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Pinus heldreichii is a high-altitude coniferous tree species naturaly occurring in small and disjuncted populations in the Balkans and southern Italy. The aim of this study was to assess diversity and composition of fungal communities in living needles of P. heldreichii specifically focusing on fungal pathogens. Sampling was carried out at six diff...
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... were taken at six sites that were situated in four mountain areas, representing the P. heldreichii distribution range in Montenegro (Figure 1). The sampling sites were Orjen (ORJ), Prekornica (PRE), Kučka Korita (KKO), Kučka korita North (KKN), Kuči MT (KMT) and Prokletije (PRO) (Figure 1, Table 1). ...Context 2
... following criteria were used for identification: sequence coverage >80%; 94-97% similarity to genus level and >98% similarity to species level. Sequences deviating from these criteria were identified only to a high taxonomic rank and were given unique names as shown in Table 3 and Table S1. Representative sequences of fungal nonsingletons are available from GenBank under accession numbers MT241905-MT242268. ...Context 3
... 3. Relative abundance of the 25 most common fungal taxa detected in needles of Pinus heldreichii at six sites in Montenegro. Sites are as in Table 1. ...Context 4
... of fungal ITS2 rDNA from 150 needle samples, PacBio sequencing and quality filtering resulted in 31,829 high quality reads. Sequence assembly and BLASTn analyses showed that the fungal community in the phyllosphere of P. heldreichii was composed of 375 fungal taxa (Table 2, Table S1). Nonfungal taxa and singletons were excluded. ...Context 5
... 79 (21.0%) Basidiomycota and 1 (0.2%) Mortierellomycotina. Identification at least to genus level was successful for 254 (67.7%) out of 375 fungal taxa (Table S1), and those represented 78.5% of all fungal sequences. ...Context 6
... of fungal ITS2 rDNA from 150 needle samples, PacBio sequencing and quality filtering resulted in 31,829 high quality reads. Sequence assembly and BLASTn analyses showed that the fungal community in the phyllosphere of P. heldreichii was composed of 375 fungal taxa (Table 2, S1). Nonfungal taxa and singletons were excluded. ...Context 7
... 79 (21.0%) Basidiomycota and 1 (0.2%) Mortierellomycotina. Identification at least to genus level was successful for 254 (67.7%) out of 375 fungal taxa (Table S1), and those represented 78.5% of all fungal sequences. ...Context 8
... most common fungal pathogens of pine needles were S. polyspora (8.9%), C. niveum (5.5%), Neocatenulostroma germanicum (2.3%), Allantophomopsiella pseudotsugae (2.1%) and Cenangium acuum (0.7%) (Table 3). A pine needle pathogen, Dothistroma septosporum (0.25%) was also detected, but at lower relative abundance (Table S1). The other detected pathogenic species that are mainly known as wound pathogens of deciduous Information on the 25 most common fungal taxa representing 79.0% of all fungal sequences is in Table 3. ...Context 9
... most common fungal pathogens of pine needles were S. polyspora (8.9%), C. niveum (5.5%), Neocatenulostroma germanicum (2.3%), Allantophomopsiella pseudotsugae (2.1%) and Cenangium acuum (0.7%) (Table 3). A pine needle pathogen, Dothistroma septosporum (0.25%) was also detected, but at lower relative abundance (Table S1). The other detected pathogenic species that are mainly known as wound pathogens of deciduous trees and/or agricultural crops, were Phaeomoniella 2814_15 (3.0%), Ramoconidiophora euphorbiae (1.5%), Collophorina sp. ...Context 10
... detected fungal endophytes that are known to produce antimicrobial metabolites were Phaeosphaeria pontiformis (4.4%), Microsphaeropsis olivacea (2.2%), Lachnellula calyciformis (2.0%) and Mollisia ligni (0.7%). Ubiquitous saprotrophs were rare (Table S1). ...Context 11
... detected fungal endophytes that are known to produce antimicrobial metabolites were Phaeosphaeria pontiformis (4.4%), Microsphaeropsis olivacea (2.2%), Lachnellula calyciformis (2.0%) and Mollisia ligni (0.7%). Ubiquitous saprotrophs were rare (Table S1). The community composition of the phyllosphere fungi varied among different sites (Figure 4). ...Context 12
