Keeping the Culture Young!

[omsource]

Hey Crew,

Just starting this post up so we can share ideas on keeping our precious cultures young and vigorous.

Most experienced cultivators have techniques to keep their mushroom strains young and vigorous in the

petri dish state.

This are quite relevant techniques, as if the petri dishes are subcultured again and again and again making the cultures "older" they face a possibility of degeneration. *it's similar to a copy of a copy of a copy, the copies get worse and worse as you go along.

Degeneration presents itself in many ways, from inactive mycelium, fluffy mycelium instead of ryzomorphic or "strandy", mutations in the mushroom fruitbodies (usually negative yet sometimes beneficial) or even the lack of ability to fruit in general.

If anyone has any teks on keeping the culture young lets share em!

Cheers,

OmsOurce

[Amazonian]

G'day Omsource. I thought keeping several prints of an original strain and then going back to use spores every now and then would keep a strain fairly 'young' for a while ? Or people could swap Mycelium of the same strain and mix it with their old strain?

Edited October 20, 2010 by Amazonian

[watertrade]

keeping cultures young and vigorous is boring, It is the only drawback of having a culture collection! you have to keep them alive.

this is the best way i have found and have only lost one culture.

I don't really have a technique but what I have always done. have master cultures stored which I go back to to make fresh plates so rather than go from plate1 to plate2 to plate3 to plate 4

its more like storage 1 to plate 2, storage 1 to plate 2 etc. this keeps your cultures genetically young.

how do I store them?

3 ways-

Agar chunks in distilled water in technoplast tubes x 3

Cardboard pellets in technoplast tubes x 3 +

agar slants

every couple of years you can make new masters but I find they last quite a while, some are 5 years and still viable and vigorous.

of course not all strains cope with storage like this - most of the edibles do but some don't really like it. I store the tubes in a cupboard ( no space in the fridge! )

and most important of all! give cultures away to people so if you lose it your friends can give it back. In the last few years there has been a huge increase in the number of cultures available in Australia, only 5 or so years ago it was hard to get a handful of the common edibles. I would guess there would be 35ish individual species doing the rounds at the moment.

[Hyphal]

and most important of all! give cultures away to people so if you lose it your friends can give it back. In the last few years there has been a huge increase in the number of cultures available in Australia, only 5 or so years ago it was hard to get a handful of the common edibles. I would guess there would be 35ish individual species doing the rounds at the moment.

 

Thats great advice..

I have just found out that possibly my whole spore collection which was stored in a storage unit, in a small wooden box, double bagged may be un-viable. So far 4 different individual prints have been useless.

I have used a perfectly healthy print after 3 years storage in a mook so I wasn't expecting this - it feels like I have I have done them a dis-service...

So I'd like to add, that you should keep your prints in the fridge.

[omsource]

awesome ideas,

I've heard about keeping a master culture to whip up

some petri dishes that are then used to inoculate spawn as you mentioned watertrade.

the only confusion that I have around that concept is that

the master culture would be used up quite quickly especially

if you are pumping out the grain spawn in a commercial grow operation.

Would you simply let your agar chunks regrow before storing them,

or only take a small amount of mycelium to transfer?

Amazonian your technique is definitly valid, but if you store

prints instead of the actual mycelium then upon spore germination

and hybridization you would end up with a different strain.

That doesnt mean that the new strain would be worse, but it's definitely

possible.

One fairly experienced mycologist recommends isolating a single spore and

germinating into a petri dish culture, then matching it up with another

compatible spore- petri dish.

when you want to make up a dish for spawn production simply mate the two spore

cultures in a new dish and away you go.

Your cultures would be only 1 transfer old every time!

Whatdoyathink?

maybe a bit more effort than it's worth

but the culture library is invaluable!

[Amazonian]

if you store

prints instead of the actual mycelium then upon spore germination

and hybridization you would end up with a different strain.

 

I thought it would be like a pod of seeds, they are all going to produce the same tree? I am still learning. .

