New methods for detecting and enumerating fungal spores of plant  pathogens

R. Kennedy; A.J. Wakeham

doi:10.17221/10317-pps

New methods for detecting and enumerating fungal spores of plant pathogens

Plant Protection Science ◽

10.17221/10317-pps ◽

2002 ◽

Vol 38 (SI 1 - 6th Conf EFPP 2002) ◽

pp. S38-S42

Author(s):

R. Kennedy ◽

A.J. Wakeham

Keyword(s):

Plant Pathogens ◽

Fungal Spores ◽

Fungal Spore ◽

Airborne Spores ◽

Suction System ◽

Spore Trap ◽

New Methods ◽

Fungal Plant Pathogens ◽

Specialist Knowledge ◽

User Friendly

Information on the presence or absence of airborne spores or other particles would be useful in an increasing number of areas including agriculture. Traditional methods used for detecting and enumerating of airborne spores of fungal plant pathogens are time consuming and require specialist knowledge. Some spore types (e.g. ascospores) are difficult to differentiate using these methods. To facilitate this, new methods, which can be used to accurately differentiate fungal spore types, are required. A Burkard 7-day volumetric spore trap used in combination with an immunofluorescence test has been developed to detect and quantify field-trapped ascosporic inoculum of Mycosphaerella brassicicola (the ringspot pathogen of brassicas). This test has also been found useful in the validation of more rapid user-friendly immunoassay based trapping procedures. A microtiter immunospore trapping device, which uses a suction system to directly trap air-particulates by impaction into microtiter wells, has been used successfully for the rapid detection and quantification of ascosporic inoculum of M. brassicicola. The system shows potential for the rapid field-detection of airborne ascosporic inoculum of the ringspot pathogen.

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A New Method To Monitor Airborne Inoculum of the Fungal Plant Pathogens Mycosphaerella brassicicola andBotrytis cinerea

Applied and Environmental Microbiology ◽

10.1128/aem.66.7.2996-3003.2000 ◽

2000 ◽

Vol 66 (7) ◽

pp. 2996-3003 ◽

Cited By ~ 40

Author(s):

R. Kennedy ◽

A. J. Wakeham ◽

K. G. Byrne ◽

U. M. Meyer ◽

F. M. Dewey

Keyword(s):

Plant Pathogens ◽

Collection Efficiency ◽

Correlation Coefficients ◽

Fungal Spore ◽

Sampling Period ◽

Enzyme Linked Immunosorbent Assay ◽

Suction System ◽

Spore Trap ◽

Brussels Sprouts ◽

Fungal Plant Pathogens

ABSTRACT We describe a new microtiter immunospore trapping device (MTIST device) that uses a suction system to directly trap air particulates by impaction in microtiter wells. This device can be used for rapid detection and immunoquantification of ascospores ofMycosphaerella brassicicola and conidia of Botrytis cinerea by an enzyme-linked immunosorbent assay (ELISA) under controlled environmental conditions. For ascospores ofM. brassicicola correlation coefficients (r 2) of 0.943 and 0.9514 were observed for the number of MTIST device-impacted ascospores per microtiter well and the absorbance values determined by ELISA, respectively. These values were not affected when a mixed fungal spore population was used. There was a relationship between the number of MTIST device-trapped ascospores of M. brassicicola per liter of air sampled and the amount of disease expressed on exposed plants ofBrassica oleracea (Brussels sprouts). Similarly, when the MTIST device was used to trap conidia of B. cinerea, a correlation coefficient of 0.8797 was obtained for the absorbance values generated by the ELISA and the observed number of conidia per microtiter well. The relative collection efficiency of the MTIST device in controlled plant growth chambers with limited airflow was 1.7 times greater than the relative collection efficiency of a Burkard 7-day volumetric spore trap for collection of M. brassicicola ascospores. The MTIST device can be used to rapidly differentiate, determine, and accurately quantify target organisms in a microflora. The MTIST device is a portable, robust, inexpensive system that can be used to perform multiple tests in a single sampling period, and it should be useful for monitoring airborne particulates and microorganisms in a range of environments.

