scholarly journals Differentiation and Anaerobiosis in Standing Liquid Cultures of Streptomyces coelicolor

2003 ◽  
Vol 185 (4) ◽  
pp. 1455-1458 ◽  
Author(s):  
Geertje van Keulen ◽  
Henk M. Jonkers ◽  
Dennis Claessen ◽  
Lubbert Dijkhuizen ◽  
Han A. B. Wösten
Keyword(s):  
Metabolic Pathways ◽  
Minimal Medium ◽  
Streptomyces Coelicolor ◽  
Anaerobic Growth ◽  
Liquid Cultures ◽  
Air Interface ◽  
Solid Agar ◽  
Aerial Hyphae ◽  
Agar Media ◽  
Surface Liquid

ABSTRACT Streptomyces coelicolor differentiates on solid agar media by forming aerial hyphae that septate into spores. We here show that differentiation also occurs in standing liquid minimal media. After a period of submerged growth, hyphae migrate to the air interface, where they become fixed by a rigid reflecting film. Colonies that result from these hyphae form sporulating aerial hyphae. In addition, submerged hyphae in the liquid minimal medium may attach to the surface. Liquid standing cultures easily become anoxic only 1 to 2 mm below the surface. Yet, biomass increases, implying the existence of metabolic pathways supporting anaerobic growth.

scholarly journals Streptomyces coelicolor Genes ftsL and divIC Play a Role in Cell Division but Are Dispensable for Colony Formation

2007 ◽  
Vol 189 (24) ◽  
pp. 8982-8992 ◽  
Author(s):  
Jennifer A. Bennett ◽  
Rachel M. Aimino ◽  
Joseph R. McCormick
Keyword(s):  
Cell Division ◽  
Double Mutant ◽  
Minimal Medium ◽  
Streptomyces Coelicolor ◽  
Bacterial Cell Division ◽  
Transmission Electron ◽  
Aerial Hyphae ◽  
Mutant Strains ◽  
High Sucrose ◽  
Insertion Mutations

ABSTRACT We have characterized homologues of the bacterial cell division genes ftsL and divIC in the gram-positive mycelial bacterium Streptomyces coelicolor A3(2). We show by deletion-insertion mutations that ftsL and divIC are dispensable for growth and viability in S. coelicolor. When mutant strains were grown on a conventional rich medium (R2YE, containing high sucrose), inactivation of either ftsL or divIC resulted in the formation of aerial hyphae with partially constricted division sites but no clear separation of prespore compartments. Surprisingly, this phenotype was largely suppressed when strains were grown on minimal medium or sucrose-free R2YE, where division sites in many aerial hyphae had finished constricting and chains of spores were evident. Thus, osmolarity appears to affect the severity of the division defect. Furthermore, double mutant strains deleted for both ftsL and divIC are viable and have medium-dependent phenotypes similar to that of the single mutant strains, suggesting that functions performed by FtsL and DivIC are not absolutely required for septation during growth and sporulation. Alternatively, another division protein may partially compensate for the loss of both FtsL and DivIC on minimal medium or sucrose-free R2YE. Finally, based on transmission electron microscopy observations, we propose that FtsL and DivIC are involved in coordinating symmetrical annular ingrowth of the invaginating septum.

TEM processing procedure for a bacillus organism grown on solid media

1990 ◽  
Vol 48 (3) ◽  
pp. 624-625
Author(s):  
Jane Payne ◽  
Philip Coudron
Keyword(s):  
Electron Microscopy ◽  
Room Temperature ◽  
Fume Hood ◽  
Vacuum Aspiration ◽  
Solid Media ◽  
Transmission Electron ◽  
Solid Agar ◽  
Processing Procedure ◽  
Agar Media ◽  
Rat Bone

This transmission electron microscopy (TEM) procedure was designed to examine a gram positive spore-forming bacillus in colony on various solid agar media with minimal artifact. Cellular morphology and organization of colonies embedded in Poly/Bed 812 resin (P/B) were studied. It is a modification of procedures used for undecalcified rat bone and Stomatococcus mucilaginosus.Cultures were fixed and processed at room temperature (RT) under a fume hood. Solutions were added with a Pasteur pipet and removed by gentle vacuum aspiration. Other equipment used is shown in Figure 3. Cultures were fixed for 17-18 h in 10-20 ml of RT 2% phosphate buffered glutaraldehyde (422 mosm/KgH2O) within 5 m after removal from the incubator. After 3 (30 m) changes in 0.15 M phosphate buffer (PB = 209-213 mosm/KgH2O, pH 7.39-7.41), colony cut-outs (CCO) were made with a scalpel.

Effects of growth phase and the developmentally significant bldA-specified tRNA on the membrane-associated proteome of Streptomyces coelicolor

Microbiology ◽  
2005 ◽  
Vol 151 (8) ◽  
pp. 2707-2720 ◽  
Author(s):  
Dae-Wi Kim ◽  
Keith F. Chater ◽  
Kye-Joon Lee ◽  
Andy Hesketh
Keyword(s):  
Minimal Medium ◽  
Streptomyces Coelicolor ◽  
Regulatory Gene ◽  
Transmembrane Helix ◽  
Morphological Differentiation ◽  
Hypothetical Proteins ◽  
Membrane Proteome ◽  
Protein Mass ◽  
Associated Proteins ◽  
Transporter Systems

