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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
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Growth Inhibitory Compound Discussion (BLIS-Chitinase) 8
#27525216 - 10/31/21 10:37 AM (2 years, 2 months ago) |
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Growth Inhibitory Compound Discussion (BLIS-Chitinase) ChitinasesQuote:
As chitin is a component of the cell walls of fungi and exoskeletal elements of some animals (including mollusks and arthropods), chitinases are generally found in organisms that either need to reshape their own chitin or dissolve and digest the chitin of fungi or animals. Chitinivorous organisms include many bacteria(Aeromonads, Bacillus, Vibrio, among others), which may be pathogenic or detritivorous. They attack living arthropods, zooplankton or fungi or they may degrade the remains of these organisms
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Chitinases have the ability of chitin digestion that constitutes a main compound of the cell wall in many of the phytopathogens such as fungi.
Characterization of a chitinase with antifungal activity
Bacteriocins
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Bacteria produce and excrete a versatile and dynamic suit of compounds to defend against microbial competitors and mediate local population dynamics. These include a wide range of broad-spectrum non-ribosomally synthesized antibiotics, lytic enzymes, metabolic by-products, proteinaceous exotoxins, and ribosomally produced antimicrobial peptides (bacteriocins). Most bacteria produce at least one bacteriocin
Bacteriocins from the rhizosphere microbiome
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6963780/
It's notable that many of the infected cultures did not exhibit any morphological traits and where white, some possesed rings, others fluffy mycelium with exudates. With all white-colored colonies having villous texture, these white cultures were less virulent and lacked the presence of conidia. So even under the scope we wouldn't likely be able to identify.
This mentions one mentions inhibitory zones:
"There was no zone of inhibition, found in this interaction."
https://www.semanticscholar.org/paper/Isolation-and-identification-of-Mycogone-causing-in-Kouser-Shah/2c5b2af21be4a7c1f98ea282998646638af280db#paper-header
This seems like an odd mention. I'd like to find out more about culture morphology that includes zones of inhibition.
"Chitinases of pathogenic fungi not only play vital roles in spore germination, septum formation, cell division, and morphogenesis, but the enzymes are also important in the host interaction . In addition to degradation of the host fungal cell wall, chitinases also inhibit hyphae growth and bud tube elongation.
https://www.frontiersin.org/articles/10.3389/fmicb.2020.596719/full
I am convinced that scalloped edges are morphological identifiers for some inhibitory substance like chitinases or BLIS (bacteriocin-like inhibitory substances), I refuse to believe that inhibitory zones are healthy growth patterns.
 
The left hand plate was used to produce the following:
 
Front and back of the same plate . These plates are heavily scalloped, floccose and thicken toward the margin which prevents light from passing through the culture.
Inhibitory zones are used to test the strength of both antibiotics and fungicides:
Antifungal properties of Chitinases:

