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In this paper, we report the influence of pine, oak and spruce forest litter on the growth and ectomycorrhizal (ECM) formation of Scots pine seedlings after the first growing season in a bare-root forest nursery. The mixture of collected forest litters and humus were used to obtain a 20-cm mulching layer on the prepared seedbeds. The concentrations of all nutrients and the C/N ratio of growth media were significantly higher in forest litter treatments than in negative control represented by mineral soil without litter. Addition of each forest litter type significantly enhanced pine seedling height and root-collar diameter compared to negative control. A significant positive influence on dry mass of stem, needles, roots and total dry mass of the seedling has been found only for pine litter. Based on molecular identification, seven ECM fungal taxa (Wilcoxina mikolae, Suillus luteus, Cenococcum geophilum, Meliniomyces bicolor, Laccaria laccata, unidentified Atheliaceae, unidentified Ascomycetes) were distinguished in the observed mycorrhizal communities. Each forest litter type significantly increased the total number of mycorrhizal tips and ECM fungal diversity compared to the control soil. However, results showed a lack of significant differences in species composition and relative abundance of ECM fungi between different litter types. Such result suggests that forest litter has not been a key source of inoculum for tested fungal species, as root systems of all pine seedlings from different litter types were dominated by a few nursery- adapted ECM fungi, probably originating from natural air-borne inoculum. Our data rather indicate that forest litter considerably improves environmental conditions for development of ECM fungi previously present in the nursery soil. Therefore, any of the forest litter types used in our studies may be able to promote planting stock quality on a small scale in the nursery phase.

Ectomycorrhizae are obligate symbiotic structures of forest tree roots and are susceptible to environmental changes. Ectomycorrhizae of Scots pine (Pinus sylvestris L.) trees grown in forests influenced by different levels of air pollutants were investigated. Total numbers of mycorrhizal root tips in the soil horizons and the frequency of mycorrhizal morphotypes were studied as indicators of ectomycorrhizal status. The studies were conducted in two comparable young plantations in western Poland with high and low pollution exposure and in two mature forest ecosystems in southern Poland differing in pollution exposure. At the young Scots pine plantation with high pollution exposure (Luboti), numbers! of mycorrhizal root tips in the soil layer 0-5 cm were, lower during three growing seasons than at the cleaner site (Kornik). In the soil layer 15-30 cm the total number pf mycorrhizal root tips was found slightly higher at the polluted than at the control site. There was no difference yin the frequency of the seven mycorrhizal morphotypes found at the two young plantations. At the heavily polluted mature forest stand (Niepolomice Forest) the total number of mycorrhizal root tips was lower in zones of severe pollution levels than in the less polluted zones. Moreover, the average number of mycorrhizae in the Niepolornice Forest was lower than in the moderately polluted forest ecosystem (Ratanica catchment). The mycorrhizaidiversityof the mature pines in the heavily polluted forest, was reduced (four morphotypes) as compared to the moderately polluted forest (eight morphotypes). The relationship between industrial pollution and mycorrhizal status vas more pronounced at the mature Scots pine stands thanat the young plantations. [ABSTRACT FROM AUTHOR]

