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OfflineWorkmanV
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Isolation of mycelium from dried mushrooms - Fruiting update
    #3762775 - 02/10/05 04:10 PM (19 years, 3 months ago)



Too bad there is so much condensation on these plates, but you can still get the idea.

Update 2/18/05

Well, that nice white mycelium isn't mushroom mycelium after all. A quick look under the scope revealed these odd looking structures.



They are the conidia of a species of Pestalotia, a genus of fungi that often causes disease in plants.

Pestalotiopsis link for more information.

It is a bummer and a bit embarrassing but such is the nature of science. I will redo the experiment with agar dosed with benlate fungicide to inhibit sporulating molds and allow the mushroom spores to germinate. Back to the drawing board.

Edited by Workman (05/02/05 03:33 PM)

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OfflineKevin
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #3762907 - 02/10/05 04:36 PM (19 years, 3 months ago)

Nice man.  :thumbup:


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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #3763421 - 02/10/05 06:06 PM (19 years, 3 months ago)

Since a gill fragment was taken, couldn't this be the germination of spores from a dried specimen versus the isolation of viable mycelium from a dried specimen?

Regardless though, this is good for people to know.

Joshua


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OfflineWorkmanV
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Joshua]
    #3763907 - 02/10/05 07:23 PM (19 years, 3 months ago)

Hmmm, yeah I can see how that is a bit confusing. I guess I should have said, "isolation of viable mycelium from spores attached to the gills of a dried mushroom".

Thanks for pointing that out.


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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #3764060 - 02/10/05 07:51 PM (19 years, 3 months ago)

Awesome picture workman.

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InvisibleYidakiMan
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: dmc_]
    #3764370 - 02/10/05 09:56 PM (19 years, 3 months ago)

Very cool. I am going to use this to get a culture of V. volvacea.


Did you use any antibiotic or hydrogen peroxide?

Edited by YidakiMan (02/10/05 09:56 PM)

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OfflineFluxburn
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: YidakiMan]
    #3764820 - 02/11/05 12:01 AM (19 years, 3 months ago)

wow workman I wish I knew similar to you so I could be your assistant, to bad for me.


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OfflineWorkmanV
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: YidakiMan]
    #3766614 - 02/11/05 12:56 PM (19 years, 3 months ago)

Oh yes, I used Gentamycin sulfate antibiotic in the agar. Sorry, I should have mentioned that before. I posted that image pretty late in the evening and it seems I left a lot of information out.


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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #3768116 - 02/11/05 06:54 PM (19 years, 3 months ago)

I am always glad to see your work!

Joshua


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OfflineWorkmanV
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Re: Isolation of mycelium from dried mushrooms - Contamination update [Re: Workman]
    #3806580 - 02/20/05 12:15 PM (19 years, 2 months ago)

Oops, edited the top post to reveal a tactical error.


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Re: Isolation of mycelium from dried mushrooms - Contamination update [Re: Workman]
    #3807219 - 02/20/05 03:21 PM (19 years, 2 months ago)



those are funky-looking thingies, workman, if unwanted. the antennae-thingies make them look like they're insects  :rolleyes:


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Offlinedebianlinux
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Joshua]
    #3827423 - 02/24/05 12:59 PM (19 years, 2 months ago)

Quote:

Joshua said:
I am always glad to see your work!

Joshua




agreed, i always ogle in amazement

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OfflineRogerRabbitM
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: debianlinux]
    #3837982 - 02/26/05 04:40 PM (19 years, 2 months ago)

Any updates?
When I first did this, I only used dry tissue from the stipe, to avoid getting spores. I wanted to make sure it was the mycelium recovering, not spores germinating.

Those are some cool looking cells, even if a contaminant. May I ask what type camera you're using?

The antenna looking things are known as cilia. They are used to create movement of the fluids around the cell.


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OfflineWorkmanV
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: RogerRabbit]
    #3842777 - 02/27/05 06:04 PM (19 years, 2 months ago)

2 days ago I put the gill tissue from several specimens on malt agar with benlate and gentamycin. Nothing to report yet. But I will update this post when something happens.

The camera is an old Sony Mavica FD87.


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OfflineMycelium5150
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #3843362 - 02/27/05 07:48 PM (19 years, 2 months ago)

you still have that mavica! Isnt that the one that copies pics to floppies


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OfflineWorkmanV
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Mycelium5150]
    #3846830 - 02/28/05 01:12 PM (19 years, 2 months ago)

Yeah, the FD stands for Floppy disk. I have a Kodak DX4330 as well but the Kodak is just very handy to knock out web quality pics.


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OfflineWorkmanV
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #3892742 - 03/09/05 12:58 PM (19 years, 2 months ago)

The benlate (10mg/liter) gentamycin sulfate (100mg/liter) attempt worked great. No signs of molds or bacteria at all. A vast difference compared to the earlier attempt with only gentamycin.



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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #3892792 - 03/09/05 01:11 PM (19 years, 2 months ago)

Are there any domestic stores that sell benlate? Feed stores?

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OfflineCyber
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: YidakiMan]
    #3892816 - 03/09/05 01:18 PM (19 years, 2 months ago)

I looked up benlate and found all kinds of stuff, Pulled from the market in 2001 because of birth defects! Kids born with out eyes, missing limbs, etc. Where did you get it?

