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Immuno-gluorescence study of five zygomycetous fungi with two chitin-binding probes

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dc.contributor.author Sahai, Amarpal S. en_US
dc.date.accessioned 2009-07-09T18:38:33Z
dc.date.available 2009-07-09T18:38:33Z
dc.date.issued 1993-07-09T18:38:33Z
dc.identifier.uri http://hdl.handle.net/10464/1908
dc.description.abstract A polyclonal antiserum was prepared against a purified microsomal chitinase isolated from the fungus Choanephora cucurbitarum. Indirect immunofluorescence was used to localize chitinase at various developmental stages of five zygomycetous fungi and during abiotrophic mycoparasite interaction with a susceptible and resistant host. This was compared to localization of oligomers of N-acetylglucosamine with the lectin wheat germ agglutinin (WGA). Dotimmunoblot and Western blot techniques revealed that the anti-serum reacted strongly with the antigen from which it was derived. Cross reactivity of the antiserum was found with WGA and another chitin binding lectin, Phyto/acca americana agglutinin (PAA). Immuno-fluorescence results showed the direct involvement of chitinase in spore swelling, germination, sporangium development and response during mechanical injury. There appeared to be no involvement of chitinase during apical hyphal growth or new branch initiation in any of the fungi tested despite mild proteolysis and permeabilization of the cell surface prior to labelling. Binding with WGA revealed similar patterns of fluorescence to that of chitinase localization but differed by showing fluorescence and therefore chitin localization at the apex and new branch initiation when tested at different developmental stages. There was no difference between chitinase localization and binding with WGA in a susceptible host and resistant host challenged with the mycoparasite, Piptocephalis virginiana. Differences in binding ability of antichitinase and lectin WGA suggests that the latter is not a suitable indicator for indirect localization of the lytic enzyme, chitinase. en_US
dc.language.iso eng en_US
dc.publisher Brock University en_US
dc.subject Immunofluorescence. en_US
dc.subject Zygomycetes. en_US
dc.subject Chitin. en_US
dc.subject Fungi. en_US
dc.title Immuno-gluorescence study of five zygomycetous fungi with two chitin-binding probes en_US
dc.type Electronic Thesis or Dissertation en_US
dc.degree.name M.Sc. Biological Sciences en_US
dc.degree.level Masters en_US
dc.contributor.department Department of Biological Sciences en_US
dc.degree.discipline Faculty of Mathematics and Science en_US


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