Search Results (14)

The diversity of foliar fungal endophyte communities was examined in three economically and ecologically important pine species in Scotland: Scots pine, Corsican pine and lodgepole pine. Two plantation sites comprising all three species were selected in climatically contrasting parts of Scotland and were sampled in late spring by collecting healthy needles from two age classes. Surface sterilisation was carried out before obtaining cultures of fungal isolates, and representatives of common sterile morphotypes were sequenced to determine taxonomic placement. Overall relative proportions of the dominant taxa across sites, tree species and needle age classes were as follows: Anthostomella spp. (52%), Lophodermium seditiosum (17%) and Desmazierella acicola (7%). Many other less frequent taxa were recovered. The results agreed with previous endophyte studies in that the combined effects of site and tree species produced unique endophytic fungal assemblages. Phylogenetic analyses revealed potential sub-species variation associated with site in Anthostomella pinea. Our findings point to the potential naturalisation of European fungal endophytic species (e.g., Anthostomella spp.) in Scottish pine plantations, particularly in association with Corsican pine. Full article

Multispectral imagery from unmanned aerial vehicles (UAVs) can provide high-resolution data to map tree mortality caused by pests or diseases. Although many studies have investigated UAV-imagery-based methods to detect trees under acute stress followed by tree mortality, few have tested the feasibility and accuracy of detecting trees under chronic stress. This study aims to develop methods and test how well UAV-based multispectral imagery can detect pine needle disease long before tree mortality. Multispectral images were acquired four times through the growing season in an area with pine trees infected by needle pathogens. Vegetation indices (VIs) were used to quantify the decline in vitality, which was verified by tree needle retention (%) estimated from the ground. Results showed that several VIs had strong correlations with the needle retention level and were used to identify severely defoliated trees (<75% needle retention) with 0.71 overall classification accuracy, while the accuracy of detecting slightly defoliated trees (>75% needle retention) was very low. The results from one study area also implied more defoliation observed from the UAV (top view) than from the ground (bottom view). We conclude that using UAV-based multispectral imagery can efficiently identify severely defoliated trees caused by needle-cast pathogens, thus assisting forest health monitoring. Full article

Antimicrobial properties of the new strains of micro-organisms isolated from natural sources of various ecological niches in the Moscow region and the Republic of Tatarstan were studied. Antifungal activity of isolates was detected in a test culture of toxin-producing microscopic fungi that can cause animal and plant diseases: Aspergillus flavus, Candida albicans, Fusarium oxysporum and Penicillium spp. Of the 46 studied micro-organisms of genera Bacillus, Lactobacillus, Lactococcus and Streptomyces isolates, there are four strains (Bacillus subtilis, Lactobacillus plantarum, Propionibacterium freudenreichii and Streptomyces spp.) that showed an ability to produce biologically active metabolites with a pronounced antimicrobial potential against phytopathogenic fungi metabolites. Based on the selected four strains, a Bacterial product LRV composition has been created. Scots pine, pedunculate oak and small-leaved linden seedlings with single and double foliar treatment and Bacterial product LRV at a concentration of 10 mL/L led to an increase in the growth of the aboveground part by 31.8, 51.9 and 25.4%, respectively, and the underground part by 25.0, 37.2 and 25.7%, respectively, compared to the control. The weight of seedlings at the end of the study exceeded the control variant by an average of 26.0, 44.0 and 78.0%, respectively. Plant protection Bacterial product LRV use did not have a significant effect on the group of molds that caused the powdery mildew and Schütte disease damage to trees. The Biological product LRV provided plant protection from fungal diseases caused by Lophodermium pinastri Chev. and Microsphaera alphitoides. Full article

