissues across both seasons. Berry assays provided a rapid initial assessment of isolate virulence but did not fully reflect pathogen behavior in woody tissue under field conditions. Biological treatments using native bacterial strains (Pseudomonas sp. AMCR2b, GcR15a, and Rhodococcus sp. PU4) significantly reduced lesion severity in V. vinifera under field conditions, although efficacy varied by tissue type and season. Biocontrol effects were generally more stable in lignified arms, and under high disease pressure, only the most robust strains maintained consistent protection, in some cases matching or surpassing the efficacy of the fungicide tebuconazole. These results show that both pathogenicity and biocontrol performance against Botryosphaeria dieback in V. vinifera under field conditions are strongly influenced by tissue type and season, supporting bacterial biocontrol as a sustainable component of integrated disease management in vineyards. Full article

Fig (Ficus carica L.), a deciduous fruit tree that belongs to the Moraceae family, is cultivated worldwide as an important fruit crop for raw and processed foods. Quantitative real-time PCR (RT-qPCR) is a widely used method in F. carica to elucidate expression of genes related to various physiological responses. However, no studies have identified appropriate reference genes for RT-qPCR normalization in F. carica. In this study, 12 genes were selected from the F. carica genome as candidate reference genes for normalizing target gene expression. All candidate genes exhibited high amplification efficiency and specificity in the absence of primer dimers or extra PCR amplicons. The expression levels of the candidate genes were measured in three different plant tissues (fruit, leaf, and stem) under fungal pathogen infection using RT-qPCR. Their expression stabilities were evaluated using four computational algorithms: geNorm, Normfinder, delta-Ct, and BestKeeper. The RefFinder program was also used to calculate the geometric mean of the stability rankings obtained from these algorithms. The comprehensive ranking revealed that FcYLS8, FcPP2A, and FcAP2M were the most stable reference genes under biotic stress in the fruits, leaves, and stems, respectively. In contrast, traditional reference genes such as FcACT2, FcEF-1α, FcGAPDH, FcUBC21, and FcUBQ5 exhibited relatively low expression stability in all tested tissues. This study identified and validated stable reference genes for RT-qPCR normalization in F. carica, thus providing a valuable resource for accurate gene expression studies under biotic stress and highlighting the importance of validating reference genes to ensure reliable and reproducible RT-qPCR analysis. Full article

Litter decomposition dynamics are largely governed by microbial interactions. While the involvement of endophytic fungi in early-stage decomposition and microbial succession is well established, their species-specific contributions to decomposer community assembly remain insufficiently understood. This study investigated the effects of single-strain endophytic colonization using dominant species (Tubakia dryina, Tubakia dryinoides, Guignardia sp.) and rare species (Neofusicoccum parvum, Penicillium citrinum) on Quercus acutissima leaf decomposition through a controlled field experiment in a karst ecosystem. Endophytes accelerated decomposition rates across treatments but paradoxically reduced transient CO₂ emissions, linked to intensified microbial carbon and phosphorus limitations in late stages. Contrary to expectations, decomposition efficiency was governed by endophytic fungal species traits rather than colonization abundance, with rare species outperforming dominant taxa. Endophytes induced significant fungal community restructuring, reducing Ascomycota while enriching lignin-degrading Basidiomycota, but minimally affected bacterial composition. Co-occurrence networks revealed endophyte-driven fragmentation of microbial connectivity, with only two keystone fungal hubs (Trechispora sp. and Russula carmesina) identified compared to natural communities. Endophytic colonization improved fungal community assembly, mediated by an increase in lignin-degrading Basidiomycota and the suppression of pathogenic Leotiomycetes lineages. Our findings demonstrate that endophytes hierarchically regulate decomposer communities through phylogenetically conserved fungal interactions, prioritizing functional trait selection over competitive dominance, thereby stabilizing decomposition under nutrient constraints. This mechanistic framework advances predictions of litter decay dynamics in forest ecosystems undergoing microbial community perturbations. Full article