Soil fungi play an indispensable role in the functioning of terrestrial habitats. Most landscape-scale studies of soil fungal diversity try to identify the fungal taxa present at a study site and define the relationships between their abundance and environmental factors. The specific spatial distribution of these fungi over the site, however, is not addressed. Our study's main objective is to propose a novel approach to landscape-scale mapping of soil fungi distribution using next generation sequencing and geographic information system applications. Furthermore, to test the proposed approach and discuss its performance, we aimed to conduct a case study mapping the spatial distribution of soil fungi on the Wielka Żuława island. The case study was performed on the Wielka Żuława island in northern Poland, where soil samples were collected every 100 m in an even grid. The fungal taxa and their relative abundance in each sample were assessed using the Illumina platform. Using the data obtained for the sampled points, maps of soil fungi spatial distribution were generated using three common interpolators: inverted distance weighted (IDW), B-spline, and ordinary Kriging. The proposed approach succeeded in creating maps of fungal distribution on Wielka Żuława. The most abundant groups of soil fungi were Penicillium on the genus level, Aspergillaceae on the family level, and ectomycorrhizal fungi on the trophic group level. Ordinary Kriging proved to be the most accurate at predicting relative abundance values for the groups of fungi significantly spatially autocorrelated at the sampled scale. For the groups of fungi not displaying spatial autocorrelation at the sampled scale, IDW provided the most accurate predictions of their relative abundance. Although less accurate at predicting exact relative abundance values, B-spline performed best in delineating the spatial patterns of soil fungi distribution. The proposed approach to landscape-scale mapping of soil fungi distribution could provide new insights into the ecology of soil fungi and terrestrial ecosystems in general. Producing maps of predicted fungal distribution in landscape-scale soil fungi diversity studies would also facilitate the reusability and replicability of the results. Outside the area of research, mapping the distribution of soil fungi could prove helpful in areas such as agriculture and forestry, nature conservation, and urban planning., (© 2023. The Author(s).)

An uncontrolled, natural episode of flooding with waters contaminated with As-rich pyrite (FeAsS) particles caused serious ecological damage leading to necrosis of plants growing in a fresh wet meadow located in an area characterized by unique geological structures rich in arsenopyrites. One of the few plant species capable of surviving this event was Salix aurita L., which grew in numbers in the analyzed area, but individual plants were affected differently by toxic flooding. No significant phenotypic changes (Group I), through partial leaf and/or stem necrosis (Group II) up to necrosis of the whole parental plant and root suckers (Group III), were observed for various willow clumps. These varied phenotypic responses of S. aurita to As-rich sediments were compared with the biochemical status of the foliage of willow trees, and with their rhizosphere physiological parameters. Our in situ study revealed that the biochemical status of leaves reflects the phenotypic damage incurred by adult willows growing in their natural environment and affected by the flooding. In leaves of willows with increasingly negative phenotypic changes (Groups I → II → III) as well as increasing levels of reactive oxygen species, malondialdehyde and decreased levels of glutathione and thiol groups were detected. Phytochelatins, commonly considered major As chelators, were not detected in S. aurita leaves. Despite a decrease in the size of leaves with the intensity of tree damage, all leaves expressed a normal level of leaf pigments. Phenotypic changes observed for particular willow clumps were only partly related to soil As levels. Moreover, As and S (but not Fe) foliar levels were related but did not correspond strictly with foliar biochemical features, or with soil As levels, soil pH or soil microbial activity, with the latter two drastically decreased in the rhizospheres of willows from Groups II and III., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

A large proportion of ongoing malaria parasite transmission is attributed to low-density subclinical infections not readily detected by available rapid diagnostic tests (RDTs) or microscopy. Plasmodium falciparum gametocyte carriage is subclinical, but gametocytemic individuals comprise the parasite reservoir that leads to infection of mosquitoes and local transmission. Effective detection and quantification of these carriers can help advance malaria elimination strategies. However, no point-of-need (PON) RDTs for gametocyte detection exist, much less one that can perform noninvasive sampling of saliva outside a clinical setting. Here, we report on the discovery of 35 parasite markers from which we selected a single candidate for use in a PON RDT. We performed a cross-sectional, multi-omics study of saliva from 364 children with subclinical infection in Cameroon and Zambia and produced a prototype saliva-based PON lateral flow immunoassay test for P. falciparum gametocyte carriers. The test is capable of identifying submicroscopic carriage in both clinical and nonclinical settings and is compatible with archived saliva samples., (Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

