Search Results (16)

The wood decay fungi Cytospora sorbicola and Calosphaeria pulchella severely threaten the worldwide cultivation of sweet cherry trees (Prunus avium L.). Both fungi cause similar symptoms, including vascular necrosis, which leads to branch and twig dieback. In advanced stages of the disease, cankers are visible on tree branches and trunks. The sweet cherry is the most widely planted fruit tree in Chile, with 74,000 hectares in 2023. According to the planted surface, the predominant sweet cherry varieties are Lapins, Santina, Regina, and Bing. Variety-dependent susceptibility studies on Cyt. sorbicola and Cal. pulchella are lacking. The main entry points for wood necrosis-causing fungi are pruning wounds; therefore, we evaluated the aggressiveness of Cyt. sorbicola and Cal. pulchella in one-year-old sweet cherry plants. Santina and Lapins showed the lowest necrotic lesion caused by Cyt. sorbicola (13.6 and 14.31 mm, respectively), followed by Bing (19.51 mm) and Regina (26.14 mm). All plants infected by Cyt. sorbicola showed shoot blight regardless of the variety. In addition, there was a varying susceptibility to Cal. pulchella, with Lapins (21.6 mm), Bing (22.83 mm), Santina (27.62 mm), and Regina (30.8 mm) showing increasing levels of observed necrosis. The lesion caused by Cal. pulchella was more significant than that observed for Cyt. sorbicola, regardless of the cherry tree genotype. We identified each fungal growth from the wood necrosis progression area using two independent novel PCR-HRM strategies based on the ITS fungal region, which allowed us to differentiate each pathogen of interest individually or simultaneously. This study demonstrates different levels of susceptibility of sweet cherry tree genotypes to wood-degrading pathogens, emphasizing the need to include these factors in phytosanitary management programs. Full article

Twig blight disease is the primary disease that affects the production of Myrica rubra in China. It was reported that exogenous brassinolide (BL) can improve disease resistance in plants. Here, we examined the effects of exogenous BL on disease resistance, chlorophyll contents, antioxidant enzyme activity, ROS accumulation, and key gene expression of M. rubra to analyze the mechanism of BR-induced resistance of twig blight disease in M. rubra. The results demonstrated that 2.0 mg·L⁻¹ of BL could significantly lessen the severity of twig blight disease in M. rubra. Exogenous BL increased the contents of chlorophyll a, chlorophyll b, carotenoids, and total chlorophyll. Moreover, exogenous BL also significantly enhanced the activity of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), and decreased malondialdehyde (MDA) content and reactive oxygen species (ROS) accumulation in leaves, such as H₂O₂ and O₂^·−. Additionally, exogenous BL dramatically up-regulated the expression of pathogenesis-related (PR) genes such as MrPR1, MrPR2, and MrPR10, as well as important genes such as MrBAK1, MrBRI1, and MrBZR1 involved in brassinosteroid (BR) signaling pathway. The transcriptome analysis revealed that a total of 730 common differentially expressed genes (DEGs) under BL treatment were found, and these DEGs were primarily enriched in four Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Based on these findings, nine important candidate genes related to the resistance of twig blight disease under BL treatment were further identified. In this study, we elucidated the effects of exogenous BL on enhancing the resistance of M. rubra to twig blight disease and preliminary analyzed the potential mechanism of resistance induction, which will provide a crucial foundation for the management and prevention of twig blight disease in M. rubra. Full article

Sclerotinia sclerotiorum (Lib.) de Bary (1884) is a fungal plant pathogen with worldwide distribution and a varying host range from different botanical families. It can cause damage to a large variety of crops such as sunflower, soybean, dry bean, canola, some vegetables, and ornamental plants. This article reports the occurrence of twig blight on the forsythia plant from the NE region of Romania. The disease was observed on Forsythia × intermedia Zab. plants from the Arboretum Park of the Iasi University of Life Sciences (IULS), located in Iasi City, Romania. Infected tissue was investigated through morphological characteristics using Sanger sequencing. Genomic DNA was extracted from the isolate obtained from naturally infected plants, and the ribosomal internal transcribed spacer region was amplified using the ITS1, ITS2, and LSU D1 and D2. Based on the results of this study, molecular and morphological data suggest that Forsythia twig blight can be caused by S. sclerotiorum. Constant monitoring of Sclerotinia sclerotiorum across multiple hosts and time intervals will reduce potential spread and future economic losses in cultivated species. Full article

