A New Method To Monitor Airborne Inoculum of the Fungal Plant Pathogens
            <i>Mycosphaerella brassicicola</i>
            and
            <i>Botrytis cinerea</i>

Kennedy, Roy; Wakeham, Alison; Byrne, K. G.; Meyer, U. M.; Dewey, F. M.

doi:10.1128/aem.66.7.2996-3003.2000

Cited by 58 publications

(55 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…These include immunological and DNA and polymerase chain reaction (PCR)-based methods. Immunological detection of Mycosphaerella brassicicola from spore traps has been developed to rationalize pesticide applications within crop protection programs (42). However, pathogen discrimination is still dependent on spore production, which may occur at a late stage in development in the disease cycle and by which stage prophylactic intervention may not be a management option.…”

mentioning

confidence: 99%

Development of Nested Polymerase Chain Reaction Detection ofMycosphaerellaspp. and Its Application to the Study of Leaf Disease inEucalyptusPlantations

Glen¹,

Smith²,

Langrell³

et al. 2007

Phytopathology®

View full text Add to dashboard Cite

Glen, M., Smith, A. H., Langrell, S. R. H., and Mohammed, C. L. 2007. Development of nested polymerase chain reaction detection of Mycosphaerella spp. and its application to the study of leaf disease in Eucalyptus plantations. Phytopathology 97:132-144.Mycosphaerella leaf disease (MLD) is a serious disease of two of the major eucalypt species grown in temperate regions worldwide, Eucalyptus globulus and E. nitens. More than 30 species of Mycosphaerella have been reported on eucalypts worldwide. Accurate, rapid, and early discrimination of Mycosphaerella spp. causing crown damage to E. globulus and E. nitens will assist the development of sustainable management strategies. This study describes the development, and incorporation in a nested polymerase chain reaction (PCR) approach, of specific primers for the detection and identification of Mycosphaerella spp. commonly reported from leaf lesions of E. globulus and E. nitens in Australia. Primer design was assisted by sequence alignment and phylogenetic analysis of 165 nonredundant sequences from the nuclear ribosomal DNA internal transcribed spacer regions of Mycosphaerella and related species. Phylogenetic analysis revealed very high sequence similarity for two taxon groups, Mycosphaerella grandis and M. parva, and M. vespa, M. ambiphylla, and M. molleriana, and primers were designed to differentiate each of the two groups. Three other species, M. cryptica, M. nubilosa, and M. tasmaniensis, were distinct and distinguished by species-specific primers. In double-blind trials, the detection test accurately and rapidly identified Mycosphaerella spp. in cultures and discriminated against other pathogens that co-occur in or on Eucalyptus leaves, thereby verifying its reliability. The detection test has an internal amplification control in the first-round PCR with fungal-specific primers to raise confidence in test results, particularly to highlight negative results due to PCR inhibition. When applied to DNA extracted from leaf or stem samples either as multiple or single lesions, it detected and identified up to five Mycosphaerella spp. or taxon groups in both positively identified and in young (putative) MLD lesions. The samples were 20 mm 2 or larger in surface area and were collected while undertaking disease rating assessments in an experimental investigation of Eucalyptus plantations and regrowth forest. Using nested PCR detection, Mycosphaerella spp. were positively identified in 2 days, 1 to 5 months earlier than by classical methods, demonstrating the potential application of this detection test to the early discrimination of MLD components in ecological, epidemiological, and genetic investigations.

show abstract

mentioning

confidence: 99%

Development of Nested Polymerase Chain Reaction Detection ofMycosphaerellaspp. and Its Application to the Study of Leaf Disease inEucalyptusPlantations

Glen¹,

Smith²,

Langrell³

et al. 2007

Phytopathology®

View full text Add to dashboard Cite

show abstract

“…The trap was tested in wind tunnel experiments with Alternaria brassicae conidia utilizing a monoclonal antibody against germinating conidia of this pathogen (36,37). More recently, a new particle-trapping device, the microtiter immunospore trap, was developed by the Burkard Manufacturing Co., Rickmansworth, UK (19). The device uses a suction system to collect airborne particles directly into the wells of a microtiter plate.…”