[omsource]

I thought it would be like a pod of seeds, they are all going to produce the same tree? I am still learning. .

 

Yup, I'm with ya there!

[Hyphal]

Yup, I'm with ya there!

 

You will still grow the same species, but you will just need to isolate to get a good fruiting culture.

[MagikF]

Yups, it's will definitely be a different mix of genetics if you use multispore prints instead of an isolation from a petri dish. Think of it as test tube babies where a bunch of eggs are mixed with a bunch of sperms. What you get would be fraternal twins instead of identical whereas a petri dish with made from a clone would product identical twins as they are split from the same genetic material.

[Amazonian]

You will still grow the same species, but you will just need to isolate to get a good fruiting culture.

 

Of course, but i didnt think there would be variation. Lets say you had a good fruiting strong strain from a particular print. If you took prints from that batch of mushrooms, wouldnt you just get the same good fruits over and over again or are we talking 'clone like' cultures .

[MagikF]

If you take have a good fruiting clone and then take prints from that, you would still have genetic variation as the spores are a mix of genetics. You would need to either need to use the exact same piece of myc from the isolate on agar, or you would need to clone from the fruit body but once you use spores, it's all a crap shoot as to what kind of genetics you will get.

[Hyphal]

If you take have a good fruiting clone and then take prints from that, you would still have genetic variation as the spores are a mix of genetics. You would need to either need to use the exact same piece of myc from the isolate on agar, or you would need to clone from the fruit body but once you use spores, it's all a crap shoot as to what kind of genetics you will get.

 

Exactly...

[omsource]

If you take have a good fruiting clone and then take prints from that, you would still have genetic variation as the spores are a mix of genetics. You would need to either need to use the exact same piece of myc from the isolate on agar, or you would need to clone from the fruit body but once you use spores, it's all a crap shoot as to what kind of genetics you will get.

 

Good point!

Here's the big question tho,

If you have an old culture with the possibility of degeneration coming on,

and you fruit it, then take a tissue sample from the mushrooms...is this new culture a fresh "1st plate" culture with strong vigourous "young" cells.....or will the tissue sample still be considered "old" with the possibility of degeneration or senecense?

It all comes down to cell division in the end, so I'd put my money on the fresh tissue culture still being considered old, but I could definitely be wrong.

[Hyphal]

Bloody good question, I wandered that myself. I agree that it will most probably still be considered 'old' as well...

[punkin]

Considering that the fruit is just another piece of mycelium, i can't see how there could be any difference between the tired myc in the plate and the slightly more tired myc in the fruit.

[omsource]

Considering that the fruit is just another piece of mycelium, i can't see how there could be any difference between the tired myc in the plate and the slightly more tired myc in the fruit.

 

Damn! Well I guess there's no easy out in this little predicament other than ongoing maintenance of the master cultures!

[MagikF]

The tissue sample you take from the fruit will be considered a part of the master culture as it is the exact same genetically as the one on the plate. The only way it would be considered a level removed or a generation below would be if you did another isolation off one of the better fruiting bodies with spores to further isolate better genetics. As to degradation or senecense, that might occur as with an living organism that survives longer. You can probably keep a master culture for many years in optimum condition. As to preserving the line, you can always keep picking the fruit(s) with the trait you most want and further isolate. Might take 3 or 4 generations but you'll have what you want.

[yaguareté]

keeping cultures young and vigorous is boring, It is the only drawback of having a culture collection! you have to keep them alive.

this is the best way i have found and have only lost one culture.

I don't really have a technique but what I have always done. have master cultures stored which I go back to to make fresh plates so rather than go from plate1 to plate2 to plate3 to plate 4

its more like storage 1 to plate 2, storage 1 to plate 2 etc. this keeps your cultures genetically young.

how do I store them?