We studied mycorrhizal associations of North American Carya laciniosa and Carya cordiformis trees, successfully acclimated to local habitat conditions of the historic Kórnik Arboretum in Poland, in order to better understand mycorrhizal host range extensions in new environments. The root systems of Carya seedlings (1-3 years old), regenerated under a canopy of mature hickory trees, were analyzed using microscopic, morphological, and molecular techniques. Our results, for the first time, indicate that C. laciniosa and C. cordiformis have both arbuscular and ectomycorrhizal associations. In the cleared and stained roots of both Carya species, typical structures of arbuscular mycorrhizae (vesicles, arbuscules, hyphal coils, and intercellular nonseptate hyphae) were detected. On the basis of ITS rDNA sequencing, 40 ectomycorrhizal fungal taxa were revealed, with 25 on C. laciniosa and 19 on C. cordiformis. Only four fungal species (Cenococcum geophilum sensu lato, Russula recondita, Xerocomellus cisalpinus, Humaria hemisphaerica) were shared by both Carya species. The high number of infrequent fungal taxa found, as well as the calculated richness estimator, indicates that the real ectomycorrhizal community of C. laciniosa and C. cordiformis is probably richer. The ability of the exotic Carya species to form arbuscular and ectomycorrhizal linkages with native fungi could be a factor in the successful establishment of these tree species under the conditions of Kórnik Arboretum.

Abies alba (Mill.) is an important forest tree species, native to the mountainous regions of Europe but has been also widely introduced in the lowlands outside its native range. Like most forest tree species, A. alba forms obligate mutualisms with ectomycorrhizal (ECM) fungi. This investigation sought to examine ECM fungal communities of A. alba when the species grows 400 km north of its native range in the region of Pomerania in Poland. We surveyed for ECM fungi by sampling live roots from four mature forest stands where the A. alba component ranged from 20 to 100%. Ectomycorrhizal fungal symbionts were identified based on morphotyping and sequencing of the internal transcribed spacer (ITS) of nuclear ribosomal DNA (rDNA). Thirty-five ECM fungal taxa were distinguished on root tips of A. alba from all tested stands with 22 to 27 ECM fungal taxa in the individual stand. The diversity and similarity metrics revealed a lack of statistical differences in the structure of the ECM fungal community between stands varying in overstory tree composition. Cenococcum geophilum was the most common fungal species at all investigated A. alba stands, with an abundance of 50 to 70%. The ECM community was characterized by the lack of Abies-specific fungal symbionts and a rich and diverse suite of host-generalist mycobionts that seem to be sufficient for successful growth and development of A. alba outside of its native range.

Non-native tree species have been widely planted or have become naturalized in most forested landscapes. It is not clear if native trees species collectively differ in ectomycorrhizal fungal (EMF) diversity and communities from that of non-native tree species. Alternatively, EMF species community similarity may be more determined by host plant phylogeny than by whether the plant is native or non-native. We examined these unknowns by comparing two genera, native and non-native Quercus robur and Quercus rubra and native and non-native Pinus sylvestris and Pinus nigra in a 35-year-old common garden in Poland. Using molecular and morphological approaches, we identified EMF species from ectomycorrhizal root tips and sporocarps collected in the monoculture tree plots. A total of 69 EMF species were found, with 38 species collected only as sporocarps, 18 only as ectomycorrhizas, and 13 both as ectomycorrhizas and sporocarps. The EMF species observed were all native and commonly associated with a Holarctic range in distribution. We found that native Q. robur had ca. 120% higher total EMF species richness than the non-native Q. rubra, while native P. sylvestris had ca. 25% lower total EMF species richness than non-native P. nigra. Thus, across genera, there was no evidence that native species have higher EMF species diversity than exotic species. In addition, we found a higher similarity in EMF communities between the two Pinus species than between the two Quercus species. These results support the naturalization of non-native trees by means of mutualistic associations with cosmopolitan and novel fungi.

