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00:43 BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology Silent film 2017

The down- and up-shift responses on an SM* gel pad of the fluorescent marker strain BIB1126, which carries a PrapA-mCherry promoter fusion that reports on sporulation timing.

A few cells that initiated sporulation early were tracked with a yellow arrow, while some representative cells that delayed sporulation were followed with a green arrow. The latter strongly induce mCherry expression. After sporulation is complete, the marker nicely distinguishes between early and late spores. Spore revival was then induced by the addition of L-alanine. All spores that grew out in response to induction with L-alanine are circled. They all show low fluorescence.
  • Published: 2017
  • Publisher: BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology
  • Language: Silent film
00:33 BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology Silent film 2017

Heterochronic sporulation response of B. subtilis on an SM* gel pad and the subsequent spore revival upon addition of L-alanine.

The strain BIB1019 carries PtrpE*-mCherry and PspoIIE-gfp reporters. After 90 h of starvation, spores have been released from the mother cell. To visualize the distribution of early and late spores prior to the nutrient up-shift, false colored frames were included with early and late spores denoted in yellow and green, respectively. Note that most spores geminated but only early spores succeed in resuming vegetative growth. Spores that grew out upon nutrient stimulation are circled in red.
  • Published: 2017
  • Publisher: BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology
  • Language: Silent film
00:33 BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology Silent film 2017

C-terminal mCherry tagged Ald expression from its native promoter

The down- and up-shift responses of strain BIB1423, which expresses fluorescently tagged Ald-mCherry from its native promoter (Pald-ald-mCherry). At the onset of sporulation, cells that sporulate early have higher fluorescence levels than cells that delay sporulation due to dilution. The fluorescence is carried from the progenitors into the developing spore. Upon spore release, fluorescence drops due to unknown reasons and then remains stable. Early spores show higher fluorescence than late spores. All spores that grew out in response to L-alanine stimulation were circled. They all show higher levels of fluorescence compared to non-outgrowing spores.
  • Published: 2017
  • Publisher: BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology
  • Language: Silent film
00:32 BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology Silent film 2017

Effect of KinA induction on sporulation and spore revival of the Pald Ald-mCherry reporter strain

Left: KinA was induced at t=0 resulting in accelerated sporulation. All KinA-induced spores are highly fluorescent and most are capable of outgrowth in response to L-alanine. Right: The induction of KinA was delayed (t=20h) and induced in the progenitor cells of late spores. While the highly fluorescent early spores grew out none of the resulting lowly fluorescent, late but KinA-induced spores grew out. This shows that KinA induction does not alter the spore properties per se.
  • Published: 2017
  • Publisher: BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology
  • Language: Silent film
00:24 BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology Silent film 2017

Effect of decelerating sporulation via RapA overexpression

Down- and up-shift responses of a mutant colony of strain BIB1330 (Phyperspank-rapA) and a WT colony BIB1126 (PrapA-mCherry). Both strains were starved in co-culture on a SM* gel pad that was supplemented with IPTG to induce RapA in the mutant strain and decelerate its sporulation.
  • Published: 2017
  • Publisher: BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology
  • Language: Silent film
00:03 BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology Silent film 2017

Up-shift response of spores of the fluorescent marker strain PrapA-mCherry upon exposure to L-alanine.

Spores were generated by the Sterlini-Mandelstam protocol in liquid shake-flask culture, and show a broadly heterogenous distribution of fluorescence. Spores were then spotted onto an SM gel pad and spore revival was induced by adding L-alanine.
  • Published: 2017
  • Publisher: BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology
  • Language: Silent film
00:34 BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology Silent film 2017

Effect of Ald re-programming on B. subtilis spore revival

Down- and up-shift responses of strain BIB1300, which harbors a PrapA-mCherry promoter fusion that reports on sporulation timing and in which Ald expression can be controlled by an IPTG-inducible promoter. Cells were spotted onto an SM* gel pad. At the indicated time IPTG was added to the pad. This results in the expression of Ald in the progenitor cells of late spores. After sporulation was complete, spore revival was induced with L-alanine. The spores that will grow out are circled. Note that red-fluorescent late spores are able to grow out successfully.
  • Published: 2017
  • Publisher: BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology
  • Language: Silent film
00:27 BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology Silent film 2017

Effect of an ald gene knock-out on spore revival

Down- and up-shift responses of a mutant colony of strain BIB1416 (Δald , PtrpE-mCherry) and a non-fluorescent WT colony BIB224. Both strains were starved in co-culture on an SM* gel pad and show comparable sporulation dynamics. After sporulation is complete, spore revival was induced with L-alanine. Both mutant and WT spores show comparable germination. However, only WT spores grew out while the mutant spores did not.
  • Published: 2017
  • Publisher: BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology
  • Language: Silent film
00:22 BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology Silent film 2017

Effect of accelerating sporulation via overexpression of KinA

Down- and up-shift responses of a mutant colony of strain BIB1332 (Pspank-kinA) and a WT colony BIB1126 (PrapA-mCherry). Both strains were starved in co-culture on a SM* gel pad, which was supplemented with IPTG to induce KinA in the mutant strain and accelerate its sporulation. After sporulation was complete, spore revival was induced with L-alanine.
  • Published: 2017
  • Publisher: BioQuant Center of the University of Heidelberg (BioQuant), Max-Planck-Institute for Terrestrial Microbiology
  • Language: Silent film
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