Section: Discussionmentioning

confidence: 99%

“…The device uses a suction system to collect airborne particles directly into the wells of a microtiter plate. Kennedy et al (19) demonstrated the utility of this device for detecting airborne spores of M. brassicicola and B. cinerea by ELISA. We found in this study that P. pachyrhizi urediniospores captured in a passive air sampler could be quickly and easily identified.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

An Immunofluorescence Assay to Detect Urediniospores of Phakopsora pachyrhizi

et al. 2008

View full text Add to dashboard Cite

An indirect immunofluorescence spore assay (IFSA) was developed to detect urediniospores of Phakopsora pachyrhizi, utilizing rabbit polyclonal antisera produced in response to intact nongerminated (SBR1A) or germinated (SBR2) urediniospores of P. pachyrhizi. Both antisera were specific to Phakopsora spp. and did not react with other common soybean pathogens or healthy soybean leaf tissue in enzyme-linked immunosorbent assay (ELISA). SBR1A and SBR2 bound to P. pachyrhizi and P. meibomiae urediniospores were detected with goat anti-rabbit Alexa Fluor 488-tagged antiserum using a Leica DM IRB epifluorescent microscope with an I3 blue filter (excitation 450 to 490 nm, emission 515 nm). The assay was performed on standard glass microscope slides; double-sided tape was superior to a thin coating of petroleum jelly both in retaining spores and in immunofluorescence. The IFSA was used to confirm the identity of P. pachyrhizi urediniospores captured on glass slides from passive air samplers from Georgia, Kentucky, and Ohio during 2006.

show abstract

“…These techniques have been successfully applied to detect bacteria and fungi in air samples (Schmechel et al 2003;Speight et al 1997;Nugent 1997;Kennedy et al 2000). Given the nature of these detection techniques, liquid and filter material are preferable sampling media.…”

Section: Introductionmentioning

confidence: 99%

Performance Characteristics of the Aerosol Collectors of the Autonomous Pathogen Detection System (APDS)

Mainelis

Masquelier

Makarewicz

et al. 2005

Aerosol Science and Technology

View full text Add to dashboard Cite

This research analyzes the physical performance characteristics of the aerosol collectors of the autonomous pathogen detection system (APDS) that was recently developed by the Lawrence Livermore National Laboratory. The APDS is capable of continuous and fully autonomous monitoring for multiple airborne threat organisms and can be used as part of a monitoring network for urban areas and major public gatherings. The system has already been successfully tested with airborne Bacillus anthracis and Yersinia pestis biowarfare agents. The APDS aerosol collection system consists of a PM-style cap to remove large particles and a low-pressure drop virtual impactor preconcentrator positioned in front of a wettedwall cyclone. The aerosol collectors operate at flow rates as high as 3750 l/min and collect airborne particles into 4 ml of liquid for subsequent detection. In our tests we determined the overall collection efficiency of the system by measuring the difference between inlet and outlet particle concentrations. The tests were performed with polydisperse oleic acid and monodisperse polystyrene latex (PSL) particles (0.6-3.1 µm), and for three values of the major air flow rates in the virtual impactor (1760, 2530, and 3300 l/min), two values of the product, or cyclone, flow rates (375 and 450 l/min), and two different volumes of collection liquid (4 and 6 ml). We found that the cutoff size (d 50 ) of the entire collection system varied from 1.5 to 2.0 µm when collecting PSL particles, with 3.1 µm PSL particles being collected with efficiency of approximately 85%. When collecting oleic acid particles the d 50 of the entire system varied from 1.1 to 1.6 µm. The concentration rates of the aerosol collection system were found to increase with increasing overall collection flow rate and approached one million per minute at the highest tested flowrates. Such high concentrating rates and high air sample volumes make the APDS collection system highly suitable for detecting low concentrations of airborne pathogens.

show abstract

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

Cited by 58 publications

References 20 publications

Development of Nested Polymerase Chain Reaction Detection ofMycosphaerellaspp. and Its Application to the Study of Leaf Disease inEucalyptusPlantations

Development of Nested Polymerase Chain Reaction Detection ofMycosphaerellaspp. and Its Application to the Study of Leaf Disease inEucalyptusPlantations

An Immunofluorescence Assay to Detect Urediniospores of Phakopsora pachyrhizi

Performance Characteristics of the Aerosol Collectors of the Autonomous Pathogen Detection System (APDS)

Contact Info

Product

Resources

About