Applying dimorphic yeasts as model organisms to study mycelial growth: Part 1. Experimental investigation of the spatio-temporal development of filamentous yeast colonies

ABSTRACTThe bacterial cell wall maintains cell shape and protects against bursting by turgor. A major constituent of the cell wall is peptidoglycan (PG), which is continuously modified to enable cell growth and differentiation through the concerted activity of biosynthetic and hydrolytic enzymes. Streptomycetes are Gram-positive bacteria with a complex multicellular life style alternating between mycelial growth and the formation of reproductive spores. This involves cell wall remodeling at apical sites of the hyphae during cell elongation and autolytic degradation of the vegetative mycelium during the onset of development and antibiotic production. Here, we show that there are distinct differences in the cross-linking and maturation of the PGs between exponentially growing vegetative hyphae and the aerial hyphae that undergo sporulation. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis identified over 80 different muropeptides, revealing that major PG hydrolysis takes place over the course of mycelial growth. Half of the dimers lacked one of the disaccharide units in transition-phase cells, most likely due to autolytic activity. The deacetylation of MurNAc to MurN was particularly pronounced in spores and strongly reduced in sporulation mutants with a deletion ofbldDorwhiG, suggesting that MurN is developmentally regulated. Altogether, our work highlights the dynamic and growth phase-dependent changes in the composition of the PG inStreptomyces.IMPORTANCEStreptomycetes are bacteria with a complex lifestyle and are model organisms for bacterial multicellularity. From a single spore, a large multigenomic multicellular mycelium is formed, which differentiates to form spores. Programmed cell death is an important event during the onset of morphological differentiation. In this work, we provide new insights into the changes in the peptidoglycan composition and over time, highlighting changes over the course of development and between growing mycelia and spores. This revealed dynamic changes in the peptidoglycan when the mycelia aged, with extensive peptidoglycan hydrolysis and, in particular, an increase in the proportion of 3-3 cross-links. Additionally, we identified a muropeptide that accumulates predominantly in the spores and may provide clues toward spore development.

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Spatio-Temporal Development of Supercavitation Over an Impulsively Launched Projectile

Volume 2: Fora ◽

10.1115/fedsm2006-98115 ◽

2006 ◽

Author(s):

C. J. Weiland ◽

P. P. Vlachos

Keyword(s):

High Speed ◽

Temporal Development ◽

Two Phase ◽

Time Resolved ◽

Gas Gun ◽

Supercavitating Flow ◽

Image Velocimetry ◽

Spatio Temporal ◽

Asymmetric Vortices ◽

First Time

Supercavitation inception and formation was studied over blunt projectiles. The projectiles were fired using a gas gun method. In this method, projectiles are launched under the action of expanding detonation gases. Both qualitative and quantitative optical flow diagnostics using high speed digital imaging were used to analyze the spatio-temporal development of the supercavitating flow. For the first time, quantification of the supercavitation was achieved using Time Resolved Digital Particle Image Velocimetry (TRDPIV) detailing the two phase flow field surrounding the translating projectiles and the gas vapor bubble. Experimental results indicate that the supercavity forms at the aft end of the projectile and travels forward along the direction of projectile travel. The impulsive start of the projectile generates two asymmetric vortices which are shed from the blunt nose of the projectile. The vortices interact with the moving cavity and subsequently deform. This interaction is believed to directly contribute to the instabilities in the flight path.

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