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Applied and Environmental Microbiology, November 2008, p. 6774-6781, Vol. 74, No. 21
0099-2240/08/$08.00+0     doi:10.1128/AEM.01233-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Time-Lapse Microscopy of Streptomyces coelicolor Growth and Sporulation {triangledown} ,{dagger}

Vinod Jyothikumar, Emma J. Tilley, Rashmi Wali, and Paul R. Herron*

Strathclyde Institute of Pharmacy and Biomedical Science, University of Strathclyde, Royal College, 204 George Street, Glasgow G1 1 XW, United Kingdom

Received 3 June 2008/ Accepted 25 August 2008

Bacteria from the genus Streptomyces are among the most complex of all prokaryotes; not only do they grow as a complex mycelium, they also differentiate to form aerial hyphae before developing further to form spore chains. This developmental heterogeneity of streptomycete microcolonies makes studying the dynamic processes that contribute to growth and development a challenging procedure. As a result, in order to study the mechanisms that underpin streptomycete growth, we have developed a system for studying hyphal extension, protein trafficking, and sporulation by time-lapse microscopy. Through the use of time-lapse microscopy we have demonstrated that Streptomyces coelicolor germ tubes undergo a temporary arrest in their growth when in close proximity to sibling extension sites. Following germination, in this system, hyphae extended at a rate of ~20 µm h–1, which was not significantly different from the rate at which the apical ring of the cytokinetic protein FtsZ progressed along extending hyphae through a spiraling movement. Although we were able to generate movies for streptomycete sporulation, we were unable to do so for either the erection of aerial hyphae or the early stages of sporulation. Despite this, it was possible to demonstrate an arrest of aerial hyphal development that we suggest is through the depolymerization of FtsZ-enhanced green fluorescent protein (GFP). Consequently, the imaging system reported here provides a system that allows the dynamic movement of GFP-tagged proteins involved in growth and development of S. coelicolor to be tracked and their role in cytokinesis to be characterized during the streptomycete life cycle.

* Corresponding author. Mailing address: Strathclyde Institute of Pharmacy and Biomedical Science, University of Strathclyde, Royal College, 204 George Street, Glasgow G1 1 XW, United Kingdom. Phone: 44-141-5482531. Fax:. 44-141-5484924. E-mail: paul.herron{at}strath.ac.uk

{triangledown} Published ahead of print on 12 September 2008.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, November 2008, p. 6774-6781, Vol. 74, No. 21
0099-2240/08/$08.00+0     doi:10.1128/AEM.01233-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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