1887

Abstract

, the chestnut blight fungus, can be infected by virulence-attenuating mycoviruses of the family . Previous studies have led to the hypothesis that the hypovirus-infected phenotype is partly due to metabolic changes induced by the viral infection. To investigate this, we measured the metabolic rate and respiration of colonies grown on solid medium. These experiments supported historical observations of other fungal species done in liquid cultures that the metabolic rate steadily declines with age and differentiation of the mycelium. Hypovirus infection increased metabolic rate in the youngest mycelium, but a subsequent decline was also observed as the mycelium aged. By measuring both CO production and O consumption, we also observed that changes occur in carbohydrate metabolism as a result of ageing in both infected and uninfected mycelium. Mycelium on the periphery of the colony exploited fermentation pathways extensively, before transitioning to aerobic carbohydrate metabolism and finally lipid metabolism in the interior regions, despite abundant remaining glucose. However, the hypovirus affected the extent of these changes, with infected mycelium apparently unable to utilize lipid-related metabolic pathways, leading to an increased depletion of glucose. Finally, we used metabolic profifiling to determine the changes in accumulation of primary metabolites in wild-type and hypovirus-infected mycelium and found that approximately one-third of the 164 detected metabolites were affected. These results are consistent with those expected from the physiological measurements, with significant alterations noted for compounds related to lipid and carbohydrate metabolism. Additionally, we observed an increase in the accumulation of the polyamine spermidine in the presence of hypovirus. Polyamines have been implicated in antiviral responses of mammalian systems; therefore this may suggest a novel antiviral response mechanism in fungi.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.029033-0
2009-12-01
2020-11-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/155/12/3913.html?itemId=/content/journal/micro/10.1099/mic.0.029033-0&mimeType=html&fmt=ahah

