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Re: Morels [Re: trscstghst]
    #8273272 -

I have to agree with Hot Nuts. Doing a clone in this manner is just asking for failure. Besides why would you go all out to do such a thing, never knowing if your contaminated or not. Approaching cloning in this manner is risky. Using agar provides you with assurity and also gives you the opportunity to create good sterile backups. The single most important part of the procedure! I am not going to say you cannot manage it however, just why even attempt it. There is no reason I can see. You cloned it to keep something.

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Re: Morels [Re: trscstghst]
    #8274434 -

I've cloned some pretty crappy looking mushrooms from grocery stores like this too, wet, slimy, old.
Yes the rate of failure will be higher with a wild mushroom but it is a good alternate to someone not set up for agar work.
It would be especially difficult with wild morels since the flesh is soo thin.
Waterlogged you wouldn't have a chance.
Kinda a moot point anyway unless you want to seed outdoor beds.
I don't think hardly anyone has grown these inside.
I bow to your wisdom Hotnuts, good points you raised.
:bow:
:jester:


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Re: Morels [Re: Peterthinks]
    #8279400 -

Peterthinks said:
A contamed LC looks like chunky crap.
not really that hard to see if it's bad.
floating islands of green....blobs of brown...anything but nice round jelly blobs like see thru cotton balls in water is probably bad.
bad just LOOKS bad.



Sadly, this isn't the only case and (after personal failures) is why I don't inoculate dishes/LCs with spores or wild tissue

Quote:
Agar work is in a dish with gas exchange, LC is in a sealed jar with a filter for gas exchange.
Agar cultures will crawl out of a dish and contaminate themselves. LC will run out of sugar and go to sleep.
:jester:


That's true, with fast-growing cultures they can grow out the sides. In my opinion, though, if the plate gets to that point it is too old anyway. I don't know how the LC works but there is a point where hyphal density severely effects the vigor of mycelium on any other substrate.

And for gas exchange not being filtered, the standard is to wrap the dish with parafin film.

LCs are fantastic for increasing colonization and needles make inoculation convenient. However, to me, specific cultures (I clone from the wild) are too valuable to risk losing just to save a step. The context here is morels, so we've been talking about wild clones anyway :laugh:


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Re: Morels [Re: Hotnuts]
    #8293988 -

Lazlo you responded to me a while back about my way of getting the spores. You described doing it on agar but yes that way is a 50 / 50 shot,
my way increases the odds up to 80 or 90 % success with no contaminates.
It reduces the trouble of sorting out hyphae and mycelia, and seriously who wants to deal with that sort of trouble. Mathematically the odds of getting a contaminate in the spores that are ejected upward is lower much much lower I know from experience. So if you can engineer a way to get them to eject their spores sooner or on demand would greatly improve my way and save the specimens from an early transition to goo. They'd still be edible then. That's the only drawback to my way. I think it may have something to do with low vapor pressure in the air but then again from what I observed was they were in a saran wrapped berry container and sunlight did something to trigger the spore dispersal. Oh btw I figured out how to get big orange sclerotia. :wink:
And yes they do coalesce and even if they do turn a little orangish the sclerotia can have a regrowth of thick white tiny hyphae on the same sclerotia. They start out as granules and more and more until they amount to a large mass. Sometimes they start out as a large area of white compact mycelium that eventually turns yellowish then to orangish and so does the long hyphae growing down into the nutrient rich layer.
Some of the aerial hyphae gets what I would call knots like knots in your hair and it looks clumpy in a sort of way, this is sclerotia as well, there is a reason why the sclerotia love the side of the glass jar and its a good reason.

This all happened when I knocked up 2 jars on Wednesday of the last week of March 2008, prep'd the Ower way; bottom layer nutrient rich and the top layer nutrient poor.
Its just some of you guys are allowing the fungus to eat and consume
and thats all it does (if you allow it), you need to figure out how to control what it eats and how much.
Believe me nutrient poor isn't totally 100% nutrient poor but
it certainly isn't even 10% nutrient rich. You'll have to give it a faint trace amount of nutrients on top of the nutrient poor layer.
I see most of you growing it on corn kernels, grains, peat moss, dark peat moss top soil and like me even mulch. And yet these are still nutrient rich.

