Determination of airborne fungal spores of Gaziantep (SE Turkey)

The aim of this study was to present the first airborne fungal spore research results of SE of Turkey. The presence and abundance of fungal spores were investigated with a Hirst-type volumetric spore trap (Lanzoni, VPSS 2000) for 2 years between the periods January 2010 and December 2011. A total of 211,521 spores and 47 fungal taxa belonging to Anamorphic Fungi, Phyla Ascomycota and Basidiomycota were recorded. Aspergillus/Penicillium spores type, unidentified spores, spores of Myxomycota and hyphal fragments were also recorded as groups. The relationship between fungal spore counts and several meteorological parameters was examined. Cladosporium was determined as the predominant genus (56.48 %); hyphal fragments (14.94 %), Ustilago (13.96 %) and Alternaria (5.79 %) were revealed as the common fungal aerosols of Gaziantep atmosphere. With this study, the first aeromycological survey of SE of Turkey has been conducted and new information on the field of aerobiology in Turkey has been introduced.     

Deodorization of pig slurry and characterization of bacterial diversity using 16S rDNA sequence analysis

Fusarium graminearum is a filamentous fungal plant pathogen that infects major cereal crops. The fungus produces both sexual and asexual spores in order to endure unfavorable environmental conditions and increase their numbers and distribution across plants. In a model filamentous fungus, Aspergillus nidulans, early induction of conidiogenesis is orchestrated by the fluffy genes. The objectives of this study were to characterize fluffy gene homologs involved in conidiogenesis and their mechanism of action in F. graminearum. We characterized five fluffy gene homologs in F. graminearum and found that FlbD is the only conserved regulator for conidiogenesis in A. nidulans and F. graminearum. Deletion of fgflbD prevented hyphal differentiation and the formation of perithecia. Successful interspecies complementation using A. nidulans flbD demonstrated that the molecular mechanisms responsible for FlbD functions are conserved in F. graminearum. Moreover, abaA-wetA pathway is positively regulated by FgFlbD during conidiogenesis in F. graminearum. Deleting fgflbD abolished morphological effects of abaA overexpression, which suggests that additional factors for FgFlbD or an AbaA-independent pathway for conidiogenesis are required for F. graminearum conidiation. Importantly, this study led to the construction of a genetic pathway of F. graminearum conidiogenesis and provides new insights into the genetics of conidiogenesis in fungi.     

Nutrient sensing and acquisition in fungi: mechanisms promoting pathogenesis in plant and human hosts

Fungal pathogens destroy our crops and cause hazardous human infections, therefore threatening our health and food security. The ability of fungal pathogens to sense and respond to dynamic host microenvironments enables the establishment and progression of disease. Sensing nutritional cues is vital throughout fungal infection of either plants or mammals: enabling the pathogen to invade, adapt and survive in the face of host immunity. Acquiring nutrients from their host for energy, growth and repair is also essential to a fungal pathogen's success. Cell-surface proteins embedded in the fungal plasma membrane sense and transport host macro- and micronutrients, including carbon and nitrogen sources and minerals such as iron and zinc. Using examples from model crop (Fusarium graminearum, Magnaporthe oryzae and Ustilago maydis) and human (Aspergillus fumigatus, Candida albicans, Cryptococcus neoformans) pathogens we review the nutrient sensing and transporting roles of fungal cell-surface receptor, transporter and transceptor proteins, and their importance to plant and human fungal disease. We discuss how their cellular localisation, central role in cell signalling and importance to disease makes these fungal cell-surface proteins candidates in the search for new strategies to control fungal diseases, while highlighting the areas where further research is needed to make this possible.     

Predicting the risk of soybean rust in Minnesota based on an integrated atmospheric model

To minimize crop loss by assisting in timely disease management and reducing fungicide use, an integrated atmospheric model was developed and tested for predicting the risk of occurrence of soybean rust in Minnesota. The model includes a long-range atmospheric spore transport and deposition module coupled to a leaf wetness module. The latter is required for spore germination and infection. Predictions are made on a daily basis for up to 7 days in advance using forecast data from the United States National Weather Service. Complementing the transport and leaf wetness modules, bulk (wet plus dry) atmospheric deposition samples from Minnesota were examined for soybean rust spores using a specific DNA test and sequence analysis. Overall, the risk prediction worked satisfactorily within the bounds of the uncertainty associated with the use of modeled 7-day weather forecasts, with more than 65% agreement between the model forecast and the DNA test results. The daily predictions are available as an advisory to the user community through the University of Minnesota Extension. However, users must take the actual decision to implement the disease management strategy.     

