Aeromycology: studies of fungi in aeroplankton

Air is a natural environment for spores of many genera and species of fungi. Despite its small size and a significant dispersion they have a great impact on human health and different areas of our activities, such as agricultural production. The study on spores of fungi that belong to aeroplankton or bioaerosole is called aeromycology. The most frequent fungi present in the air are Cladosporium and Alternaria species. Their numbers are abundant regardless of latitude and height above the sea level and above the ground. They mostly originate from agricultural environment. Other frequently listed species of fungi, whose spores are present in the air include of Aspergillus, Penicillium, Fusarium, Sclerotinia and Ganoderma. The concentration of spores in the air strongly depends on the abundance of their formation during the studied period. This in turn relates to geobotanical region, vegetation, degree of urbanization, climatic conditions, season, current weather, wind force and direction, local microclimate, and many other factors. Changes in humidity affect the concentration of different types of fungal spores. In general they are divided to ‘dry’ (Alternaria, Cladosporium, Puccinia, Ustilago, Melampsora, Epicoccum, Drechslera) and ‘wet’ (Didymella, Fusarium, Ganoderma, Gliocladium, Leptosphaeria, Verticillium). Study of the composition of species and genera are being done using different types of spore samplers, mostly volumetric instruments. Visual identification is based on colony morphology of the fungus and the shape and size of spores. The identification at the species level is possible with molecular tools. Methods based on DNA/RNA amplification are very sensitive and accurate. They allow the identification below the species level, e.g. chemotypes, mating types or isolates with genes or alleles of interest. Aerobiological monitoring is widely used in the epidemiology of human diseases (inhalant allergies) and infections of arable crops (decision support systems for the protection of cultivated plants). Aeromycology is interconnected with such diverse areas as industrial aerobiology, bioterrorism, ecology, climatology or even speleology and cultural heritage.Powietrze jest naturalnym środowiskiem dla zarodników licznych rodzajów i gatunków grzybów. Pomimo niewielkich rozmiarów i znacznego rozproszenia mają one wielki wpływ na zdrowie ludzi i różne kierunki ich działalności, w tym w szczególności na produkcję rolniczą. Badania nad zarodnikami grzybów stanowiącymi część aeroplanktonu są przedmiotem aeromykologii. Niezależnie od szerokości geograficznej i wysokości nad poziomem morza w powietrzu szczególnie często występują grzyby z rodzajów Cladosporium i Alternaria, a ich źródłem jest najczęściej środowisko rolnicze. Innymi często notowanymi rodzajami grzybów, których zarodniki występują w powietrzu są m.in. Aspergillus, Penicillium, Fusarium, Sclerotinia i Ganoderma. Stężenie zarodników w powietrzu jest ściśle uzależnione od obfitości ich tworzenia w danym okresie, co jest pochodną regionu geobotanicznego, szaty roślinnej, stopnia zurbanizowania danej lokalizacji, warunków klimatycznych, pory roku, aktualnej pogody, siły i kierunku wiatru, lokalnego mikroklimatu i wielu innych czynników. Zmiany wilgotności powietrza wpływają na stężenie zarodników różnych rodzajów grzybów, określanych na tej podstawie jako „suche” (Alternaria, Cladosporium, Puccinia, Ustilago, Melampsora, Epicoccum, Drechslera) lub „mokre” (Didymella, Fusarium, Ganoderma, Gliocladium, Leptosphaeria, Verticillium). Badania składu rodzajowego i gatunkowego prowadzone są przy zastosowaniu różnego rodzaju chwytaczy zarodników, a identyfikacja wizualna na podstawie morfologii kolonii grzyba oraz kształtu i wymiarów zarodników uzupełniana jest obecnie przez wyjątkowo czułe metody detekcji molekularnej, specyficzne względem rodzajów, gatunków, chemotypów, a nawet składu genów i kompozycji poszczególnych alleli. Monitoring aerobiologiczny znajduje bezpośrednie wykorzystanie w epidemiologii chorób ludzi (alergologia) i roślin uprawnych (systemy wspierania decyzji w ochronie roślin uprawnych). Badania z zakresu aeromykologii znajdują zastosowanie w tak różnych kierunkach jak aerobiologia przemysłowa, bioterroryzm, ekologia, dziedzictwo kulturowe, klimatologia lub speleologia