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Role of Phosphorylation in Fungal Spore Germination

10.32747/1993.7568761.bard ◽

1993 ◽

Author(s):

Martin B. Dickman ◽

Oded Yarden

Keyword(s):

Protein Kinase ◽

Spore Germination ◽

Plant Pathogens ◽

Hyphal Growth ◽

Fungal Spore ◽

Tube Formation ◽

Kinase Gene ◽

Fungal Plant Pathogens ◽

Dependent Protein ◽

Protein Kinase Gene

Spore germination is a common and fundamental event in fungal development and in many instances an essential phase of fungal infection and dissemination. Spore germination is also critical for hyperparasites to function as biocontrol agents as well as in fermentation proceses. Our common objective is to understand the mechanisms which regulated spore germination and identify factors involved in pathogenicity related prepenetration development. Our approach is to exploit the overall similarity among filamentous fungi using both a plant pathogen (Colletotricum trifolii) and a model system that is genetically sophisticated (Neurospora crassa). The simulataneous use of two organisms has the advantage of the available tools in Neurospora to rapidly advance the functional analysis of genes involved in spore germination and development of an economically important fungal phytopathogen. Towards this we have isolated a protein kinase gene from C. trifolii (TB3) that is maximally expressed during the first hour of conidial germination and prior to any visible gene tube formation. Based on sequence similarities with other organisms, this gene is likely to be involved in the proliferative response in the fungus. In addition, TB3 was able to functionally complement a N. crassa mutant (COT-1). Pharmacological studies indicated the importance of calmodulin in both germination and appressorium differentiation. Using an antisense vector from N. crassa, direct inhibition of calmodulin results in prevention of differentiation as well as pathogenicity. Both cAMP dependent protein kinase (PKA) and protein kinase C (PKC) like genes have been cloned from C. trifolii. Biochemical inhibition of PKA prevents germination; biochemical inhibitors of PKC prevents appressorium differentiation. In order to analyze reversible phosphorylation as a regulatory mechanism, some ser.thr dephosphorylative events have also been analyzed. Type 2A and Type 2B (calcineurin) phosphatases have been identified and structurally and functionally analyzed in N. crassa during this project. Both phosphatases are essential for hyphal growth and maintenance of proper hyphal architecture. In addition, a first novel-type (PPT/PP5-like) ser/thr phosphatase has been identified in a filamentous fungus. The highly collaborative project has improved our understanding of a fundamental process in fungi, and has identified targets which can be used to develop new approaches for control of fungal plant pathogens as well as improve the performance of beneficial fungi in the field and in industry. In addition, the feasibility of molecular technology transfer in comparative mycology has been demonstrated.

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Variation in Airborne Fungal Spore Concentrations in Four Different Microclimate Regions in Romania

Notulae Botanicae Horti Agrobotanici Cluj-Napoca ◽

10.15835/nbha4129032 ◽

2013 ◽

Vol 41 (2) ◽

pp. 450 ◽

Cited By ~ 5

Author(s):

Nicoleta IANOVICI ◽

Cristina MARIA ◽

Mădălina Nicoleta RĂDUŢOIU ◽

Anca HANIŞ ◽

Dorina TUDORICĂ

Keyword(s):

