Filamentous fungi, such as <i>Aspergillus nidulans</i>, are composed of tubular, highly polarized, multinucleate cells called hyphae. Polar growth involves secretion specifically at the hyphal tip. Secretion involves intracellular transport and co-ordination of the cytoskeleton and the endomembrane system. <p>Intracellular transport is likely mediated by cytoskeletal elements, which, in fungal cells consist primarily of actin and microtubules (MTs). An <i>A. nidulans</i> strain transformed with green fluorescent protein (GFP) tagged α-tubulin was utilized in the investigation of relationship between cytoplasmic MT arrays and hyphal growth rate. <i>A. nidulans</i> MTs were observed to be long and flexuous and to run roughly parallel to the long axis of hyphae. No correlation between relative MT abundance and hyphal growth rate was observed, although non-growing hyphae had a lower relative MT abundance than growing hyphae. Actin depolymerization decreased hyphal growth rate while MT depolymerization did not. MT stabilization increased hyphal growth rate. Ethanol, the solvent in which the MT and actin inhibitors were dissolved, increased both average overall growth rate and growth rate variability for individual hyphae. Taxol appeared to interact with irradiation to decreased growth rate during imaging. <p>Golgi are involved in secretion and potentially in polar growth. An <i>A. nidulans</i> α-coatomer protein (COP)I homolog (α-COPI), tagged with GFP, was used to investigate the role(s) of fungal Golgi in polar growth. α-COPI-GFP co-localized with the known Golgi marker, α-2,6-sialyltransferase (ST), tagged with red fluorescent protein (RFP), in untreated hyphae. Based on this observation, I propose that α-COPI-GFP can be used as a proxy for fungal Golgi localization. Fungal Golgi were more abundant at hyphal tips than subapically. Fungal Golgi forward (tipward) velocity correlated with hyphal growth rate. Fungal Golgi forward velocity was, on average, approximately ten times greater than average hyphal growth rate. Actin depolymerization reduced fungal Golgi forward velocity while MT depolymerization did not. However, MT stabilization increased fungal Golgi forward velocity. <p>Polymerized MTs do not appear to be essential for hyphal growth but do appear to be involved in hyphal growth rate variability. MTs also appear to play some role in the movement of fungal Golgi. The distribution and movement of fungal Golgi is clearly related to polarity.
| Identifer | oai:union.ndltd.org:USASK/oai:usask.ca:etd-05042007-171757 |
| Date | 07 May 2007 |
| Creators | Hubbard, Michelle Anne |
| Contributors | Wang, Hong, Kaminskyj, Susan G. W., Hemmingsen, Sean M., Fowke, Larry C., Chivers, Douglas P. |
| Publisher | University of Saskatchewan |
| Source Sets | University of Saskatchewan Library |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://library.usask.ca/theses/available/etd-05042007-171757/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University of Saskatchewan or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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