The detection of Plasmodiophora brassicae using loop-mediated isothermal DNA amplification

Plasmodiophora brassicae, the cause of clubroot, is a very serious problem preventing from successful and profitable cultivation of oilseed rape in Poland. The pathogen was found in all main growing areas of oilseed rape; it also causes considerable problems in growing of vegetable brassicas. The aim of this work was to elaborate fast, cheap and reliable screening method to detect P. brassicae. To achieve this aim the Loop-mediated isothermal DNA amplification (LAMP) technique has been elaborated. The set of three primer pairs was designed using LAMP software. The detection was performed with the GspSSD polymerase, isolated from bacteria Geobacillus sp., with strand displacement activity. DNA extraction from clubbed roots obtained from farmers’ fields of oilseed rape infected by P. brassicae was done using a modified CTAB method. The reaction was performed for 60 min at 62oC. The visual detection was done using CFX96 Real Time PCR Detection System (BioRad) or Gerie II Amplicatior (Optigen). The detection with LAMP proved its usefulness; it was easy, fast and accurate and independent of plant age. The detection limit was 5 spores per 1 µl of the spore suspension, so LAMP was less sensitive than quantitative PCR tests reported in the literature. However, the method is cheap and simple, so it is a good alternative, when it comes to practical use and the assessment of numerous samples

Morphology of uredinia and urediniospores of the fungus Melampsora larici-epitea Kleb. a damaging pathogen of common osier (Salix viminalis L.) in Poland

Rust (Melampsora spp.) is a damaging disease of willows (Salix spp.), including common osier (S. viminalis L.). So far, the pathogens of this species found in Europe were identified as M. larici-epitea Kleb. or M. ribesii-viminalis Kleb. Moreover, a stem infecting form (SIF), deprived of a sexual stage in its life cycle was reported. The aim of this study was to find out which species of the rust fungi cause disease symptoms on common osier in Poland. The isolates from common osier were compared to the ones originating from its putative hybrids with trembling aspen (Populus tremula L.) and Simon's poplar (P. simonii Carr.). Fungal isolates were obtained in 2008-2010 from 15 different genotypes of willows, including seven varieties of common osier (4 Swedish and 3 Polish), two landraces of common osier and six putative hybrids with poplars. Fungal isolates originated from three experiment sites, including west (Wielkopolska and Lubuskie) and north-east (Warmia) regions of Poland. To ensure the genetic uniformity, the isolates were derived from single uredinia, obtained from natural infection conditions. In all collected samples the position of uredinia was always hypophyllous. The diameter of uredinia was measured by Sigma Scan Pro software, after inoculation of four standard genotypes, including two common osier and two willow hybrids. The studies proved that the main cause of common osier rust is Melampsora larici-epitea f. typica. All studied isolates, including the ones obtained on putative hybrids, were very similar according to the size of uredinia and the size and morphology of urediniospores. The average size of a uredinium was 1.1 mm diameter and slightly differed between the isolates (from 0.9 to 1.3 mm), depending on willow genotype, the quality of plant material used for artificial inoculations and infection conditions. The average size of a typical urediniospore was 12.4 x 10.5 µm, but the sizes varied from 9.8 to 13.2 µm. Urediniospores from common osier were slightly bigger and more oval (12.5 x 10.4 µm) as compared to spores from the putative hybrids Salix x Populus (12.3 x 10.6 µm), but the differences were statistically insignificant. The spores were ovoid, globoid or angular, evenly echinulated. The size of uredinia, as well as the size and morphology of urediniospores were in full agreement with literature data for M. larici-epitea f. typica

Country-Wide qPCR Based Assessment of Plasmodiophora brassicae Spread in Agricultural Soils and Recommendations for the Cultivation of Brassicaceae Crops in Poland