... (Basidiomycotina) were rare and their relative abundance at different sites varied between 0.03% and 6.3% (Figure 4). Table 1. The NMDS of fungal communities associated with needles of P. heldreichii ( Figure 5) showed that KKO, PRO and PRE sites clustered together and were separated from the remaining sites along axis 1. Fungal communities at the ORJ and KMT showed a closer proximity (Figure 5), while fungal communities at the KKN site differed from all remaining sites and were separated along both axis 1 and 2. Classes comprising <1.0% of relative abundance for a given set of sequences were combined and shown as other. ...Context 13
... NMDS of fungal communities associated with needles of P. heldreichii ( Figure 5) showed that KKO, PRO and PRE sites clustered together and were separated from the remaining sites along axis 1. Fungal communities at the ORJ and KMT showed a closer proximity (Figure 5), while fungal communities at the KKN site differed from all remaining sites and were separated along both axis 1 and 2. Classes comprising <1.0% of relative abundance for a given set of sequences were combined and shown as other. Sites are as in Table 1. The NMDS of fungal communities associated with needles of P. heldreichii ( Figure 5) showed that KKO, PRO and PRE sites clustered together and were separated from the remaining sites along axis 1. Fungal communities at the ORJ and KMT showed a closer proximity (Figure 5), while fungal communities at the KKN site differed from all remaining sites and were separated along both axis 1 and 2. ...Context 14
... Chaetothyriales 2814_18 and Geastrumia sp. (Table S1). Diversity 2020, 12, x (Table S1). ...Context 16
... while providing detailed and semi-quantitative information, these tools enable studying the effects of different factors on fungal diversity and community composition [29,37,38]. By using PacBio sequencing, we detected nearly 400 fungal taxa, which were associated with needles of P. heldreichii including fungi present in very small abundances (Table S1). Many fungal taxa remained unidentified, which remains a major challenge in fungal taxonomy [39]. ...Context 17
... example, there were 446 and 260 fungal taxa recorded during studies of needle-associated fungi of Pinus sylvestris in Sweden and Poland, respectively [39,42]. Furthermore, by using fungal culturing, there were 118 fungal taxa associated with needles of Pinus taeda in USA [43], 35 taxa associated with needles of Pinus halepensis [44] and P. Table 2. Sites are as in Table 1. ...Context 18
... while providing detailed and semi-quantitative information, these tools enable studying the effects of different factors on fungal diversity and community composition [29,37,38]. By using PacBio sequencing, we detected nearly 400 fungal taxa, which were associated with needles of P. heldreichii including fungi present in very small abundances (Table S1). Many fungal taxa remained unidentified, which remains a major challenge in fungal taxonomy [39]. ...Context 19
... was suggested that C. niveum is more frequently associated with P. nigra under warmer climate conitions, while Cyclaneusma minus is favoured by wet, humid, above-freezing conditions, and thus, more commonly infects P. sylvestris grown in central and northern Europe [39,50,57]. Our study is in agreement, as on P. heldreichii, C. niveum dominated the fungal community, while C. minus was recorded at a low frequency (Table S1). Interestingly, C. niveum was more common on sites with prevailing moderate than harsh enviromental conditions. ...Context 20
... accum develops predominantly on needles damaged by frosts or drought and its development is favoured by a high humidity. Similarly, Dothistroma septosporum, was recorded at a low relative abundance and only on the PRE (Table S1). Investigation of Dothistroma septosporum accomplished using PCR and species-specific primers, has demonstrated the presence of this potentially invasive pathogen across the P. heldreichii distribution range in Montenegro [17]. ...Context 21
... Materials: The following are available online at http://www.mdpi.com/1424-2818/12/5/172/s1, Table S1: Relative abundance of fungal taxa detected in needles of Pinus heldreichii at six different sites in Montenegro, Table S2: Sørensen similarity index of the phyllosphere fungal communities among the six Pinus heldreichii sampling sites. ...Citations
... Additionally, some samples from the same population have a high EO yield, while others have low EO yield. As known, P. heldreichii is distributed at high altitudes of about 1400-2200 masl and is characterized by high adaptability to extreme climates and tolerance to low temperatures [43]. A compensatory effect of P. heldreichii to extreme climatic conditions and low water supply is slowing its metabolism [44]. ...
