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OfflineThe shroomy 1
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Petri Dish Question (with pics)
    #7420047 - 09/17/07 03:02 PM (16 years, 5 months ago)

I'm trying to isolate the "Best Myc" to transfer to another dish. My problem is that my 4 dishes haven't produced any Rhizomorphic mycelia. The strain is F+. I'm quickly running out of room on the petri dish! The dishes were inoculated with a drop from a multi-spore syringe. Any ideas? What part do I transfer from the dishes?


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OfflineThe shroomy 1
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Re: Petri Dish Question (with pics) [Re: The shroomy 1]
    #7420246 - 09/17/07 03:48 PM (16 years, 5 months ago)

One Hour later....... still no response:foreheadslap:


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InvisibleJrsxt
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Re: Petri Dish Question (with pics) [Re: The shroomy 1]
    #7420252 - 09/17/07 03:51 PM (16 years, 5 months ago)

Hrmm. You could let it go a little longer, or you can just select the best looking portion.


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OfflineThe shroomy 1
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Re: Petri Dish Question (with pics) [Re: The shroomy 1]
    #7420686 - 09/17/07 05:44 PM (16 years, 5 months ago)

Only one Idea?


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InvisibleRoadkillM
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Re: Petri Dish Question (with pics) [Re: The shroomy 1]
    #7420743 - 09/17/07 05:59 PM (16 years, 5 months ago)

your gonna have to do a transfer and hope that you get some more Rhizomorphic growth on your next set of plates.

what kind of agar are you using?


tc


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OfflineThe shroomy 1
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Re: Petri Dish Question (with pics) [Re: Roadkill]
    #7420752 - 09/17/07 06:01 PM (16 years, 5 months ago)

Potato Dextrose. Do I cut from the heavy center or from the thin outer edges?


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OfflineWronguy

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Re: Petri Dish Question (with pics) [Re: The shroomy 1]
    #7420782 - 09/17/07 06:09 PM (16 years, 5 months ago)

You have some sectors that are very tomentose. If you indeed have a culture of mycelia there, you're best chance to isolate any sectors is always going to be on the tips of the growth. If you take a chunk from the center you run a higher chance of grabbing more sectors.

If you have a magnifying glass that would help you identify any sectors that aren't visible with the naked eye.


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OfflineThe shroomy 1
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Re: Petri Dish Question (with pics) [Re: Wronguy]
    #7420792 - 09/17/07 06:12 PM (16 years, 5 months ago)

Quote:

Wronguy said:
You have some sectors that are very tomentose. If you indeed have a culture of mycelia there,




Are you saying that this could be contamination?


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OfflineWronguy

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Re: Petri Dish Question (with pics) [Re: The shroomy 1]
    #7420835 - 09/17/07 06:22 PM (16 years, 5 months ago)

There is always that possibility. Can you give us a time frame from inoculation to those photos? I've personally never worked with the F+ strain, so there is a possibility the mycelia pictured is normal, but I doubt it.

The biggest issue I see here is that tomentose mycelia are not as likely to produce primordia as rhizomorphic are. I would sure hate to see you spend a lot of time and effort isolating a strain that produces little to no fruits.


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OfflineThe shroomy 1
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Re: Petri Dish Question (with pics) [Re: Wronguy]
    #7420894 - 09/17/07 06:37 PM (16 years, 5 months ago)

Inoculation was last Monday.  I did it during Monday night football:grin:  (Those pics were taken earlier today)


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Edited by The shroomy 1 (09/17/07 06:39 PM)


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OfflineWronguy

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Re: Petri Dish Question (with pics) [Re: The shroomy 1]
    #7421011 - 09/17/07 07:12 PM (16 years, 5 months ago)

That is a lot of growth for only 7 days; especially with spores. You have 3-5 days just for spore germination alone, let alone full growth. The structure to that mycelium also looks suspect.

My best guess is you appear to have the contaminate Mycelia Sterilia (White Mold). This contaminate has the appearance of whitish mycelium and is cottony. Unless someone else has another opinion, I'm betting you have a contaminate there.

I hope I'm wrong.