Clubroot is a damaging disease of oilseed rape and vegetable brassicas worldwide, caused by the soil-borne protist Plasmodiophora brassicae Wor. Due to the long life of resting spores, the assessment of the pathogen abundance in agricultural fields can serve as a guideline for disease control at the country-wide level or the regional scale. Between 2013 and 2019, we collected 431 soil samples from fields cultivated with Brassicaceae crops throughout 16 provinces of Poland. The samples were subjected to qPCR based analysis of P. brassicae DNA concentration. From these data, the spore loads and gene copies g−1 soil were calculated and used to produce an assessment of the current clubroot risk potential at a country-wide and regional scale. The country-wide map, showing the spread of the pathogen in agricultural soils, was made using ArcGis software package implementing the interpolation with the Inverse Distance Weight method. The calculation of gene copies specific to P. brassicae helped to formulate the recommendations for farmers in respect to the cultivation guidelines. It showed a high risk of yield losses in defined regions of north, south-west and central Poland and an urgent need to undertake intensive preventative measures

Pseudothecial maturation and ascospore release of Leptosphaeria maculans and L. biglobosa in south-east Poland

Stem canker of brassicas is a severe disease of oilseed rape in Australia, Canada and Europe, including Poland. The disease is caused by Leptosphaeria maculans and L. biglobosa - two pathogens belonging to the class Dothideomycetes. The species differ in pathogenicity, but they have identical shape and size of fruiting bodies and spores of the generative and vegetative stages. Both pathogens are often found together in infected tissues of oilseed rape plants. The main goal of the experiments was to measure the rate of pseudothecial maturation and to monitor ascospore concentration of L. maculans and L. biglobosa in air samples. The paper is the first investigation on the generative stage development of these two species in south-east Poland. The studies were done for three consecutive years (2005-2007), for six most important months in pathogen development and plant infection, including 3 months in the spring (March - May) and 3 months in the autumn (September - November). The stage of pseudothecial maturation was assessed visually, based on the development of asci and ascospores. Monitoring of spore concentration in the air was performed using a Hirst-type 7-day volumetric trap. It was proved that differences in pseudothecial maturation rate in south-east Poland, encompassing the climatic regions of the Carpathian Foothills and Cracow, do not exceed two weeks within one season. The first and the highest ascospore concentration dates depended on weather conditions in a particular season. The total number of spores during the studied seasons varied from 9 to 12 spores/m3, which was from 70 to 90 times lower than the average from five other monitoring sites around Poland. The short exposition to spore showers and very small concentrations of L. maculans and L. biglobosa ascospores in air samples were the most probable reasons for relatively small damage of oilseed rape crops by stem canker in the south-east part of Poland

The origin of pine pollen grains captured from air at Calypsobyen, Svalbard

Spitsbergen is the largest island in the Svalbard Archipelago (Norway) that has been permanently populated. The harsh Arctic climate prevents development of large vascular plants such as trees. A two-year aerobiological survey was conducted within the framework of two consecutive polar expeditions (2014 and 2015) in Spitsbergen (Calypsobyen, Bellsund). The air quality was measured continuously from June/July to August using a 7-day volumetric air sampler, Tauber trap and moss specimens. Collected air samples and gravimetric pollen deposits were processed following transfer to sterile laboratory conditions and analyzed with the aid of light microscopy. Days when pine pollen grains were detected in the air were selected for further analysis. Clusters of back-trajectories, computed using the Hybrid Single Particle Lagrangian Integrated Trajectory model in combination with ArcGIS software as well as the Flextra trajectory model, showed the movement of air masses to the sampling location at Hornsund, and thus indicated the likely origin of pollen grains. The GlobCover 2009 and CORINE Land Cover 2012 datasets were employed to establish the distribution of coniferous forests in the areas of interest. Conclusions were drawn based on the analyses of the circulation of air masses, using visualization of global weather conditions forecast to supercomputers. For the first time we have demonstrated that pine pollen grains occurring in pine-free Spitsbergen, could originate from numerous locations, including Scandinavia, Iceland, Siberia and northern Canada. Pollen grains were transported via air masses for distances exceeding ~2000 km. Both air samples and gravimetric pollen deposits revealed the same pattern of Pinus pollen distribution