Botryosphaeria dieback, a significant grapevine trunk disease (GTD) caused by various pathogens, represents a serious threat to viticulture. Biocontrol emerges as a promising sustainable alternative to chemical control, aligning toward environmentally friendly viticultural practices. This study evaluated the in vitro, in vivo, and in situ biocontrol potential of Chilean native bacteria isolated from wild flora and endophytic communities of grapevine against Neofusicoccum parvum. In vitro biocontrol assays screened 15 bacterial strains at 10, 22, and 30 °C, identifying four Pseudomonas strains with >30% mycelial growth inhibition. In diffusible agar and double plate assays, plant growth-promoting bacteria AMCR2b and GcR15a, which were isolated from native flora, achieved significant inhibition of N. parvum growth, with reductions of up to ~50% (diffusible agar) and up to ~46% (double plate). In vivo experiments on grapevine cuttings revealed that strains AMCR2b and GcR15a inhibited mycelial growth (17–90%); younger grapevines (1–5 years) were more susceptible to N. parvum. In situ trials using Vitis vinifera L. cv. Cabernet Sauvignon and Sauvignon Blanc demonstrated higher fungal susceptibility in Sauvignon Blanc. These results highlight the potential of Pseudomonas sp. AMCR2b and GcR15a to be effective biocontrol agents against GTDs at a wide range of temperatures, contributing to sustainable viticulture. Full article

During 2021 and 2022, a field investigation was conducted in Istria, Croatia, searching for trees exhibiting signs of Botryosphaeria dieback. Samples of symptomatic trees were collected from 26 different locations and analysed. Isolates that morphologically corresponded to species from the Botryosphaeriaceae family were selected, and detailed morphological characterisation and molecular identification of the isolates were conducted. Based on morphological characteristics and phylogenetic analysis using the internal transcribed spacer (ITS), beta-tubulin (TUB2), and translation elongation factor 1-alpha (TEF1-α) regions, six species of fungi from the Botryosphaeriaceae family were identified: Botryosphaeria dothidea (Moug. ex Fr.) Ces. & De Not.; Diplodia mutila (Fr.) Fr.; Diplodia seriata De Not.; Dothiorella iberica A.J.L. Phillips, J. Luque & A. Alves; Dothiorella sarmentorum (Fr.) A.J.L. Phillips, Alves & Luque; and Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips. This is the first report of D. mutila, Do. sarmentorum, and Do. iberica causing Botryosphaeria dieback on olive trees in Croatia, and the first study investigating the resistance of Croatian olive varieties to species from the Botryosphaeriaceae family. Pathogenicity testing of selected isolates and assessment of variety resistance were conducted on four different olive varieties, namely Buža, Istarska bjelica, Leccino, and Rosinjola, using representative isolates of the mentioned species. The most aggressive species was found to be N. parvum. Olive varieties exhibited differences in susceptibility depending on the fungus they were infected with. Full article

Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense (Foc), significantly affects the productivity of the banana crop in the field. Currently, there are no effective control measures available, and therefore, there is an urgent need to develop novel strategies to control the spread of the disease. Biological control is a promising strategy for the management and control of the disease. The aim of this study was to identify fungal endophytes associated with Moringa (Moringa oleifera), Neem tree (Azadirachta indica) and Lavender (Lavandula angustifolia) and their antifungal activities against Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4). We isolated 69 fungal endophytes from different tissues of M. oleifera, A. indica and L. angustifolia and screened for antifungal activity against Foc TR4. Six fungal endophytes exhibited highest antagonistic activity against Foc TR4 based on dual-culture assays. Based on morphology and ITS gene sequence analysis, the selected six endophytes were identified to be related to Alternaria alternata (MB7 and NR3), Neofusicoccum parvum (LB1), Fusarium oxysporum (LR1), Talaromyces amestolkiae (MB14) and Alternaria tenuissima (NB6). The culture filtrates of the six fungal endophytes (LB1, LR1, MB7, MB14, NB6 and NR3) exhibited more than 50% inhibition of mycelial growth of Foc TR4 in vitro and were producers of β-1,3-glucanase. The six fungal endophytes showed biocontrol efficacy against Fusarium wilt in pot experiments. The findings from this study demonstrate that fungal endophytes LB1, LR1, MB7, MB14, NB6 and NR3 should be explored as biocontrol agents and biofertilizers in banana production. Full article