References

  1. Allen T. D., Nuss D. L. 2004; Linkage between mitochondrial hypovirulence and viral hypovirulence in the chestnut blight fungus revealed by cDNA microarray analysis. Eukaryot Cell 3:1227–1232
    [Google Scholar]
  2. Allen T. D., Dawe A. L., Nuss D. L. 2003; Use of cDNA microarrays to monitor transcriptional responses of the chestnut blight fungus Cryphonetria parasitica to infection by virulence-attenuating hypoviruses. Eukaryot Cell 2:1253–1265
    [Google Scholar]
  3. Bachrach U. 2007; Antiviral activity of oxidized polyamines. Amino Acids 33:267–272
    [Google Scholar]
  4. Bideaux C., Alfenore S., Cameleyre X., Molina-Jouve C., Uribelarrea J. L., Guillouet S. E. 2006; Minimization of glycerol production during the high-performance fed-batch ethanolic fermentation process in Saccharomyces cerevisiae, using a metabolic model as a prediction tool. Appl Environ Microbiol 72:2134–2140
    [Google Scholar]
  5. Buck K. 1986 Fungal Virology – an Overview. Boca Raton, FL: CRC Press;
    [Google Scholar]
  6. Carlisle M. J., Watkinson S. C., Gooday G. W. 2001 The Fungi , 2nd edn. San Diego: Academic Press;
    [Google Scholar]
  7. Chen B., Choi G. H., Nuss D. L. 1994; Attenuation of fungal virulence by synthetic infectious hypovirus transcripts. Science 264:1762–1764
    [Google Scholar]
  8. Choi G. H., Nuss D. L. 1992; Hypovirulence of chestnut blight fungus conferred by an infectious viral cDNA. Science 257:800–803
    [Google Scholar]
  9. Cochrane V. W. 1958 Physiology of Fungi New York: Wiley;
    [Google Scholar]
  10. Crabtree H. G. 1929; Observations on the carbohydrate metabolism of tumours. Biochem J 23:536–545
    [Google Scholar]
  11. Dawe A. L., Nuss D. L. 2001; Hypoviruses and chestnut blight: exploiting viruses to understand and modulate fungal pathogenesis. Annu Rev Genet 35:1–29
    [Google Scholar]
  12. De Deken R. H. 1966; The Crabtree effect: a regulatory system in yeast. J Gen Microbiol 44:149–156
    [Google Scholar]
  13. Dixon M. 1952 Manometric Methods as Applied to the Measurement of Cell Respiration and Other Processes , 3rd edn. London: Cambridge University Press;
    [Google Scholar]
  14. Ferreira M. E., Colombo A. L., Paulsen I., Ren Q., Wortman J., Huang J., Goldman M. H., Goldman G. H. 2005; The ergosterol biosynthesis pathway, transporter genes, and azole resistance in Aspergillus fumigatus . Med Mycol 43:Suppl 1S313–S319
    [Google Scholar]
  15. Fleet G. H., Heard G. M. 1993; Yeast growth during fermentation. In Wine Microbiology and Biotechnology pp 27–54 Chur, Switzerland: Harwood Academic Publishers;
    [Google Scholar]
  16. Foster J. W. 1949 Chemical Activities of Fungi New York: Academic Press;
    [Google Scholar]
  17. Gentile A. C. 1954; Carbohydrate metabolism and oxalic acid synthesis by Botrytis cinerea . Plant Physiol 29:257–261
    [Google Scholar]
  18. Gessaman J. A., Nagy K. A. 1988; Energy metabolism: errors in gas-exchange conversion factors. Physiol Zool 61:507–513
    [Google Scholar]
  19. Gheshlaghi R., Scharer J. M., Moo-Young M., Douglas P. L. 2007; Metabolic flux analysis for optimizing the specific growth rate of recombinant Aspergillus niger . Bioprocess Biosyst Eng 30:397–418
    [Google Scholar]
  20. Golinski M. R., Boecklen W. J., Dawe A. L. 2008; Two-dimensional fractal growth properties of the filamentous fungus Cryphonectria parasitica: the effects of hypovirus infection. J Basic Microbiol 48:426–429
    [Google Scholar]
  21. Hansen D. R., Van Alfen N. K., Gillies K., Powell W. A. 1985; Naked dsRNA associated with hypovirulence of Endothia parasitica is packaged in fungal vesicles. J Gen Virol 66:2605–2614
    [Google Scholar]
  22. Hawker L. E. 1950 Physiology of Fungi London: Oxford University Press;
    [Google Scholar]
  23. Jewett M. C., Hofmann G., Nielsen J. 2006; Fungal metabolite analysis in genomics and phenomics. Curr Opin Biotechnol 17:191–197
    [Google Scholar]
  24. Jin Y., Bok J. W., Guzman-de-Pena D., Keller N. P. 2002; Requirement of spermidine for developmental transitions in Aspergillus nidulans . Mol Microbiol 46:801–812
    [Google Scholar]
  25. Koffler H., Emerson R. L., Perlman D., Burris R. H. 1945; Chemical changes in submerged penicillin fermentations. J Bacteriol 50:517–548
    [Google Scholar]
  26. Lin H., Lan X., Liao H., Parsley T. B., Nuss D. L., Chen B. 2007; Genome sequence, full-length infectious cDNA clone, and mapping of viral double-stranded RNA accumulation determinant of hypovirus CHV1-EP721. J Virol 81:1813–1820
    [Google Scholar]
  27. McCabe P. M., Van Alfen N. K. 2001; Molecular basis of symptom expression by the Cryphonectria hypovirus. In dsRNA Genetic Elements: Concepts and Applications in Agriculture, Forestry, and Medicine pp 125–144 Edited by Tavantzis S. M. Boca Raton, FL: CRC Press;
    [Google Scholar]
  28. Melvin R. G., Van Voorhies W. A., Ballard J. W. 2007; Working harder to stay alive: metabolic rate increases with age in Drosophila simulans but does not correlate with life span. J Insect Physiol 53:1300–1306
    [Google Scholar]
  29. Moss B. J., Kim Y., Nandakumar M. P., Marten M. R. 2008; Quantifying metabolic activity of filamentous fungi using a colorimetric XTT assay. Biotechnol Prog 24:780–783
    [Google Scholar]
  30. Murayama S. Y., Negishi Y., Umeyama T., Kaneko A., Oura T., Niimi M., Ubukata K., Kajiwara S. 2006; Construction and functional analysis of fatty acid desaturase gene disruptants in Candida albicans . Microbiology 152:1551–1558
    [Google Scholar]
  31. Newhouse J. R., Hoch H. C., MacDonald W. L. 1983; The ultrastructure of Endothia parasitica. Comparison of a virulent with a hypovirulent isolate. Can J Bot 61:389–399
    [Google Scholar]
  32. Nuss D. L. 2005; Hypovirulence: mycoviruses at the fungal–plant interface. Nat Rev Microbiol 3:632–642
    [Google Scholar]
  33. Pedneault K., Angers P., Gosselin A., Tweddell R. J. 2006; Fatty acid composition of lipids from mushrooms belonging to the family Boletaceae. Mycol Res 110:1179–1183
    [Google Scholar]
  34. Pieckenstain F. L., Garriz A., Chornomaz E. M., Sanchez D. H., Ruiz O. A. 2001; The effect of polyamine biosynthesis inhibition on growth and differentiation of the phytopathogenic fungus Sclerotinia sclerotiorum . Antonie Van Leeuwenhoek 80:245–253
    [Google Scholar]
  35. Shu P. 1953; Fermentation, oxygen uptake in shake flask fermentations. J Agric Food Chem 1:1119–1123
    [Google Scholar]
  36. Thomson J. M., Gaucher E. A., Burgan M. F., De Kee D. W., Li T., Aris J. P., Benner S. A. 2005; Resurrecting ancestral alcohol dehydrogenases from yeast. Nat Genet 37:630–635
    [Google Scholar]
  37. Van Voorhies W. A., Ward S. 1999; Genetic and environmental conditions that increase longevity in Caenorhabditis elegans decrease metabolic rate. Proc Natl Acad Sci U S A 96:11399–11403
    [Google Scholar]
  38. Van Voorhies W. A., Khazaeli A. A., Curtsinger J. W. 2004; Testing the “rate of living” model: further evidence that longevity and metabolic rate are not inversely correlated in Drosophila melanogaster . J Appl Physiol 97:1915–1922
    [Google Scholar]
  39. Van Voorhies W. A., Melvin R. G., Ballard J. W., Williams J. B. 2008; Validation of manometric microrespirometers for measuring oxygen consumption in small arthropods. J Insect Physiol 54:1132–1137
    [Google Scholar]
  40. Walsberg G., Wolf B. 1995; Variation in the respiratory quotient of birds and implications for indirect calorimetry using measurements of carbon dioxide production. J Exp Biol 198:213–219
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.029033-0
Loading
/content/journal/micro/10.1099/mic.0.029033-0
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error