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OfflineAnnoA
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Culture storage technique
    #758830 - 07/19/02 11:32 AM (14 years, 4 months ago)

There is an interesting technique for long term storage of mushroom cultures in distilled, sterile water.

http://www.shroomery.org/index/par/23543

I stored several cultures this way 12 months ago.

2 days ago I recovered a few cultures by placing them on agar.

Several are already showing signs of recovery, but one really bursted into growth.
This is a P. ostreatus I got from Egghead.




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OfflineDinoMyc
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Re: Culture storage technique [Re: Anno]
    #759363 - 07/19/02 03:14 PM (14 years, 4 months ago)

nice
have you tried any other techniques with these samples to compare with?
thanks for letting everyone know of your success.


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Edited by DinoMyc (07/19/02 03:15 PM)


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OfflineSuntzu
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Re: Culture storage technique [Re: Anno]
    #761405 - 07/20/02 09:28 AM (14 years, 4 months ago)

Hey Anno, would you mind posting a list of the species that did/did not survive the dH2O storage?

I have had great success with cubes and pans, all the pleurotus species seem to do fine with it. Reishi, maitake, cordyceps as well. . .but there are a couple that don't seem to take to it. I lost my stropharia r.a., nearly lost my panellus. Might be cooincidence, but those are both fairly slow-growing, at least on the media I was using. It would be nice to see if some species are better stored on slants.


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OfflineStampede
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Re: Culture storage technique [Re: Suntzu]
    #761510 - 07/20/02 10:09 AM (14 years, 4 months ago)

Yes i read that on another board .. but i'am planning to put it to use have a limited selection of edibles i would like to keep around for a while .. Great Info .. and how many days is P oyster been on there?


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OfflineAnnoA
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Re: Culture storage technique [Re: Suntzu]
    #766053 - 07/21/02 10:40 PM (14 years, 4 months ago)

Day 5 after revival:

Growing very good:
P. ostreatus
P. djamor

Growing:
P. cornucopiae

Not growing:
H. erinaceus
P. nameko


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OfflineAnnoA
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Re: Culture storage technique [Re: Suntzu]
    #766071 - 07/21/02 10:46 PM (14 years, 4 months ago)

What would be important to add...
I stored the cultures in water 2 diferent ways:

1.) I cut small agar cubes out of the petri dish and transfered those cubes to the water slant.
2.) I scraped the mycelium of the perti dish surface and transfered the mycelium strands in the water slant..

This revival trial is with the small cubes, I?m yet to try this with the strands.


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OfflineDinoMyc
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Re: Culture storage technique [Re: Anno]
    #766521 - 07/22/02 06:46 AM (14 years, 4 months ago)

doesnt nameko have issues with other storage techniques?
perhaps I am confuseing it with another.


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OfflineAnnoA
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Re: Culture storage technique [Re: DinoMyc]
    #2320506 - 02/10/04 05:01 AM (12 years, 9 months ago)

I don't know, but I think it's time to place some cultures on agar again.


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Offlineragadinks
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Re: Culture storage technique [Re: Anno]
    #2320637 - 02/10/04 08:36 AM (12 years, 9 months ago)

When I look at the date of the postings it must have been quite some time since you have stored them in dH20 ?
I am curious about the results.
Maybe we could make a list for the FAQ how long which strain was stored in dH20 or other medium ?


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OfflineAnnoA
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Re: Culture storage technique [Re: Anno]
    #2971295 - 08/06/04 10:40 AM (12 years, 3 months ago)

What is other poeple's experience on this?


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InvisibleYidakiMan
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Re: Culture storage technique [Re: Anno]
    #2971556 - 08/06/04 12:13 PM (12 years, 3 months ago)

I put most of my cultures in dh2O several months ago. I didn't reanimate any of them, against my better judgement because they are all still alive on agar. But I did send some to ATWAR and she said that they have not reanimated yet. I know that I did pick the good ones that had visible fractured hyphae.

Before school starts I'm going to make more. Currently I only have them stored in one place... again, against my better judgement.


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Re: Culture storage technique [Re: Anno]
    #2971855 - 08/06/04 01:53 PM (12 years, 3 months ago)

The only and first time I revived some mycelium from the dH2o long-term storage was a shitake strain after 6 month after the mycelium on the agar plate in the fridge had died.
It worked very well ...


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Re: Culture storage technique [Re: ragadinks]
    #2974872 - 08/07/04 09:43 AM (12 years, 3 months ago)

I found following mail from Joe Kish here:

Quote:

To Rene, "Perf. Fungi E."
Your negative assumptions concerning the storage technique
article were made without even reading it. There lies the problem.
The statistics that you do not have available were contained in the
article's References.
One Reference that appeared in Applied Microbiology, Aug. 1974,
by M.R.McGinnis, A.A.Padhye, and L.Ajello described the tests
that were performed. "Four hundred and seventeen isolates of 147
species belonging to 66 genera of filamentous fungi, yeasts, and
aerobic actinomycetes were maintained in sterile distilled water at
room temperature over periods ranging from 12 to 60 months in
four independent experiments. Of the 417 cultures, 389 (93%)
survived storage." The References are obtainable from your
University library.
Joe Kish




He says that in a test 93% from 417 tested cultures survived in dH2O over a period from 12 to 60 months.
It's a pity that I cannot find the paper on the net, since it would be interesting which cultures ecactly have been tested.


