Supplemental Videos of “Phenotypic memory in Bacillus subtilis links dormancy entry and exit by a spore quantity-quality tradeoff”
10
2017
688
4 Minuten 58 Sekunden
10 Ergebnisse
00:32
54Mutlu, Alper et al.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.
2017Max-Planck-Institute for Terrestrial Microbiology et al.
00:24
41Mutlu, Alper et al.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.
2017Max-Planck-Institute for Terrestrial Microbiology et al.
00:22
44Mutlu, Alper et al.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.
2017Max-Planck-Institute for Terrestrial Microbiology et al.
00:33
77Mutlu, Alper et al.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.
2017Max-Planck-Institute for Terrestrial Microbiology et al.
00:34
48Mutlu, Alper et al.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.
2017Max-Planck-Institute for Terrestrial Microbiology et al.
00:03
81Mutlu, Alper et al.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.
2017Max-Planck-Institute for Terrestrial Microbiology et al.
00:43
68Mutlu, Alper et al.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.
2017Max-Planck-Institute for Terrestrial Microbiology et al.
00:33
169Mutlu, Alper et al.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.
2017Max-Planck-Institute for Terrestrial Microbiology et al.
00:27
48Mutlu, Alper et al.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.
2017Max-Planck-Institute for Terrestrial Microbiology et al.
00:43
58Mutlu, Alper et al.2017Max-Planck-Institute for Terrestrial Microbiology et al.