Urban Areas ◽

Fungal Spores ◽

Pollen Grains ◽

Fungal Spore ◽

Western Country ◽

Ambrosia Artemisiifolia ◽

Habitat Characteristics ◽

Airborne Spores ◽

Spores And Pollen ◽

Airborne Fungal Spore

Airborne fungal counts and types were examined in four regions from Romania. The aim of our study was to determine the concentrations of Alternaria, Cladosporium, Drechslera-type, Epicoccum and Nigrospora atmospheric fungal spores in urban areas: Braşov, Bucharest, Cluj-Napoca and Timişoara. Other objective of this study was to examine the relationships between atmospheric fungal spores and pollen. Few studies have confirmed the high frequency and concentrations for some of these spore types in Romania. The sampling sites differed in habitat characteristics, such as vegetation and microclimate. Airborne spores’ sampling was carried out using the Lanzoni volumetric sampler and during August 2008. The totals of airborne spores recorded in this study show a considerable variation. Cladosporium and Alternaria have been identified as the most abundant and frequent fungal aeroallergens in Timişoara and Bucharest. Drechslera-type, Epicoccum, and Nigrospora had very low values of spore count in all sites. Observations show that seven types of pollen appear regularly and few of these are in great abundance. Fungi are present in the air in concentrations considerably in excess of that of pollen grains. Ambrosia artemisiifolia is the most important pollen allergen in all of the sampled areas discussed here, especially in western country (Timişoara).

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Sensitivity of Bacterial and Fungal Plant Pathogens to the Lytic Peptides, MSI-99, Magainin II, and Cecropin B

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.2002.15.7.701 ◽

2002 ◽

Vol 15 (7) ◽

pp. 701-708 ◽

Cited By ~ 45

Author(s):

Ali R. Alan ◽

Elizabeth D. Earle

Keyword(s):

Phytophthora Infestans ◽

Plant Pathogens ◽

Fungal Spores ◽

Alternaria Solani ◽

Leaf Disk ◽

Cecropin B ◽

Fungal Plant Pathogens ◽

Magainin Ii ◽

Lytic Peptides

In vitro and leaf disk assays of bacterial and fungal plant pathogens were conducted using three cationic lytic peptides, MSI-99, magainin II (MII), and cecropin B (CB). Growth of bacterial organisms was retarded or completely inhibited by low concentrations of these lytic peptides. The peptides also significantly reduced germination of fungal spores and growth of mycelia; however, higher concentrations of peptides were needed to inhibit fungal growth compared with those needed to inhibit bacteria. The relative efficacy of the peptides depended on the microorganism tested, but CB was the most inhibitory to the majority of the bacteria and fungi assayed. MSI-99, a synthetic derivative of MII with increased positive charge, showed equal or two- to fivefold higher antibacterial activity compared to MII in the in vitro assays. MSI-99 was also superior to MII against the oomycete, Phytophthora infestans but was slightly inferior to MII in assays with the true fungi, Penicillium digitatum and Alternaria solani. In the leaf disk assays, pretreating spores of Alternaria solani and Phytophthora infestans with the peptides at concentrations as low as 10 μg per ml led to significant reductions in the size of early blight lesions and prevented development of any late blight lesions on tomato leaf disks. Our results from in vitro and leaf disk assays suggest that MSI-99 can be used as a transgene to generate tomato lines with enhanced resistance to bacterial and fungal diseases of this crop.

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Inhibition of fungal spore germination by gramicidin S and its potential use as a biocontrol against fungal plant pathogens

Letters in Applied Microbiology ◽

10.1111/j.1472-765x.1986.tb01534.x ◽

1986 ◽

Vol 3 (1) ◽

pp. 5-7 ◽

Cited By ~ 6

Author(s):

THELMA MURRAY ◽

FIONA C. LEIGHTON ◽

B. SEDDON

Keyword(s):

Spore Germination ◽

Plant Pathogens ◽

Fungal Spore ◽

Gramicidin S ◽

Fungal Plant Pathogens ◽

Potential Use

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The study of the effects of airborne fungal spores in allergic and pathogenic infections in Akoko environment, Ondo State, Nigeria

World Journal of Biology Pharmacy and Health Sciences ◽

10.30574/wjbphs.2020.3.3.0054 ◽

2020 ◽

Vol 3 (3) ◽

pp. 030-042

Author(s):

Essien Benjamin Christopher

Keyword(s):

