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Invisibleshymanta
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Is this normal mycelium?
    #5640844 - 05/17/06 04:28 AM (17 years, 10 months ago)

Is this normal mycelium?

Multi-spore on PDA inoculated on 5/6. I don't know if it is a contam, but I've never seen mycelium look like this.

Thoughts?

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OfflineRogerRabbitM
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Re: Is this normal mycelium? [Re: shymanta]
    #5641123 - 05/17/06 08:14 AM (17 years, 10 months ago)

Looks like multispore inoculation on agar. You have a ton of 'strains' going there. Make up more dishes and begin transferring the best looking mycelium in very small pieces to new plates. Isolate for rhizomorphs.

The pictures below are from a strain isolation cycle. The first picture is from a small swipe of spores on each section of the plate, and the first batch of transfers away from the original colony ten days later. The second picture is from two or three days after the first, and shows the very first signs of rhizomorphic mycelium. They should be transferred as soon as noticed. Don't allow petri dishes to grow fully out. The third picture is after the second transfer and shows a fully isolated rhyzomorphic strain in the six o'clock position on the dish. The other two sections on the last dish still need another transfer. From starting the spores on agar until the last picture which is ready to inoculate grains, was just under three weeks. Speed is critical, so make transfers fast and get your final product ready as soon as possible.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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Invisibleshymanta
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Re: Is this normal mycelium? [Re: RogerRabbit]
    #5645199 - 05/18/06 04:18 AM (17 years, 9 months ago)

That's good. This dish came from a batch that contaminated 9 out of 12. I'll have to look over my notes and see what I did differently for that batch.

My normal procedure for isolation is to grow out a specimen and clone the best looking fruit. In your opinion, how does that compare to doing transfers on agar to isolate as it pertains time, quality, and health of the strain?

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OfflineRogerRabbitM
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Re: Is this normal mycelium? [Re: shymanta]
    #5645365 - 05/18/06 07:20 AM (17 years, 9 months ago)

By growing out a flush and cloning, you're still relying on the luck of the draw from multispore to get your strain. When you isolate on agar, you can get 20 or more good, rhizomorphic isolates from a single swipe of spores or two drops of solution from a syringe.

I grow out all isolates that look good, then pick the best ones and keep them. By labeling the petri dishes and keeping them in the refrigerator, I can go back to the original culture for years into the future once I see which fruits best or has other qualities I desire. If you clone, you already have one full generation towards senescence. By transferring the best strains from petri dishes to test tubes, they can be stashed away in the refrigerator for long term storage. It's possible to have 20 or more year old cultures with no more than a few weeks of cell division time on them.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Invisibleshymanta
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Re: Is this normal mycelium? [Re: RogerRabbit]
    #5649131 - 05/19/06 04:43 AM (17 years, 9 months ago)

Quote:

RogerRabbit said:
By growing out a flush and cloning, you're still relying on the luck of the draw from multispore to get your strain.




It seems to me that it would be the other way around. By cloning you could choose form the gene pool visibly expressed (fruited) and you would be guaranteed a fruiting strain. As I understand it, rhyzomatic mycelium should fruit, but I would be pissed to waist substrate on a strain the didn't fruit.

Would being able to see genetic character in addition to the time saved be altogether not worth it? ...verses making several transfers to isolate and having to test for fruitablity?

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OfflineRogerRabbitM
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Re: Is this normal mycelium? [Re: shymanta]
    #5649244 - 05/19/06 07:07 AM (17 years, 9 months ago)

Not really. It's not hard to fruit out ten to twenty small trays to find the best isolate. Once you do, that same isolate can produce awesome flushes of super potent mushrooms for the rest of your life. By going back to the master cultures in your refrigerator, you eliminate that first generation's worth of cell division and have a young culture to work with forever, if you're careful storing it.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Invisibleshymanta
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Re: Is this normal mycelium? [Re: RogerRabbit]
    #5652516 - 05/20/06 07:37 AM (17 years, 9 months ago)

What substrate do you use to test an isolate and what size trays? Just one more quick question, what about cultures that fruit in cold conditions? Is a refrigerator cold enough to stop their growth, too?

Thanks, RR and sorry this thread turned out not to be anything about contamination. lol. Perhaps a mod can put it somewhere more appropriate.

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OfflineRogerRabbitM
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Re: Is this normal mycelium? [Re: shymanta]
    #5652585 - 05/20/06 08:27 AM (17 years, 9 months ago)

Yes. The refrigerator is 35F to 45F which is cool enough to keep mycelium for the most part dormant. Even cold weather fruiting species grow out their mycelial network in the summer months when temperatures are much warmer.

I test strains by fruiting in a small tray spawned with one quart of rye and one quart of pasteurized manure. This results in very fast colonization so I can quickly determine the keepers from the tossers.
RR


--------------------
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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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