Pathogenesis-related (PR) proteins are important in plant pathogenic resistance and comprise 17 families, including the PR4 family, with antifungal and anti-pathogenic functions. PR4 proteins contain a C-terminal Barwin domain and are divided into Classes I and II based on the presence of an N-terminal chitin-binding domain (CBD). This study is the first to isolate two PR4 genes, PaPR4-a and PaPR4-b, from Picea asperata, encoding PaPR4-a and PaPR4-b, respectively. Sequence analyses suggested that they were Class II proteins, owing to the presence of an N-terminal signal peptide and a C-terminal Barwin domain, but no CBD. Tertiary structure analyses using the Barwin-like protein of papaya as a template revealed structural similarity, and therefore, functional similarity between the proteins. Predictive results revealed an N-terminal transmembrane domain, and subcellular localization studies confirmed its location on cell membrane and nuclei. Real-time quantitative PCR (RT-qPCR) demonstrated that PaPR4-a and PaPR4-b expression levels were upregulated following infection with Lophodermium piceae. Additionally, PaPR4-a and PaPR4-b were induced in Escherichia coli, where the recombinant proteins existed in inclusion bodies. The renatured purified proteins showed antifungal activity. Furthermore, transgenic tobacco overexpressing PaPR4-a and PaPR4-b exhibited improved resistance to fungal infection. The study can provide a basis for further molecular mechanistic insights into PR4-induced defense responses. Full article

Picea asperata, a common tree species in the subalpine areas of Li County, Sichuan Province, China, is susceptible to Lophodermium piceae. Remote sensing has the advantages of large-scale, fast information acquisition, and low cost, which can overcome the shortcomings of ground survey. Hence, we used Landsat 8 satellite multi-spectral images and forest resource distribution data to investigate and analyze this forest disease at a large scale. Firstly, we extracted the spatial distribution information of Picea asperata and chose a temporal sequence indicator to establish a regression model and obtained a significantly negative correlation between the damage degree of plants and the change rate of normalized difference vegetation index (NDVI). Accordingly, the investigation results of the disease have good consistency with the ground survey data in spatial distribution and damage degree. On this basis, a temporal regression analysis was performed by combining the remote sensing investigation results with climate variables, and canonical correspondence analysis (CCA) was utilized in the spatial comprehensive analysis of Lophodermium piceae with terrain, soil and forest stand factors. Conclusively, this study effectively coped with the difficulties in full investigation and analysis of Lophodermium piceae in ecologically fragile subalpine areas of Western Sichuan. It is of important reference value in the early warning and monitoring of this disease, and also provides objective and reliable information support for ecological restoration and management planning in the Wenchuan earthquake-stricken areas. Full article

Thaumatin-like proteins (TLPs) are involved in the plant defense response against pathogens, and most of them exhibit antifungal activity. However, the role of TLPs in pathogen-induced defense responses in spruce is not fully understood. In this study, four TLP genes encoding thaumatin-like protein, designated as PlTLP1–4, were isolated and identified from Picea likiangensis needles. Sequence analysis showed that PlTLP1, PlTLP3, and PlTLP4 contained 16 conserved cysteine residues, while PlTLP2 had only 10 conserved cysteine residues. qPCR analysis showed that PlTLPs were expressed in all tissues tested, PlTLP1, PlTLP3, and PlTLP4 had the highest expression levels in young fruits, while PlTLP2 had the highest expression levels in roots. In addition, the expression levels of four PlTLPs were significantly upregulated during infection by Lophodermium piceae. Four recombinant PlTLPs expressed in Escherichia coli exhibited obvious β-1,3-glucanase activity. The antifungal activity assay showed that four recombinant PlTLPs had significant inhibitory effects on the mycelial growth of L. piceae, Fusarium proliferatum, Botrytis cinerea, and Roussoella doimaesalongensis. Microscopic observation revealed that the recombinant PlTLP1–4 induced the morphological changes of the mycelia of L. piceae, and the recombinant PlTLP2 and PlTLP3 induced the morphological changes of the mycelia of F. proliferatum and R. doimaesalongensis, while all the recombinant PlTLPs had no obvious negative effect on the morphology of B. cinerea mycelium. These results suggest that PlTLP genes may play an important role in the defense response of P. likiangensis against L. piceae invasion. Full article