Cryptomeria japonica D. Don is a coniferous tree species widely grown in southern China for its high ornamental value. Recently, during disease surveys in China, a symptom of dieback occurred on C. japonica in Nanjing, Jiangsu Province, China. A total of 130 trees were surveyed and more than 90% showed the same symptom. The crowns of affected trees were brown when viewing from a distance, and the bark showed no difference from the healthy ones. In this study, 157 isolates were isolated from the 3 affected plants of C. japonica, and based on the living culture on PDA, the fungal isolates were preliminarily divided into 6 groups. Thirteen representative isolates were selected for the pathogenicity test, and seven of them showed obvious pathogenicity on C. japonica, causing stem basal canker. These isolates were identified based on comparisons of the DNA sequences of the internal transcribed spacer regions (ITS), partial translation elongation factor 1-alpha (tef1), β-tubulin (tub2), and DNA-directed RNA polymerase II subunit (rpb2) and combined with their morphological characteristics. Results showed that these seven isolates belong to two taxa in Neofusicoccum, including a species new to science. The new species, Neofusicoccum cryptomeriae, was hereby described and illustrated. The other species was N. parvum. Both species were pathogens of stem basal canker of Cryptomeria japonica. Full article

Agarwood is the most expensive non-construction wood product in the world. As a therapeutic agent, agarwood can cure some diseases, but few studies have been carried out on the antagonistic abilities of endophytic fungi associated with agarwood. Agarwood is mainly found in the genus Aquiaria. The objectives of this study are to understand the antimicrobial activities and their potential as biocontrol agents of the endophytic fungi of Aquilaria sinensis. First, fresh samples of A. sinensis were collected from Yunnan and Guangdong Provinces in 2020–2021, and the endophytic fungi were isolated and identified to genus level based on the phylogenetic analyses of the Internal Transcribed Spacer (ITS) region. In this bioassay, 47 endophytic strains were selected to check their bioactivities against three bacterial pathogens viz. Erwinia amylovora, Pseudomonas syringae, and Salmonella enterica; and three fungal pathogens viz. Alternaria alternata, Botrytis cinerea, and Penicillium digitatum. The antibiosis test was carried out by the dual culture assay (10 days), and among the 47 strains selected, 40 strains belong to 18 genera viz. Alternaria, Annulohypoxylon, Aspergillus, Botryosphaeria, Colletotrichum, Corynespora, Curvularia, Daldinia, Diaporthe, Fusarium, Lasiodiplodia, Neofusicoccum, Neopestalotiopsis, Nigrospora, Paracamarosporium, Pseudopithomyces, Trichoderma, Trichosporon and one strain belongs to Xylariaceae had antimicrobial activities. In particular, Lasiodiplodia sp. (YNA-D3) showed the inhibition of all the bacterial and fungal pathogens with a significant inhibition rate. In addition, the strains viz; Curvularia sp. (GDA-3A9), Diaporthe sp. (GDA-2A1), Lasiodiplodia sp. (YNA-D3), Neofusicoccum sp. (YNA-1C3), Nigrospora sp. (GDA-4C1), and Trichoderma sp. (YNA-1C1) showed significant antimicrobial activities and are considered worthy of further studies to identify individual fungal species and their bioactive compounds. This study enriches the diversity of endophytic fungi associated with agarwood, and their potential antagonistic effects against bacterial and fungal pathogens. Full article

In this research, we aimed to isolate and identify native strains of Trichoderma spp. with potential activity against avocado pathogens (Neofusicoccum parvum, Colletotrichum gloeosporioides, Diaporthe sp., and Phomopsis perseae). Strains of Trichoderma spp. were isolated from roots and soil obtained from avocado orchards from different regions of Mexico. Twenty-five Trichoderma spp. strains were isolated, of which six (TSMICH7, TSMICH8, TRMICH9, TSMICH10, TSMICH15, and TRJAL25) showed greater antagonistic capacity in vitro (>80%) against avocado pathogens. After 96 h, the antagonistic strain undergoes a thickening of hyphae, while the phytopathogen tends to thin, except for Diaporthe sp., which tends to thicken. The characterization of these strains was carried out through morphological observations and the amplification and sequencing of rDNA fragments (ITS regions), as well as the translation elongation factor 1-alpha (Tef 1-α), achieving the identification of Trichoderma harzianum. However, in the in vivo evaluation (applying directly to the avocado fruit), the TSMICH7 strain maintained considerably high effectiveness (>90%) against the four phytopathogens tested, mainly with P. perseae, N. parvum, and Diaporthe sp., increasing the activity of glucanases and chitinases. Therefore, T. harzianum could be used as a biological control agent to inhibit post-harvest pathogens in avocados, thus avoiding significant losses of this fruit of international importance. Full article