Pinus heldreichii Christ. (Bosnian pine), a Tertiary relict and Balkan sub-endemic, has not been comprehensively studied for its essential oil (EO) profile and bioactivity of its different plant parts. This study aimed to determine the EO yield, composition and antimicrobial activity from different parts of P. heldreichii at three different populations (mountains) in Bulgaria. Furthermore, the study assessed the antioxidant activities of plant tissue, including leaves (needles), twigs wood, male and female cones. The EOs yield from different plant parts ranged from 0.09 % (leaves) to 0.74 % (wood of twigs), with monoterpenes being the predominant class. Limonene, α-pinene, β-caryophyllene, germacrene D, β-pinene, and β-myrcene were detected in the EO extracted from all analyzed trees. However, these compounds were not found in the EO extracted from all plant parts of the same trees. Four chemical groups (chemotypes) were identified for EO from twigs, and three chemotypes were identified for EO from leaves. The chemotypes were based on the percent ratio of the main EO constituents (>5 %). Leaves tissue showed the highest values in terms of polyphenols and flavonoids, as well as higher ABTS radical scavenging activity, while the highest antimicrobial activity against Staphylococcus aureus subsp. aureus was seen in the EOs obtained from twigs. This is the first study to identify several chemotypes based on leaf and twigs EO of P. heldreichii distributed in Bulgarian flora. Furthermore, the EO of twigs tips (TT), male cones (MC), and wood of one-two-year-old twigs (WT) of the same trees were reported for the first time. The total polyphenol, flavonoid content, and radical scavenging activity of tissues of annual twigs wood and biennial twigs wood, leaf tissue, MC tissue, and the twigs tips tissue is also reported for the first time in the accessible literature. These findings highlight the potential of P. heldreichii to provide EOs with varying compositions and bioactivities, making them suitable for nutraceutical, pharmacological, and potentially food additive applications. Furthermore, the identification of chemotype accessions in this study suggests their selection for the development of new forest crop as a source for natural products with desirable composition and bioactivity.
... For example, the fungus was found to be endophytic in twigs of Scots pine in Germany, where it was isolated from 23% of samples (Bußkamp et al., 2020). This fungus has also been isolated as an endophyte of other tree and shrub species in Europe (Ellis & Ellis, 1997;Hormazabal et al., 2005;Lazarević & Menkis, 2020;Ruszkiewicz-Michalska & Mułenko, 2003). Microsphaeropsis olivacea was also reported to be the causal agent of the brown spine rot of Camelthorn (Alhagi maurorum Medik.) in Iran (Razaghi & Zafari, 2016). ...
In Poland's pine forests, mistletoe ( Viscum album subsp. austriacum ) abundance has increased over the past 20 years. This mistletoe infestation has decreased the growth and vigour of Scots pine trees. In this study, we surveyed the culturable fungi from healthy and diseased V . album subsp. austriacum leaves from two stands in Poland. In total, 63 distinct species were identified, 99.8% of which belonged to the phylum Ascomycota . The community compositions of fungi in mistletoe leaves were similar in healthy and diseased leaves as Alternaria alternata , Aureobasidium pullulans , Botryosphaeria visci , Fusarium paeoniae and Microsphaeropsis olivacea were consistently found in leaves of all symptom types. The most frequently isolated fungus from asymptomatic leaves was M . olivacea , followed by A . alternata , A . pullulans and Hypoxylon rubiginosum . In comparison, the most frequently isolated fungi from leaves with rusty‐brown necrotic spots were (in decreasing order) M . olivacea , B . visci , F . paeoniae and A . alternata , while the most frequently isolated fungi from leaves with black or dark brown spots were M . olivacea , A . alternata , A . pullulans , Epicoccum layuense and F . paeoniae . This study was the first comprehensive report showing that certain fungal species may be pathogens of V . album subsp. austriacum in Poland. This study was also the first report of F . paeoniae , F . juglandicola , Diaporthe vacuae and Heterotruncatella spartii from V . album , and the first report of D . vacuae and H . spartii in Poland.