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. Full article

The induction of defense responses in Pinus sylvestris L. seedlings by methyl jasmonate (MeJA) was investigated in three experiments. Two different MeJA application methods were tested, and induction of defense responses was assayed by seedling inoculation with Heterobasidion annosum (Fr.) Bref. and Lophodermium seditiosum Minter, Staley and Millar. In the first experiment, five-year-old P. sylvestris ramets of one clone were directly treated with MeJA, followed by inoculation with H. annosum. In the second experiment, open-pollinated Scots pine seedlings were treated with MeJA by direct spraying and vaporization, and inoculation with H. annosum was done using a slightly modified protocol. In the third experiment, open-pollinated Scots pine seedlings were treated with MeJA by vaporization and inoculated with L. seditiosum. Direct application of MeJA induced seedling mortality, and in some cases, decreased resistance to inoculation with H. annosum. Application of MeJA by vaporization was less stressful for seedlings, and resulted in increased resistance to both H. annosum and L. seditiosum. In addition, an unforeseen Neodiprion sertifer (Geoffroy) and Hylobius abietis L. infestation provided anecdotal evidence of the efficacy of MeJA in inducing resistance to insect pests as well. Further studies are required on the induction of resistance to additional diseases and pests. Induced resistance could be used as a possible protective mechanism for Scots pine seedlings prior to planting during reforestation of stands to increase vitality and survival. Full article

In Northern Europe, climate change may facilitate the prevalence of Scots pine, yet also promote the spread of pathogens attacking this species. A common biotic risk for Scots pine in nurseries and young stands is Lophodermium needle cast, primarily caused by Lophodermium seditiosum, which negatively affects the survival and growth of saplings. Reduced tree growth has been observed several years after damage by Lophodermium needle cast. However, for decision-making in protection or resistance breeding, an estimate of financial loss is important. Thus, the study aimed to assess the long-term influence of Lophodermium needle cast on the growth and financial value of Scots pine stands. The effect of needle cast damage during the sixth growing season on growth at the age of 17 years was evaluated in a control-crossed Scots pine progeny trial, and the results indicated a significantly negative effect on the height and diameter of the trees. A significant family effect also existed on the severity of the needle cast damage. Long-term simulations indicated that severely damaged Scots pines had a reduced equivalent annual annuity (EAA) of almost 100% at the final harvest. More intensive early management to reduce stand density could partly compensate for losses caused by needle cast. A higher EAA for the most resistant group of trees regardless of the stand management scenarios suggests an economically justified potential for improved resistance. Strong negative (−0.62 to −0.70) correlations of height and survival with the proportion of severely affected trees at the family mean level implies that resistant genotypes can be selected along with improved growth in progeny trials, which are affected by needle cast. Full article

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 different sites in Montenegro, where 2-4 year-old living needles of P. heldreichii were collected. Following DNA isolation, it was amplified using ITS2 rDNA as a marker and subjected to high-throughput sequencing. Sequencing resulted in 31,831 high quality reads, which after assembly were found to represent 375 fungal taxa. The detected fungi were 295 (78.7%) Ascomycota, 79 (21.0%) Basidiomycota and 1 (0.2%) Mortierellomycotina. The most common fungi were Lophodermium pinastri (12.5% of all high-quality sequences), L. conigenum (10.9%), Sydowia polyspora (8.8%), Cyclaneusma niveum (5.5%), Unidentified sp. 2814_1 (5.4%) and Phaeosphaeria punctiformis (4.4%). The community composition varied among different sites, but in this respect two sites at higher altitudes (harsh growing conditions) were separated from three sites at lower altitudes (milder growing conditions), suggesting that environmental conditions were among major determinants of fungal communities associated with needles of P. heldreichii. Trees on one study site were attacked by bark beetles, leading to discolouration and frequent dieback of needles, thereby strongly affecting the fungal community structure. Among all functional groups of fungi, pathogens appeared to be an important component of fungal communities in the phyllosphere of P. heldreichii, especially in those trees under strong abiotic and biotic stress. Full article