It takes 2 weeks for even the smallest granules of sclerotia to form at 64.4 °F (yeah yeah I know you read "The Farm" and their crap and even that female mycologist's copy of the same parameters and that is just it,
they are feeding you what they want you to believe; hey gives them more time to be potential capitalists) but from spore to sclerotia can and does happen in 2 weeks tops. And it depends on the depth of your nutrient poor layer, it should be at least I believe 3.5 cm or 1.5 inch in depth, so finally ask yourself is that nutrient poor layer truly and truly nutrient poor? Btw make sure you moisten it before you pressure cook it, actually there needs to be a sufficient level of moisture the fungus will soak it up, especially in the 3 inch nutrient rich layer.

co·a·lesce  (k-ls)
intr.v. co·a·lesced, co·a·lesc·ing, co·a·lesc·es
1. To grow together; fuse.
2. To come together so as to form one whole; unite: The rebel units coalesced into one army to fight the invaders. See Synonyms at mix.
[Latin coalscere : co-, co- + alscere, to grow, inchoative of alere, to nourish; see al-2 in Indo-European roots.]
coa·lescence n.
coa·lescent adj.


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If you wanna ride, don't ride the white horse, if you wanna ride, ride the white pony

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Re: Morels [Re: AcidHorse]
    #8294053 -

Btw I found 2 handfuls of southwestern Ohio black morels and even the pokeweed hasn't started growing. Saw Dutchman's Breeches and Trillium plants. Hmm and there was a slight cold snap, lows in the 30s. My first time finding black sponges, in the past I've only found snakeheads and yellows/tans. Its funny though snakeheads should be up right at this time.
hmmmmm peculiar. Found a black growing in the dense roots of the Dutchman's too. Which makes me think morels aren't symbiotic but are facultatively parasitic. Its possible their degree and level of parasiticness is low or weak so some plants aren't phased by them. I've never seen morels benefit a plant in any way. Also I tried seedlings along with a jar of completely colonized morel mycelium and the damn stuff killed the seedlings! Turned the tips of the roots a dark brown and stems of the seedlings became weak and top heavy. Whoa. so are they parasitic? I wonder. And another thing one guy was asking about haploid and whatnot, well if you get spores into a substrate; That will be a multispore
situation, maybe some inbred toothless hillbillies it will begat, but if you can do it singly with a micro-pipette be my guest. One of those will run you at least 300$ to 3000+ $$$$ especially if you get the digital ones.
Man, today I feel like a ninja.


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If you wanna ride, don't ride the white horse, if you wanna ride, ride the white pony

Edited by AcidHorse (04/16/08 11:04 PM)

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Re: Morels [Re: AcidHorse]
    #8294149 -

Its funny I have success with morel development yet I can't get my shiitake or king oysters to grow. Wouldn't that be funny I get morels to grow but can't get shiitake or oysters to grow.

--to the doctorate graduate mycologist--

And the answer to your assumption is no, I know about shiitake primordia, yeah its funny they look ironically like morel sclerotia, well at least with this variation of shiitake. The golden ones. The whole jar is a rusty reddish color of discolored mycelium and hyphae, with some whitish growths that will tan . unlike morel mycelia and hyphae which there are some clear differences.
And absolutely everything is super sterile, flame sterilized, matching vapor pressure with the total sea-level atmospheric pressure and beyond, so it gets cooked. And bleach is my best friend too.
So it seems that this shiitake may indeed be a warm weather species or variation. Need the 80s.
Oysters is another mystery, spores sometimes other times no spores, spores germinate in the container, sample the spores and get freakin trichoderma.
Time for Rubik's Cube!


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If you wanna ride, don't ride the white horse, if you wanna ride, ride the white pony

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Re: Morels [Re: AcidHorse]
    #8297330 -

you should buy a camera id like to see what work you have done :thumbup:


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We play on earth to pass the time :foreheadslap:

Over-population the root of all Evil-brings the Elites Closer to the gates.

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Re: Morels [Re: makaveli8x8]
    #8300820 -

So, how about somebody posting their own work on morels here with pictures to back up some of this back-and-forth arguing about the "better" method.

I'm a firm believer in agar first, LC second (if at all- not suitable for all people).

I'm looking at using agar for cloning morels from a patch that is still producing a couple. It would be nice to see some well-documented write-ups.

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Re: Morels [Re: MycoAu]
    #8300895 -

For indoor cultivation or "patent replication research", spend the extra effort in finding Morel rufobrunnea. I'm still looking for this, but I have a spy in San Francisco on the lookout. For outdoor cult, stick with whats local. If morels don't grow locally, go back to step 1.

