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The Protistan Cell Surface pp 142–163Cite as

Characterization of zoospore and cyst surface structure in saprophytic and fish pathogenic Saprolegnia species (oomycete fungal protists)

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The importance of the surface structure and chemistry in zoospores and cysts of oomycetes is briefly reviewed and the organelle systems associated with encystment described. The surface structure and chemistry of primary and secondary zoospores and cysts of Saprolegnia diclina (a representative saprophytic species) and S. parasitica (a representative salmonid fish pathogen) were explored using the lectins concanavilin A (Con A and wheat germ agglutinin (WGA) and monoclonal antibodies (MAbs) raised against a mixed zoospore and cyst suspension of S. parasitica. The binding of lectins and antibodies to spores was determined using immunofluorescence microscopy with fluorescein isothiocyanate-labelled probes and with electron microscopy with gold-conjugated probes applied to spore suspensions post-fixation. In both species Con A, which is specific for glucose and mannose sugars, bound to both the surface of primary and secondary zoospores (the surface glycocalyx) and their cyst coats and readily induced zoospore encystment. The binding to the cysts appeared to be mainly associated with the matrix material released from the primary and secondary encystment vesicles and which appeared to diminish with time. No binding to germ tube walls was observed with this lectin. The MAb labelling showed a generally similar binding pattern to the primary and secondary cysts to that observed with Con A, although the binding to zoospores was more variable. Primary zoospores bound the antibodies but secondary zoospores appeared less reactive. It is suggested that the MAbs share a common epitope with one or more of the Con A-binding components. In both species WGA, which is specific for amongst other things the sugar N-acetyl glucosamine, bound to localised apical patches on the primary zoospores. This lectin also binds to the ventral groove region of secondary zoospores of S. diclina, which were induced to encyst by this lectin. In contrast secondary zoospores of S. parasitica were not induced to encyst by the addition of WGA and showed a patchy dorsal binding with this lectin. WGA also binds to both the inner wall of discharged primary cysts and the young germ tube walls of both species. These observations are discussed both in relation to other oomycete spores and to their possible functional and ecological significance

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Abbreviations

BSA:

bovine serum albumin

Con A:

Concanavalin A

DBA:

Dolichos biflorus agglutinin

ELISA:

enzyme-linked immunosorbent assay

EM:

electron microscope

EV:

encystment vesicles

FCS:

foetal calf serum

FITC:

Fluorescein isothiocyanate

FV:

peripheral fibrillar vesicles

G + F 0.2%:

glutaraldehyde and 2.0% formaldehyde primary fixative solution

2G 2%:

glutaraldehyde primary fixative

LM:

light microscopy

MAbs:

monoclonal antibodies

LPV:

large peripheral vesicles

PBS:

phosphate buffered saline

PCV:

flattened peripheral cisternae

PEV:

primary encystment vesicle

PIPES:

piperazine-N,Nl-bis(2-ethane sulfonic acid)

PNA:

Ricinus communis agglutinin

RAM-FITC/Au10–20 :

Fluorescein isothiocyanate/gold (10 or 20 nm) labelled rabbit anti-mouse immunoglobulin

RCA:

Ricinus communis agglutinin

SEM:

scanning electron micrograph

SBA:

soybean agglutinin

SEV:

secondary encystment vesicles

TEM:

transmission electron micrograph

UEA I:

Ulex europaeus agglutinin

WGA:

wheat germ agglutinin

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Authors and Affiliations

  1. Department of Biological and Nutritional Sciences, Agriculture Building, The University of Newcastle upon Tyne, Newcastle upon Tyne, NE1 7RU, UK

    A. W. Burr & G. W. Beakes

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  1. A. W. Burr
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  2. G. W. Beakes
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Editors and Affiliations

  1. School of Botany, University of Melbourne, Parkville, Victoria, Australia

    Richard Wetherbee  & Jeremy D. Pickett-Heaps  & 

  2. Provasoli-Guillard Center for Culture of Marine Phytoplankton, Bigelow Laboratory for Ocean Sciences, West Boothbay Harbor, Maine, USA

    Robert A. Andersen

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Burr, A.W., Beakes, G.W. (1994). Characterization of zoospore and cyst surface structure in saprophytic and fish pathogenic Saprolegnia species (oomycete fungal protists). In: Wetherbee, R., Pickett-Heaps, J.D., Andersen, R.A. (eds) The Protistan Cell Surface. Springer, Vienna. https://doi.org/10.1007/978-3-7091-9378-5_8

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