Phenylalanine ammonia-lyase (PAL) is the crucial enzyme of the phenylpropanoid pathway, which plays an important role in plant disease resistance. To understand the function of PAL in Picea asperata, in this study, the full-length cDNA sequence of the PAL gene from this species was isolated and named PaPAL. The gene contains a 2160-bp open reading frame (ORF) encoding 720 amino acids with a calculated molecular weight of 78.7 kDa and a theoretical isoelectric point of 5.88. The deduced PaPAL protein possesses the specific signature motif (GTITASGDLVPLSYIA) of phenylalanine ammonia-lyases. Multiple alignment analysis revealed that PaPAL has high identity with other plant PALs. The tertiary structure of PaPAL was predicted using PcPAL from Petroselinum crispum as a template, and the results suggested that PaPAL may have a similar function to that of PcPAL. Furthermore, phylogenetic analysis indicated that PaPAL has a close relationship with other PALs from the Pinaceae species. The optimal expression condition of recombinant PaPAL in Escherichia coli BL21 (DE3) was 0.2 mM IPTG (isopropyl β-D-thiogalactoside) at 16 °C for 4 h, and the molecular weight of recombinant PaPAL was found to be approximately 82 kDa. Recombinant PaPAL was purified and exhibited high PAL activity at optimal conditions of pH 8.6 and 60 °C. Quantitative real-time PCR (qRT-PCR) showed the PaPAL gene to be expressed in all tissues of P. asperata tested, with the highest expression level in the needles. The PaPAL gene was induced by the pathogen (Lophodermium piceae), which caused needle cast disease, indicating that it might be involved in defense against needle cast disease. These results provide a basis for understanding the molecular mechanisms of the PAL gene in the process of P. asperata disease resistance. Full article

Fungi play an essential role in recovering the quality and fertility of soil. There is a limited understating of the complex response of fungal diversity to different organic materials in clay loam soil. Here, we report the response of soil fungi toward the short-term application of manure (M), sugarcane straw (S), and sugarcane straw plus manure (MS), including no organic material control (CK) at two different time points (50 and 100 days after application). Illumina sequencing was used to examine the fungal communities. Our results reveal a significant shift among the soil fungal community structure associated with each organic material application. After both time points, amendments—especially M and MS—decreased the fungal richness and stimulated the copiotrophic fungal group (Ascomycota) compared to the control soil (CK) and S-amended soil. On the contrary, as compared to the M and MS-amended soils, the CK and S-amended soils at both time points increased the fungal richness and stimulated the oligotrophic fungal groups. Organic material use, especially M and MS, showed variable results regarding pathogenic fungi enhancing the abundance of Lophodermium and Cercophora and decreasing Fusarium. Concerning the abundance of plant-beneficial fungi, Mortierella was reduced, and Podospora was increased by M and MS input. FUNGuild showed that the amendment of organic materials efficiently declined the abundance of endophytes and plant pathogens, but also enhanced the animal pathogens in terms of abundance with respect to CK at two time points. This study could be useful to provide a novel understanding of the management of soil-borne pathogens by organic amendments for the sustainable production of short-term crops. Full article

Needle pathogens of larch (Larix spp.) in the Nordic countries are under-studied. Their incidence in Finland tends to be low and local, and this may be a function of enemy release, since species of larch were introduced to the region. Here, the ecology and incidence of larch needle pathogens and the abiotic factors that also affect larch in northern Finland are reviewed. Field observations and related laboratory analyses during the past 35 years have mainly been obtained near the Kivalo Research Area within the Arctic Circle, Finnish Lapland. The relatively recent introduction of Hypodermella laricis is a primary focus. This pathogen is not only new to Nordic countries, but can cause severe outbreaks, defoliation and crown-thinning in the canopies of all ages of most planted larch species worldwide. Symptoms of H. laricis clearly differ from those of Mycosphaerella laricina; the latter has affected Larix sibirica at high latitudes for decades. The effects of Meria laricis, Lophodermium laricinum, various rust fungi, and wind and frost are also discussed. Full article

Chitosan has become a promising biological agent for disease control and plant growth promotion. The objective of this study was to assess the effects of chitosan, applied as an active ingredient of Beta-chikol (Poli-Farm, Łowicz, Poland), to control damping-off and Lophodermium needle cast on Scots pine seedlings growing in field conditions. Beta-chikol was used for seed treatment and as a foliar spray at recommended rates and concentrations. For each experimental variant (chitosan, fungicides, unprotected), inventories of seedlings were performed, after germination and again after six weeks. In the aboveground parts of seedlings, the concentration of endogenous salicylic acid was determined by HPLC. At the end of the growing season, seedling growth parameters were determined. Beta-chikol used as foliar spray limited infection by the damping-off fungi but was ineffective when used as a seed treatment. Lophodermium needle cast was not observed during the study period. After the application of Beta-chikol, the concentration of salicylic acid did not increase. The application of Beta-chikol enhanced all growth parameters under investigation. Our results indicate the possibility of using chitosan in the form of Beta-chikol to stimulate plant growth and protect pine seedlings against parasitic damping-off in forest nurseries. Full article