Search Results (17)

Pinus species are extensively abundant in Europe and, as pioneer trees, prominently influence local ecology. However, pine forests in Lithuania, Montenegro, and Ukraine have been significantly damaged by pine bark beetles (Tomicus sp.), which are closely associated with ophiostomatoid and other pathogenic fungi. This study aimed to identify the diversity of ophiostomatoid and other fungi associated with Tomicus sp. in these three countries. Fungi were isolated from beetles and identified. High-throughput sequencing of ITS2 rDNA yielded 285,828 reads, of which 91,141 high-quality reads were retained, representing 561 fungal operational taxonomic units (OTUs). The most important groups of fungi included ophiostomatoids, yeasts, and plant pathogens. While the fungal communities associated with Tomicus spp. were influenced more by environmental factors than by beetle species, the presence of known pathogens such as Ophiostoma spp. indicates that Tomicus spp. could play a significant role in dispersing harmful fungi. Although the virulence of these fungi may vary, their association with potentially pathogenic species suggests that Tomicus spp. may contribute to forest health decline, especially if environmental conditions or host susceptibility change. Full article

Pine wilt disease (PWD) is a devastating disease that occurs worldwide and affects conifers infested by the pine wood nematode (PWN, Bursaphelenchus xylophilus). PWD has caused serious economic and ecological losses in China. The mechanism of disease outbreak is complex, with the associated fungi, specifically ophiostomatoid fungi, thought to play an essential role. However, few ophiostomatoid fungal associates of PWD have been accurately identified. In the present study, we isolated fungi from bark beetles collected from Pinus thunbergii infested by the pine wood nematode on Laoshan Mountain, Shandong province. Three ophiostomatalean fungi were identified and assigned to Graphilbum and Ophiostoma based on phylogenetic analyses and comparison of morphological and cultural features, namely Gra. laoshanense sp. nov., Gra. translucens, and O. ips. This study increases the understanding of the diversity of ophiostomatoid fungi associated with PWD and provides resources for parsing this complex disease. Full article

Phytopathogenic ophiostomatoid fungi play an important role in pine wilt disease (PWD), caused by the pinewood nematode (PWN), since they begin proliferating once the pine hosts decay and can serve as a food source for the PWN. In a recent study, the ophiostomatoid fungi that are associated with naturally infected Pinus pinaster were profiled and cultured. To understand the influence of volatiles that are commonly emitted by pines on fungal growth, the present work aimed at analyzing the influence of α-pinene, β-pinene and trans-β-caryophyllene on a Leptographium isolate. The volatiles promoted fungal growth in the first 24 h, but lost their effect after 48 or 72 h, probably due to compound volatilization. After 5 days, the fungal growth was comparable to that of control cultures, except for α-pinene, which appeared to slightly inhibit fungal growth. Profiling the influence of volatile organic compounds on the PWD complex can contribute to a better understanding of the chemical communication that is occurring between its different intervenients. Full article

Forest decline is a significant issue affecting critical ecosystem processes worldwide. Here, we describe mortality in Pinus sylvestris L. monitored trees caused by the inhabitation of pine bark beetles (Tomicus minor Hart.) in symbiosis with ophiostomatoid fungi (Ophiostoma piceae (Munch) H. et P. Sydow) infection in the forest-steppe of central Siberia. Stem sap flow (Q) and stem diameter fluctuations (dRc) were monitored in eight pine trees during seven consecutive growing seasons (2015–2021). In addition, microcore sampling every ten days allowed the determination of stem wood formation in monitored trees in the 2021 growing season. During 2020 and 2021, two cases of Q termination were recorded among the monitored trees, with microcores revealing no cambium formation. Thus, the seasonal Q onset matches the beginning of the beetle dispersal period when they attack and inhabit tree stems. The decline of circumferential stem size began 10–12 days after Q onset, during the massive inhabitation of beetles into the stems. The disturbance of Q in trees occurred in 21–23 days, and total cessation of Q was observed 23–26 days after the Q onset at the beetle’s egg development phase. The timing of dRc disturbance and Q cessation observed directly coincides with the beetle life cycle. Thus, the phenology of pine trees and T. minor beetles is driven by seasonal weather conditions, particularly the cumulative air temperature (>0 °C). Full article

