CBS-KNAW fungal biodiversity centre
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Centraalbureau voor Schimmelcultures

Utrecht, The Netherlands

Fungal Physiology

This group is a collaboration between the CBS Fungal Biodiversity Centre and Utrecht University, Chair of Microbiology. Information on projects of this group at Utrecht University can be found at: http://www.bio.uu.nl/microbiology/

Fungal physiology is the basis of biotope and global dispersion of fungal species. It determines the nutrients it can use, the environmental conditions it can endure and its competitive position in its ecosystem. The ability of fungi to survive in every known biotope, both natural and man-made, relies in part on their capacity to use a wide range of carbon sources. In nature, many fungi degrade polymeric carbon sources (e.g. polysaccharides, proteins, lignin) to use the monomeric components as carbon source. However, the available carbon sources vary strongly in nature, both between biotopes and in time. While some fungi have become specialists that focus on specific carbon sources or specific biotopes, other have are more generalists that can grow in many biotopes and use a large variety of carbon sources. Differences in physiology may therefore also reflect species boundaries.

Degradation of polymeric carbon sources occurs extracellularly by a broad range of enzymes, of which the production is tightly controlled by a network of regulators. This enables fungi to produce an enzyme mixture that is tailored specifically for the available carbon sources at any given time. The released monomeric compounds are transported into the cell and metabolized through a large variety of metabolic pathways. These pathways are often co-regulated with the extracellular enzymes that release the compounds entering the pathways, resulting in a highly complex regulatory and metabolic network. To study fungal physiology in relation to natural substrates it is therefore necessary to address all these aspects of fungal biology: production of extracellular enzymes, metabolic pathways and regulators controlling the fungal response to the substrates present in the environment.

Fungal biodiversity with respect to polysaccharide degradation
Enzymes involved in polysaccharide degradation are among the best studied proteins in biology. Many genes encoding these enzymes have been cloned and biochemical evidence for the function of many of the corresponding proteins has been described. These enzymes can be divided into families based on amino acid motifs in their sequence resulting in a comprehensive classification that is available online through www.cazy.org. With the availability of more and more genome sequences, this database has grown exponentially and a high demand for accurate function prediction of CAZy-related genes has arisen. Analysis of fungal genome sequences with respect to polysaccharide degradation enables determination of the hydrolytic potential of fungi. In collaboration with the scientists responsible for the CAZy database, Bernard Henrissat and Pedro Coutinho, we have shown that this hydrolytic potential of fungi applied specifically to plant polysaccharides correlates very well to the ability of the fungus to grown on polysaccharides and can explain (in part) the biotope of a specific species. As we are adding more species to our comparative analysis, the differences between the species with respect to growth profile and biotope become more clear and can often be predicted based on their genome sequence.

Improved understanding of fungal physiology in relation to natural carbon sources
Very little is known about physiology of fungi in relation to natural carbon sources as most physiological studies have been performed on defined media under laboratory conditions or in bioreactors. To improve our understanding of fungal physiology under natural conditions, in depth studies with model organisms are required to provide the knowledge base for biodiversity studies. Two model organisms have been chosen to study these processes. Both organisms have publicly available genome sequences, transcriptomics and proteomics technology, are readily transformed allowing gene introduction or inactivation and have a long history of research:

- Aspergillus niger is one of the best studied organisms with respect to degradation of natural carbon sources and is one of the most important industrial fungi in the world. Many genes and their corresponding enzymes involved in polysaccharide degradation have been studied in detail and 5 regulators related to this process have already been characterized. Currently, three genome sequences are available for A. niger as well as genome sequences of 7 other Aspergillus species, with several more in process.

- Magnaporthe grisea, also known as the rice blast fungus is one of the most important plant pathogenic fungi worldwide. Its main impact is on rice production, causing an annual crop loss that could feed millions of people. It contains many of the systems related to plant polysaccharide degradation that have been studied in A. niger, but initial results suggest significant differences between the two species.

Detailed studies of the regulatory systems, extracellular functions and metabolic pathways related to natural carbon sources in these two fungi will provide additional targets for biodiversity studies in a larger range of fungi.

Interaction between fungal species in the environment
In their natural environment, fungal species co-exist with other organisms (e.g. other fungi, bacteria). Some fungal species may compete for nutrients while others may benefit from each other. Very little is known about the relationship between fungal species and how they affect each other’s physiology. Studying the ability of fungal species to co-exist, identifying competitive, synergistic and parasitic relationships, and determining the effect of co-cultivation on degradation of natural carbon sources will provide the first data into the overall physiology of fungal populations. To this end, fungal species will be co-cultivated on a range of substrates and analysis of their physiology will be performed at the level of growth and morphology, cell biology, gene expression, metabolism, production of extracellular enzymes and degradation of the natural substrates. Combination of model fungi will be used as well as combinations of fungal strains isolated from the same environment.

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