Detection of arbuscular mycorrhizal fungal spores in the air across different biomes and ecoregions

Aerial dispersal of fungal spores is common, but the role of wind and air movement in dispersal of spores of arbuscular mycorrhizal (AM) fungi is largely unknown. Several studies have examined the possibility of AM fungal spores being moved by wind vectors without observing spores taken from the air environment. For the first time this study observed the presence of AM fungal spores in the air. The frequency of AM fungal spores in the air was determined in six North American biomes composed of 18 ecoregions. Multiple samples were taken from both the air and the soil at each location. AM fungal spores were found in high abundance in the soil (hundreds of spores per gram of soil), however, they were rarely found in the air (most samples contained no AM fungal spores). Furthermore, only the Glomus morphotype was found in the air, whereas spores in the soil were taxomomically more diverse (Glomus, Acaulospora, Gigaspora, Scutellospora morphotypes were observed). The proportion of Glomus spores in the air relative to Glomus spores in the soil was highest in more arid systems, indicating that AM fungi may be more likely to be dispersed in the air in such systems. Nonetheless, the results indicate that the air is not likely a dominant mode of dispersal for AM fungi.     

Natural folding of airborne fungal spores: a mechanism for dispersal and long-term survival?

Analysis of numerous air samples has indicated that dormant, viable fungal spores are highly present, which suggests that aerial dispersion is important for fungi. Whereas the majority of the spores may travel only very short distances, there is indication that a notable number of them cover much longer distances. Harmomegathy is a terminology coined by Wodehouse (1935) describing the natural folding of pollen to accommodate controlled and reversible water loss. Here, we discuss evidence that this concept may also apply to airborne fungal spores that face similar challenges and have to survive periods of drought and low temperatures while retaining viability to germinate after deposition upon a suitable moist substrate. In fact, (air)dried conidia, appear collapsed, survive for much longer times compared to spores in liquid, that deteriorate in time. This indicates that for some types of fungal spores, true dormancy is reached in the desiccated state. For these airborne spores this might be regarded as a pre-adaptation that supports long-distance transport of viable cells through air. We state that spores are naturally folded during transport in air if the humidity is low enough. We hypothesize that this is a pre-adaptation supporting release, dispersal and survival of airborne spores. Moreover, the smaller size of dry naturally-folded spores may also be relevant, e.g. for the opportunistic pathogenic fungus Aspergillus fumigatus reduced spore size supports deposition within the alveoli in the lung.     

Characterisation of the phagocytic uptake of Aspergillus fumigatus conidia by macrophages

Aspergillus fumigatus is an opportunistic fungal pathogen responsible for severe, life-threatening infections in immunocompromised patients. Airborne conidia are the infectious agent and can reach the lower parts of the respiratory system. In the lung, phagocytes represent the first line of defence. Resident macrophages are able to track down, engulf and kill the invading spores. Phagocytosis of the conidia is therefore a prerequisite for their efficient elimination. Using human and murine macrophages we analysed the phagocytic uptake of A. fumigatus conidia. We found that conidial phagocytosis is an actin-depending process that additionally requires myosin motor, phosphoinositide-3-phosphate kinase and tyrosine kinase activity. Both broad range tyrosine kinase inhibitors and inhibitors that specifically block src kinases had a strong impact on the conidial uptake. Immunofluorescence data demonstrate the recruitment of tyrosine-phosphorylated proteins to the vicinity of engulfed conidia. Uptake of the conidia was accompanied by a strong and local reorganisation of the actin cytoskeleton, whereas no prominent reorganisation was apparent for the microtubules. Both confocal immunofluorescence and electron microscopic data revealed the presence of large ruffle-like structures engaged in the uptake of conidia. This suggests that the internalisation of A. fumigatus spores can be mediated by a process resembling macropinocytosis, which is furthermore supported by the detection of intracellular conidia within spacious vacuoles. Taken together, our data provide new insights into the internalisation of A. fumigatus spores by macrophages, a key process in the early immune defence against an Aspergillus infection.     