Effects of meteorological factors on the levels of <Emphasis Type="Italic">Alternaria</Emphasis> spores on a potato crop

Alternaria solani Soraeur produces early blight in Solanum tuberosum L., leading to significant agricultural losses. The current study was carried out on the extensive potato crop situated in north-western of Spain during 2007, 2008 and 2009. In this area potato crops are the most important source of income. In this work we used a Hirst-type volumetric spore-trap for the aerobiological monitoring of Alternaria spores. The highest spore concentrations were recorded during the 2009 cycle (10,555 spores), and the lowest concentrations were recorded during the 2008 cycle (5,471 spores). Over the 3 years of study, the highest concentrations were registered during the last stage of the crop. The aim of the study was to observe the influence of meteorological factors on the concentration of Alternaria spores, which can lead to serious infection and early blight. Prediction of the stages during which a crop is particularly vulnerable to infection allows for adjustment of the application of fungicide and is of environmental and agricultural importance. For this reason, we tested three models (P-Days, DD and IWP) to predict the first treatment and decrease the negative effect that these spores have on potato crops. The parameter that showed the most significant correlation with spore concentrations was minimum temperature. We used ARIMA (autoregressive integrated model of running mean) time-series models to determine the forecast. We considered weather data as predictor variables and the concentration of spores on the previous day as the fixed variable.     

Impact of disease frequency and host density on pollination and transmission of an African anther-smut fungus

The vast majority of flowering plants rely solely on insects for pollination. A number of pathogens have evolved mechanisms to exploit these close associations and use pollinators as vectors of infective propagules. Factors that affect pollinator movements and successful pollination may in turn also influence successful transmission of fungal spores. Here we investigate the effect of host density and the frequency of diseased Oxalis lanata individuals infected by the anther-smut fungus, Thecaphora capensis, on the likelihood of receiving pollen and fungal spores. Specifically, we determined the numbers of spores and pollen grains deposited on stigmatic surfaces of selected flowers under natural and standardized conditions where host density and disease frequency varied. The effect of host flower density and diseased flower frequency on pollen and spore transfer was variable under natural conditions and these factors interacted significantly. However, an increase in host density and disease frequency significantly influenced pollen and spore deposits under standardized conditions. The effect of host density was, however, not linear and an optimal flower density for pollen and fungal spore transmission was found. Similar to other systems of vector-borne disease, the transmission of anther-smut of Oxalis lanata is more frequency-dependent than density-dependent. This study represents a first step towards understanding the disease transmission process of T. capensis on Oxalis and lays the foundation for future comparative studies between this and other systems to develop and test general hypotheses of disease dynamics in vector-borne disease transmission systems.     

The contribution of aerobiology to agriculture

Aerobiology is the science which studies the atmospheric dispersion of biological materials and their impact on the environment and organisms. Therefore aerobiology covers many fields of basic and applied sciences and one of these is agriculture. The study of the dispersion of phytopathogenic spores was one of the first applications of aerobiology together with the study of pollinic aeroallergens in medicine, when aerobiology began to interest scientists more than 60 years ago. At the moment aerobiology could help agriculture in different ways, being able to detect and diffuse large quantities of information useful to optimize the production of many crops. The airborne particulate considered important in agriculture includes pollen, spores and insects; thus, when aerobiology is spoken of as being applied to agriculture, we understand the involvement of many sectors such as agronomy, plant pathology, mycology, botany, enthomology and others.     

Effects of wind speed and direction on monthly fluctuations of <Emphasis Type="Italic">Cladosporium</Emphasis> conidia concentration in the air

This study determined the relationship between airborne concentration of Cladosporium spp. spores and wind speed and direction using real data (local wind measured by weather station) and modelled data (air mass flow computed with the aid of HYbrid Single Particle Lagrangian Trajectory model). Air samples containing fungal conidia were taken at an urban site (Worcester, UK) for a period of five consecutive years using a spore trap of the Hirst design. A threshold of ≥6000 s m^?3 (double the clinical value) was applied in order to select high spore concentration days, when airborne transport of conidia at a regional scale was more likely to occur. Collected data were then examined using geospatial and statistical tools, including circular statistics. Obtained results showed that the greatest numbers of spore concentrations were detected in July and August, when C. herbarum, C. cladosporioides and C. macrocarpum sporulate. The circular correlation test was found to be more sensitive than Spearman’s rank test. The dominance of either local wind or the air mass on Cladosporium spore distributions varied between examined months. Source areas of this pathogen had an origin within the UK territory. Very high daily mean concentrations of Cladosporium spores were observed when daily mean local wind speed was v _s ≤ 2.5 m s^?1 indicating warm days with a light breeze.     