It's really easy to scrape tissue away from the pits of a dried morel. It's very easy to isolate away from contaminants. In my experience, its likely that the morel fungus will overgrow the plate before contaminants will germinate underneath the morel hyphae. Darkness is very important, in my experience it seems to limit the number of phenotypes to two or three.

For expansion, I like to do it the same way I do Hericium. Two plates to 600ml of water in the blender, poured 20ml at a time into master quart jars. Each quart goes into a large spawn bag that is layered bottom half topsoil top half sawdust. I can't really tell you about sclerotia because I haven't checked yet. They were made around Jan 1st, intended for planting in late spring or fall. Oh yea, the species is M. deliciosa.

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Re: Morels [Re: UrbanFungi]
    #8301214 -

I have some samples of Morel rufobrunnea but this species reportedly doesn't even form sclerotia and that was my experience as well. I thought I was doing something wrong but this seems to be just the nature of this species.


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Re: Morels [Re: Workman]
    #8308605 -

That is interesting. Maybe Kuo is correct that Mills worked with M. rufobrunnea. It makes a ton of sense considering San Diego is supposedly the place he honed his skill where he "was able to grow morels on his kitchen windowsill". I can't cite that but I read it in one of Mill's many newspaper bios available online.

That would mean the entire chapter in the patent about sclerotia formation is the misinformation or counterintelligence designed to throw people off.

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Re: Morels [Re: Peterthinks]
    #8310411 -

I have some on agar right now in my schools micro lab. I'll take pics when I go in tomorrow.


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Re: Morels [Re: Ubermensch]
    #8310455 -

I have 2 isolates of the Euro and American black and neither of the 2 produce sclerotia anymore. Minimal at best. I have no idea why, as they were both golden when I first started the 2 on agar. I've got 2 yellow's that i'm going to work with to see what's up with them.

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Re: Morels [Re: Hotnuts]
    #8311326 -

Uber, here's a couple pics of one of my M. esculenta clones on agar. (At least I hope it's morel mycelium. This is my first Morel cloning attempt, so I'm very open to comments/suggestions/pointing-out-that-I'm-growing-a-pretty-mold.)



This are pictures of the same culture/clone a few days apart. (I have other pictures of different samples of the same mushroom cloned- though some of the others are definitely contaminated with bacteria.)

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Re: Morels [Re: MycoAu]
    #8312268 -

HA!

See everyone!? Just like I said in my post out there.

You guys take pictures from 10 feet away, LIKE that is going to help me to help you.
You need a super close up, non-microscopic but at least magnified enough to see the pattern of hyphal growth.


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If you wanna ride, don't ride the white horse, if you wanna ride, ride the white pony

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Re: Morels [Re: makaveli8x8]
    #8312278 -

hehe camera? hmmm, that would blow the cover of my substrate


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If you wanna ride, don't ride the white horse, if you wanna ride, ride the white pony

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Re: Morels [Re: AcidHorse]
    #8312286 -

IMO, that looks good to me. Transfer to new plates right now and then await to see if any sclerotia fire up. Keep the transfer dishes in darkness, room temps.

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Re: Morels [Re: Hotnuts]
    #8312308 -

my morel culture will turn a very distinct brown after a few weeks, like squirrel fur, so if yours does that then that might be a way to indicate.

also if you have access to a microscope you might be able to id it that way to


--------------------
We were sent to hell for eternity :hellfire: Ø:omgawesome:h®
We play on earth to pass the time :foreheadslap:

Over-population the root of all Evil-brings the Elites Closer to the gates.

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Re: Morels [Re: makaveli8x8]
    #8314382 -

AcidHorse, chill out a minute. I'm still working on learning to take micros. Plus, I didn't have time to set up everything for microscope pics- just posted some I already had. I figured that the people who've actually cultured this mushroom would be able to tell morel mycelium "from 10 ft away". I know they aren't great shots, but I'll fix that later.

I am glad you mentioned the use of the microscope for pics though. It's something I'll have to work on when I get the time.

Makaveli- does the mycelium need to run out of space/nutrients for the color change to occur? Or is it just a matter of time/age?

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Re: Morels [Re: MycoAu]
    #8314739 -

Visible sclerotia is the key to an easy i.d. for Morel culturing. Keep those plates in darkness after you've made a transfer. I wouldn't trust those P1 plates at all for being completely clean. After that give them a few weeks in total darkness to see what the deal is.

Edited by Hotnuts (04/22/08 02:57 PM)

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