First record on epizootics of Entomophthora grylli on grasshopper in Indian subcontinent: pathogen city and bio control potential on Oxya velox

A naturally-occurring fungus called Entomophthora grylli was for the first time isolated from two species of grasshopper Oxya velox and Oxya vicinia in Jammu and Kashmir, India. The epizootic was confined along Indo–Pak border between 74 degrees 24′ and 75 degrees 18′, East longitude and 32 degrees 50′ and 33 degrees 30′ North latitude. The fungus proved to be highly pathogenic and the natural mortality was significantly influenced by the population density which increased from 26.00 to 73.60 over the period of epizootics. On the basis of the available literature this appears to be the first record from the Indian sub-continent. While many infected grasshoppers apparently produced neither conidia nor resting spores, the cadavers were found to be full of hyphal bodies and resting spores towards the end of epizootics. These resting spores or their germ tubes were not invasive as such but if provided a saturated environment for a week, they start germinating, resulting in germ conidia which were able to induce dermal pathogencity. Further, it was observed that the disease could not be transmitted to healthy individuals by ingestion. However, the intra-haemocoel infectivity of fresh resting spores, germinated resting spores, and germ conidia proved to be highly pathogenic as they resulted in 81.4% grasshopper mortality. Although E. grylli is fastidious, it is possible to multiply it on a large scale as protoplasts which are infective upon injection in their hosts. However, the lack of a cell wall renders them very fragile, and they are neither infective upon application to the insect's cuticle nor upon ingestion. In the present study, a method on delivery of pathogen through “sticky molasses pan trap” was developed for inducing infection in grasshoppers in a paddy nursery which would facilitate its use as a bioinsecticide, analogous to other entomopathogenic fungi.     

Eicosanoid biosynthesis inhibitors influence mortality of Pieris brassicae larvae co-injected with fungal conidia

Influence of fungal species (conidia spores) on mortality of Pieris brassicae larvae differed when injected into the larvae. The effects of B. bassiana (ARSEF-1151) were expressed in a conidial dose-dependent manner on mortality of the larvae. An increased and faster mortality of the larvae followed B. bassiana (ARSEF-1151) infection when the spores were co-injected with the eicosanoid biosynthesis inhibitors (dexamethasone, naproxen, phenidone, esculetin). These compounds express different modes of action. These lethal effects were reversed when dexamethasone was injected with eicosanoid precursor arachidonic acid (20:4n-6). Nodulation is the predominant cellular reaction to bacterial and fungal injection in insects. Injection of each of five fungal species showed that nodulation intensity varies according to infecting fungal species. These findings support the idea that virulent effects of entomopathogenic, fungal species can be increased when P. brassicae immune systems are suppressed.     

Modeling the Influence of Rainfall and Temperature on the Phenology of Infection of Gypsy Moth, Lymantria dispar, Larvae by the Fungus Entomophaga maimaiga

A computer model driven by daily maximum-minimum temperature and rainfall records was developed to investigate the influence of weather on times when gypsy moth larvae, Lymantria dispar, can become infected by the fungal pathogen Entomophaga maimaiga. In the model, gypsy moth eggs are hatched and neonates are exposed primarily to germinating resting spores in the soil during the spring. Risk of infection is related to forest floor moisture. Larval and fungal development follows a degree-day model. When larvae become 4th instars, they can again become infected by resting spores because they hide in the litter during daylight hours. If rain falls when infected caterpillars die, the fungus sporulates, producing conidia. The number of conidia produced is assumed to directly influence the probability of infection of other larvae. The model was run using weather records from 1990 through 1992. Predicted times of infection were compatible with estimated fungal recruitment rates and changes in field disease prevalence rates. Assumptions about infection mechanisms in the model are discussed as they relate to the real world.     

Cell cycle and cell death are not necessary for appressorium formation and plant infection in the fungal plant pathogen <Emphasis Type="Italic">Colletotrichum gloeosporioides</Emphasis>

Background  

In order to initiate plant infection, fungal spores must germinate and penetrate into the host plant. Many fungal species differentiate specialized infection structures called appressoria on the host surface, which are essential for successful pathogenic development. In the model plant pathogen Magnaporthe grisea completion of mitosis and autophagy cell death of the spore are necessary for appressoria-mediated plant infection; blocking of mitosis prevents appressoria formation, and prevention of autophagy cell death results in non-functional appressoria.     