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OfflineAnnoA
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Cyber]
    #3892821 - 03/09/05 01:19 PM (19 years, 2 months ago)

Before 2001 in the store I guess.

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Offlineshirley knott
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Anno]
    #3892827 - 03/09/05 01:20 PM (19 years, 2 months ago)

lol. i'm sure it is still manufactured, or a related chemical is, if it proved useful (as clearly it does)


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OfflineRogerRabbitM
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: shirley knott]
    #3894872 - 03/09/05 08:20 PM (19 years, 2 months ago)

From looking at that last pic, It looks like you rolled the dry tissue into the agar, thus several points of growth emerging. They appear to be growing together seamlessly, as opposed to forming sectors which is what one would expect if it were spores germinating. It looks to me like the dry tissue has begun to grow again. Is that your take as well?


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OfflineWorkmanV
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: RogerRabbit]
    #3897957 - 03/10/05 01:38 PM (19 years, 2 months ago)

I did streak dried gill tissue over the surface using sterile forceps to spread out the spores and then leave the tissue on the agar at the end of the streak. Typically the tissue mass contaminates but spores left by the streaking usually germinate cleanly. I doubt there is anything but spores producing mycelium but there is no way of knowing. There are several mycelium types growing.

I did get a 10 pound bag of benlate as a lawn fungicide just before it was pulled from the market. It should last awhile using only 10mg amounts at a time.


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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #3903241 - 03/11/05 01:50 PM (19 years, 2 months ago)

If you think there might be a contamination issue. Simply pour another layer of hot agar on top, completely covering the growth you have going. A whole new layer. The mycelium will crawl through to the top of the new layer of agar and will leave behind the contaminates. Then lightly scrape some of the new growth and transfer to a whole new dish. That should clean up your culture.

Edited by hotnutz (03/11/05 01:52 PM)

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Offlinered914
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Hotnuts]
    #3926536 - 03/16/05 04:24 PM (19 years, 2 months ago)

Hotnutz..
Just how hot would the agar be? sounds like a great plan.

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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #3928811 - 03/17/05 12:40 AM (19 years, 2 months ago)

Quote:

Workman said:
I did streak dried gill tissue over the surface using sterile forceps to spread out the spores and then leave the tissue on the agar at the end of the streak.  Typically the tissue mass contaminates but spores left by the streaking usually germinate cleanly.  I doubt there is anything but spores producing mycelium but there is no way of knowing.  There are several mycelium types growing.




But a clean culture COULD be cultured out of this...........right???. (You would know!!)

I think its time for me to get back on the agar and get some experiments running.

There HAS to be a way to go from dried specimen to clean culture, no matter the contaminates.  :thumbup:

Pure cultures have been cultured from wild prints which have been exposed to any and everything outside, yet are still cultured out. I've done it myself.

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OfflineWorkmanV
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: MrMaddHatter]
    #3930088 - 03/17/05 11:06 AM (19 years, 2 months ago)

Right, any obvious mycelium can be isolated away from contaminates. Sometimes it takes more than one attempt.


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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #3935090 - 03/18/05 10:25 AM (19 years, 2 months ago)

I tried putting questionable spores in agar, and covering it with two sheets of boiled newspaper after cooling. The mycelium grew out to the outside layer. Peal the outer layer off to a fresh dish of agar. It appears clean. It would appear that contams do not like to be covered. I have also moved to the wide mouth jam jars to get away from the agar quickly drying out as the newspaper wicks off water fast. I tried spores alone in newspaper, but it works for oysters but not cubes, I guess they need more nutes.

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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #4123798 - 05/02/05 02:55 PM (19 years, 21 days ago)

Results of attempted fruiting of above plate on grass seed cased with peat/calcium carbonate.

Day 11 after casing, incubated at 78F




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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #4123890 - 05/02/05 03:21 PM (19 years, 21 days ago)

Nice!!! :bow:


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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #4128899 - 05/03/05 05:35 PM (19 years, 20 days ago)

:thumbup: Very nice work!

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OfflineWorkmanV
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #4133174 - 05/04/05 02:45 PM (19 years, 19 days ago)

Its coming along.



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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: Workman]
    #4133438 - 05/04/05 03:41 PM (19 years, 19 days ago)

its nice to know that you are working to bring new strains that no one else has to the market


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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: tahoe]
    #4163283 - 05/11/05 06:48 PM (19 years, 12 days ago)

why do you use grass seeed?


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OfflineWorkmanV
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: xburn]
    #4163630 - 05/11/05 08:01 PM (19 years, 12 days ago)

Ah, thats a good question. I use it because it seems to work well with a wide range of species. It is my first choice for specimens with unknown growth parameters.


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Offlinexburn
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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: xburn]
    #4165753 - 05/12/05 09:07 AM (19 years, 11 days ago)

oh i see, workman i do wonder how would i get into doing the work that you do. I am facsinated nore by the mycology than the drugs and stuff.


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Re: Isolation of viable mycelium from dried mushrooms - Pictorial [Re: xburn]
    #4172955 - 05/13/05 10:14 PM (19 years, 9 days ago)

This is awesome workman. Great work, you obviously know your stuff :smile: I'm very impressed and interested in this. I look forward to seeing what they turn out like.

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Re: Isolation of mycelium from dried mushrooms - Fruiting update [Re: Workman]
    #27530262 - 11/04/21 02:55 PM (2 years, 6 months ago)

amazing, this has me hopeful

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