Val d’Agri is an important orchard area located in the Basilicata Region (Southern Italy). A phenomenon affecting cv. “Golden Delicious” apples which lead to tree death has been observed in the past several years in this area. This phenomenon has already been detected in about 20 hectares and is rapidly expanding. The symptoms observed were “scaly bark” and extensive cankers, mainly located in the lower part of the trunk, associated with wood decay. Dead plants ranged from 20% to 80% and, in many cases, trees were removed by farmers. In order to identify the causes of this phenomenon, investigations were started in autumn/winter 2019. In order to determine the possible causal agents, fungal and bacterial isolations, from symptomatic tissues, were performed in laboratory. Bacterial isolations gave negative results, whereas pure fungal cultures (PFCs) were obtained after 3–4 passages on potato dextrose agar (PDA) media. Genetic material was extracted from each PFC and amplified by PCR using three pairs of primers: ITS5/4, Bt2a/Bt2b and ACT-512F/ACT-783R. The amplicons were directly sequenced, and nucleotide sequences were compared with those already present in the NCBI GenBank nucleotide database. All isolated fungi were identified based on morphological features and multilocus molecular analyses. Neofusicoccum parvum, Diaporthe eres and Trametes versicolor were most frequently isolated, while Pestalotiopsis funerea, Phomopsis spp. and Diaporthe foeniculina were less frequently isolated. All nucleotide sequences obtained in this study have been deposited into the EMBL database. Pathogenicity tests showed that N. parvum was the most pathogenic and aggressive fungus, while Phomopsis sp. was demonstrated to be the less virulent one. All the investigated fungi were repeatedly reisolated from artificially inoculated twigs of 2-year-old apple trees, cv. “Golden Delicious”, and subsequently morphologically and molecularly identified. The role played by the above-mentioned fungi in the alterations observed in field is also discussed. Full article

Botryosphaeriaceae represents a diverse family of fungi with a cosmopolitan distribution and a wide host range. We studied species diversity and overlap of Botryosphaeriaceae on selected tree species of Anacardiaceae in disturbed (farming or forestry) and undisturbed (isolated and/or protected) ecosystems in the Limpopo and Mpumalanga provinces, South Africa. The disturbed sites resided at Tshikundamalema and Tshipise in Limpopo and the undisturbed sites at Nwanedi and the Mapungubwe National Park in Limpopo and the Kruger National Park in Mpumalanga. Asymptomatic branches were collected from Mangifera indica, Sclerocarya birrea and Lannea schweinfurthii trees in 2017 and 2018. Eleven species were identified using a multi-gene sequencing approach, including Diplodia allocellula, Dothiorella brevicollis, Do. dulcispinae, Do. viticola, Lasiodiplodia crassispora, L. exigua, L. gonubiensis, L. mahajangana, Neofusicoccum parvum, Oblongocollomyces sp. 1 and Oblongocollomyces sp. 2. Ten of the 11 species were identified in undisturbed ecosystems (eight species being unique), while only three species were identified in disturbed ecosystems (one species being unique). Two species were generalists on trees in disturbed and undisturbed ecosystems. Lasiodiplodia mahajangana was the most dominant species as it occurred on the three tree species of Anacardiaceae. Isolates of N. parvum occurred on both S. birrea (a native species) and M. indica (a non-native species) that occurred adjacent to each other in disturbed ecosystems, confirming the ability of this invasive pathogen to cross-infect native and non-native hosts and its abundance in human-disturbed environments. The findings from this study confirm the lack of host specificity for most species of Botryosphaeriaceae. The results also indicate that disturbance through human activity, such as clear-cutting, selective cutting and land-use changes, negatively influences the diversity of the Botryosphaeriaceae. Full article