Sterile males of Ceratitis capitata (Diptera: Tephritidae) as disseminators of Beauveria bassiana conidia for IPM strategies

Sterile Mediterranean fruit fly, Ceratitis capitata (Wied.), males were evaluated as vectors to spread Beauveria bassiana (Bals) conidia to wild C. capitata populations under field conditions. The inoculated sterile males were released by air, using the chilled adult technique over 7000 ha of coffee growing in Chimaltenango, Guatemala, Central America. The impact of releases was determined using dry traps baited with a food attractant. The effects of these releases on Apis mellifera, Linnaeus (honey bee), Hypothenemus hampei, Ferrari (coffee berry borer) and the parasitic mite Varroa destructor (Oudeman) were also evaluated. Inoculated sterile males were able to transmit fungal spores to 44% of the wild C. capitata flies captured in traps, which likely were infected through intra- and intersexual interactions during leks, mating or mating attempts. There was no transmission of the fungal spores to non-target insect species such as coffee berry borer, honey bees or varroa. We conclude that sterile males of Mediterranean fruit fly inoculated with B. bassiana can act as effective vectors of conidia to wild populations, constituting a safe, environmentally friendly and selective alternative for suppressing the medfly under a Sterile Insect Technique-based IPM approach.     

球孢白僵菌对烟蚜茧蜂生命参数及控害效果的影响

在室内评价了球孢白僵菌对烟蚜茧蜂生命参数及控害效果的影响。分别在烟蚜茧蜂寄生桃蚜后不同时间进行高剂量(1900孢子/mm2)接菌,检测蚜虫感病率和寄生蜂形成的僵蚜率及僵蚜出蜂率。结果表明,球孢白僵菌对僵蚜率和僵蚜出蜂率的影响随接菌时间不同而变化。在烟蚜茧蜂寄生前1d、寄生当天和寄生后3d接菌,蚜虫感病率分别为59.6%、56.2%和34.8%;与对照相比,僵蚜率分别下降94%、59%和47%,僵蚜出蜂率分别减少83%、54%和49%。在寄生后5d或7d接菌,僵蚜率和僵蚜出蜂率不受明显影响,但蚜虫感病率降低到8.2%以下。对蚜尸内白僵菌菌体含量检测表明,随着烟蚜茧蜂寄生后接菌时间的推移,菌体数量迅速下降。寄生蜂寄生后5d或7d接菌,蚜尸内几乎检测不到菌体。直接喷雾接菌烟蚜茧蜂,成蜂寿命缩短4d左右,且81.8%的蜂尸受白僵菌感染。接菌后的寄生蜂对蚜虫寄生率几乎无影响,但寄生蜂在蚜虫体内的存活时间缩短了27.8%。     

Conidia of the nematophagous fungus Drechmeria coniospora adhere to but barely infect Acrobeloides buetschilii

Abstract Conidia of the endoparasitic fungus Drechmeria coniospora were able to adhere to the nematode Acrobeloides buetschilii ; approximately 70% of the nematodes were carrying spores 16 h after their addition to the fungal culture. Young nematodes were preferentially affected; adhesion mainly took place in the head region of the animal. Full infection, characterized by colonization of the nematode body and production of conidiophores and conidia, was observed, albeit at low frequencies. In case of failure, a second adhesive knob was often formed on the germinated spore. The low frequencies of infection are not related to a loss of virulence, due to prolonged in vitro culturing of the fungus. Fresh spores, obtained from cultures which were passed several times through the host, did not show higher infection frequencies.     