3 ways-

Agar chunks in distilled water in technoplast tubes x 3

Cardboard pellets in technoplast tubes x 3 +

agar slants

every couple of years you can make new masters but I find they last quite a while, some are 5 years and still viable and vigorous.

of course not all strains cope with storage like this - most of the edibles do but some don't really like it. I store the tubes in a cupboard ( no space in the fridge! )

and most important of all! give cultures away to people so if you lose it your friends can give it back. In the last few years there has been a huge increase in the number of cultures available in Australia, only 5 or so years ago it was hard to get a handful of the common edibles. I would guess there would be 35ish individual species doing the rounds at the moment.

 

Just wondering what a technoplast tube is?

Is it a screw lid test tube?

thanks

[themushroombloke]

I'm going to say that I first hand have learnt the answer to this question as i was given one petri dish of p.pulmonorious and I put it into three grain jars. That I then turned into 10 from each jar (30) when I tried expanding on the second generation the spawn seemed to slow down. I had too much spawn to put into substrates so I made some outdoor patches. But still I had spawn, So once that slow to colonise spawn was fully colonised I made up some bags those have only just fruited and the yield was woeful and it was slow to colonise the sugar cane bagess. Then I have cloned a fruiting body from this slow to colonise bags and it took a whil to spring to life on the agar but it worked, so I transfered to grain and the lazy bastard has only jumped onto one or two grains in like 5 days. I have also left some grain spawn purposely to die in a jar to see what it does... well mine went fluffy (apparently this happens when spawn gets old). When I took the old/dead spawn out of the jar it had a horrible smell it had a yellow tinge and when I broke up the grain it was chalky, not that super white fluffy, you get when you bash your grains.

So I can conclusively say: Look after your culture library otherwise it's like having sex with your cousin!! lots of retarded mushrooms.

cheers

themushroombloke

[Zen Peddler]

Always clone and keep them in the fridge. There are some cultures doing the rounds that are still performing great after maybe 10 years,

[tripsis]

I'm with WT, share what you have and it will be the safest way to preserve the culture. This is especially true in Australia as getting cultures here is difficult and comes with inherent risks.

Beyond that, actually storing the cultures for prolonged periods of time can be done in a number of ways. I use all three methods that WT mentioned, but would like to look into storage in liquid paraffin. Cultures stored in this way can last 15 years or more at room temperature. Of course, not all cultures can be preserved over a long period of time. For example, the Agarikon (Laricifomes officinalis), degenerates in culture after a while, losing its original genetic integrity. Perhaps with species like this, log cultures are a better bet, an idea from gecko on AE.

One method I've not yet tried, but sound promised, is storage of dried mycelium. This can be either by storing mycelium of dried agar, or dried grains, sawdust, etc. From what I've read and heard, it works and might do so indefinitely. It may not work with all species, but it has potential.

Edited November 8, 2010 by tripsis

[Zen Peddler]

Psilocybe azurescens obviously doesnt degenerate given the fact that cultures have been around in OZ since 01 are still performing as well if not better given recent posts on this forum.

[Hyphal]

Psilocybe azurescens obviously doesnt degenerate given the fact that cultures have been around in OZ since 01 are still performing as well if not better given recent posts on this forum.

 

That doesn't mean that those cultures are ten years old though? A fresh print can be taken at any time and used to grow a fresh culture...

[shruman]

I know it performs well 'naturalized', no idea bout on agar though, I'm sure it would still be kicking around in the bottom of someones fridge. Plates would be cool to see.

Do'nt people say naturalized leaps off much better onto substrates than spawn from a culture.

Which begs the question?, What happens when you have an outdoor patch, it fruits drops spores on the patch spores germinate & mycelium grows?, does it knit up become part ot the patch & add new genetics to the patch or what exactly?

Senescence, might aswell spell it correctly for searchability

Yeah I dunno seems only to be a problem for you sterile nuts...

Mushrooms apear to enjoy the rigours of life maybe they feel like little prisoners in their sterile cells.

Enoki have been propogated asexualy for how long?

We too should work as a mycelial network, share what ya got.

[Zen Peddler]

I doubt anyone that has grown azurescens in australia has done it from print.

Edited December 12, 2010 by Zen Peddler BlueGreenie