Grapevine trunk diseases (GTDs) are the most widespread fungal diseases, affecting grapevines in all the major growing regions of the world, and their complete eradication is still not possible. Aiming to search alternatives to avoid the spread and high incidence of these diseases, the present work intended to molecularly identify the grapevine endophytic community, the phytopathogenic fungi associated with GTDs in vineyards within the Alentejo region, and to test potential antagonist microorganisms as biological control candidates against GTDs-associated fungi. Grapevine endophytic community showed a wide variety of fungi in GTDs’ asymptomatic and symptomatic plants, nine of them previously described as GTDs-associated fungi. GTDs prevalent fungi identified in symptomatic plants were Diaporthe sp., Neofusicoccum sp., and H. viticola. Almost all these fungi were also detected in asymptomatic plants, which shows the importance of investigating the interactions of fungal communities and confirms the need for early diagnosis of these diseases. Direct inhibition antagonism tests were performed among identified endophytes and GTDs phytopathogenic fungi, and all the endophyte fungi showed potential as biocontrol agents. Our findings suggest that endophytes are promising candidates for their use in biological control due to their antagonistic activity against the mycelia growth of some GTDs-associated fungi. Full article

As key players in biotic stress response of plants, jasmonic acid (JA) and its derivatives cover a specific and prominent role in pathogens-mediated signaling and hence are promising candidates for a sustainable management of phytopathogenic fungi. Recently, JA directed antimicrobial effects on plant pathogens has been suggested, supporting the theory of oxylipins as double gamers in plant-pathogen interaction. Based on these premises, six derivatives (dihydrojasmone and cis-jasmone, two thiosemicarbazonic derivatives and their corresponding complexes with copper) have been evaluated against 13 fungal species affecting various economically important herbaceous and woody crops, such as cereals, grapes and horticultural crops: Phaeoacremonium minimum, Neofusicoccum parvum, Phaeomoniella chlamydospora, Fomitiporia mediterranea, Fusarium poae, F. culmorum, F. graminearum, F. oxysporum f. sp. lactucae,F. sporotrichioides, Aspergillus flavus, Rhizoctonia solani,Sclerotinia spp. and Verticillium dahliae. The biological activity of these compounds was assessed in terms of growth inhibition and, for the two mycotoxigenic species A. flavus and F. sporotrichioides, also in terms of toxin containment. As expected, the inhibitory effect of molecules greatly varied amongst both genera and species; cis-jasmone thiosemicarbazone in particular has shown the wider range of effectiveness. However, our results show that thiosemicarbazones derivatives are more effective than the parent ketones in limiting fungal growth and mycotoxins production, supporting possible applications for the control of pathogenic fungi. Full article

Artemisia is a plant genus highly studied for its medicinal applications. The studies on the associated fungal endophytes are scarce. Ten plants specimens of Artemisia thuscula from Tenerife and La Palma were sampled to isolate the endophytic fungi. Identification of the endophytic fungi was based on morphology, Internal Transcribed Spacer (ITS) and Large Subunit (LSU) regions sequencing and indicates 37 fungal species affiliated to 25 fungal genera. Colonization rate varied among plants (CR = 25% to 92.11%). The most dominant colonizers found were Alternaria alternata (CF = 18.71%), Neofusicoccum sp. (CF = 8.39%) and Preussia sp. (CF = 3.23). Tendency for host specificity of most endophytic fungal species was observed. Sorensen–Dice index revealed that of 45 cases in the matrix, 27 of them were of zero similarity. Further, only one case was found to have 57% similarity (TF2 and TF7) and one case with 50% similarity (TF1 and TF4). The rest of the cases had values ranging between 11% and 40% similarity. Diversity indices like Brillouin, Margalef species richness, Simpson index of diversity and Fisher’s alpha, revealed plants from La Palma with higher values than plants from Tenerife. Three nutrient media (i.e., potato dextrose agar―PDA, lignocellulose agar―LCA, and tomato juice agar―V8) were used in a case study and revealed no differences in terms of colonization rate when data was averaged. Colonization frequency showed several species with preference for nutrient medium (63% of the species were isolated from only one nutrient medium). For the phylogenetic reconstruction using the Bayesian method, 54 endophytic fungal ITS sequences and associated GenBank sequences were analyzed. Ten orders (Diaporthales, Dothideales, Botryosphaeriales, Hypocreales, Trichosphaeriales, Amphisphaeriales, Xylariales, Capnodiales, Pleosporales and Eurotiales) were recognized. Several arrangements of genera draw the attention, like Aureobasidium (Dothideales) and Aplosporella (Botryosphaeriales) which are clustered with a recent ancestor (BS = 0.97). Full article