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OfflineThe shroomy 1
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Re: Petri Dish Question (with pics) [Re: The shroomy 1]
    #7421061 - 09/17/07 07:28 PM (16 years, 5 months ago)

Well, I will say this, I inoculated 8 pint jars of WBS with the same syringe. It took about 5 days for the jars to get at 25% colonization, I shook the shit out of them and they recovered extremely well! This is a pic of one of the jars. (I just took this pic about 10 min ago).


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OfflineThe shroomy 1
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Re: Petri Dish Question (with pics) [Re: The shroomy 1]
    #7421067 - 09/17/07 07:30 PM (16 years, 5 months ago)

These syringes are from Ralphsters.  I use a homemade flowhood.  I have never had a contaminated syringe from Ralph.:rockon:


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OfflineWronguy

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Re: Petri Dish Question (with pics) [Re: The shroomy 1]
    #7421470 - 09/17/07 09:08 PM (16 years, 5 months ago)

Your grains look good in the pic, but is the growth rhizomorphic (strandy) or tomentose (cottony)? There is documented correlation between cottony mycelia and poor fruiting, if it fruits at all. If the syringe you've inoculated your grains with is the same as the PDA dish, you're really going to have to wait it out to see what happens.

Based on your plate though, I don't see a very good strain there. The only thing you can really do is make transfers from the outer edges of the plate onto new plates and see if you get any appreciable rhizomorphic sectoring. Once you've made your transfer and can see sectoring, transfer it again, and so on.

Strain isolation is such a pain in the ass! You really could spend multiple hours working with a single set of dikaryons only to find that they do not produce mushroom fruits. You may get lucky and hit the nail on the head first go around.

Bottom line is you have to try before anyone, myself included, can make a good call on what you have. I hope this helps you out a bit.


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OfflineThe shroomy 1
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Re: Petri Dish Question (with pics) [Re: Wronguy]
    #7421563 - 09/17/07 09:39 PM (16 years, 5 months ago)

It has. Thank you so much. The jars do have some Rhizo growth, but it's mostly Tomentose.


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Invisibledumbfounded1600
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Re: Petri Dish Question (with pics) [Re: The shroomy 1]
    #7421925 - 09/17/07 11:10 PM (16 years, 5 months ago)

I wouldn't even fuck with it...I just threw out a jar because the mycelium that was forming at the top PF Tek was fuzzy and the strains of rhizomorph was very light colored and not as white as I think is white....Its like the mold that comes from WBS when you don't sterilize it....like a white cob web mold....If it doesnt have that smell of fresh mushrooms I'd toss it and start over

then again wtf do I know I havent even cultivated a single mushroom yet lol


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OfflineThe shroomy 1
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Re: Petri Dish Question (with pics) [Re: dumbfounded1600]
    #7422078 - 09/18/07 12:07 AM (16 years, 5 months ago)

Quote:

dumbfounded1600 said:
I wouldn't even fuck with it...I just threw out a jar because the mycelium that was forming at the top PF Tek was fuzzy and the strains of rhizomorph was very light colored and not as white as I think is white....Its like the mold that comes from WBS when you don't sterilize it....like a white cob web mold....If it doesnt have that smell of fresh mushrooms I'd toss it and start over

then again wtf do I know I havent even cultivated a single mushroom yet lol


LOL! Thanks.. I think I'll see this one through. I'll update in a few days.


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Offlinexaxphaanes
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Re: Petri Dish Question (with pics) [Re: The shroomy 1]
    #7422258 - 09/18/07 02:02 AM (16 years, 5 months ago)

For some reason since it is all cottony it looks like mold hope not though.


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OfflineRogerRabbitM
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Re: Petri Dish Question (with pics) [Re: xaxphaanes]
    #7422590 - 09/18/07 07:10 AM (16 years, 5 months ago)

The reason you don't see rhizomorphs early on is because there's so many substrains active, they cover each other up. It usually takes two or three transfers to begin to see rhizomorphic growth.

I'd transfer a tiny piece from each sector to new dishes. I usually make the first transfers as soon as the spores begin to germinate, when the total growth is the size of a dime or less. Repeat the process a few times. Don't try to isolate 'one' rhizomorphic growth. Instead, isolate as many as you can, and then fruit each one. Once you determine the best isolate, you can store it in master culture slants to use for years to come without degradation.
RR


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