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WS-5995 B, an antifungal agent inducing differential gene expression in the conifer pathogen Heterobasidion annosum but not in Heterobasidion abietinum

Abstract

The mycorrhization helper bacterium Streptomyces sp. AcH 505 inhibits Norway spruce root infection and colonisation by the root and butt rot fungus Heterobasidion annosum 005 but not by the congeneric strain Heterobasidion abietinum 331 because of higher sensitivity of H. annosum 005 towards the AcH 505-derived naphthoquinone antibiotic WS-5995 B. Differences in antibiotic sensitivity between two isolates belonging to two species, H. annosum 005 and H. abietinum 331, were investigated by comparative gene expression analysis using macroarrays and quantitative RT-PCR after WS-5995 B, structurally related mollisin and unrelated cycloheximide application. Treatment with 25 µM WS-5995 B for 2 h resulted in a significant up-regulation of expression of inosine-5′-monophosphate dehydrogenase, phosphoglucomutase and GTPase genes, while the expression of genes encoding for thioredoxin and glutathione dependent formaldehyde dehydrogenase was down-regulated in the sensitive fungal strain. No differential expression in the tolerant strain was detected. Application of WS-5995 B at higher concentrations over a time course experiment revealed that H. annosum 005 and H. abietinum 331 responded differently to WS-5995 B. The fungal gene expression levels depended on both the concentration of WS-5995 B and the duration of its application. The WS-5995 B-unrelated cycloheximide caused highly specific changes in patterns of gene expression. Our findings indicate considerable variations in response to bacterial metabolites by the isolates of the conifer pathogen.

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Acknowledgements

We are indebted to Margret Ecke for technical assistance in microbial cultures, Dr. Julia Riedlinger for WS-5995 B preparation, and Dr. Karlheinz Seifert for providing purified mollisin. We thank Dr. Kari Korhonen and Dr. Franz Oberwinkler for Heterobasidion isolates. We further thank Åke Olson, Magnus Karlsson and Jan Stenlid for making available plasmid clones in the present macroarray study. This study was supported by the German Research Foundation (D.F.G.; graduate college ‘Infection Biology’; N.A.L.), the Swedish Research Council for Agricultural Sciences and Forestry (FORMAS), the Carl Tryggers Stiftelse (CTS, Sweden) and by Maj and Tor Nessling Foundation.

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Correspondence to Nina A. Lehr.

Electronic Supplementary Material

Below is the link to the electronic supplementary material.

Supplementary Table 1

Complete data set presenting fold changes of genes differentially expressed after macroarray analysis of H. annosum 005 liquid culture supplemented with 25 µM WS 5995 B (DOC 511 kb)

Supplementary Table 2

Fold changes determined by quantitative real-time RT-PCR ± standard deviation of H. annosum 005 (H. an) and H. abietinum 331 (H. ab). A concentration series of WS-5995 B (50, 100, 150 µM) and a time course (0.5, 2, 6 h) have been performed (DOC 157 kb)

Supplementary Fig. 1

The influence of WS-5995 B concentration and longevity of WS-5995 B treatment on Cytochrome P450 gene expression in Heterobasidion abietinum 331 and in Heterobasidion annosum 005. The fold changes of the relative gene expression levels are indicated at 50, 100 and 150 µM WS-5995 B after 0.5, 2, and 6 h treatment. aCytochrome P450 gene expression in Heterobasidion abietinum 331. bCytochrome P450 gene expression in Heterobasidion annosum 005 (DOC 68 kb)

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Lehr, N.A., Adomas, A., Asiegbu, F.O. et al. WS-5995 B, an antifungal agent inducing differential gene expression in the conifer pathogen Heterobasidion annosum but not in Heterobasidion abietinum . Appl Microbiol Biotechnol 85, 347–358 (2009). https://doi.org/10.1007/s00253-009-2254-7

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Keywords

  • Heterobasidion
  • Plant pathogen
  • Macroarray
  • Gene expression
  • Antibiotic
  • Streptomyces