prog detail - International Mycological Association
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S37IS2 - 0486<br />
Visualization of the endocytic pathway and endosomal structures in the filamentous fungus Aspergillus<br />
oryzae<br />
Y Higuchi, T Nakahama, JY Shoji, M Arioka, K Kitamoto<br />
Department of Biotechnology, The University of Tokyo, Tokyo, Japan<br />
Endocytosis is an important process for cellular activities. However, in filamentous fungi, the existence of endocytosis<br />
has been so far elusive. In this study, we used AoUapC-EGFP, the fusion protein of a putative uric acid-xanthine<br />
permease with EGFP (enhanced green fluorescent protein) in the filamentous fungus Aspergillus oryzae, to examine<br />
whether the endocytic process occurs or not. Upon the addition of ammonium into the medium the fusion protein was<br />
internalized from the plasma membrane. The internalization of AoUapC-EGFP was completely blocked by sodium<br />
azide, cold, and cytochalasin A treatments, suggesting that the internalization possesses the general features of<br />
endocytosis. These results demonstrate the occurrence of endocytosis in filamentous fungi. Moreover, we discovered<br />
the endosomal compartments that appeared upon the induction of endocytosis. The endosomal compartments<br />
displayed intermittent and bidirectional movement longitudinally along the hyphae, in a microtubule-dependent<br />
manner. Effects of the deletion of the motor proteins will be also included in the presentation.<br />
S37IS3 - 0727<br />
Network structyure and dynamics of fungal mycelia<br />
D P Bebber 1, J Hynes 2, P R Darrah 1, L Boddy 2, M D Fricker 1<br />
1. University of Oxford, Oxford, UK<br />
2. University of Cardiff, Wales, UK<br />
Many physical phenomena, from road systems to the internet, can be modelled as networks. Network analyses have<br />
revealed important properties of, and commonalities among, diverse experimental structures. The fungal mycelium is<br />
a transport network that competes in a complex and changing environment. The architecture of the network<br />
continuously adapts to local nutritional or environmental cues, damage or predation, through growth, branching,<br />
fusion and regression. We investigate whether mycelial network architecture optimizes resource capture, exploration,<br />
translocation, or defence.<br />
We use image analysis techniques to digitize the growing mycelia of cord-forming saprotrophic fungi, and to assign<br />
transport capacities and physical resiliences to the cords (Fig. 1). These models are then analysed using a variety of<br />
network-based statistics. We compare modelled transport capacity and routing with real values derived from<br />
scintillation imaging of radiolabelled mycelia, and compare in silico attack with responses of real mycelia to<br />
experimental attack by fungivorous collembola.<br />
As the mycelium grows, some cords thicken to increase transport capacity, such that the network’s effective diameter<br />
remains static even as its physical size increases. We term this property a ‘physiological small world’ effect. Differential<br />
cord thickening also increases the resilience of the network to attack, by protecting a central core component at the<br />
expense of weaker peripheral cords. The network responds to actual physical damage by increasing redundancy in<br />
transport routes.<br />
Our analyses have revealed that the fungal mycelium is a self-organised network that balances transport and<br />
defence. Further investigation of variation in mycelial structure among fungal species may reveal trade-offs between<br />
properties such as exploration and resilience, and suggest adaptations to different ecological niches. Network theory<br />
provides a new and exciting way of understanding fungal biology and ecology.<br />
S37PS1<br />
Advanced microscopic imaging coupled with x-ray absorption spectroscopy to characterise fungal metal<br />
and mineral transformations<br />
Geoffrey M. Gadd 1, John Charnock 2, Andrew D. Bowen 1 and Marina Fomina 1<br />
1 Division of Environmental and Applied Biology, Biological Sciences Institute, College of Life Sciences, University of<br />
Dundee, Dundee, DD1 4HN, UK 2 SRS Daresbury Laboratory, Daresbury, Warrington, WA4 4AD, UK<br />
In this study, scanning electron microscopy (SEM)-based techniques were used to study metal-mineral transformations<br />
by fungi, especially the formation of mycogenic minerals, while X-ray absorption spectroscopy (XAS) was used to<br />
determine the speciation of metals within biomass. It was found that fungal-mineral interactions at the microscale<br />
level could be successfully studied using different SEM approaches which preserved living fungal microstructures and<br />
the microenvironment where the precipitation of mycogenic minerals occurred. Environmental scanning electron<br />
microscopy (ESEM) in the wet mode was the best method for observing such interactions in their natural<br />
microenvironment; X-ray element mapping demonstrated sequestration and localization of metals. Cryo-SEM allowed<br />
the observation of both interior and exterior morphology and appeared to preserve the complex structure of<br />
ectomycorrhizal roots better than other microscopic techniques. The amorphous state or poor crystallinity of metal<br />
complexes within the biomass and relatively low metal concentrations makes the determination of the metal<br />
speciation a challenging problem but this can be overcome by using synchrotron-based element-specific XAS<br />
techniques. Here, we exposed fungi and ectomycorrhizas to a variety of copper-, zinc- and lead-containing minerals.<br />
XAS revealed that oxygen ligands (phosphate, carboxylate) played a major role in toxic metal coordination within<br />
fungal biomass during the accumulation of mobilized toxic metals. The use of state-of-the-art SEM techniques and<br />
XAS has provided new information about the role of fungi in metal-mineral transformations and their importance in<br />
“geomycological” processes.<br />
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