Walnut dieback can be caused by several fungal pathogenic species, which are associated with symptoms ranging from branch dieback to fruit necrosis and blight, challenging the one pathogen–one disease concept. Therefore, an accurate and extensive description of the walnut fungal pathobiome is crucial. To this end, DNA metabarcoding represents a powerful approach provided that bioinformatic pipelines are evaluated to avoid misinterpretation. In this context, this study aimed to determine (i) the performance of five primer pairs targeting the ITS region in amplifying genera of interest and estimating their relative abundance based on mock communities and (ii) the degree of taxonomic resolution using phylogenetic trees. Furthermore, our pipelines were also applied to DNA sequences from symptomatic walnut husks and twigs. Overall, our results showed that the ITS2 region was a better barcode than ITS1 and ITS, resulting in significantly higher sensitivity and/or similarity of composition values. The ITS3/ITS4_KYO1 primer set allowed to cover a wider range of fungal diversity, compared to the other primer sets also targeting the ITS2 region, namely, GTAA and GTAAm. Adding an extraction step to the ITS2 sequence influenced both positively and negatively the taxonomic resolution at the genus and species level, depending on the primer pair considered. Taken together, these results suggested that Kyo set without ITS2 extraction was the best pipeline to assess the broadest fungal diversity, with a more accurate taxonomic assignment, in walnut organs with dieback symptoms. Full article

Bayberry twig blight caused by the ascomycete fungus Pestalotiopsis versicolor is a devastating disease threatening worldwide bayberry production. However, the molecular basis underlying the pathogenesis of P. versicolor is largely unknown. Here, we identified and functionally characterized the MAP kinase PvMk1 in P. versicolor through genetic and cellular biochemical approaches. Our analysis reveals a central role of PvMk1 in regulating P. versicolor virulence on bayberry. We demonstrate that PvMk1 is involved in hyphal development, conidiation, melanin biosynthesis, and cell wall stress responses. Notably, PvMk1 regulates P. versicolor autophagy and is essential for hyphal growth under nitrogen-depleting conditions. These findings suggest the multifaceted role of PvMk1 in regulating P. versicolor development and virulence. More remarkably, this evidence of virulence-involved cellular processes regulated by PvMk1 has paved a fundamental way for further understanding the impact of P. versicolor pathogenesis on bayberry. Full article

Blossom and twig blight, caused by Monilinia spp., is the main disease in apricot trees. In this study, we installed transparent rain shelters in apricot orchards to study their influence on the modification of the microclimate at the level of the tree canopy and on the reduction in moniliosis damage in twigs. Rain shelters significantly reduced the leaf wetness time measured within the foliage compared to the unsheltered trees (a reduction of between 43% and 67%). However, very few differences were observed in the daily averaged air temperature (up to 6%) and daily averaged air relative humidity (up to 1%). In the first experiment, on the apricot variety Bergarouge^® (CEP Innovation, Lyon, France), moniliosis damage on twigs in the absence of phytosanitary protection was reduced by up to 62% for the trees provided with rain protection compared to the trees that did not receive rain shelters. A second experiment, involving five apricot tree varieties, made it possible to verify that fungicide protection could be reduced for the trees protected by rain covers, reducing moniliosis damage on twigs compared to full fungicide protection combined without rain protection. Finally, a third experiment comprising two apricot tree varieties has shown that in organic orchards, rain protection provides protection against moniliosis (twig blight) that is equivalent to an organic farming fungicide protection programme based on the use of copper sulphate and calcium polysulphide. Full article