Ectomycorrhizal (ECM) communities of mature trees and regenerating seedlings of a non-native tree species Pinus mugo grown in a harsh environment of the coastal region of the Curonian Spit National Park in Lithuania were assessed. We established three study sites (S1, S2, and S3) that were separated from each other by 15 km. The ECM species richness was rather low in particular for mature, 100-year-old trees: 12 ectomycorrhizal taxa were identified by molecular analysis from 11 distinguished morphotypes. All 12 taxa were present on seedlings and on mature trees, with between 8-11 and 9-11 taxa present on seedlings and mature trees, respectively. Cenococcum geophilum dominated all ECM communities, but the relative abundance of C. geophilum mycorrhizas was nearly two times higher on seedlings than on mature trees. Mycorrhizal associations formed by Wilcoxina sp., Lactarius rufus, and Russula paludosa were also abundant. Several fungal taxa were only occasionally detected, including Cortinarius sp., Cortinarius obtusus, Cortinarius croceus, and Meliniomyces sp. Shannon's diversity indices for the ECM assemblages of P. mugo ranged from 0.98 to 1.09 for seedling and from 1.05 to 1.31 for mature trees. According to analysis of similarity, the mycorrhizal communities were similar between the sites (R = 0.085; P = 0.06) and only slightly separated between seedlings and mature trees (R = 0.24; P < 0.0001). An incidental fruiting body survey that was conducted weakly reflected the below-ground assessment of the ECM fungal community and once again showed that ECM and fruiting body studies commonly supply different partial accounts of the true ECM fungal diversity. Our results show that P. mugo has moved into quite distinct habitats and is able to adapt a suite of ECM symbionts that sufficiently support growth and development of this tree and allow for natural seedling regeneration.

In this paper, we report the effect of Scots pine genotypes on ectomycorrhizal (ECM) community and growth, survival, and foliar nutrient composition of 2-year-old seedlings grown in forest bare-root nursery conditions in Lithuania. The Scots pine seeds originated from five stands from Latvia (P1), Lithuania (P2 and P3), Belarus (P4), and Poland (P5). Based on molecular identification, seven ECM fungal taxa were identified: Suillus luteus and Suillus variegatus (within the Suilloid type), Wilcoxina mikolae, Tuber sp., Thelephora terrestris, Cenococcum geophilum, and Russuloid type. The fungal species richness varied between five and seven morphotypes, depending on seed origin. The average species richness and relative abundance of most ECM morphotypes differed significantly depending on pine origin. The most essential finding of our study is the shift in dominance from an ascomycetous fungus like W. mikolae in P2 and P4 seedlings to basidiomycetous Suilloid species like S. luteus and S. variegatus in P1 and P5 seedlings. Significant differences between Scots pine origin were also found in seedling height, root dry weight, survival, and concentration of C, K, Ca, and Mg in the needles. The Spearman rank correlation coefficient revealed that survival and nutritional status of pine seedlings were positively correlated with abundance of Suilloid mycorrhizas and negatively linked with W. mikolae abundance. However, stepwise multiple regression analysis showed that only survival and magnesium content in pine needles were significantly correlated with abundance of ECM fungi, and Suilloid mycorrhizas were a main significant predictor. Our results may have implications for understanding the physiological and genetic relationship between the host tree and fungi and should be considered in management decisions in forestry and ECM fungus inoculation programs.

Poplars are among the few tree genera that can develop both ectomycorrhizal (ECM) and arbuscular (AM) associations; however, variable ratios of ECM/AM in dual mycorrhizal colonizations were observed in the roots of a variety of poplar species and hybrids. The objective of our study was to analyze the effect of internal and external factors on growth and dual AM and ECM colonization of poplar roots in three 12-15-year-old common gardens in Poland. We also analyzed the abundance of nonmycorrhizal fungal endophytes in the poplar roots. The Populus clones comprised black poplars (Populus deltoides and P. deltoides x Populus nigra), balsam poplars (Populus maximowiczii x Populus trichocarpa), and a hybrid of black and balsam poplars (P. deltoides x P. trichocarpa). Of the three sites that we studied, one was located in the vicinity of a copper smelter, where soil was contaminated with copper and lead. Poplar root tip abundance, mycorrhizal colonization, and soil fungi biomass were lower at this heavily polluted site. The total mycorrhizal colonization and the ratio of ECM and AM colonization differed among the study sites and according to soil depth. The influence of Populus genotype was significantly pronounced only within the individual study sites. The contribution of nonmycorrhizal fungal endophytes differed among the poplar clones and was higher at the polluted site than at the sites free of pollution. Our results indicate that poplar fine root abundance and AM and ECM symbiosis are influenced by environmental conditions. Further studies of different site conditions are required to characterize the utility of poplars for purposes such as the phytoremediation of polluted sites.