Epidemiology of allergic disease: the fungi

Summary In indoor aerial environments and in particular that of homes, the number and type of fungal spores present in the air depend, more than on air exchange with the outside atmosphere, on the presence of sources or dispersion within the building itself. The concentration of spores is half that normally recorded outside the building, withAspergillus, Penicillium, Cladosporium, Alternaria, Mucor, Rhizopus andOidiodendron as the dominating genera. Aerosporological investigations carried out in homes should ideally be done at night, with the exposition of cultural dishes. The repetitive isolation of the same fungus may be particularly important in the etiology of allergic manifestation in man. Particular environments, if insufficiently aerated, with high levels of relative humidity and insufficient illumination, may become ecological niches particularly well suited to fungi, both during its latent form or during a phase of rapid development. Together with rapidly developing fungi, for instanceMucor andRhizopus, slow developing moulds such asPenicillium, Aspergillus andTrichoderma may also be found, characterized however by a production of a great number of easily dispersed conidia. Etiological research into the allergic manifestations of patients continuously inhaling spores responsible for particular allergies appears to be insufficiently developed.     

植物病原真菌的菌核研究进展

真菌菌核是相关真菌在特殊环境下由营养体菌丝交织和聚集形成的具有抵御恶劣环境能力的休眠结构,在真菌生活史及病原真菌的病害循环中具有重要的生物学和生态学意义。许多引起严重植物病害的病原真菌能够形成菌核,这类真菌通过菌核度过逆境。菌核在适宜条件下萌发形成子囊盘和孢子或菌丝,造成植物的侵染。本文从形成菌核的真菌种类、生物学特性、黑色素及分子生物学等方面进行综述,旨在为深入研究真菌菌核及其在真菌学、植物病理学及药用真菌学等领域应用提供研究思路和参考依据。     

Study of airborne fungal spores in Madrid, Spain

The concentration of fungal spores in the atmosphere of Madrid was recorded and analyzed for the year 2003. Airborne spores were sampled continuously with a Hirst-type spore trap located on the roof of a building of the School of Pharmacy, at about 8 m above ground level. Correlation between the mean daily spore concentrations and meteorological variables were explored by means of Spearman’s correlation analyses. Seventy spore types were identified, of which the most numerous were Cladosporium, Aspergillaceae (conidia), Coprinus, Agaricales (basidiospores), Ustilago (teliospores) and Pleospora (ascospores). These six types of spores represented more than 70% of the total. Cladosporium represented 41% of the total fungal spores, while Ustilago spores, the concentrations of which in May and June exceeded 47% of the monthly total spore count, constituted the second most important group. Spores reached their highest concentrations in the spring months, and in the autumn, mainly in October. A␣positive significant correlation was found between airborne spore counts and temperature and relative humidity. The results provide a picture of the spectrum of airborne fungal spores present in the atmosphere of Madrid and of the `peak' periods of their presence. Future studies will provide more detailed information on the seasonal dynamics of the spores most frequently found in the air as well as on the extent to which atmospheric conditions influence their release, dispersion and sedimentation processes.     

The cytotoxic properties of a plant lipid transfer protein involve membrane permeabilization of target cells

AIMS: To determine whether Ha-AP10, a member of the plant lipid transfer proteins (LTPs) family produces a direct cytotoxic effect on fungal cells mediated by membrane permeabilization. LTPs can inhibit fungal growth and are considered members of the ubiquitous class of antimicrobial peptides. However, the way they exert their effects on target cells is not yet understood. METHODS AND RESULTS: Viability assays demonstrate that Ha-AP10 acts as a fungicidal compound but no harmful effect is observed on plant cells. Liposome leakage assays show that the protein induces a moderate release of fluorescent probes encapsulated in model membranes, indicating its ability to interact with phospholipids. Using a fluorescent indicator of damage at the membrane level, we demonstrate that Ha-AP10 is able to induce the permeabilization of intact fungal spores in a dose-dependent manner. CONCLUSION: The results presented here demonstrate the permeabilization of fungal spores caused by Ha-AP10. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first demonstration of fungal membrane damage by an LTP, giving a clue to elucidate the basis of its antimicrobial properties.     

