The preparation and use of a fluorescent antibody reagent for Botrytis cinerea grown on glass slides

Preece, T.F.; Cooper, Dorothy J.

doi:10.1016/s0007-1536(69)80163-4

Cited by 14 publications

(7 citation statements)

References 8 publications

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“…La redierdie de methodes moins brutales est un objectif souvent difficile a concilier avec la pratique. Un travail d'extraction a partir du seul mycelium supprime generalement cette phase de concentration, a condition de disposer d'un materiel fongique suffisant (BURRELL et al 1966, PREECE et COOPER 1969. Cependant on se prive du milieu de culture qui contient des elements exogones pouvant presenter un interet certain.…”

Section: Discussionunclassified

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Etude sérologique de quelques Botrytis

Olivier¹

1979

Journal of Phytopathology

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Section: Discussionunclassified

“…Differents protocoles d'immunisations sont utilises dans les travaux ant^rieurs. La voie sous-cutan^e, avec adjuvant, a foumi ^ PREECE et COOPER (1969) des titres equivalents a ceux que nous avons obtenus (-1/64). PREECE (1971) signale l'injection hebdomadaire par voie intraveineuse.…”

Section: Discussionunclassified

Etude sérologique de quelques Botrytis

Olivier¹

1979

Journal of Phytopathology

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“…Serological techniques for the detection and identification of fungi, recently reviewed by Preece and Cooper (1969), have not been widely used but have obvious potential. From our preliminary experiments with E. armeniacae two points of interest emerge:…”

Section: Discussionmentioning

confidence: 99%

“…Results of the immunofluorescence experiments indicate that the anti-mycelium serum contained antibodies to intracellular antigens, but the presence of antibodies to cell-wall antigens was difficult to assess owing to non-specific fluorescence of cell walls, a problem also encountered with other fungi (Preece and Cooper 1969). If intracellular antigens (mainly proteins) are more immunogenic than cell-wall antigens (mainly polysaccharides) there may be immunogenic competition within the animal.…”

Section: Discussionmentioning

confidence: 99%

The Serological Properties of Eutypa Armeniacae Mycelium and Ascospores

Francki¹,

Carter²

1970

Aust. Jnl. Of Bio. Sci.

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In epidemiological studies of the apricot vascular pathogen Eutypa armeniacae Hansf. & Carter in this laboratory (Moller and Carter 1965;Carter and Moller 1967) some diagnostic problems have been encountered. The pathogen is sufficiently variable in culture to be confused with other Ascomycetes, such as species of Valsa and Cryptovalsa which may colonize dead vascular tissue below pruning wounds which have been invaded by E. armeniacae. It is also difficult to differentiate E. armeniacae ascospores from those of related Ascomycetes caught in spore traps during wet weather.The preparation of antisera to antigens of E. armeniacae ascospores and mycelium and their possible use in identification of the fungus are the subjects of this communication. Materials and MethodsPerithecial stromata of E. armeniacae were soaked in water for I hr and attached to the lid of a closed container for several hours to collect the liberated ascospores. The ascospores were then suspended in a small quantity of distilled water and the concentration estimated by haemacytometer. A quantity of the suspension containing 2-5 X 107 ascospores was then emulsified with an equal volume of Freund's complete adjuvant and injected subcutaneously into several sites on the flank of a rabbit. Two further injections containing similar quantities of spores were administered 1 and 5 weeks after the initial injection. The rabbit was bled through an ear vein 1 week after the final injection and several more times during the month following.E. armeniacae was cultured on Czapek-Dox agar and after 2-3 weeks of growth the hyphae were scraped off the agar, suspended in 0 ·14M N aCI, and ground in a glass tissuehomogenizer. Microscopic examination of the suspension showed the presence of small pieces of hyphae. This material was emulsified with an equal volume of adjuvant and used to immunize a rabbit as described above for the ascospore antigen.Serological tests with mycelial antigens were carried out by the double-diffusion precipitation technique (Crowle 1961) using 0·75% agar in 0·02M phosphate buffer, pH 7,2, containing 0'14M NaCl and 0·02% sodium azide. Agglutination tests using ascospores as antigens were carried out on microscope slides. Immunofluorescence tests with both mycelium and ascospores were carried out by the indirect method (Weller and Coons 1954). Smears of the antigens were prepared by drying suspensions on microscope slides which were then irrigated with rabbit antiserum for 15 min. The smears were washed thoroughly with 0 ·14M NaCl and irrigated with fluorescein isocyanate-labelled sheep anti-rabbit globulin serum for 15 min and again washed with 0 ·14M NaCl. The smears were mounted in glycerol and examined in a Leitz Ortholux microscope equipped with both phase-contrast and ultraviolet illumination.* Manuscript

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“…(Preece &' Cooper 1969) and Phytophthora cinnamomi Rands (Malajczuk et aI. 1975 1978 and unpublished), so a modification of the method of Malajczuk et aI.…”

Section: Introductionmentioning

confidence: 99%

An immunofluorescent counting method for tremorgenic penicillia

Wyatt

Menna

1981

New Zealand Journal of Agricultural Research

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Antibodies produced against tremorgenic strains of Penicillium crustosum and Penicilliumjanczewskii, conjugated indirectly with fluorescein, were used to stain mycelium of pure cultures of moulds and also hyphae present in natural substrates.There was strong fluorescent staining of strains of nine of 11 Penicillium species known to produce tremorgens and a variable staining reaction of two. Strains of seven of 11 non-tremorgenic Penicillium species did not react, three stained weakly or variably and one stained strongly. None of the isolates of 17 other mould genera examined, including tremorgenic strains of Aspergillus spp., was reactive. Fluorescent hyphae were seen in preparations of soil suspensions and plant washings from which reactive Penicillia were isolated by culture on dilution plates.

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The preparation and use of a fluorescent antibody reagent for Botrytis cinerea grown on glass slides

Cited by 14 publications

References 8 publications

Etude sérologique de quelques Botrytis

Etude sérologique de quelques Botrytis

The Serological Properties of Eutypa Armeniacae Mycelium and Ascospores

An immunofluorescent counting method for tremorgenic penicillia

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