Between 2018 and 2021, several mastic trees (Pistacia lentiscus var. Chia) sampled in the field and the nursery of the Chios Mastiha Growers Association (CMGA) were analyzed to determine the cause of dominant diseases. Symptoms included defoliation, leaf, and twig blight, wilting and/or apoplexy of trees and apoplexy of young hardwood cuttings. Moreover, brown discoloration had also been observed on older woody parts of the trees such as branches and tree trunks. Several pathogens have been isolated and identified as the causing agents. Neopestalotiopsis and Alternaria species were isolated consistently from necrotic tissues of mastic trees (branches, twigs, and leaves) in the field and the nursery. All fungal isolates’ pathogenicity was confirmed by applying Koch’s postulates on young mastic trees under glasshouse conditions. Fungal pathogens were identified by sequence analyses of the ITS, β-tubulin, and histone gene regions. Alternaria species were analyzed further by sequencing the endopolygalacturonase (endoPG) and the Alternaria major allergen (Alta1) genes. More specifically, the isolates were identified as Neopestalotiopsis clavispora, Alternaria arborescens, and A. alternata based on morphological features and sequence analyses. This is the first report of N. clavispora, A. arborescens, and A. alternata on P. lentiscus var. Chia. Full article

Pistachio crops have a great economic potential, as their global production has increased dramatically over the past few decades. Therefore, it is important to maintain the healthy phytosanitary status of pistachio crops. In a Chilean pistachio orchard, a dieback of the trees was observed, with blighting of twigs and severe necrosis in the trunk and twigs. Bacterial isolation, pathogenicity tests and molecular characterization were conducted to determine the causal agent of the observed disease. The bacterial isolation and analysis of 16S rRNA gene led to the identification of Pantoea genus bacteria. Pathogenicity tests carried out on fruits inoculated with Pantoea isolates induced large necrosis seven days post-inoculation. Further inoculations were carried out on pruning cuttings and on the trunk of 18-month-old pistachio plants. Thirty-one weeks after inoculation, necrotic lesions were observed in the wood of pistachio plants. Sequence analysis of housekeeping genes enabled the isolated bacterium to be identified as Pantoa agglomerans, and to verify its role as the causal agent of the pistachio dieback with necrotic lesions. This is the first report of an occurrence of P. agglomerans inducing dieback in pistachio. Full article

Olive trees are infected and damaged by Botryosphaeriaceae fungi in various countries. The botryosphaeriaceous fungus Neofusicoccum mediterraneum is highly aggressive and is a major concern for olive groves in Spain and California (USA), where it causes ‘branch and twig dieback’ characterized by wood discoloration, bark canker, and canopy blight. During surveys of olive groves in Apulia (southern Italy), we noticed that—in some areas—trees were heavily affected by severe branch and twig dieback. In addition, chlorosis and the appearance of red-bronze patches on the leaf preceded the wilting of the foliage, with necrotic leaves persisting on the twigs. Given the severity of the manifestation in zones also subject to olive quick decline syndrome (OQDS) caused by Xylella fastidiosa subsp. pauca, we investigated the etiology and provide indications for differentiating the symptoms from OQDS. Isolation from diseased wood samples revealed a mycete, which was morphologically and molecularly identified as N. mediterraneum. The pathogenicity tests clearly showed that this fungus is able to cause the natural symptoms. Therefore, also considering the low number of tested samples, N. mediterraneum is a potential causal agent of the observed disease. Specifically, inoculation of the twigs caused complete wilting in two to three weeks, while inoculation at the base of the stem caused severe girdling wedge-shaped cankers. The growth rate of the fungus in in vitro tests was progressively higher from 10 to 30 °C, failing to grow at higher temperatures, but keeping its viability even after prolonged exposure at 50 °C. The capacity of the isolate to produce catenulate chlamydospores, which is novel for the species, highlights the possibility of a new morphological strain within N. mediterraneum. Further investigations are ongoing to verify whether additional fungal species are involved in this symptomatology. Full article