https://www.researchgate.net/figure/Antifungal-Activity-of-Chitinases-against-Various-Fungi-The-fungi-used-are-as-follows_fig4_7145317
Considering that Mycogone, Aspergillus, Trichoderma, Bacteria, etc, can produce Chitinases; all bacteria can produce at least one form of bacteriocin. It seems reasonable to suspect that these compounds will be found on our plates and certainly found in grain processed for spawn production, furthermore it seems reasonable to suspect that these compounds if present would effect morphology by preventing growth of the host organism where these products are found.
I believe that many of the positive visual identifiers for generally stressed grain masters may be the result of these compounds, or similar compounds; live vegetative bacteria may not be present but their byproducts may very well be. Chitinases, BLIS, or some similar mechanism seems reasonable, it also appears to be reasonable to state that symptom severity will reflect stage of infection, virulence etc.
Edited by Stipe-n Cap (12/24/22 12:46 PM)
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Myco-Parasite Discussion [Re: sandman420]
#27525304 - 10/31/21 12:29 PM (2 years, 2 months ago) |
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Off to a good start, guys. I reposted my earlier comments from general discussion for ease of reference.
The pic set directly under wet bubble heading, photo D exhibits the very typical flocculant mycelium I have experienced with scalloped plates.
Edited by Stipe-n Cap (10/31/21 12:48 PM)
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Yes, but I clearly dont think thats the end of the conversation.
And yes, there are times when visually similar traits will manifest on plates that do not represent an infection, however there is clearly something else going on, I have enough experience to know when my plates are infected.
Edited by Stipe-n Cap (10/31/21 12:59 PM)
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Are you trained in microscopy to identify such things? I only ask because it seems germane.
To be clear I'm not attempting or suggesting a black or white approach to something as complicated as microbiology.
the first link outlined the lack of observable conidia:
"On PDA chlamydospores were not observed on Hp2 and Hp9, similarly, conidia were not observed for H2, Hp8, and Hp9 . The isolates with brown colony morphology produced more conidia compared to the isolates with white colony morphology."
I'm willing to wager that a laymen may not be able to positively ID mycoparasites under some condiotions, at least.
I cannot comment on your plates, I don't have any experience with your cultures, I borrowed you pic as an example of what to expect.
I should probably remove the picture. (Removed)
I think that its reasonable to assume that there are:
1. Mycoparasites growing alongside our plate cultures;
2. These mycoparasites produce metabolites;
3. Pathogenic parasitic fungi produce Chitinases ( both Mycogone perniciosa and Trichoderma);
4. Chitinases effect culture morphology, including visible inhibatory zones:
"Chitinases of pathogenic fungi not only play vital roles in spore germination, septum formation, cell division, and morphogenesis, but the enzymes are also important in the host interaction . In addition to degradation of the host fungal cell wall, chitinases also inhibit hyphae growth and bud tube elongation."
https://www.frontiersin.org/articles/10.3389/fmicb.2020.596719/full
I beleive these statements to be reasonable and supported by evidence, cleary there will be some overlap in morphological features that apply to cultures that may be healthy.
Edited by Stipe-n Cap (10/31/21 01:46 PM)
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Myco Parasites [Re: Kizzle]
#27526120 - 11/01/21 08:39 AM (2 years, 2 months ago) |
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The information linked above does show mycogone growing on PDA as a pure culture, however when it came time to infect the host mushroom they inoculated the casing layer, which is definitely an important bit of information that they didn't culture them side by side. Having said that mycogone is clearly capable of growing on agar and the sources don't specifically mention that the parasite is incabale of growing with the host mycelium.
More info is needed to confirm that it doesn't parasitize growing mycelium, do you have a link?
Same here, I'm just a glorified monkey trying to stumble around in the dark to find out what's going on with mycelium at a microscopic level, without a microscope
I know next to nothing about mycoparasites, so the best way to educate myself is to get this conversation going so that we can all engage in a group exploration of the subject through discussion.
Edited by Stipe-n Cap (11/01/21 08:49 AM)
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Quote:
sandman420 said: Also it just Must be contagious to the spore print. Surely you've all got a swab or print that will grow nothing but fucked up "white mold" off looking myclium or wont fruit or bubbles or wahtnot. It just must be from the print.
This has been my only experience with what I presume to be mycoparasites, always from swab or print. If what we're experiencing is a mycogone, this mechanism of infection seem most likely due to the organisms preference for infecting basidiocarp and basidium. This makes sense to me.
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The spore print > agar > grain > fruit > spore print > on and on life-cycle does NOT EXIST in commercial cultivation and therefore is not part of any studies on this I have seen.
Good point.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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We shouldn't be married to the idea that it's even mycogone, there's clearly something going on but it seems like mycogone isn't a perfect fit for a lot of the issues that we're seeing, not to say that it isn't an issue at times.
I'm attempting to move to Vancouver Island, atm. When I get out there I know people with connections at UVic, I'm hoping going to make some friends with actual mycologists over there.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Mycoparasite/Mycovirus Discussion [Re: sandman420]
#27526709 - 11/01/21 06:11 PM (2 years, 2 months ago) |
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I agree, viruses are definitely a great topic of discussion as well. Too bad about the pay wall.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Mycoparasite/Mycovirus Discussion [Re: sandman420]
#27526736 - 11/01/21 06:33 PM (2 years, 2 months ago) |
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I'd definitely read it. I wish someone would make mycology video lectures available online.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Nice find, sandman.
Here is an interesting photo posted on the shroomery discord server, the cultivator claims that it had been damaged by uv irradiation:

This occurred after 63mJ of UV-C.
Just an interesting anecdote.
I'll go over that info tomorrow, I just gave it a quick scan.
Edited by Stipe-n Cap (11/05/21 06:03 PM)
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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No idea, I don't know much about the application of UV.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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The only way I'm going to get any satisfaction is to purchase some scopes and then get to work.
I'd like to purchase some Trichoderma from a grow shop, inoculate some plates and watch how the mycelium interacts at the contact boundary.
I'm not sure how to get my hands on other mycoparasites but I'm sure that it's possible.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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This is my experience as well, nof fruits, no pins.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Growth Inhibitory Compound Discussion (BLIS-Chitinase) [Re: GandalfTheWhite]
#28007081 - 10/19/22 08:57 PM (1 year, 3 months ago) |
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Classic floccose appearance with scalloped edge, I don't personally even touch those unless I plan on scoping it; considering I don't yet own a scope....do you own one?
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