Collaborative project anyone?

[ian]

I have a cunning plan for a project. 

Go outside right now and grab the first dog you find, ignore the screaming owner and go get another dog. Have a good look at them. Chances are they look very different - but hey, they're the same species - that's domestication for you. Many of those traits - the floppy ears and relatively short muzzle compared to the wolf aren't things we selected for - they came along for the ride.

Let's domesticate physarum! Surely it's domesticated? I hear you say ( yes, I have very sharp hearing - and good imagination). But no - we usually propagate physarum by asexual means - cloning basically, so we don't see any variation and there are few strains in circulation. Our slime moulds need to get it on..

I've dabbled and I got some notable changes - a white physarum ( didn't live long), a very dark orange and a much paler yellow. We could build on that.. 

Here's the idea - select one for stronger colouring, one for paler and see what comes along with those traits - we make spores, grow into new slimes and choose the best. Physarum has the useful trait of being basically immortal so it's easy to keep ancestors going for back breeding (or even gene sequencing if someone gives us a shovel full of cash).

Breeding is easy - add spores to water and oats, leave for a bit, pour onto non sterile agar and wait a few weeks - low effort and colour is easy to select for, this really is a low bar for commitment - and it might just produce something publishable. I'm raising initial spores at the moment 

Who's with me? 

[Benny Fulmer]

as soon as mine arrives i would love to join you on this! i may also receive assistance from the southwest usa group in receiving another sclerotia for (hopefully) more genetic variation. this might also be a good incentive for people to go out and find their own specimens to bring into the fold...liven up the gene pool a little

[ian]

Cool, we'll need spores for this - I have some drying at the moment, they take a few weeks of sitting around dry before they'll hatch. Do you have access to agar? My current method is squash sporocarp in 1ml water, add one oat flake, leave dark for two weeks and spot out onto agar - colonies appear over the next 7 - 20 days. You can do it on paper but colonies take longer to form.

The idea would be to post out spores and sclerotia of A2 ( the very orange) and jm ( the pale) and start by selfing - raise from a single parent sporocarp then select from the next generations.

Crossing can be done properly - if you can be bothered to sit with a microscope and pipette collecting single spores ( takes ages and has a low success rate so only did jm) but I figured for a cross we just mix two batches of spores and hope for the best. As long as we raise and save sclerotia we can always unravel the genetic mess later.

Still bashing out details - any ideas are most welcome 

[Evan Brown]

I'm working on raising one of the sclerotia I got from you up using the concentric rings of oats method on agar agar. I'm going to let that one form sporangia and start from there.  

[ian]

Cool, it's easier to deal with sporanigia if you get the slime off the food when it's big enough. They wander around a lot when they're starving and exposed to light. For my small scale goes I've used clear topped honey tubs - nice and escape proof. For the big trays, I have no idea to stop escapes - I'm trying wiping the rim of the tray with detergent.

I left some in a clear sealed bag once it made spores - that might be a possible route for larger scales - harvesting would just be crush the bag around a bit and sieve.

They have a dormancy period of a few weeks btw.

[Evan Brown]

I didn't know about the dormancy period, thanks for the info! I'm not sure if it's just my search engine, but it seems there's not a ton of information about raising plasmodium from spores. When I obtain the spores how would I go about using them?  Let them sit around for a couple weeks on a shelf and then just add to water with bacteria? do they need a dry area to form a plasmodium or will that emerge from liquid in a tube?  I was planning on using something similar to a honey tub to obtain sporangia and probably just a test tube to propagate the spores.

[Eleanor Relf]

Are you hoping to become the keeper of the Physarum strain library?  I read they have been used for research on cell cycle genetics so presumably that was with colour markers or something? Also, presumably there could be funding available for gene sequencing if they are useful for such things and it means not reinventing the wheel everytime someone wants to do any research.  I would love to join in with this, but I sense I would need a microscope and cash for plates, agar etc, which I don't have at the moment, so I watch with interest, but I might go and forage for some wild slimes in a shoddy unscientific manner and keep this in mind if anything interesting happens. :) 

[ian]

Hi

I'm hoping to stimulate some innovation and interest in treating these like any other species we bring into cultivation. I'd like to curate and store cultures, when fun things start cropping up I'd have some weight to put behind apply for sequencing cash.

It's also about setting up zero tech methods - these things manage it in the wild all by themselves, the image of a sterile white lab with containment cabinets and incubators puts people off and isn't needed for physarum.

I'm raising amoeba by dipping an oat flake in spores then adding it to water - the amoeba feed off the bacteria on the decaying oat. Making plasmodia is a question of dotting out onto agar or damp paper and waiting, overcrowding and starvation push them into cell fusion. A microscope helps in spotting the young slimes but it's not essential and nothing needs to be sterile. As the agar's non nutrient, vegan 'gelatin' is all you need.  

[Eleanor Relf]

But if you introduce two strains of P. polycephalum plasmodia, will they fuse?  And then what would happen to the expression of your colour genes? If this isn't known, then it's best to keep them separate, no? Especially as you could get the same 'look' via different gene combinations...  Hence the need for some procedures to prevent cross-contamination of strains.  Having seen how eager they are to explore (I'm guessing even more eager at the amoeboid stage) I can't see a way to do that without sterile lab conditions.  Do they revert to amoebae from plasmodia or do they always complete the cycle through sporangia etc?

[Eleanor Relf]

Oh. I've just noticed on another post - John Holden says Physarum polycephalum isn't native in the wild in the UK. So I can't just go and find one to be useful to this project. I'd have to obtain one from someone (Ian ;) ) ... Is that correct?

[Evan Brown]

I'm told you can still find them in the wild, but generally yes. Ian got me started and he's been an absolute wealth of information.

[ian]

I've seen references to polycephalum in the wild in the uk but given that there impossible to id with iud spores I think any id has viewed with a caveat