Level ofFusarium infection in wheat spikelets related to location and number of inoculated spores

The flowering time is the most susceptible period for primary infection of wheat heads byFusarium spp. During this period spores can be deposited into the opened wheat florets where they may later cause infections. We quantitatively explored the relationship between variables related to the flowering process and the infection level byFusarium graminearum in single spikelets. We imitated open (chasmogamous) and closed (cleistogamous) flowering by injecting well-defined amounts of spores into and between wheat florets. Applying the spores between the florets resulted in weaker disease symptoms and significantly lower amounts ofFusarium mycotoxins. With larger numbers of spores, the disease symptoms became more pronounced and the mycotoxin amounts per spikelet increased significantly. Our results indicate that the probability of primary infection is approximately proportional to the number of spores reaching the open florets during the flowering process. The breeding of wheat lines which flower partially or completely cleistogamously might reduce theFusarium susceptibility in wheat.     

Summer airborne mycoflora of Timişoara (Romania) and relationship to meteorological parameters

The present aeromycological investigation was undertaken to study atmospheric fungal spores in Timi?oara (western Romania). This study was carried out using a Hirst type volumetric sampler. The study revealed the existence of a rich airborne mycoflora. The atmospheric fungal spores were classified and evaluated into three groups (‘major’, ‘minor’ and ‘sporadic’) depending upon their catch percentage in the air. Cladosporium/Fusarium/Leptosphaeria-group, Alternaria, Helminthosporium airborne fungal spores and airborne fungal fragments regularly recorded (frequency 100% of days). Cladosporium accounted for 81.09% of the outdoor fungal spores. The airborne fungal fragments have been identified as abundant in our geographic area. Spearman’s correlations were applied to meteorological parameters and airborne fungal spore concentrations. In addition, correlations were calculated between the fungal spore concentrations and the meteorological variables from the previous day. A total of eleven weather factors were selected for this investigation. Following Spearman’s correlations, I identified two patterns of behaviour: most of the airborne fungal spores prefer cloudiness, lower near-surface soil temperature, lower atmospheric pressure, higher relative humidity and precipitation (pattern A) while other spore concentrations favour increased sunshine, higher near-surface soil temperature and dry conditions (pattern B). The behaviour of some fungal spores during the warm season has proven unclear (pattern C). This study demonstrates the need for investigations throughout the year and the evaluation with complementary statistical methods, regarding the correct interpretation of airborne mycoflora relationships with meteorological parameters.     

Recent advances in genes involved in secondary metabolite synthesis,hyphal development,energy metabolism and pathogenicity in Fusarium graminearum (teleomorph Gibberella zeae)

The ascomycete fungus, Fusarium graminearum (teleomorph Gibberella zeae), is the most common causal agent of Fusarium head blight (FHB), a devastating disease for cereal crops worldwide. F. graminearum produces ascospores (sexual spores) and conidia (asexual spores), which can serve as disease inocula of FHB. Meanwhile, Fusarium-infected grains are often contaminated with mycotoxins such as trichothecenes (TRIs), fumonisins, and zearalenones, among which TRIs are related to the pathogenicity of F. graminearum, and these toxins are hazardous to humans and livestock. In recent years, with the complete genome sequencing of F. graminearum, an increasing number of functional genes involved in the production of secondary metabolites, hyphal differentiation, sexual and asexual reproduction, virulence and pathogenicity have been identified from F. graminearum. In this review, the secondary metabolite synthesis, hyphal development and pathogenicity related genes in F. graminearum were thoroughly summarized, and the genes associated with secondary metabolites, sexual reproduction, energy metabolism, and pathogenicity were highlighted.     

A comparison of seasonal changes in deposition of spores of Erysiphe graminis on different trapping surfaces

Vertical sticky cylinder traps were as useful as a volumetric trap for detecting seasonal changes in numbers of Erysiphe spores from barley crops. Changes in spore catches by both traps also resembled changes in numbers of pustules developing on initially healthy barley seedlings exposed in the same crop. Seasonal changes in catches on cylinders exposed at some distance from any crop differed from those on cylinders just above barley crops. Cylinder traps are cheap, simple and quicker to scan than volumetric traps and seem likely to be acceptable alternatives to them for much work on cereal mildew and other air-dispersed pathogens, particularly for rautine trapping at many sites.     