The Role of Victim’s Resilience and Self-Esteem in Experiencing Internet Hate

Despite the growing prevalence of research on Internet hate, little is still known about the psychological factors that differentiate those who are negatively affected by being subjected to Internet hate and those who are not affected at all or only to a small degree. In the present studies, we aimed to verify if resilience and self-esteem could be predictors of such responses. A total of 60 public figures (politicians, athletes, and artists; 46.7% women) and 1128 ordinary Internet users (25.1% women) participated in the study. Participants completed The Brief Resilience Scale, The Self-Esteem Scale, and The Internet Hate Concern Scale, which was created for this study, and determined how often they experience hate online. The results showed that the public figures experience Internet hate more often but were less concerned with it than the ordinary Internet users, who received online hate less often, but were more worried about it. In both groups, high self-esteem and high resilience were negative predictors of greater concern with received online hate. Our study is the first step to understanding what makes the difference between people who cope well and are not particularly concerned, and people who are greatly affected by received Internet hate

Fungal Biodiversity and Their Role in Soil Health

Soil health, and the closely related terms of soil quality and fertility, is considered as one of the most important characteristics of soil ecosystems. The integrated approach to soil health assumes that soil is a living system and soil health results from the interaction between different processes and properties, with a strong effect on the activity of soil microbiota. All soils can be described using physical, chemical and biological properties, but adaptation to environmental changes, driven by the processes of natural selection, are unique to the latter one. This mini review focuses on fungal biodiversity and its role in the health of managed soils as well as on the current methods used in soil mycobiome identification and utilization next generation sequencing (NGS) approaches. The authors separately focus on agriculture and horticulture as well as grassland and forest ecosystems. Moreover, this mini review describes the effect of land-use on the biodiversity and succession of fungi. In conclusion, the authors recommend a shift from cataloging fungal species in different soil ecosystems towards a more global analysis based on functions and interactions between organisms

Infestation of Polish Agricultural Soils by Plasmodiophora Brassicae Along The Polish-Ukrainian Border

There has been a rapid, worldwide increase in oilseed rape production that has resulted in enormous intensification of oilseed rape cultivation, leading to tight rotations. This in turn, has caused an accumulation of pests as well as foliar and soil-borne diseases. Recently, clubroot has become one of the biggest concerns of oilseed rape growers. Clubroot is caused by the soil-borne protist Plasmodiophora brassicae Woronin. The pathogen may be present in groundwater, lakes, and irrigation water used in sprinkling systems. It can be easily transmitted from one field to another not only by water, but also by soil particles and dust transmitted by wind and on machinery. The aim of our overall study was to check for P. brassicae infestation of Polish agricultural soils. This paper presents the 2012 results of a study performed along the Polish-Ukrainian border in two provinces: Lublin (Lubelskie Voivodeship) and the Carpathian Foothills (Podkarpackie Voivodeship), in south-east Poland. Monitoring was done in 11 counties, including nine rural and two municipal ones. In total, 40 samples were collected, out of which 36 were collected from fields located in rural areas and four from municipal areas, with two per municipal region. Each sample was collected at 8-10 sites per field, using a soil auger. The biotest to detect the presence of P. brassicae was done under greenhouse conditions using seedlings of the susceptible Brassicas: B. rapa ssp. pekinensis and the Polish variety of oilseed rape B. napus cv. Monolit. Susceptible plants grown in heavily infested soils produced galls on their roots. A county was regarded as free from the pathogen, if none of the bait plants became infected. The pathogen was found in three out of 40 fields monitored (7.5%) in the Carpathian Foothill region. The fields were located in two rural counties. The pathogen was not found in Lublin province, and was also not detected in any of the municipal counties. The detection with a biotest was fully confirmed by PCR-based molecular detection of P. brassicae DNA in soil samples

Trichoderma spp. – application and prospects for use in organic farming and industry

Fungi of the genus Trichoderma are a very large group of microorganisms that play a significant role in the environment. They use a variety of mechanisms to colonise various ecological niches. Several Trichoderma spp. positively affect plants by stimulating plant growth, and protecting plants from fungal and bacterial pathogens. They are used in biological plant protection as biofungicides as well as in bioremediation. Members of the genus Trichoderma are also utilised in various industry branches – mainly in the production of enzymes, antibiotics, and other metabolites, but also of biofuel. Moreover, the genus Trichoderma comprises edible and medicinal mushrooms, but also the pathogens of humans. Currently, Trichoderma has entered the genomic era and parts of genome sequences are publicly available. This is why, Trichoderma fungi have the potential to be used for human needs to an even greater extent than before. Nevertheless, further studies are needed to increase the efficiency and safety of the application of these fungi