In this study, we present the detailed molecular investigation of the ectomycorrhizal (ECM) community of Quercus petraea and Quercus robur seedlings grown in bare-root forest nurseries. In all tested oak samples, mycorrhizal colonization was nearly 100%. Morphological observation and molecular investigations (sequencing of fungal ITS rDNA) revealed a total of 23 mycorrhizal taxa. The most frequent and abundant fungal taxa were Hebeloma sacchariolens, Tuber sp., and Peziza sp.; from the detected fungal taxa, 20 were noted for Q. petraea and 23 for Q. robur. Depending on the nursery, the species richness of identified ECM fungal taxa for both oak species ranged from six to 11 taxa. The mean species richness for all nurseries was 5.36 and 5.82 taxa per Q. petraea and Q. robur sample, respectively. According to the analysis of similarity, ECM fungal communities were similar for Q. petraea and Q. robur (R = 0.019; p = 0.151). On the other hand, detected fungal communities were significantly different between nurseries (R = 0.927; p < 0.0001). Using the Spearman rank correlation, it was determined that the ectomycorrhizal diversity (in terms of richness, the Shannon diversity, evenness, and Simpson dominance indices) is significantly related to the soil parameters of each nursery. We conclude that individual nursery may be considered as separate ecological niches that strongly discriminate diversity of ECM fungi.

We report the effects of pine and oak litter on species composition and diversity of mycorrhizal fungi colonizing 2-year-old Pinus sylvestris L. seedlings grown in a bare-root nursery in Lithuania. A layer of pine or oak litter was placed on the surface of the nursery bed soil to mimic natural litter cover. Oak litter amendment appeared to be most favorable for seedling survival, with a 73% survival rate, in contrast to the untreated mineral bed soil (44%). The concentrations of total N, P, K, Ca, and Mg were higher in oak growth medium than in pine growth medium. Relative to the control (pH 6.1), the pH was lower in pine growth medium (5.8) and higher in oak growth medium (6.3). There were also twofold and threefold increases in the C content of growth medium with the addition of pine and oak litter, respectively. Among seven mycorrhizal morphotypes, eight different mycorrhizal taxa were identified: Suillus luteus, Suillus variegatus, Wilcoxina mikolae, a Tuber sp., a Tomentella sp., Cenococcum geophilum, Amphinema byssoides, and one unidentified ectomycorrhizal symbiont. Forest litter addition affected the relative abundance of mycorrhizal symbionts more than their overall representation. This was more pronounced for pine litter than for oak litter, with 40% and 25% increases in the abundance of suilloid mycorrhizae, respectively. Our findings provide preliminary evidence that changes in the supply of organic matter through litter manipulation may have far-reaching effects on the chemistry of soil, thus influencing the growth and survival of Scots pine seedlings and their mycorrhizal communities.

The trace metal contents in fruiting bodies of ectomycorrhizal (ECM) fungi, symbiotic partners of Scots pine, were studied on three sites situated in west-central Poland. Elements were determined by atomic absorption spectrometry in 123 samples of 16 species. The study explored the differences in metal accumulation in relation to site, fungal species, age and part of the fruiting body and results were related to metal content in soil and plant material (roots and needles). Soil analysis revealed that results were obtained under environmental conditions not subject to strong anthropogenic pressure. Median metal concentrations did not differ disparately between sites, although the concentrations of each of the tested metals in the individual species varied to a large extent. Extremely high levels of Al with a large bioconcentration factor (BCF) were found in sporocarps of Thelephora terrestris. The spread between the highest and the lowest concentration (max/min) was very wide in Al, Cd and Pb and these elements may be considered to be absorbed preferentially by fruiting bodies of some species whereas Fe, Mn and Zn, with relatively low values of max/min, are normally absorbed by the majority of fungi. There was no clear relationship between caps and stipes in metal content. However, a tendency to higher metal concentration in the caps was observed. The metal content in young and older fruiting bodies of five different fungi was species dependent. In order to estimate the degree of accumulation of each element by plant and mushrooms, bioconcentration factors (BCFs) were calculated. In plant material (roots and needles), highest values of BCFs were noted for essential metals, like Zn and Mn. Lead showed a definite exclusion pattern (BCF below 1). In fruiting bodies of tested fungi, especially in Amanita muscaria, cadmium was the most intensively accumulated metal. Lead was excluded by plants but was accumulated or excluded by fungi depending on the species. The significance of mycological observation and quantification of the metal content in monitoring of the forest ecosystem is discussed.

