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 Terry M
Stranger in a Strange Land
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 Re: Elm Oysters  [Re: Mycelio]
#15700755 - 01/22/12 08:14 AM (1 year, 3 months ago) |
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The basic bio 101 species rules are pretty simple for us monokaryotes (critters with one nucleus per cell) who create new strains (races, subspecies, lines, etc.) only via sexual reproduction of our gametes. Gametes are haploid cells with one set of chromosomes (1 unique complement of DNA), like eggs and sperm that combine to make a new diploid cell ( 2 sets of chromosomes, one from poppa and one from momma). If two organisms can breed AND have fertile offspring, they're in the same species. If they can't have offspring, or the offspring aren't fertile (horse + donkey = infertile mule), they're not of the same species.
The confusion for me comes because mating of mycelium doesn't have to be a pure gamete meets gamete process. It can happen between mycelium of the same individual organism. Mycelia can have multiple nuclei. They also can have way more than 2 sexes. So sexual reproduction among fungi is not a cut and dried thing.
What is "sexual compatibility" as applied to mycelia which may be polyploid, and are capable of essentially fucking themselves? The rules for creation of zones of aversion between cultures require some thinking, I think. There's a paper called "INTERACTIONS OF RECOMBINED MYCELIA OF THE FUNGUS FOMITOPSIS PINICOLA (SOW. EX FR.) KARST. ON PDA MEDIUM," by Vladimir Lazarev, GENETIKA, Vol. 34, No. 1, 21-32, 2002. It addresses this in part, but I need to read it again because its conclusions were by no means clear to me.
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Mycelio
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Re: Elm Oysters [Re: Terry M]
#15700819 - 01/22/12 09:04 AM (1 year, 3 months ago) |
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Wow, that is quite an interesting paper. Let me add a direct link:
http://www.dgsgenetika.org.rs/abstrakti/vol34_no1_rad03.pdf
It's fascinating to see some numbers. I never expected the rate of fusion compatibility to be so low.
Unfortunately, I think there are two types of compatibility, first the ability for hyphal fusion and exchange of nuclei, independent of mating type, just like tissue compatibility for organ transplants.
Sexual compatibility seems to be another story, dependent on mating type and likely some other factors, as from time to time sterile, spore less mushrooms are found in nature, where fruiting happens, but meiosis in the basidia or asci fails.
Carsten
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Terry M
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Quote:
GroboClone said:
Has anyone come across studies with DNA analysis?
I'm down to compare the different cultures floating around.
Just need to be pointed in the right direction and I'll take it from there.
Gives me a good excuse to try out the new (to me) PCR machine. 
Here are a bunch of existing H. ulmarius DNA sequences:
http://www.ebi.ac.uk/ena/data/view/Taxon:71891
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Terry M
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Re: Elm Oysters [Re: Terry M]
#15701741 - 01/22/12 02:33 PM (1 year, 3 months ago) |
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Carsten, there's something basic from this paper that I'm just not understanding. Maybe you can explain it to me.
Why did the author use "fragments of the hymenal layer (0.5 x 0.5 cm) representing the inoculum ..."? Couldn't taking bits of a hymenal layer include lots of spores, thus generate a polyspore, multi-strain culture? It would grow out into a bunch of sectors. Do you choose one sector? Then why not just clone some of the trama, which would give a monospore culture directly?
If polyspore cultures were directly and purposely used rather than purified monospore cultures, who knows which strain would "win out" in a mycelium confrontation on agar? Wouldn't this just introduce uncontrolled variables that would make comparison of different innocula pairings less meaningful?
I don't get it.
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Mycelio
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Re: Elm Oysters [Re: Terry M]
#15703188 - 01/22/12 07:43 PM (1 year, 3 months ago) |
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Right, on a second look, we have to be careful, interpreting data from this paper.
The author could have taken trama samples, most likely representing a single, dikaryotic strain, but then several up to all of his strains might have been identical.