... Many of them are epiphytic and then become endophytic by entering the internal tissues. Some phyllosphere fungi could turn to pathogens (Behnke-Borowczyk et al. 2019;Lazarevic and Menkis 2020), while others have antagonistic capacities and influence plant performance (Bashir et al. 2022). Yeasts are the major fungal epiphytes, among which Cryptococcus, Sporobolomyces and Rhodotorula are the commonly occurring genera (Glushakova and Chernov 2004). ...
Around plants, from interior to exterior, belowground to aboveground, they together comprise an intricate microbial ecosystem. Diverse microbial life inhabits it. Compared with the attention on rhizosphere microbiome (e.g., plant growth promoting rhizobacteria and soil borne pathogens), the current knowledge of phyllosphere mycobiome is still limited. In this chapter, we aim to provide a synthesis of current knowledge on phyllosphere mycobiome, including foliar endophytic fungi and epiphytic fungi from giant trees to dwarf shrubs, and from uncharted tropical rainforests to rural farmlands. Their high diversity and essential ecological functions are reviewed. We highlight the current knowledge about fungal biogeographic patterns, temporal dynamics, and community assembly processes in phyllosphere at the different temporal and spatial scales. Their environmental drivers, population sources as well as interactions with host plants and other plant-associated microbes are fully discussed. Considering the deep impacts of global change on the entire earth system, we further summarize the responses and potential feedbacks of phyllosphere mycobiome to several global change factors. By accumulating the knowledge of phyllosphere mycobiome, conserving their diversity, and utilizing their functions, we will be better to deal with the global environmental issues and rebuild a healthier plant planet in a sustainable way.KeywordsEndophytic fungiEpiphytic fungiPhyllosphere mycobiomeBiogeographic patternCommunity assemblyBiotic interactionsEcological functions
... The needle cast caused by L. seditiosum is a far more serious problem in North and Central Europe than in South Europe (Lazarev et al. 2007). The occurrence of L. seditiosum is not limited by ecological conditions, but pines are more susceptible in humid areas and mountainous areas, according to the experience from the mountainous areas of Montenegro, and taiga and forest-steppe areas in central Siberia (Lazarev et al. 2007;Lazarević and Menkis 2020;Sheller et al. 2020). ...
... To construct phylogenetic relationships and assess species boundaries within the genus, the Sanger sequencing method of different DNA regions (ITS nrDNA, LSU nrDNA, Actin) was used (Sokolski et al. 2004;Gao et al. 2013;Koukol et al. 2015;Salas-Lizana and Oono 2018). More recently, the diversity of fungal communities in pine needles was assessed using DNA metabarcoding analysis (Millberg et al. 2015;Behnke-Borowczyk et al. 2019;Lazarević and Menkis 2020). ...
Lophodermium species are well known to be among the dominant endophytic fungi colonizing pine needles. The occurrence of two species with different life strategies on different Pinus trees across Slovakia was detected by PCR using species-specific primers. In general, the commensalistic species L. pinastri was more frequent than parasitic L. seditiosum. There were no significant differences observed between urban and natural environments, but natural environments have more frequent colonization by L. pinastri and less frequent were trees negatively tested to Lophodermium colonization. Among the most frequently sampled trees were non-native P. nigra and native P. sylvestris. A significant difference in Lophodermium incidence was detected between these two species in a natural environment, with increased frequency of L. pinastri on native Scots pine. In addition to one clade of L. pinastri, culture based species identification confirmed the presence of L. corconticum and an undescribed clade of L. seditiosum that is not sensitive to used PCR primers for the species detection.