Blueberries (Vaccinium corymbosum) are widely cultivated worldwide and largely consumed due to their known antioxidant and medicinal properties. Although Diaporthe species have been documented in Portugal as causal agents of blueberry twig blight and dieback, there is still scarce information on the species that cause these symptoms. Moreover, Diaporthe vaccinii, recently synonymized with D. eres, has been considered a concern to blueberry production worldwide. However, the current knowledge about its impact on blueberries remains unclear. The diversity of Diaporthe species associated with diseased blueberry plants were assessed through a national survey. A multilocus sequence analysis of the rDNA internal transcribed spacer (ITS) region, the translation elongation factor 1-alpha (tef1-α), β-tubulin (tub2), calmodulin (cal) and histone 3 (his3) genes unveiled the presence of Diaporthe ambigua, D. amygdali, D. crousii, D. foeniculina, D. hybrida, D. leucospermi, D. malorum and D. rudis. Moreover, all species were fully characterized based on a detailed morphological description. Diaporthe amygdali, D. hybrida, D. leucospermi and D. malorum are reported for the first time on diseased blueberries in Portugal. Results show that D. eres exhibited a high level of intraspecific variability within isolates, given that the strain CBS 160.32 might be a minor pathogen on blueberry plants, whereas CAA829 was revealed to be the most aggressive. Overall, this study also demonstrates that Diaporthe amygdali and D. eres may be two of the most aggressive species to blueberry plants. This study improves our understanding of the Diaporthe species and it’s causing of dieback and twig blight on Portuguese blueberry orchards. Additionally, the identification of these pathogens represents crucial information for blueberry producers to apply appropriate phytosanitary measures, as well as offering new insights into the potential pathogenicity of D. eres on this host. Full article

The opportunistic pathogen Sphaeropsis sapinea (≡Diplodia sapinea) is one of the most severe pathogens in Scots pine, causing the disease Diplodia tip blight on coniferous tree species. Disease symptoms become visible when trees are weakened by stress. Sphaeropsis sapinea has an endophytic mode in its lifecycle, making it difficult to detect before disease outbreaks. This study aims to record how S. sapinea accumulates in trees of different health status and, simultaneously, monitor seasonal and age-related fluctuations in the mycobiome. We compared the mycobiome of healthy and diseased Scots pines. Twigs were sampled in June and September 2018, and filamentous fungi were isolated. The mycobiome was analyzed by high-throughput sequencing (HTS) of the ITS2 region. A PERMANOVA analysis confirmed that the mycobiome community composition significantly differed between growth years (p < 0.001) and sampling time (p < 0.001) but not between healthy and diseased trees. Sphaeropsis sapinea was the most common endophyte isolated and the second most common in the HTS data. The fungus was highly abundant in symptomless (healthy) trees, presenting in its endophytic mode. Our results highlight the ability of S. sapinea to accumulate unnoticed as an endophyte in healthy trees before the disease breaks out, representing a sudden threat to Scots pines in the future, especially with increasing drought conditions experienced by pines. Full article

In the past decade Colletotrichum gloeosporioides, C. karstii, and Alternaria alternata represent emerging fungal pathogens on citrus in the Mediterranean basin. Selection of tolerant Citrus germplasm offers evaluable long-term solution and should be considered as promising alternative to limit synthetic fungicide application to manage Alternaria and Colletotrichum infections in Citrus groves. In this study, the high variability of pre-harvest disease symptoms among 37 Citrus accessions was investigated in Italy in the most representative Sicilian production districts covering 1500 ha and including 20 homogeneous areas over an eleven-year survey period (2010–2020). Early fruit drop, brown spot, and anthracnose on fruit and leaves, pre-harvest fruit drop, and twig blight and defoliation associated to Alternaria and Colletotrichum spp. were identified on oranges, mandarins, and lemons. Comprehensively, first extensive data obtained herein on field susceptibility within “Tarocco” blood orange group to above disease symptoms clearly indicate as “Tarocco Nucellare 57-1E-1,” “Tarocco Tapi,” “Tarocco Sant’Alfio,” and “Tarocco Catania” accessions should be preferred to remaining Tarocco ones. A broad degree of tolerance or susceptibility was also observed within other Citrus group as it happens for tolerant lemon “Femminello Zagara Bianca,” thus demonstrating a putative resource for further studies to employ in a breeding program for genetic improvement of Citrus. Full article