Plant Pathogens ◽

Fungal Spores ◽

Fungal Spore ◽

Relevant Information ◽

Significant Difference ◽

Airborne Fungal Spores ◽

The Mean ◽

High Concentrations ◽

Airborne Fungal Spore ◽

June July

Fungal spores and hyphal fragments have been associated with out-door allergens and constitute human, animal and plant pathogens with long history of epidemiology. Airborne fungal spores of Akoko environment during the months of October 2016 to September 2017 were trapped and analyzed palynologicaly to determine the genera with allergic and pathogenic implications. Very high concentrations of these aerospora were documented. Out of 35 fungal spore type identified, the most commonly documented genera were species of Nigrospora, Endophragmiella, Ustilago, Botryodiplodia, Curvularia, Pithomyces, Corynespora and Venturia among others. The availability of these airborne fungal spore types is not only a reflection of their degree of abundance in the atmosphere, but an indication of the availability of host plants and other spore sources in the region. Statistical analysis shows that there was significant difference in the mean monthly fungal spore recorded. Multiple comparisons (using DMRT) showed that the mean fungal spores recorded in the month of October was significantly different (P<0.05) from that recorded in the month of July but not significantly different from those recorded for other months. Seasonal variation showed that the highest mean monthly fungal spore abundance were more from June - July and October - December due to higher sporulation activities by the fungi. This study would provide relevant information that could be useful in monitoring the frequency and intensity of fungal allergies and other pathogenic disease conditions of plants, animals and humans in the study environment and proffer adequate measures for safety health and environment.

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An advanced slit-type volumetric spore trap for monitoring bioaerosols; new methods for identifying fungal spores

Australasian Plant Pathology ◽

10.1071/ap04047 ◽

2004 ◽

Vol 33 (3) ◽

pp. 393 ◽

Cited By ~ 1

Author(s):

Heike G. Neumeister-Kemp ◽

Aaron Maxwell ◽

Peter C. Kemp ◽

Bernie Dell ◽

Giles E. St J. Hardy

Keyword(s):

Fungal Spores ◽

Spore Trap ◽

New Methods

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Variability and Geographical Origin of Five Years Airborne Fungal Spore Concentrations Measured at Saclay, France from 2014 to 2018

Remote Sensing ◽

10.3390/rs11141671 ◽

2019 ◽

Vol 11 (14) ◽

pp. 1671 ◽

Cited By ~ 3

Author(s):

Roland Sarda-Estève ◽

Dominique Baisnée ◽

Benjamin Guinot ◽

John Sodeau ◽

David O’Connor ◽

...

Keyword(s):

Geographical Origin ◽

Fungal Spores ◽

Fungal Spore ◽

Agricultural Practice ◽

Receptor Model ◽

Atmospheric Variability ◽

Atmospheric Concentration ◽

Spore Trap ◽

Temperature And Precipitation ◽

Atmospheric Concentrations

Airborne fungal spores (AFS) represent the major fraction of primary biological aerosol particles (PBAPs), and they are studied worldwide largely due to their important role within the Earth system. They have an impact on climate and human health, and they contribute to the propagation of diseases. As their presence in the air depends largely on studied ecosystems, a spore trap was used to monitor their atmospheric concentrations from 2014 to December 2018 in Saclay, a suburban area in the megacity of Paris. The main objective of this work was: (1) to understand the atmospheric variability of AFS in relation to different variables such as meteorological factors, agricultural practice, and (2) to identify their geographical origin by using a source receptor model. During our period of observation, 30 taxa have been identified under a light microscope. In order of importance, Ascospores, Cladosporium, Basidiospores, Tilletiopsis, Alternaria were found to be the most abundant types respectively (50.8%, 33.6%, 7.6%, 1.8%, and 1.3%) accounting for 95% of the atmospheric concentrations. We observed a general decrease associated with a strong interannual variability. A bimodal seasonal cycle was identified with a first maximum in July and a second in October. The main parameters driving the atmospheric concentration are temperature and precipitation. The daily variability is strongly activated by successive periods of hot weather and rainfall, multiplying the concentration by a factor of 1000 in less than 12 hours. Results from the source receptor model ZeFir point out unambiguous different origins of AFS due to specific sources impacting the observation site. Our study also indicated that a hydrological stress has a direct effect on the daily concentrations. This last point should be taken into account for every stressed ecosystem studied in a global warming context. This is particularly important for Mediterranean areas where water is a key control of the growth and dispersion of fungal spores.

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