... Coniothyrium olivaceum) was the most common pathogen in shoots (Table 4). Although it can occur as an endophyte [98,99], it was also shown to cause brown spine rot in colonised tree tissues [100]. It has a broad host range and geographical distribution, suggesting that it can adapt to a variety of conditions. ...
Climate change, which leads to higher temperatures, droughts, and storms, is expected to have a strong effect on both health of forest trees and associated biodiversity. The aim of this study was to investigate the diversity and composition of fungal communities associated with the functional tissues and rhizosphere soil of healthy-looking Picea abies to better understand these fungal communities and their potential effect on tree health in the process of climate change. The study sites included 30 P. abies stands, where needles, shoots, roots, and the rhizosphere soil was sampled. DNA was isolated from individual samples, amplified using ITS2 rRNA as a marker and subjected to high-throughput sequencing. The sequence analysis showed the presence of 232,547 high-quality reads, which following clustering were found to represent 2701 non-singleton fungal OTUs. The highest absolute richness of fungal OTUs was in the soil (1895), then in the needles (1049) and shoots (1002), and the lowest was in the roots (641). The overall fungal community was composed of Ascomycota (58.3%), Basidiomycota (37.2%), Zygomycota (2.5%), Chytridiomycota (1.6%), and Glomeromycota (0.4%). The most common fungi based on sequence read abundance were Aspergillus pseudoglaucus (7.9%), Archaeorhizomyces sp. (3.6%), and Rhinocladiella sp. (2.0%). Pathogens were relatively rare, among which the most common were Phacidium lacerum (1.7%), Cyphellophora sessilis (1.4%), and Rhizosphaera kalkhoffii (1.4%). The results showed that the detected diversity of fungal OTUs was generally high, but their relative abundance varied greatly among different study sites, thereby highlighting the complexity of interactions among the host trees, fungi, and local environmental conditions.
... In recent years the study of fungal endophyte communities has been revolutionised with the development of metabarcoding based on amplification of ITS sequences from DNA extracted from individual samples, including in pines (Peršoh, 2013;Bullington and Larkin, 2015;Millberg et al., 2015;Taudiere et al., 2018, Lazarević andMenkis, 2020). This technique has the ability to detect unculturable and slow-growing taxa, can signal the presence of taxa at very low frequency (Thomas et al., 2016;Zinger et al., 2019), and with suitable caution (Amend et al., 2010), can be used to estimate the relative frequencies of taxa from counts of amplicon number. ...
To determine the role of environmental and host genetic factors in shaping fungal endophyte communities we used culturing and metabarcoding techniques to quantify fungal taxa within healthy Scots pine (Pinus sylvestris) needles in a 7-y old provenance-progeny trial replicated at three sites. Both methods revealed a community of ascomycete and basidiomycete taxa dominated by the needle pathogen Lophodermium seditiosum. Differences in fungal endophyte taxon composition and diversity indices were highly significant among trial sites. Within two sites, fungal endophyte communities varied significantly among provenances. Furthermore, the communities differed significantly among maternal families within provenances in 11/15 and 7/15 comparisons involving culture and metabarcoding data respectively. We conclude that both environmental and host genetic variation shape the fungal endophyte community of P. sylvestris needles.
... The isolation of the total DNA was completed using the cetyltrimethylammonium bromide (CTAB) method, after freeze drying and homogenising the needles. Amplification of the DNA using ITS2 rDNA as a marker, the PacBio amplicon sequencing and bioinformatics followed the method in a study by Lazarević and Menkis (2020). The fungal taxa were taxonomically identified using the GenBank database and the Blastn algorithm. ...
... The host range includes the genera Thuja, Abies, Tsuga, Larix, Picea and Pinus (Talgo et al. 2010). It was also found on needles of high-altitude pines in the Balkan region (Lazarević & Menkis 2020), showing that it has adapted to a broad host range and different environmental conditions. ...
... & Crous (1.3%) ( Table 1). Neocatenulostroma germanicum is a recently described fungus causing needle blight on pines in Lithuania, Ukraine and Poland (Markovskaja et al. 2016;Behnke-Borowczyk et al. 2019), but was also detected on needles of Pinus heldreichii H. Christ in Montenegro (Lazarević & Menkis 2020). The occurrence of N. germanicum on Abies was not known before, thereby the present study provides new information on its host range. ...