Bark beetles maintain symbiotic associations with a diversity of microbial organisms, including ophiostomatoid fungi. Studies have frequently reported the role of ophiostomatoid fungi in bark beetle biology, but how fungal symbionts interact with host chemical defenses over time is needed. We first investigated how inoculations by three fungal symbionts of mountain pine beetle affect the terpene chemistry of live lodgepole pine trees. We then conducted a complimentary laboratory experiment specifically measuring the host metabolite degradation by fungi and collected the fungal organic volatiles following inoculations with the same fungal species on lodgepole pine logs. In both experiments, we analyzed the infected tissues for their terpene chemistry. Additionally, we conducted an olfactometer assay to determine whether adult beetles respond to the volatile organic chemicals emitted from each of the three fungal species. We found that all fungi upregulated terpenes as early as two weeks after inoculations. Similarly, oxygenated monoterpene concentrations also increased by several folds (only in logs). A large majority of beetles tested showed a strong attraction to two fungal species, whereas the other fungus repelled the beetles. Together this study shows that fungal symbionts can alter host defense chemistry, assist beetles in overcoming metabolite toxicity, and provide possible chemical cues for bark beetle attraction. Full article

During a routine survey conducted in July 2021, several dead and dying Ulmus macrocarpa trees were observed in an urban forest located in the Gyeonggi Province of South Korea. The trees had symptoms of wilt with yellowing and browning of leaves, and, in most cases, the trunks of dying trees were infested by bark beetles. Isolations were made from small pieces of wood taken from dying trees, and beetles were collected from the infested stems. Fungal isolates and the beetles were identified using DNA sequence-based phylogenies and morphology, respectively. The results revealed that the fungus was Ophiostoma novo-ulmi, the causal agent of Dutch elm disease, and the associated bark beetle was Scolytus jacobsoni. This study provides the first record of Dutch elm disease in South Korea and suggests that a nationwide survey for the disease should be undertaken. Full article

The European spruce bark beetle, Ips typographus (L.), is a major pest of Norway spruce. During outbreaks, the beetles can colonize moderately stressed trees via mass attacks mediated by aggregation pheromones, while at endemic population levels, beetles infest trees with impaired defenses. I. typographus introduces ophiostomatoid fungi into the phloem, which can support host colonization. Low-density fungal infections are locally contained by hypersensitive wound reactions; larger necrotic lesions indicate lower tree resistance. Here, we made links between drought stress, susceptibility to fungal infections, and the attractiveness of spruce for host-searching I. typographus males. We sampled bark cores from roofed, non-roofed and untreated control trees of a rainfall exclusion field site. Drought stress was assessed using pre-dawn twig water potentials, and tree defenses were assessed using inoculations with Grosmannia penicillata. Subsequently, we performed Petri dish arena choice tests in the lab, where male beetles could choose between the bark samples of differentially stressed trees. We found that the attractiveness of bark cores increased with drought stress and the extent of hypersensitive wound reactions to fungal infection. Furthermore, beetles stayed longer in those Petri dish sections with the sample of their final choice. The bioassays provide evidence for the primary attraction of male I. typographus to tissues of Norway spruce and preference of beetles for stressed trees. Full article

Pinus armandii (P. armandii) is extensively abundant in western China and, as a pioneer tree, and prominently influences local ecology. However, pine forests in this region have been significantly damaged by Dendroctonus armandi (D. armandi) infestations, in close association with ophiostomatoid fungi. This study aimed to identify the diversity of ophiostomatoid fungi associated with D. armandi infesting P. armandii in western China. A total of 695 ophiostomatoid fungal strains were isolated from 1040 tissue pieces from D. armandi galleries and 89 adult beetles at four sites. In this study, based on multiloci DNA sequence data, as well as morphological and physiological characteristics, seven species belonging to five genera were identified including three known species, Esteyea vermicola, Graphium pseudormiticum and L. wushanense, two novel taxa, Graphilbum parakesiyea and Ophiostoma shennongense, and an unidentified Ophiostoma sp. 1. A neotype of Leptographium qinlingense. Ophiostoma shennongense was the dominant taxon (78.99%) in the ophiostomatoid community. This study provides a valuable scientific theoretical basis for the occurrence and management of D. armandi in the future. Full article