Selection is required for efficient Cas9-mediated genome editing in Fusarium graminearum

Genome engineering using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated nucleases, such as Cas9 (CRISPR-associated protein 9), are revolutionising molecular biology. In this study, we established a Cas9-based genome editing system in Fusarium graminearum, a highly destructive fungal pathogen of cereal crops. Although the molecular toolkit of F. graminearum is well developed compared to other fungi, Cas9-mediated engineering offers a number of potential benefits, such as the ability to create marker free mutants in this species. Here we have used a codon-optimised Cas9 nuclease and dual ribozyme-based expression of a single guide RNA (sgRNA) to induce mutations. Cas9-mediated mutations were identified through a fungicide resistance-based phenotypic screen, which selects for null mutations in the FgOs1 gene encoding an osmosensor histidine kinase. In the absence of selection, however, mutations were identified at very low frequency. Examination of the mutant alleles identified suggests that, a microhomology-mediated end joining (MMEJ) DNA repair pathway is likely to be the predominant process involved in erroneous repairing of Cas9-induced double-stranded breaks in F. graminearum.     

Functional evaluation of a homologue of plant rapid alkalinisation factor (RALF) peptides in Fusarium graminearum

The cereal infecting fungus Fusarium graminearum is predicted to possess a single homologue of plant RALF (rapid alkalinisation factor) peptides. Fusarium mutant strains lacking FgRALF were generated and found to exhibit wildtype virulence on wheat and Arabidopsis floral tissue. Arabidopsis lines constitutively overexpressing FgRALF exhibited no obvious change in susceptibility to F. graminearum leaf infection. In contrast transient virus-mediated over-expression (VOX) of FgRALF in wheat prior to F. graminearum infection, slightly increased the rate of fungal colonisation of floral tissue. Ten putative Feronia (FER) receptors of RALF peptide were identified bioinformatically in hexaploid wheat (Triticum aestivum). Transient silencing of two wheat FER homoeologous genes prior to F. graminearum inoculation did not alter the subsequent interaction outcome. Collectively, our VOX results show that the fungal RALF peptide may be a minor contributor in F. graminearum virulence but results from fungal gene deletion experiments indicate potential functional redundancy within the F. graminearum genome. We demonstrate that virus-mediated over-expression is a useful tool to provide novel information about gene/protein function when results from gene deletion/disruption experimentation were uninformative.     

The impact of urban and forest fires on the airborne fungal spore aerobiology

The occurrence of airborne fungal spores is driven by several biogeographic and climatic factors. In addition, the occurrence of fires near fungal ecosystems seems to play an important role on the aerobiology of fungal spores. Wildfires are prevalent throughout the world and particularly so in several South European countries with Mediterranean climate. The present survey aimed at analysing the impact of urban and forest fires on the airborne fungal spore content of Madeira Island. Data suggest that after a fire occurrence, the concentration of fungal spores tends to increase in the air, peaking on the 10th day after a fire event. It is likely that fire-induced convections promote release of fungal spores from their natural habitat and that the soil heating conditions could trigger the germination of fungi colonizing post-fire plant residues and soil. It is also possible that the registered low precipitation and wind intensity could help increase the concentration of fungal spores in the atmosphere during 1–2 weeks. These findings need further research, not only at a regional but also at a larger scale, in order to clarify the specific impact of fires on such biological particles and the possible consequences on public health.     

Potential impact of climate change on fungal distributions: analysis of 2 years of contrasting weather in the UK

The impact of climate change on fungal growth and spore production is less well documented than for allergenic pollen grains, although similar implications for respiratory tract diseases in humans occur. Fungal spores are commonly described as either “dry” or “wet” according to the type of weather associated with their occurrence in the air. This study examined the distribution of selected fungal spores (Alternaria spp., Cladosporium spp., Didymella spp., Epicoccum spp., Leptosphaeria spp. and rusts) occurring in the West Midlands of UK during 2 years of contrasting weather. Spore specimens were collected using a 7-day volumetric air sampler and then analysed with the aid of light microscopy. Distributions of spores were then studied using normality tests and Mann–Whitney U test, while relationships with meteorological parameters were investigated using Spearman’s rank test and angular-linear correlation for wind direction analysis. Our results showed that so-called wet spores were more sensitive to the weather changes showing statistically significant differences between the 2 years of study, in contrast to “dry” spores. We predict that in following years we will observe accelerated levels in allergenic fungal spore production as well as changes in species diversity. This study could be a starting point to revise the grouping system of fungal spores as either “dry” or “wet” types and their response to climate change.