Previous proteomic analyses of Streptomyces coelicolor by two-dimensional electrophoresis and protein mass fingerprinting focused on extracts from total cellular material. Here, the membrane-associated proteome of cultures grown in a liquid minimal medium was partially characterized. The products of some 120 genes were characterized from the membrane fraction, with 70 predicted to possess at least one transmembrane helix. A notably high proportion of ABC transporter systems was represented; the specific types detected provided a snapshot of the nutritional requirements of the mycelium. The membrane-associated proteins did not change very much in abundance in different phases of growth in liquid minimal medium. Identification of gene products not expected to be present in membrane protein extracts led to a reconsideration of the genome annotation in two cases, and supplemented scarce information on 11 hypothetical/conserved hypothetical proteins of unknown function. The wild-type membrane proteome was compared with that of a bldA mutant lacking the only tRNA capable of efficient translation of the rare UUA (leucine) codon. Such mutants are unaffected in vegetative growth but are defective in many aspects of secondary metabolism and morphological differentiation. There were a few clear changes in the membrane proteome of the mutant. In particular, two hypothetical proteins (SCO4244 and SCO4252) were completely absent from the bldA mutant, and this was associated with the TTA-containing regulatory gene SCO4263. Evidence for the control of a cluster of function-unknown genes by the SCO4263 regulator revealed a new aspect of the pleiotropic bldA phenotype.

Aeromonas hydrophila in Shellfish Growing Waters: Incidence and Media Evaluation

1989 ◽  
Vol 52 (1) ◽  
pp. 7-12 ◽  
Author(s):  
CARLOS ABEYTA ◽  
STEPHEN D. WEAGANT ◽  
CHARLES A. KAYSNER ◽  
MARLEEN M. WEKELL ◽  
ROBERT F. STOTT ◽  
...  
Keyword(s):  
Aeromonas Hydrophila ◽  
Environmental Samples ◽  
Enterotoxin Production ◽  
Beef Extract ◽  
Solid Agar ◽  
Agar Media

Levels of Aeromonas hydrophila determined for the shellfish growing area of Grays Harbor, Washington, ranged from 3 to 4600/100 g in oysters and from 3 to 2400/100 ml in water. Of isolates tested, 80% produced a hemolysin, a trait reported to correlate with enterotoxin production and pathogenicity. Two enrichment broths, Tryptic Soy Broth with ampicillin (TSBA) and Modified Rimler Shotts Broth (MRSB) were compared in combination with three solid agar media: Rimler Shotts (RS), Peptone Beef Extract Glycogen (PBG), and MacConkey's (MCA) agars. TSBA was far superior to MRSB in isolating this species from the environmental samples tested.

Effect of ascorbate on oxygen uptake and growth of Escherichia coli B

1988 ◽  
Vol 34 (6) ◽  
pp. 822-824 ◽  
Author(s):  
Holly E. Richter ◽  
Jacek Switala ◽  
Peter C. Loewen
Keyword(s):  
Escherichia Coli ◽  
Oxygen Uptake ◽  
Electron Acceptor ◽  
Minimal Medium ◽  
Anaerobic Growth ◽  
Rich Medium ◽  
Terminal Electron Acceptor ◽  
Subsequent Change ◽  
Escherichia Coli B

The addition of ascorbate to aerobically growing cultures of Escherichia coli B caused only a short pause in growth and no subsequent change in the rate or extent of growth. The effect of ascorbate on oxygen uptake varied from inhibition in minimal medium to stimulation in rich medium. Cyanide-resistant growth and oxygen uptake were stimulated by ascorbate. Both the rate and extent of anaerobic growth were stimulated in proportion to the amount of ascorbate added when fumarate was the terminal electron acceptor. Ascorbate had no effect on any aspect of anaerobic growth in the absence of a terminal electron acceptor or in the presence of nitrate.

scholarly journals Dimerization of the RamC Morphogenetic Protein of Streptomyces coelicolor

2004 ◽  
Vol 186 (5) ◽  
pp. 1330-1336 ◽  
Author(s):  
Michael E. Hudson ◽  
Justin R. Nodwell
Keyword(s):  
Protein Kinase ◽  
Streptomyces Coelicolor ◽  
Kinase Domain ◽  
Terminal Protein ◽  
Amino Terminal ◽  
Morphogenetic Protein ◽  
Aerial Hyphae ◽  
Multiple Copies ◽  
Defective Allele

ABSTRACT RamC is required for the formation of spore-forming cells called aerial hyphae by the bacterium Streptomyces coelicolor. This protein is membrane associated and has an amino-terminal protein kinase-like domain, but little is known about its mechanism of action. In this study we found that the presence of multiple copies of a defective allele of ramC inhibits morphogenesis in S. coelicolor, consistent with either titration of a target or formation of inactive RamC multimers. We identified a domain in RamC that is C terminal to the putative kinase domain and forms a dimer with a Kd of ∼0.1 μM. These data suggest that RamC acts as a dimer in vivo.

scholarly journals Developmental-Stage-Specific Assembly of ParB Complexes in Streptomyces coelicolor Hyphae

2005 ◽  
Vol 187 (10) ◽  
pp. 3572-3580 ◽  
Author(s):  
Dagmara Jakimowicz ◽  
Bertolt Gust ◽  
Jolanta Zakrzewska-Czerwinska ◽  
Keith F. Chater
Keyword(s):  
Green Fluorescent Protein ◽  
Developmental Stage ◽  
Enhanced Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Streptomyces Coelicolor ◽  
Green Fluorescent Protein Fusion ◽  
Protein Fusion ◽  
Chromosome Partitioning ◽  
Aerial Hyphae ◽  
Green Fluorescent

ABSTRACT In Streptomyces coelicolor ParB is required for accurate chromosome partitioning during sporulation. Using a functional ParB-enhanced green fluorescent protein fusion, we observed bright tip-associated foci and other weaker, irregular foci in S. coelicolor vegetative hyphae. In contrast, in aerial hyphae regularly spaced bright foci accompanied sporulation-associated chromosome condensation and septation.

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