Ophiostomatoid fungi are known for their associations with bark beetles, and some species are important sources of tree diseases. Ceratocystiopsis is a genus of the ophiostomatoid fungi in order Ophiostomatales. The shortage of DNA barcodes for many species in this genus has resulted in the presence of many unnamed cryptic species. In this study, Ceratocystiopsis subelongati sp. nov. associated with Ips subelongatus infesting Pinus sylvestris var. mongolica in Inner Mongolia, China, was identified and described based on phylogenetic inference of multi-gene DNA sequences and morphological characteristics. The species is characterized by a hyalorhinocladiella- to sporothrix-like asexual state and an optimal growth temperature of 30 °C. Artificial inoculation tests in the field showed that it is mildly pathogenic to five-year-old larch trees, the main host of I. subelongatus. It is also the first described Ceratocystiopsis species associated with I. subelongatus in China. This discovery should provide new avenues for studying the symbiosis between bark beetles and ophiostomatoid fungi. Full article

Drought-induced stress and attacks by bark beetle Ips sexdentatus currently result in a massive dieback of Pinus sylvestris in eastern Ukraine. Limited and fragmented knowledge is available on fungi vectored by the beetle and their roles in tree dieback. The aim was to investigate the fungal community vectored by I. sexdentatus and to test the pathogenicity of potentially aggressive species to P. sylvestris. Analysis of the fungal community was accomplished by combining different methods using insect, plant, and fungal material. The material consisted of 576 beetles and 96 infested wood samples collected from six sample plots within a 300 km radius in eastern Ukraine and subjected to fungal isolations and (beetles only) direct sequencing of ITS rDNA. Pathogenicity tests were undertaken by artificially inoculating three-to-four-year-old pine saplings with fungi. For the vector test, pine logs were exposed to pre-inoculated beetles. In all, 56 fungal taxa were detected, 8 exclusively by isolation, and 13 exclusively by direct sequencing. Those included nine ophiostomatoids, five of which are newly reported as I. sexdentatus associates. Two ophiostomatoid fungi, which exhibited the highest pathogenicity, causing 100% dieback and mortality, represented genera Graphium and Leptographium. Exposure of logs to beetles resulted in ophiostomatoid infections. In conclusion, the study revealed numerous I. sexdentatus-vectored fungi, several of which include aggressive tree pathogens. Full article

Pine wilt disease (PWD) is a complex disease that severely affects the biodiversity and economy of Eurasian coniferous forests. Three factors are described as the main elements of the disease: the pinewood nematode (PWN) Bursaphelenchus xylophilus, the insect-vector Monochamus spp., and the host tree, mainly Pinus spp. Nonetheless, other microbial interactors have also been considered. The study of mycoflora in PWD dates back the late seventies. Culturomic studies have revealed diverse fungal communities associated with all PWD key players, composed frequently of saprophytic fungi (i.e., Aspergillus, Fusarium, Trichoderma) but also of necrotrophic pathogens associated with bark beetles, such as ophiostomatoid or blue-stain fungi. In particular, the ophiostomatoid fungi often recovered from wilted pine trees or insect pupal chambers/tunnels, are considered crucial for nematode multiplication and distribution in the host tree. Naturally occurring mycoflora, reported as possible biocontrol agents of the nematode, are also discussed in this review. This review discloses the contrasting effects of fungal communities in PWD and highlights promising fungal species as sources of PWD biocontrol in the framework of sustainable pest management actions. Full article

Advancing climate change is affecting the health and vitality of forests in many parts of the world. Europe is currently facing spruce bark beetle outbreaks, which are most often caused by wind disturbances, hot summers, or lack of rainfall and are having a massive economic impact on the forestry sector. The aim of this research article was to summarize current scientific knowledge about the structure and physical and mechanical properties of wood from bark beetle-attacked trees. Spruce stands are attacked by a number of beetles, of which Ips typographus is the most common and widespread in Central Europe. When attacking a tree, bark beetles introduce ophiostomatoid fungi into the tree, which then have a greater effect on the properties of the wood than the beetles themselves. Fungal hyphae grow through the lumina of wood cells and spread between individual cells through pits. Both white rot and brown rot fungi are associated with enzymatic degradation of lignin or holocellulose, which is subsequently reflected in the change of the physical and mechanical properties of wood. Wood-decay fungi that colonize wood after infestation of a tree with bark beetles can cause significant changes in the structure and properties of the wood, and these changes are predominantly negative, in the form of reducing modulus of rupture, modulus of elasticity, discolouration, or, over time, weight loss. In certain specific examples, a reduction in energy consumption for the production of wood particles from beetle-attacked trees, or an increase in surface free energy due to wood infestation by staining fungi in order to achieve better adhesion of paints or glues, can be evaluated positively. Full article