The expression and impact of antifungal grooming in ants

Parasites can cause extensive damage to animal societies in which many related individuals frequently interact. In response, social animals have evolved diverse individual and collective defences. Here, we measured the expression and efficiency of self-grooming and allo-grooming when workers of the ant Formica selysi were contaminated with spores of the fungal entomopathogen Metarhizium anisopliae. The amount of self-grooming increased in the presence of fungal spores, which shows that the ants are able to detect the risk of infection. In contrast, the amount of allo-grooming did not depend on fungal contamination. Workers groomed all nestmate workers that were re-introduced into their groups. The amount of allo-grooming towards noncontaminated individuals was higher when the group had been previously exposed to the pathogen. Allo-grooming decreased the number of fungal spores on the surface of contaminated workers, but did not prevent infection in the conditions tested (high dose of spores and late allo-grooming). The rate of disease transmission to groomers and other nestmates was extremely low. The systematic allo-grooming of all individuals returning to the colony, be they contaminated or not, is probably a simple but robust prophylactic defence preventing the spread of fungal diseases in insect societies.     

蓝莓根系复合共生体及其根区土壤中AM真菌调查

蓝莓Vaccinium uliginosum是欧石南菌根（ericoid mycorrhiza,ERM）真菌典型的寄主植物,但同时也可与丛枝菌根（arbuscular mycorrhiza,AM）真菌和深色有隔内生真菌（dark septate endophytes,DSE）共生形成复合共生体。本研究旨在调查和评价不同栽培体制下蓝莓成年树花果期根系共生体发育状况及其根区土壤中AM真菌资源分布状况,以期为优质蓝莓栽培管理提供理论依据和技术基础。从青岛佳沃蓝莓基地采集暖棚、冷棚和露地3种方式栽培的9-10年生‘蓝丰’、‘奥尼尔’和‘公爵’蓝莓的根系及根区土样,观察测定根系共生体着生数量、根区土壤中AM真菌孢子数量和菌种组成。结果表明,所有栽培方式下供试品种蓝莓根系均形成ERM、AM和DSE结构及其复合共生体;其中,AM着生数量最多,其次是ERM,DSE侵染率最低;复合共生体中则呈现ERM+AM>ERM+DSE>ERM+AM+DSE;蓝莓复合共生体着生数量、AM真菌侵染率、丛枝着生率及孢子数量等不同种植方式下呈现暖棚>冷棚>露地,不同品种呈现‘蓝丰’>‘公爵’>‘奥尼尔’,而ERM和DSE侵染率也呈现上述变化趋势。依据AM真菌形态特征,供分离鉴定获得5属11种AM真菌,以暖棚栽培条件下分离获得的AM真菌数量最多,‘蓝丰’根区土壤中分布的AM真菌属种最多。暖棚内成年树花果期蓝莓根系共生体发育健全,AM真菌种类和孢子数量较多,可能有利于提高蓝莓的产量、改善果实品质和抗逆性。     

Nature and bioactivities of endolichenic fungi in Pseudocyphellaria sp., Parmotrema sp. and Usnea sp. at Hakgala montane forest in Sri Lanka

Aim:  The aim of this study was to investigate the nature and bioactivities of endolichenic fungi in three abundant lichens, Pseudocyphellaria sp., Usnea sp. and Parmotrema sp. in the lower elevation of Hakgala montane forest in Sri Lanka.
Methods and Results:  Endolichenic fungal strains, fungi that live asymptomatically in the lichen thallus, much the same way as endophytic fungi live within healthy plant tissues, were isolated from three abundant lichen species, Pseudocyphellaria sp., Usnea sp. and Parmotrema sp., at Hakgala montane forest in Sri Lanka, using the surface sterilization method. Nine endolichenic fungal strains were isolated from Parmotrema sp. and Usnea sp. separately, while 11 endolichenic fungi were recovered from the lichen Pseudocyphellaria sp. Isolation of endolichenic fungus Chrysosporium sp. 2 was common to all three lichen species. Substrate utilization patterns and antifungal activities of eight endolichenic fungal species were evaluated and the results revealed that all the test fungi were able to produce at least one enzyme to utilize the test substrates. Nigrospora sp., Chrysosporium sp. 1 and 2 and Cladosporium sp. showed antifungal activities on growth of some selected plant pathogenic fungi.
Conclusions:  Endolichenic fungal strains (29) were isolated from the lichens Parmotrema sp., Usnea sp. and Pseudocyphellaria sp. in Sri Lanka. Chrysosporium sp. 2 was common in all three lichens. Some of these endolichenic fungal strains showed antifungal activities against common plant pathogenic fungi and they are capable of utilizing the substrates by producing specific enzymes.
Significance and Impact of the Study:  The diversity and prevalence of the endolichenic fungi have not been studied extensively and this is the first report of isolation and identification of endolichenic fungi in lichens available in Sri Lanka.     

