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Invisiblecosmicaug
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Mushroom presumptively identified as Macrocybe titans (Orlando area)
    #23559493 - 08/20/16 09:42 AM (7 years, 5 months ago)

Is there still any interest in this mushroom from people? I see that Saturdayraining, who started the thread, Macrocybe titans as a food crop, was last active here three years ago.

I recently had access to what I believe was Macrocybe titans. I'm at the Familab makerspace where someone brought in the mushroom yesterday and gave me access to it for a while. Though it's too early to tell, I might soon be able to produce living cultures. This was found in the Orlando area. Spore print is white.

I took a number of tissue samples which are sitting on plates of potato flake dextrose agar right now. I even tried to spread a few spores on one plate (let's see if it grows anything other than mold). If all that fails, I did freeze a small sample for the purpose of still having tissue available to me for doing PCR with ITS primers on it at some later date.

There were no signs of growth on the plate when I checked yesterday but it was only late Wednesday that I inoculated the plates so I think there's still a reasonable chance that I might get something (I think expecting clear growth in less than 48 hours might be unrealistic).



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Offlineanthiawe
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #23559596 - 08/20/16 10:22 AM (7 years, 5 months ago)

very interesting, and possible to cultivate. keep updates coming!


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Invisiblecosmicaug
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: anthiawe]
    #23563161 - 08/21/16 02:38 PM (7 years, 5 months ago)

I'm really glad I tried so many plates. One is clearly showing signs of life though it has not jumped to the agar surface yet.

There are a couple of bacterially contaminated plates (probably Bacillus subtilis type stuff). There are a couple of them with something fungal not originating on the mushroom piece which I suspect to be contaminants.

The spore plate shows nothing. I consider that good news. It means I probably did not pick up mold or bacteria. If there's anything living that has emerged from spores I probably wouldn't be seeing it yet.


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Invisiblecosmicaug
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #23623989 - 09/07/16 10:07 PM (7 years, 4 months ago)

So here's where I am at.

The spore plate contaminated and was discarded. It's something we've been having a lot but we just changed our setup (real HEPA filter from Fungi Perfecti) and I am hoping it will make a big difference.

Two weeks ago, the cloned plate that was showing growth also showed contamination and I subcultured it just in time.

One plate worked. That plate looked good on Sunday morning.

Yesterday, it was further spreading but some other fungus started coming in from the edge. I subcultured into six other plates and I'm hoping it will not have company in those.






In the pictures you can see the intruder from above in the upper right part of the plate. It is a little more evident in the photograph from below. I remain optimistic about getting good plates at the end of this.


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Invisiblecosmicaug
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #23635212 - 09/11/16 10:09 AM (7 years, 4 months ago)



The six plates I made on Tuesday look good so far.



The intent is to soon do PCR of ITS primers on both my frozen sample and also on some material from one of those plates so as to establish continuity.


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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #23635839 - 09/11/16 01:47 PM (7 years, 4 months ago)

Interesting!


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Invisiblecosmicaug
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #23738168 - 10/14/16 06:58 PM (7 years, 3 months ago)

I'm beginning to see difficulties with growing and maintaining this mushroom.

I attempted to start some millet spawn on the 3rd of October. I did break things up last Saturday. I really should have checked up on it again sooner because it has overcolonized and I think it might even be pinning which I did not want.



I also have found out that all of a sudden all of my Macrocybe titans Petri dishes look like crap. It's no longer contamination and it's probably not senescence. It's more like they have been triggered to fruit and are trying to pin.


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Invisiblecosmicaug
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #23743873 - 10/16/16 08:44 PM (7 years, 3 months ago)

Pictured here is a tub with half a brick of coco coir that has been soaked inoculated with half of the spawn bag. It's a very generous inoculation rate since I wanted the colonization to be very fast to avoid contamination issues. If I remember correctly, this inoculation happened Saturday morning. I will probably add more plant matter in a day or two (including the other half of the coco coir brick). For scale, the bottom of the tub is about 12"x19".




There's another attempt at a test grow at the lab in a pressure cooked oven bag (not pictured). There, late on Friday evening, I used the other half of the spawn to inoculate a whole coco coir brick since I was more confident of cleanliness. I suspect that even with that being half the inoculation rate that I used at home it would still would be considered a very generous inoculation rate.


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Invisiblecosmicaug
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #23837122 - 11/15/16 10:20 PM (7 years, 2 months ago)

Considering the possibility that Seaweed Solution Laboratories PDA agar mix might be crap and unsuitable for culturing mushrooms. Will probably test sometime against stuff that worked for me in the past.