Abies concolor was introduced to Europe in the 19<sup>th</sup> century and commonly planted as an ornamental tree. In 2018, after several very warm and dry vegetation seasons, which has likely caused abiotic stress in the trees, a sudden dieback of A. concolor was observed in the city of Aranđelovac in Serbia. The external symptoms were the rapid discoloration of needles in a larger part of the crown and eventually the dieback of the trees. The aim of this study was to identify the possible agents of damage by analysing the fungal communities associated with symptomatic needles using high-throughput sequencing as no symptoms of damage were seen on the branches or stems. Symptomatic needles were collected from the lower branches of five standing trees. Half of the needles were surface sterilised and the remaining were left unsterilised. DNA was extracted, amplified using ITS2 rDNA as a marker and sequenced. Quality filtering showed the presence of 6 191 high quality reads, which, following a clustering analysis, was found to represent 79 non-singletons fungal taxa. The most common fungi in both surface sterilised and unsterilised needles were the pathogens Cytospora friesii (59.7%) and Sydowia polyspora (20.6%). The results demonstrated that C. friesii and S. polyspora can cause a rapid decline of A. concolor trees subjected to abiotic stress.
... PacBio sequencing of 190 needle samples from four sampling sites revealed 261 OTUs, of which L. conigenum, Phacidiaceae sp., Capnodiales sp. and D. septosporum prevailed. The genus Lophodermium is especially diverse on the needles of pine and these species mostly function as endophytes and saprobes [1][2][3]52], except the pathogen L. seditiosum. ...
... Lophodermium conigenum was also one of the three most abundant species on 1-yearold needles of Corsican black pine in Corsica in a study based on metabarcoding of the ITS2 rDNA region, being present in all three of the investigated sites, although with variable abundance [3]. In another study utilizing metabarcoding of ITS2, needles of Pinus heldreichii H. Christ harboured L. conigenum as the second most prevalent species in six different sites in Montenegro mountains [52]. These authors concluded that L. conigenum was primarily associated with trees in good health, being one of the most dominant species in sites with moderate growth conditions and healthy trees, but with low abundance in areas where beetle attacks and necrotic lesions (agent unknown) on needles occurred. ...
... These authors concluded that L. conigenum was primarily associated with trees in good health, being one of the most dominant species in sites with moderate growth conditions and healthy trees, but with low abundance in areas where beetle attacks and necrotic lesions (agent unknown) on needles occurred. It is noteworthy that Lazarevic and Menkis [52] pursued the entire spectrum of needleassociated fungi and thus pooled together 2-4-year-old needles, both healthy looking and symptomatic ones, as a single sample per each experimental tree, this complicating direct comparisons with other studies. Based on a fungal isolation approach, L. conigenum was the most frequently isolated fungal OTU from symptomless needles of P. sylvestris in Finland [1]. ...
The relationship between the ecological success of needle pathogens of forest trees and species richness of co-inhabiting endophytic fungi is poorly understood. One of the most dangerous foliar pathogens of pine is Dothistroma septosporum, which is a widely spread threat to northern European forests. We sampled two Pinus sylvestris sites in Estonia and two in Norway in order to analyse the relations between the abundance of D. septosporum and overall fungal richness, specific fungal species composition, time of season, needle age and position in the canopy. In both countries, the overall species richness of fungi was highest in autumn, showing a trend of increase with needle age. The overall species richness in the second-year needles in Estonia and third-year needles in Norway was similar, suggesting that a critical colonization threshold for needle shed in P. sylvestris is breached earlier in Estonia than in Norway. The fungal species richness in P. sylvestris needles was largely affected by Lophodermium conigenum. Especially in older needles, the relative abundance of L. conigenum was significantly higher in spring compared to summer or autumn. The timing of recruitment and colonization mechanisms of different foliage endophytes are shortly discussed.