Coffee agroforestry systems could reconcile agricultural and environmental objectives. While pests and diseases can reduce yield, their interactions with shade and nutrition have been rarely researched, and are particularly lacking in perennial systems. We hypothesized that intermediate shade levels could reduce coffee pests while excess shade could favor fungal diseases. We hypothesized that organic rather than mineral fertilization would better synchronize with nutrient uptake and higher nutrient inputs would be associated with reduced pest and disease damage due to higher plant vigor, yet effects would be less obvious in shaded plots as coffee growth would be light-limited. Using three-year-old trees of Coffea canephora var. Robusta (robusta coffee) in the Ecuadorian Amazon, we compared a full-sun system with four shading methods creating different shade levels: (1) Myroxylon balsamum; (2) Inga edulis; (3) Erythrina spp.; or, (4) Erythrina spp. plus Myroxylon balsamum. Conventional farming at either (1) moderate or (2) intensified input and organic farming at (3) low or (4) intensified input were compared in a split-plot design with shade as the main plot factor and farming practice as the sub-plot factor. The infestation of the following pests and disease incidences were evaluated monthly during the dry season: brown twig beetle (Xylosandrus morigerus), coffee leaf miner (Leucoptera coffeella), coffee berry borer (Hypothenemus hampei), anthracnose disease (Colletotrichum spp.), thread blight (Pellicularia koleroga), and cercospora leaf spot (Cercospora coffeicola). Coffee berry borer and brown twig beetle infestation were both reduced by 7% in intensified organic treatments compared to intensified conventional treatments. Colonization of coffee berry borer holes in coffee berries by the entomopathogenic fungus Beauveria bassiana was also assessed. Brown twig beetle infestation was significantly higher under full sun than under Inga edulis, yet no other shade effects were detected. We demonstrate for the first time how intensified input use might promote pest populations and thus ultimately lead to robusta coffee yield losses. Full article

Bayberry is an important fruit tree native to the subtropical regions of China. However, a systematic twig blight disease caused by Pestalotiopsis versicolor and P. microspora, resulted in the death of the whole tree of bayberry. The main variety Dongkui is highly sensitive to the twig blight disease, but the variety Zaojia is very highly resistant to the disease. Therefore, it is very necessary to clear the difference between resistant and susceptible varieties in response to the fungal infection. In this paper, we investigated the response of resistant and susceptible bayberry cultivars to infection of twig blight pathogen by histological observation and gibberellin signaling pathway-related genes expression. Microscopic observation revealed the difference in the infection process between resistant and susceptible varieties. The results of frozen scanning electron microscopy showed that the Pestalotiopsis conidia were shrunk, the mycelium was shriveled and did not extend into the cells of resistant cultivars, while the conidia were full and the top was extended, the mycelia was normal and continued to extend to the cells of a susceptible cultivar. Indeed, the medulla cells were almost intact in resistant cultivar, but obviously damaged in susceptible cultivar after inoculation of the main fungal pathogen P. versicolor conidia, which is earlier germinated on sterile glass slide than that of a hard plastic slide. The quantitative real-time PCR results showed a significant difference between resistant and susceptible cultivars in the expression of gibberellin signaling pathway-related genes in leaves and stems of bayberry, which is closely related to infection time, the type of genes and varieties. Overall, this study provides a clue for our understanding of the resistance mechanism of bayberry against the twig blight disease. Full article

Bayberry is an attractive, nutritious, and popular fruit in China. The plant fungal pathogen Pestalotiopsis versicolor XJ27 is the causative agent of bayberry twig blight disease, which severely affects bayberry production. Traditional control techniques, such as chemical fungicides, are being used to control this disease; however, these techniques cause environmental and health hazards. In this study, we screened sweet potato rhizospheric bacteria with biocontrol potentials against P. versicolor XJ27, the bayberry twig blight pathogen. Ten isolates showed inhibition; Bacillus siamensis S3 and Bacillus tequilensis S5 showed the highest fungal growth inhibition. The antagonistic bacterial culture suspensions of S3 and S5 inhibited the mycelial growth by 82.9% and 76.2%, respectively. Their extracellular culture filtrates had mycelial growth inhibition of 86.8% and 82.2%, respectively. In detached leaf assay, the extracellular culture filtrates of S3 and S5 inhibited the size of the leaf lesion by 82.3% and 76.2%, respectively. SEM and TEM imaging showed a severe hyphal-damaged structure caused by the antagonistic bacteria. The fungal inhibition mechanisms might employ the hydrolytic enzymes and lipopeptides produced by the bacteria. Both the S3 and S5 have chitinase and protease activity; they produce a series of lipopeptides such as surfactin, iturin, and mycosubtilin. Therefore, we can suggest these bacteria as biocontrol agents for bayberry twig blight disease as an alternative to fungicides based upon their attributes of antifungal activity. Full article