I guess he was aiming for lots of different strains, so he chose multispore cultures, transferred and selected 'a pure culture'. Not defining how and what he selected indeed raises doubts.
So, not knowing from how many and how different parent strains the hymenal samples came from plus not knowing how strain selection was performed, we do have many uncertainties here, but we can still see that the tested, already dikaryotic or heterokaryotic F. pinicola strains from different fruitbodies rarely merge, while offspring of the same parent strain often fuse.
Now we have to keep in mind that those strains came from a wild, genetically diverse population, which may behave differently than our cultivated Pleurotus strains. After generations of inbreeding and selection, the genetic variation gets reduced, resulting in more hyphal fusions between dikaryotic mycelia within a cultivated strain.
In my opinion, seeing a zone of aversion between a strain of P. ostreatus and our Pleurotus 'H.u.' won't prove anything. We could have either two incompatible strains of the same species or two different species.
Only hyphal fusion could tell us if we have compatible strains of the same species. We might have to test a lot of strains from P. ostreatus, P. pulmonarius, perhaps also P. populinus and more species, until we find a match. Hopefully those chemical reagents can help reducing the species in question.
In theory, pairings of single spore isolates should be more promising. I think monokaryotic mycelia are less picky about fusing with a possible partner than dikaryotic ones, but then creating all those monospore cultures would be much annoying work.
Carsten
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jimmyjame1
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Re: Elm Oysters [Re: Mycelio]
#15703589 - 01/22/12 08:54 PM (1 year, 3 months ago) |
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my goodness you guys
this is like the never ending rabbit hole. the science you are getting into is so mindboggling.
thanks for all your hard work! hopefully we can all get to the bottom of this H.u. confusion
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Mycelio
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Yes, who would have thought that IDing an oyster would be so confusing.
In between I remembered a simpler way for a mating study, which would only require one monokaryotic (single spore) mycelium of a mislabeled elm oyster. This should be doable by streaking spores across an agar plate with the good old zigzag pattern. The isolated mycelium could be put on grain and spawned to several small substrate bags. After full colonization they should not be able to fruit. Then either the surface would be sprayed with a spore solution from P. ostreatus, P. pulmonarius and whatever is to be tested or a fresh sporulating mushroom would be placed above it. Some hundred thousand spores would germinate in each bag and in case of sexual compatibility primordia or mushrooms should reveal success.
What do you guys think?
Carsten
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 NSF
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Re: Elm Oysters [Re: Mycelio]
#15709923 - 01/24/12 03:01 AM (1 year, 3 months ago) |
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Why would they not be able to fruit when they are cultures from many spores?
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Mycelio
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Re: Elm Oysters [Re: NSF]
#15710208 - 01/24/12 05:40 AM (1 year, 3 months ago) |
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No, I meant that from the zigzag plate, one would try to isolate a monokaryotic mycelium, growing from one spore. This should not be able to fruit until it fuses with another mycelium of the opposite mating type.
Carsten
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 GroboClone
Anthropomorphic Elephant
Registered: 01/07/11
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Re: Elm Oysters [Re: Mycelio]
#15710768 - 01/24/12 11:06 AM (1 year, 3 months ago) |
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Thanks Terry, just what I was looking for.
Some very good links there.
The idea is to run the samples on an electrophoresis gel and not have to send out for sequencing.
Fungal-specific PCR primers developed for analysis of the ITS region of environmental DNA extracts
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lipa
Registered: 07/24/07
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Re: Elm Oysters [Re: Mycelio]
#15714002 - 01/25/12 12:14 AM (1 year, 3 months ago) |
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Quote:
Mycelio said:
Yes, who would have thought that IDing an oyster would be so confusing.
In between I remembered a simpler way for a mating study, which would only require one monokaryotic (single spore) mycelium of a mislabeled elm oyster. This should be doable by streaking spores across an agar plate with the good old zigzag pattern. The isolated mycelium could be put on grain and spawned to several small substrate bags. After full colonization they should not be able to fruit. Then either the surface would be sprayed with a spore solution from P. ostreatus, P. pulmonarius and whatever is to be tested or a fresh sporulating mushroom would be placed above it. Some hundred thousand spores would germinate in each bag and in case of sexual compatibility primordia or mushrooms should reveal success.
