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Here we go. Two spore germinating methods, looked pretty much the same under microscope. One was rub oat flake in spores, add to 1ml water and leave for three days in the dark. Method two - same but using a piece of filter paper pick up spores but still one oat in 1ml. It goes cloudy very quickly and bacteria are visible in about 12 hours, many empty spore cases by day 3 and and some flagellate cells.
I already know I can get plasmodia by leaving spores on damp paper with oats for several months but that's not fast enough so I'm looking for quicker methods. I think maybe limiting food might kick the haploid cells into linking up.
I'm trying strains A2 ( deeper colour) and JM (paler)
In keeping with low tech ideas I'm measuring culture in drops - the pasteur pipettes give me about 20microlitres per drop.
Paper discs - dampened with boiled tap water. One drop of culture on each four for each strain.
Agar - 4 well trays, three trays each of one, three and six drops of culture in 2cm dia wells. One One tray tray of of mixed culture 3+3 drops
Agar chunks in petri - one drop per lump, both strains.
Well I wasn't expecting anything at just six days but...
Sorry about image quality - phone held up to 'scope eyepiece, I need to find drivers for the USB microscope.
This is from A2 2.5cm whatman paper disc.
Next step is to get it off there and into its own disk for bulking up
That oat should be ready to move tomorrow then it's bulk up, make some scletrotia and then see what this thing can do.
This looks like it might be another one - the very young plasmodia lack pigment. This one ( if it is a slime ) is from JM which is pale to start with.
Our youngster grows slowly but is getting there - the only colony so far. I've been reading papers from the 60s and 70s in which amino benzoic acids were used as supplements to encourage plasmodia. I'm now trying mushroom extract - should be able to get this to a reliable 7 day turn around, in the mean time - raising spores
Further reading - way back to 1968 now, temperature appears to be critical no other sod's mentioned that in a paper. Pausing to build an incubator.... In the meantime, raising more spores...
I've been watching this and it looks great. I'm doing the opposite now. Trying to coax some fruiting bodies into producing sporangium. Nothing as exacting as your processes, but if I get the light levels right (finding the right spot in my apartment) I might get lucky.
I've noticed as well that the cloudy young plasmodium stage is very slow, but I'm alway amazed that when it goes full on diploid the growth is practically exponential. I keep my petri dishes in sealed plastic bags as Physarum is quite the escape artist.
It does seem to take several weeks for them to really get going. This week I tried streaking out spores directly onto agar - 1% oat agar and 1% boiled yeast, neither sterile.
First colony is now about 5cm across and is looking like it's about to hit exponential phase.
Nine new colonies on the top right dish ( the agar slices in one dish )!
If all of these come up positive I'll have an accommodation problem!
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