... Согласно некоторым литературным источникам, N. germanicum отмечается на соснах, в доминирующей или сопутствующей микрофлоре которых присутствует Dothistroma spp. [31,33]. В данной работе возбудители дотистромоза (DNB) и коричневого пятнистого ожога (BSNB) генетически не были выявлены как в доминирующей, так и в сопутствующей микрофлоре исследуемых экземпляров сосны черной. ...
Red band needle blight, or Dothistroma needle blight is one of the most common and harmful diseases of pine. The causative agents of the disease are pathogenic micromycetes Dothistroma septosporum (Dorogin) M. Morelet and Dothistroma pini Hulbary. Dothistroma needle blight was firstly detected in Belarus in 2012 year, but till now information about this disease in the republic is fragmentary. The article presents the results of a survey of different pine trees, carried out in the period 2016–2020 years in botanical and dendrological gardens, forest nurseries and mini-arboretums at forestry enterprises, urban stands, nurseries of decorative plants, garden centers, for the presence of Dothistroma needle blight. The species identification of the causative agent of the disease was carried out by mycological and molecular genetic methods. In this study, Dothistroma needle blight was revealed on individual trees of Pinus mugo, P. nigra and P. ponderosa in the stands of the Central Botanical Garden of the NAS of Belarus, the dendrological garden of the Glubokoe experimental forestry enterprise, in the nurseries of decorative plants in the Grodno and Minsk regions. In the collected samples of needles, the invasive species Dothistroma septosporum was identified. The frequency of occurrence of the pathogen was 4.8–7.2 %, the proportion of observation sites in which this disease was detected at 60 %. The detection of Dothistroma needle blight on pine trees, mainly on planting material imported from abroad, indicates a transboundary route of D. septosporum entering the country. Analysis of literature data indicates the potential danger of Dothistroma needle blight for pine stands in the republic, which in turn requires the organization of regular monitoring of the disease and the development of methods to limit the spread of D. septosporum in the republic.
... Lachnellula species are reported as saprotrophs or parasites, with pathogenic species generally regarded as opportunistic pathogens causing cankers on twigs, branches, and stems of young trees subjected to stressful conditions or poor sites (Oguchi 1979;Smerlis 1973). While Lachnellula species are commonly encountered on wood, they are rarely reported as endophytes of needles (Lazarević and Menkis 2020;Moler and Aho 2018;Park et al. 2018). In this study, we describe polyketide metabolites from strains of Lachnellula calyciformis collected as endophytes from a healthy Picea rubens needle and Vaccinium angustifolium tissues. ...
... For example, Yuan et al. (2011) isolated two strains of L. calyciformis from Abies beshanzuensis needles in China and showed that one strain was a needle decomposer. Lachnellula calyciformis represented 0-8.1% of endophyte strains isolated from Pinus heldreichii needles across six sites in Montenegro (Lazarević and Menkis 2020). In previous studies of foliar endophytes of conifers, we isolated branch-colonizing fungi from surface sterilized needles, for example Coccomyces strobi on Pinus strobus (McMullin et al. 2019), Tryblidiopsis magnesii on Picea glauca (Tanney and Seifert 2019), and Pezicula sporulosa on P. rubens (McMullin et al. 2017). ...
As part of a survey of foliar endophytes of conifers in the Acadian forest, we isolated a strain of Lachnellula calyciformis from a healthy Picea rubens needle. From this fungus, the new natural products deoxylachnellulone (3) and lachnelluloic acid B (5), and the known metabolite lachnellulone (1), were isolated and structurally characterized. Isolachnellulone (2) and isodeoxylachnellulone (4) occurred as isomers resulting from the facile isomerization of lachnellulone (1) and deoxylachnellulone (3), respectively. The natural products lachnellulone (1) and deoxylachnellulone (3) were also confirmed from two strains of L. calyciformis isolated as leaf and stem endophytes of Vaccinium angustifolium in eastern Canada, representing the first record of this species from a non-conifer host. All isolated compounds displayed moderate antifungal and antibiotic activity. This study of biologically active natural products from a conifer endophyte highlights the increasing apparent complexity of their collective life histories.