What do you guys think?
Carsten
Monokaryotic oyster mycelium can fruit so that is not a fullproof idea. They make for good crosses because they usually produce nice productive fruiting cultures as a result. So Chang says.
Lipa
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Mycelio
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Re: Elm Oysters [Re: lipa]
#15714046 - 01/25/12 12:28 AM (1 year, 3 months ago) |
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Damned, I wasn't aware that oysters can do that too.
Thank you for the info!
Carsten
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solarity
mm... my favourite food
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Re: Elm Oysters [Re: Mycelio]
#15734128 - 01/29/12 05:08 PM (1 year, 3 months ago) |
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Stamets on the 2 strains on a dish thing (you guys lost me after that) it's morels but same concept
http://youtu.be/dJvwoGLxmhs
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Mmmm! Goddamn, Jimmie! This is some serious gourmet shit! Usually, me and Vince would be happy with some plain ole Agaricus right, but he springs this serious GOURMET shit on us!
TRADE Always looking for unusual commercial cultures to try. Have strong edible strains to trade.
Edited by solarity (01/29/12 05:10 PM)
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Terry M
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Just an update:
I've had lab contamination problems, so rather than keep battling them in not easily sterilizable environment, I'm making a 100% dedicated lab room. This should be operational next week, when I get my new phenolic resin topped 3'x6' lab bench. I just installed a completely washable vinyl floor, which will have waterproof silicone caulk around all the edges.
In the meantime, I've got some colonized jars of Aloha H. ulmarius in the fridge on hold. These will hopefully go into filter patch bags next week. 
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Javadog
Continuing along
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Re: Elm Oysters [Re: Terry M] 1
#15759471 - 02/04/12 11:21 AM (1 year, 3 months ago) |
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Thanks for the update Terry.
I just spawned the first of my grain quarts of JJ's H. tessulatus
to some straw sacks the other day.
It strikes me as slow to recover, but no contams yet, so
hope springs eternal.
I expanded my initial quarts and so have others that I will
be using with plain sawdust sacks. My last pass will likely
be supplemented and properly pasteurized sawdust sacks.
Take care,
JD
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Edited by Javadog (02/04/12 11:22 AM)
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jimmyjame1
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Re: Elm Oysters [Re: Javadog]
#15759485 - 02/04/12 11:29 AM (1 year, 3 months ago) |
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sounds great,
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metacohl
Mycophage
Registered: 07/03/11
Posts: 57
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Amazing thread! Thank you to everyone who has contributed to unraveling this mystery. Absolutely fascinating!
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Javadog
Continuing along
Registered: 05/03/10
Posts: 7,273
Loc: USA
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Hello all,
Check out this kit:
http://www.mushroomadventures.com/p-6-elm-oyster-mushroom-kit.aspx
It looks like they made the P.o. vs. H.u. mix-up as well.
(though one of their photos even looks like P. eryngii)
Terry, how are the Aloha H.u. doing?
My work with JJ's H.t. is going slowly. It seems that it's culture
is fighting to protect a bacterium. I may have to do a hot pour if
my latest grain efforts are as weak as the last.
Take care,
JD
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NSF
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Re: Elm Oysters [Re: Javadog]
#15881879 - 02/29/12 06:07 PM (1 year, 2 months ago) |
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I thought i'd look at some non Stamets literature, hopeful that David Arora's book would clear things up a bit, but noooo...no mention of H. ulmarius in Mushrooms Demystified. I was a little shocked.
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Mycelio
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Re: Elm Oysters [Re: NSF]
#15882123 - 02/29/12 07:02 PM (1 year, 2 months ago) |
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Don't know about that particular book, but some mycologists do not separate H. ulmarius from H. tessulatus.
Carsten
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