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Release of highly elicitor-active glucans by germinating zoospores of Phytophthora megasperma f. sp. glycinea

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Abstract

An in-vitro culture system allowing the simultaneous germination of cysts was used to study the early host-independent release of phytoalexin elicitors by Phytophthora megasperma f. sp. glycinea, a soybean pathogen. Significant elicitor activity could be detected in the culture medium as early as 2 h after germination of P.m. f. sp. glycinea, race 1, cysts. The phytoalexin elicitor was heat-stable and heterogeneous in size. The apparent molecular mass ranged from 3 to 80 kDa. Anion exchange and lectin-affinity chromatography followed by sugar analysis confirmed that the elicitor activity resided primarily in glucans. The time course of elicitor release could then be accurately monitored by means of a competitive radioligand-displacement assay using the β-glucan elicitor-binding sites of soybean (Glycine max (L.) Merr.) membranes. Linkage-composition analysis of the glucan elicitors showed that they were primarily (1 → 3)β-linked with (1 → 6)-β-branches, a composition similar to that of glucans obtained by heat release from mature mycelium but different from that of elicitors obtained by acid hydrolysis or from spontaneous autohydrolytic release by senescent cultures. The naturally released elicitors displayed a biological activity in soybean cotyledon bioassays higher than purified acid-hydrolysed glucan elicitor or than the hepta-(1 → 3, 1 → 6)-β-glucoside, the smallest known carbohydrate elicitor for soybean. The present results demonstrate that elicitor release from the pathogen and perception by the potential host can take place in this system as early as during germ-tube formation and independent of the presence of host-produced endoglucanases.

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Abbreviations

EC50 :

effector concentration necessary for halfmaximal response

GC-MS:

gas chromatography-mass spectrometry

HG-APEA:

1-[2-(4-aminophenyl)ethyl]amino-1-[hexagluco-syl]-deoxyglucitol

IC50 :

inhibitor concentration necessary for half-maximal inhibition

P.m. f. sp. glycinea :

Phytophthora megasperma f. sp. glycinea

V0 :

void volume

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Authors and Affiliations

  1. Lehrstuhl für Biochemie der Pflanzen, Biologisches Institut II, Universität Freiburg, Schänzlestrasse 1, W-7800, Freiburg, FRG

    Tom Waldmüller, Eric G. Cosio, Hans Grisebach & Jürgen Ebel

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  1. Tom Waldmüller
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  2. Eric G. Cosio
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  3. Hans Grisebach
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  4. Jürgen Ebel
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Additional information

Deceased on March 19, 1990

This work was supported by the Deutsche Forschungsgemeinschaft (SFB 206). We thank Dr. H. Mayer, C. Warth and D. Borowiak (Max-Planck-Institut für Immunbiologie, Freiburg) for helpful discussions and experienced technical assistance (glucosyl-linkage analysis).

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Waldmüller, T., Cosio, E.G., Grisebach, H. et al. Release of highly elicitor-active glucans by germinating zoospores of Phytophthora megasperma f. sp. glycinea . Planta 188, 498–505 (1992). https://doi.org/10.1007/BF00197041

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