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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #23837906 - 11/16/16 07:25 AM (7 years, 2 months ago)

Nice, good luck  :thumbup:


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Invisiblecosmicaug
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #23849520 - 11/19/16 06:21 PM (7 years, 2 months ago)

Quote:

cosmicaug said:
Considering the possibility that Seaweed Solution Laboratories PDA agar mix might be crap and unsuitable for culturing mushrooms. Will probably test sometime against stuff that worked for me in the past.



The test was simply to try to subculture a bunch of things with potato flake dextrose agar that has been shown to work in the past as well as with the Seaweed Solution Laboratories PDA. This was done Thursday so it's too early to tell but I am starting to see fuzziness on some plates. This is only happening on the potato flake agar plates and not in the Seaweed Solution Laboratories PDA.

It is my understanding that the Seaweed Solution Laboratories PDA is something that was ordered on Amazon.

We'll see what we have in a week or so.


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OfflineMattisfat
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #23850410 - 11/20/16 03:32 AM (7 years, 2 months ago)

:popcorn:


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Invisiblecosmicaug
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #23861512 - 11/23/16 06:09 PM (7 years, 2 months ago)

So enough time has elapsed to allow me to confidently state that the Seaweed Solution Laboratories PDA is not good. It may even be difficult to grow mold on it.

Honestly, if you had told me I would not have believed you (and I know there's at least one thread here on Shroomery about bad agar —now I feel bad for not taking such a thread seriously). I simply don't think of these types of organisms as being fastidious nor of PDA as something that is easily screwed up.


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Invisiblecosmicaug
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #23861754 - 11/23/16 07:34 PM (7 years, 2 months ago)

Oh, and for whatever it's worth, I now know that the Macrocybe titans cultures will survive 2 days at refrigerator temperatures. I know this because last Thursday I mixed up my plates and put some in the refrigerator without meaning to and they remain alive to date (I removed them from the refrigerator on Saturday).


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Offlinezilker
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: cosmicaug]
    #27968222 - 09/25/22 10:07 PM (1 year, 4 months ago)

I know this post is many years old but a it appears you're still frequent this site (doesn't show a "last seen" time stamp).

A friend of mine found what he believes to be a M. Titan and it's almost identical to the pics in your post.  I'm having a hard time getting the clone to grow out into a recognizable mycelium structure. I took 7 samples on 3 plates. I've been battling contamination but I've read elsewhere that this species doesn't do well with a sterile environment.

Long and short...did you have any success?  Do you have any pics of early mycelium growth that you can share?  Feel free to PM me if you'd like. Thanks.


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Invisiblecosmicaug
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Re: Mushroom presumptively identified as Macrocybe titans (Orlando area) [Re: zilker]
    #27968822 - 09/26/22 11:50 AM (1 year, 4 months ago)

Quote:

zilker said:
I know this post is many years old but a it appears you're still frequent this site (doesn't show a "last seen" time stamp).

A friend of mine found what he believes to be a M. Titan and it's almost identical to the pics in your post.  I'm having a hard time getting the clone to grow out into a recognizable mycelium structure. I took 7 samples on 3 plates. I've been battling contamination but I've read elsewhere that this species doesn't do well with a sterile environment.

Long and short...did you have any success?  Do you have any pics of early mycelium growth that you can share?  Feel free to PM me if you'd like. Thanks.



Hi.

Sorry, no updates on this. I have only my carelessness to blame. I think whatever visual records of plates I made may be here already (I really need to get a lot better at documenting things). The plates were eventually discarded by someone else as I left them unattended for a while (instead of maintaining discipline & subculturing). They probably would have been in bad shape (though it might have been nice to try to rescue).

If I remember correctly, I really had very little trouble getting something started. It was really just a matter of getting enough plates going and not using the bizarrely ineffective commercial PDA. The recipe I used as an alternative to the dodgy PDA was a variation of potato dextrose agar using commercial potato flakes (instead of boiled potatoes). I (or maybe some other source I may have read?) found it in an old journal article from some pathologist (I think it is this one). This can be bought commercially or made by you.

It's too bad that I did not do a better job. I kind of wonder if it might be possible to do a bulk outdoor grow Stropharia rugosoannulata style.

What's even worse is I had a second chance a year later & have nothing to show for it as I think the plate fates may have been the same as the first time. Sorry. I even hd the foresight to do a spore print on foil but I did not give it enough time to drop enough spores. When I tried to see if there was anything recoverable from the invisible spore print I got no growth.


Edited by cosmicaug (09/26/22 11:51 AM)


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