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Wng
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Registered: 02/16/18
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Last seen: 4 years, 13 days
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P. Mexicana var. Tampanensis Log
#25121839 - 04/07/18 08:43 PM (6 years, 9 months ago) |
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This is my first attempt at cultivating sclerotia. I ordered a tampanensis print and scraped some spores onto agar "slants" in babyfood jars.
The spores germinated alot faster than I expected, at first I thought I had contams. Here is 2 days after dropping spores on agar. You can just make out the black spore mass under the fluffy white strands

Day 4:

Day 6:

Day 7(Today): I noticed a little bit of a color change in some spots where the mycelium is densest. Almost a browning, is this a sign of contam?

Heres a shot of the agar being lit from behind:

I inoculated 8 jars of agar 1 week ago and am looking some help when I do my first transfers (petri dishes arrive in 2 days). Many of these slants are showing some darker spots. Ive never seen tampanensis mycelium before, it appears much thinner than cubensis mycelium. Is this normal?
If Im trying to isolate for a good stone producing strain, what characteristics should I be looking to isolate?






In the last photos, where the 3 jars "dripped" down to the bottom and germinated, can I isolate these parts too? or should I avoid it?
Thanks for looking
Update:
The 1st set of transfers are looking good, its been 9 days since these dishes were inoculated with mycelium from the original 8 jars.
I noticed some some browning on the surface of these transfers and noticed that the browning looked like a darker amber colored mass when I shined a light from under the dishes.
Heres a dish with some early brown coloration:

Here is the amber coloring:

I transferred a piece from the region circled in yellow in hopes that I'll get a decent stone producing sub-strain. Circled in black is where I can see some brown


Another plate with deeper amber/brown coloring:


Is this amber colored stuff indicative of sclerotia producing potential?
Edited by Wng (04/19/18 08:53 PM)
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Josex
#cheat_code


Registered: 11/13/15
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Re: P. Mexicana var. Tampanensis Log [Re: Wng]
#25121907 - 04/07/18 09:10 PM (6 years, 9 months ago) |
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The brownish spots are normal with tamps, they look fine. Looks like it's gonna be hell reaching down to take a wedge from those bottles, though.
Check this out.
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boom boom

Registered: 02/27/18
Posts: 429
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Re: P. Mexicana var. Tampanensis Log [Re: Josex]
#25122250 - 04/08/18 01:19 AM (6 years, 9 months ago) |
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Yeah, browning is normal. You actually want to isolate from brown sectors. The other thing to look for is small stones forming on the agar. If you see them developing after the plate has sat for a while, transfer them away to a new plate and that culture should produce really well. Both of these tips are per RR.
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Wng
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Registered: 02/16/18
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Re: P. Mexicana var. Tampanensis Log [Re: boom boom]
#25122329 - 04/08/18 02:59 AM (6 years, 9 months ago) |
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Cool, so you mean I should cut the wedges that have some brown on them? Would small stones appear only after the whole surface of the agar is colonized?
Circled in BLUE is the visible browning. Would I transfer only leading edges like RED? Or could I cut some of the interior, denser, browner stuff out too, as in YELLOW?
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boom boom

Registered: 02/27/18
Posts: 429
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Re: P. Mexicana var. Tampanensis Log [Re: Wng]
#25122338 - 04/08/18 03:19 AM (6 years, 9 months ago) |
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This post I'm much less sure about than the last one. One thing I can say with near certainty is that if you're getting stone production before the agar is fully colonized, then you have an incredibly good culture on your hands. But generally it will be fully colonized when you see small stones.
As far as taking the wedges, maybe let it grow out a bit more before deciding? Or you can take some transfers now, and let those transfered jars colonize for a bit. Generally with spores you want to transfer really soon after germination from what I've read. Then maybe something like if you're getting the most browning at 12-3 o'clock then take a transfer from the leading edge of 12-3 o'clock, even if the leading edge hasn't turned brown yet. That's probably what I would experiment with, but there's other posters who have already done the experimenting that can offer a more complete picture. There's a thread specifically for sclerotia producers here in mushroom cultivation where a lot of experienced stone growers hang out, if you don't get the answers you're looking for in this one.
https://www.shroomery.org/forums/showflat.php/Number/22527453
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Wng
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Re: P. Mexicana var. Tampanensis Log [Re: boom boom]
#25147022 - 04/17/18 11:34 PM (6 years, 8 months ago) |
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Its been 18 days since innoculating my 8 jar-slants with spores. 8 days ago, I took 4-6 tranfers from each jar into petri dishes.
I started noticing a heavy browning and dew developing in some of my jars. Like little water droplets.

Are droplets a sign of good stone producing genetics? Should I do a transfer from these spots with liquids? Are they just metabolites?
Thanks
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elasticaltiger
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Registered: 06/24/13
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Re: P. Mexicana var. Tampanensis Log [Re: Wng]
#25147153 - 04/18/18 01:22 AM (6 years, 8 months ago) |
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Looks like mold
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Haywire
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Re: P. Mexicana var. Tampanensis Log [Re: Wng]
#25147154 - 04/18/18 01:25 AM (6 years, 8 months ago) |
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If I were you I would make a few transfers, ensuring you have clean growth. then noc up some jars and wait. you can always clone a stone that has formed in your jar instead of spending ages on isolating from a spore print. use some coffee in your substrate and use a big jar or spawn bag to grow them in. they like a lot of growing surface. while you wait for the sclerotia, you can always keep working on your agar projects.
-------------------- Ciao mamma, guarda come mi diverto
My grows Outdoor patches
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Wng
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Re: P. Mexicana var. Tampanensis Log [Re: Haywire]
#25147192 - 04/18/18 02:02 AM (6 years, 8 months ago) |
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Roger that, I already made ~40 transfers from these jars. Could be mold, those darkspots and craters are where I cut the agar previously. Could be carbon deposits from my scalpel blade rubbing off. All the transferred petri-dishes appear clean after a week.
Heres a photo of one of the transfers:


What I was wondering about was the droplets of liquid near the middle. Ive seen some photos of sclerotia forming around these dew drops and thought they could be a good sign, or a bad one.
thanks
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Haywire
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Registered: 12/29/13
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Re: P. Mexicana var. Tampanensis Log [Re: Wng]
#25147235 - 04/18/18 03:00 AM (6 years, 8 months ago) |
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I have no input on that. I've had it happen to some plates of mine as well, but I've never had sclerotia forming on a plate. However, I have harvested big ass truffles from bags inoculated with those plates.
-------------------- Ciao mamma, guarda come mi diverto
My grows Outdoor patches
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