An outbreak of mupirocin-resistant (MuR) staphylococci was investigated in two wards of a large hospital in Warsaw, Poland. Fifty-three MuR isolates of Staphylococcus aureus, S. epidermidis, S. haemolyticus, S. xylosus, and S. capitis were identified over a 17-month survey which was carried out after introduction of the drug for the treatment of skin infections. The isolates were collected from patients with infections, environmental samples, and carriers; they constituted 19.5% of all staphylococcal isolates identified in the two wards during that time. Almost all the MuR isolates were also resistant to methicillin (methicillin-resistant S. aureus and methicillin-resistant coagulase-negative staphylococci). Seven of the outbreak isolates expressed a low-level-resistance phenotype (MuL), whereas the remaining majority of isolates were found to be highly resistant to mupirocin (MuH). The mupA gene, responsible for the MuH phenotype, has been assigned to three different polymorphic loci among the strains in the collection analyzed. The predominant polymorph, polymorph I (characterized by a mupA-containing EcoRI DNA fragment of about 16 kb), was located on a specific plasmid which was widely distributed among the entire staphylococcal population. All MuR S. aureus isolates were found to represent a single epidemic strain, which was clonally disseminated in both wards. The S. epidermidis population was much more diverse; however, at least four clusters of closely related isolates were identified, which suggested that some strains of this species were also clonally spread in the hospital environment. Six isolates of S. epidermidis were demonstrated to express the MuL and MuH resistance mechanisms simultaneously, and this is the first identification of such dual MuR phenotype-bearing strains. The outbreak was attributed to a high level and inappropriate use of mupirocin, and as a result the dermatological formulation of the drug has been removed from the hospital formulary.

Nine isolates of methicillin-resistant Staphylococcus aureus (MRSA) collected in a Warsaw hospital in 1996 were typed by phenotypic (resistograms) and genotypic (PFGE and plasmid restriction analysis-REAP) methods. Twenty-four (MRSA) strains collected in this hospital during a period of the same duration in 1992 and typed earlier using resistograms and PFGE were also typed by REAP. Comparison of typing results obtained for isolates from 1992 and 1996 showed that strains characterised by PFGE patterns of two distinct types described as specific of the two clonally related groups of Polish MRSA in a multicentre study in 1992 are continuously present in the hospital. However, MRSA strains representing PFGE patterns not observed before were also found within the collection from 1996. REAP typing has proved to have a discriminatory power similar to that of PFGE analysis. Nevertheless, due to the lack of plasmids or difficulties in plasmid DNA isolation in 3 out of 33 studied strains, the typability of REAP turned out to be lower than that of PFGE.

We have constructed a yeast mitochondrial genome containing only one group-I intron, r1, from the 21s rRNA gene and introduced this genome into a strain bearing a disruption of the suv3 gene. The presence of the r1 intron alone causes a block in respiration, while the isogenic strain containing the intronless genome is respiratory competent. Northern analysis indicates that the functional suv3 protein is necessary for the yeast cell in order to process the r1-containing transcripts: in the absence of the suv3 protein the hybridization pattern of the excised r1 intron is altered and the amount of mature 21s rRNA is 50-fold lower. We suggest that the multifunctional suv3 protein, which displays motifs of ATP-dependent RNA helicases, is necessary for the in vivo pathway leading to formation of mature 21s rRNA from the transcripts containing the r1 intron in mitochondria of Saccharomyces cerevisiae.