Alkane-grown Beauveria bassiana produce mycelial pellets displaying peroxisome proliferation,oxidative stress,and cell surface alterations

The entomopathogenic fungus Beauveria bassiana is able to grow on insect cuticle hydrocarbons, inducing alkane assimilation pathways and concomitantly increasing virulence against insect hosts. In this study, we describe some physiological and molecular processes implicated in growth, nutritional stress response, and cellular alterations found in alkane-grown fungi. The fungal cytology was investigated using light and transmission electron microscopy while the surface topography was examined using atomic force microscopy. Additionally, the expression pattern of several genes associated with oxidative stress, peroxisome biogenesis, and hydrophobicity were analysed by qPCR. We found a novel type of growth in alkane-cultured B. bassiana similar to mycelial pellets described in other alkane-free fungi, which were able to produce viable conidia and to be pathogenic against larvae of the beetles Tenebrio molitor and Tribolium castaneum. Mycelial pellets were formed by hyphae cumulates with high peroxidase activity, exhibiting peroxisome proliferation and an apparent surface thickening. Alkane-grown conidia appeared to be more hydrophobic and cell surfaces displayed different topography than glucose-grown cells. We also found a significant induction in several genes encoding for peroxins, catalases, superoxide dismutases, and hydrophobins. These results show that both morphological and metabolic changes are triggered in mycelial pellets derived from alkane-grown B. bassiana.     

球孢白僵菌对烟蚜茧蜂生命参数及控害效果的影响

在室内评价了球孢白僵菌对烟蚜茧蜂生命参数及控害效果的影响。分别在烟蚜茧蜂寄生桃蚜后不同时间进行高剂量（1900孢子/mm^{2）接菌,检测蚜虫感病率和寄生蜂形成的僵蚜率及僵蚜出蜂率。结果表明,球孢白僵菌对僵蚜率和僵蚜出蜂率的影响随接菌时间不同而变化。在烟蚜茧蜂寄生前1d、寄生当天和寄生后3d接菌,蚜虫感病率分别为59.6%、56.2%和34.8%;与对照相比,僵蚜率分别下降94%、59%和47%,僵蚜出蜂率分别减少83%、54%和49%。在寄生后5d或7d接菌,僵蚜率和僵蚜出蜂率不受明显影响,但蚜虫感病率降低到8.2%以下。对蚜尸内白僵菌菌体含量检测表明,随着烟蚜茧蜂寄生后接菌时间的推移,菌体数量迅速下降。寄生蜂寄生后5d或7d接菌,蚜尸内几乎检测不到菌体。直接喷雾接菌烟蚜茧蜂,成蜂寿命缩短4d左右,且81.8%的蜂尸受白僵菌感染。接菌后的寄生蜂对蚜虫寄生率几乎无影响,但寄生蜂在蚜虫体内的存活时间缩短了27.8%。}     

Airborne spore loads and mesoscale dispersal of the fungal pathogens causing Sigatoka diseases in banana and plantain

The mesoscale dispersal of ascospores and conidia of the fungal pathogenMycosphaerella fijiensis, causing Black Sigatoka disease in banana and plantain, was investigated under field conditions in Costa Rica. Spores were sampled daily during the period April–August 1995, using three volumetric spore traps positioned in a 5 km straight line across the floor and up one side of a valley, to an elevation of 1000 m. The number of spores in each trap suggested that spores were not being released from nearby sources, but that they may have been transported from inoculum sources up to 40 km away. The implications of this when assessing the longer-distance transport of viable spores to new infection sites are discussed.