Edited by ragadinks (08/07/04 10:13 AM)


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Re: Culture storage technique [Re: ragadinks]
    #2974885 - 08/07/04 09:56 AM (12 years, 3 months ago)

Huuh, I have found the article on the net here!

This book would also very interesting:

Quote:

Long-term storage of ectomycorrhizal basidiomycetes (Hebeloma spp.) at low temperature
M. Tibbett, F.E. Sanders, J.W.G. Cairney
Journal of Basic Microbiology
Volume 39, Issue 5-6, 1999. Pages 381-384
? 1999 WILEY-VCH Verlag Berlin GmbH, Fed. Rep. of Germany


Abstract

A method for maintaining viable cultures of ectomycorrhizal Hebeloma strains in cold liquid culture medium is described. Isolates of Hebeloma spp., collected over a wide geographic range, were stored at 2 ?C for a period of three years. All cultures survived this storage period, a greater time period and success rate than has previously been reported for the long term storage of ectomycorrhizal basidiomycetes. The method may prove useful for long-term storage of other basidiomycete genera.





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Re: Culture storage technique [Re: ragadinks]
    #2974894 - 08/07/04 10:12 AM (12 years, 3 months ago)

Here is another nice link where it says that the mycelium is grown out on water agar (which seems to have not nutrients?) and is then transferred to distilled H2O.
So no scraping mycelium of standard agar necessary anymore.

Here is the text:

Quote:

Hi Mike. We store our culture collection with water storage (but we also
have a set of everything in liquid nitrogen too!). The cultures are
actually grown on water agar (1.5% agar in water, no nutrients) and then we
cut out little plugs and drop them in sterile distilled water, so you don't
have to scrape off any mycelium. My former boss helped pioneer this
technique, and he estimates that you can store most cultures for 5 years.
We have something like 13,000 cultures of wood decay fungi that we are
storing this way with no trouble that I've heard about.

Jessie Micales, Project Leader
Center for Forest Mycology Research
Forest Products Laboratory
Madison, WI




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InvisibleYidakiMan
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Re: Culture storage technique [Re: ragadinks]
    #2975011 - 08/07/04 11:47 AM (12 years, 3 months ago)

Does the 1.5% agar solution solidify?


I might be able to talk to Tom Volk about this. He worked at Forest Products Lab for awhile before taking tenure at University of Wisconsin La Crosse.


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Re: Culture storage technique [Re: YidakiMan]
    #2975037 - 08/07/04 11:56 AM (12 years, 3 months ago)

I assume that the agar solidifies, but I am not really sure about that.

> I might be able to talk to Tom Volk about this. He worked at Forest Products Lab for awhile before taking tenure at University of Wisconsin La Crosse.
Would be really great if we could get some accurate info from someone who was involved in the experiment.


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Re: Culture storage technique [Re: ragadinks]
    #2975475 - 08/07/04 03:33 PM (12 years, 3 months ago)

dH2O is not a very good storage medium. Besides containing no nutrients the dH2O will suck out the nutrients from the mycelium.

The end result: Death by osmosis.

I suppose there are plenty of ways to get around this, but using non-distilled water would be the easiest.


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InvisibleYidakiMan
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Re: Culture storage technique [Re: fastfred]
    #2975604 - 08/07/04 04:10 PM (12 years, 3 months ago)

Quote:

fastfred said:
dH2O is not a very good storage medium. Besides containing no nutrients the dH2O will suck out the nutrients from the mycelium.
The end result: Death by osmosis.





Interesting assertion, but it's hard to argue with results.

Quote:

"Four hundred and seventeen isolates of 147
species belonging to 66 genera of filamentous fungi, yeasts, and
aerobic actinomycetes were maintained in sterile distilled water at
room temperature over periods ranging from 12 to 60 months in
four independent experiments. Of the 417 cultures, 389 (93%)
survived storage." The References are obtainable from your
University library.




Quote:

...he estimates that you can store most cultures for 5 years.
We have something like 13,000 cultures of wood decay fungi that we are
storing this way with no trouble that I've heard about.





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Re: Culture storage technique [Re: fastfred]
    #2975666 - 08/07/04 04:27 PM (12 years, 3 months ago)

fastred:
it seems that the results of some studies point out that distilled H20 is in fact a very good medium for long-term storage of mycelium since the living tissue falls dormant when there are no nutrients around.


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