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A more recent version of this article appeared on October 1, 2003
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Submitted on March 22, 2003
Revised on June 17, 2003
Accepted on June 17, 2003
1 Applied Microbiology, Biozentrum, Universitaet Basel, Klingelbergstrasse 70, 4056 Basel, Switzerland, Phone: +41-61-2671480 Fax: +41-61-2671481
* Corresponding author. E-mail address: peter.philippsen{at}unibas.ch.
* Corresponding author. E-mail address: peter.philippsen{at}unibas.ch.
* Corresponding author. E-mail address: peter.philippsen{at}unibas.ch.
We used actin staining and video microscopy to analyze the development from a spore to a young mycelium in the filamentous ascomycete Ashbya gossypii. The development starts with an initial isotropic growth phase followed by the emergence of germ tubes. The initial tip growth speed of 6-10 µm/h increases during early stages of development. This increase is transiently interrupted in response to the establishment of lateral branches or septa. The hyphal tip growth speed finally reaches a maximum of up to 200 µm/h and the tips of these mature hyphae have the ability to split into two equally fast growing hyphae. A search for A. gossypii homologues of polarisome components of the yeast S. cerevisiae revealed a remarkable size difference between Spa2p of both organisms, with AgSpa2p being double as long as ScSpa2p due to an extended internal domain. AgSpa2 colocalizes with sites of polarised actin. Using video time-lapse microscopy we show that AgSpa2p-GFP polarization is established at sites of branch initiation and then permanently maintained at hyphal tips. Polarization at sites of septation is transient. During apical branching the existing AgSpa2p-GFP polarization is symmetrically divided. To investigate the function of AgSpa2p we generated two AgSPA2 mutants, a partial deletion of the internal domain alone and a complete deletion. The mutations had an impact on the maximal hyphal tip growth speed, on the hyphal diameter and on the branching pattern. We suggest that AgSpa2p is required for the determination of the area of growth at the hyphal tip and that the extended internal domain plays an important role in this process.
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