The European spruce bark beetle (Ips typographus) has become a major forest pest in Finland in recent years. The beetle is a well-known vector of mainly ophiostomatoid fungi causing blue-stain of timber and pathogens that have the ability to amplify the insect damage. It also vectors other associated organisms, such as phoretic mites. The ecology of these mites remains poorly understood, including their associations with fungi. In this study, we considered filamentous fungi and yeasts associated with mites phoretic on I. typographus. Fungal identifications were based on DNA sequences and phylogenetic analyses of the ITS and/or partial β-tubulin gene regions. Fifteen fungal species were detected, including eight yeasts and seven filamentous fungi. Eleven percent of the beetles carried mites and of these 74% carried at least one fungal species. An average of two fungal species were carried per mite. The most commonly found filamentous fungi were Grosmannia penicillata (25%), Ophiostoma bicolor (19%), O. ainoae (12%) and O. brunneolum (12%). Of the yeast species, the most commonly found was Wickerhamomyces bisporus (47%). This study is the first to report yeasts associated with I. typographus and its phoretic mites in Finland. Majority of the filamentous fungal species found are those previously reported in association with I. typographus. The results also confirmed that many of the fungal species commonly found on I. typographus are also associated with its phoretic mites. However, the nature of the symbiosis between the mites, beetles and fungal associates remains to be understood. Full article

Research Highlights: Raffaelea quercina sp. nov. is an ophiostomatoid fungus isolated from the ambrosia beetle Platypus cylindrus. The species occurs in symptomatic Portuguese cork oak trees, (Quercus suber L.), exhibiting vegetative decline. Background and Objectives: Quercus suber L. is a species restricted to the Mediterranean basin, of special economic importance as it constitutes the crucial raw material for the cork production industry, in particular for Portugal, the world’s leading producer. Over the last three decades a progressive and alarming decline of cork oak trees has been observed across its distribution area, including Portugal. The ambrosia beetle Platypus cylindrus, commonly known as the oak pinhole borer, establishes symbiotic relationships with fungi from which it depends for survival and for oak colonization. Some of these fungi are ophiostomatoid species of the Raffaelea genus, known as ambrosia fungi associated with ambrosia beetles. Some Raffaelea species exhibit phytopathogenic activity causing wilting and/or death of trees. The objective of the present study is to identify the association between P. cylindrus and Raffaelea species in Portuguese cork oak stands showing symptoms of disease and decline. Materials and Methods: A total of 300 adult insects were collected as they emerged from cork oak logs, sampled from symptomatic trees. Axenic isolates of Raffaelea species were obtained from the beetles and their galleries in the trunks and identified based on morphological features and molecular analysis of the SSU and LSU rDNA regions. Results: Two Raffaelea species were identified, i.e., R. montetyi and a novel Raffaelea species closely related to R. canadensis. The novel species is morphologically and genetically characterized in this study, and erected as Raffaelea quercina M.L. Inácio, E. Sousa & F. Nóbrega, sp. nov. Raffaelea quercina constitutes a new phytopathogenic fungal species associated with P. cylindrus and cork oak trees exhibiting symptoms of vegetative decline. Conclusions: Raffaelea species appear to have a significant role in cork oak decline. Future research on the association between P. cylindrus and Raffaelea species, encompassing the trans-European and North African wide-range of cork oak stands, would further clarify the relationships between ambrosia beetles, associated fungi and cork oak decline, contributing to a better understanding of the phenomena and for strategies aiming to halt the continuous decline of the unique cork oak stands enclosed in the Mediterranean basin. Full article

Seedlings from four families of loblolly pine (Pinus taeda L.) were grown in capped open-top chambers and exposed to three different weekly moisture regimes for 13 weeks. Moisture regimes varied in intensity and frequency of simulated rainfall (irrigation) events; however, the total amounts were comparable. These simulated treatments were chosen to simulate expected changes in rainfall variability associated with climate change. Seedlings were inoculated with two root-infecting ophiostomatoid fungi associated with Southern Pine Decline. We found susceptibility of loblolly pine was not affected by water stress; however, one family that was most sensitive to inoculation was also most sensitive to changes in moisture availability. Many studies have examined the effects of drought (well-watered vs. dry conditions) on pine physiology and host-pathogen interactions but little is known about variability in moisture supply. This study aimed to elucidate the effects of variability in water availability, pathogen inoculation and their interaction on physiology of loblolly pine seedlings. Full article