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Stipe-n Cap


Registered: 08/04/12
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Loc: Canada
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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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rockyfungus
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Endochitinases (E.C 3.2.1.14) and exo-chitinases. The endochitinases randomly split chitin at internal sites, thereby forming the dimer di- cetylchitobiose and soluble low molecular mass multimers of GlcNAc such as chitotriose, and chitotetraose.[19] The exo- chitinases have been further divided into 2 subcategories: Chitobiosidases (E.C. 3.2.1.29),[20] which are involved in catalyzing the progressive release of di-acetylchitobiose starting at the non-reducing end of the chitin microfibril, and 1-4-β-glucosaminidases (E.C. 3.2.1.30), cleaving the oligomeric products of endochitinases and chitobiosidases, thereby generating monomers of GlcNAc.[19]
Our meds tend to target steroids, cell walls, or protein production. Went down a rabbithole of trying to find what bacteria we used to develop anti-fungals and what their mechanism of action is.
https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1007184
Best I could find. I like cell biology and microbiology...
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multifractal
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Just for the record, the picture of the culture of mine you posted here cleaned up to normal looking growths and fruited very well on bulk substrate. As I've said elsewhere, I believe this morphology to sometimes occur due to somewhat genetically distinct mycelial colonies not blending well with each other. Those areas you call "inhibitory zones" actually had mycelium growing on them but just very sparse and thin mycelium. The plates pinned nicely as well over time. So I'm not sure if you should use that photo as an example to this argument. It's very possible that similar growth patterns could at time be indicative of some sort of contamination but I'm quite sure that was not the case here based on how the culture behaved like a totally clean culture while fruiting.
This plate is a child of those:

I see the pattern not infrequently on germination plates. You can see it here on the right:
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Kizzle
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It could be determined through experimentation. Isolate the parasite and intentionally introduce it to a culture. I have yet to isolate mycogone from any mushrooms while cloning. It may occur but I haven't seen any evidence of it occurring with cubensis.
Also in all my encounters with mycogone-like fruit body deformations I've never seen any indication of a disease spreading. They usually appear in late colonization or early pinning and then normal fruit bodies follow.
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Stipe-n Cap


Registered: 08/04/12
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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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sandman420
Saint PP



Registered: 06/17/04
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no doubt, it's just that it fits the wet bubble disease expressions. The pathogens may be even a complex of some things. Mycogone p. being the pathogen in the known case. But the disease appears the same at least.
Also we should include mycoviruses in this thread. I suspect they are much more common. Very little known but there are some stuff. Mushroom Virus X and La France (? probably remember that name wrong ill double check that one lol) among an endless amount of others.
I read one report that infected fruits make 25% spores that carry the mycovirus from one type, it hink it was MVX. (actually I might not have read the report just the abstract i am waiting to get free report from authors)
edit here it is. Do I read that as 25% of of all spores contain it or 25% of infected fruits spores are infected?
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The identification of a novel Pleurotus ostreatus dsRNA virus and determination of the distribution of viruses in mushroom spores Yeo Jin Kim, Ji Yeon Kim, Ji Hye Kim, Seon Mee Yoon, Young-Bok Yoo & Se Won Yie The Journal of Microbiology volume 46, pages95–99 (2008)Cite this article
307 Accesses
10 Citations
Metricsdetails
Abstract Double-stranded RNAs and virus particles were identified in Pleurotus ostreatus strain Shin-Nong in Korea. Isometric virus particles with a diameter of 33 nm were purified, which are similar to other Pleurotus viruses reported previously. This strain contains 5 dsRNAs, 8.0, 2.5, 2.4, 2.0, and 1.8 kb in size. The virus particles contain 2 dsRNAs, designated RNA-1 (2.5 kb), and RNA-2 (2.4 kb) which is a typical pattern of Partitiviridae. A non-encapsidated dsRNA of about 8.0 kb also was identified. Partial cDNA from RNA-1 was cloned, and sequence analysis revealed that this gene codes for RdRp. The comparison of the sequence from partial cDNA clone showed 35% amino acid homology with the C-terminal end of the RdRp gene of Helicobasidum mompa virus and Rosalinia necatrix virus. Specific primers designed from the partial sequences successfully amplified RT-PCR product from the infected mycelium and a single spore culture. We used these primers to determine the pattern of distribution of viruses in spores. Of the 96 different single spore cultures generated from Shin-Nong strain, a specific RT-PCR product was identified in 25 cultures, indicating that about 26% of basidiospores contain viruses.
if anyone has access to this report please msg me. $40? You think I'm made of money?
more
Quote:
Minor Pest Title: Die-back disease (Virus)
Minor Pest Description: It causes spots in the casing soil where no mycelial growth occurs. Around these spots, mushrooms of low quality appear with long stems and dirty caps. Sometimes the only indication of a virus infection is low yield. In severe cases, a few deformed mushrooms are produced. The disease can be introduced to the farm by infected spawn. It is spread by spores and mycelium from infected mushrooms. Mushrooms affected with the virus open fast, releasing infected spores. Sometimes, mushrooms that were formed inside the casing layer come out already open. Spores from infected mushrooms are easily carried by wind, insets, on implement, clothes and hands of personnel.
Minor Pest What to do.: Observe sanitation and hygiene during growing cycle Cover the beds after spawning with a paper, which must be sprinkled with 2% formalin solution every 3-4 days in order to kill all settling spores Harvest mushrooms in proper time, not allowing them to open Clean and disinfect growing rooms after growth cycle Grow tropical mushroom (Agaricus bitorquis). It is resistant to the virus
would like access to these papers also, I feel like we could get somewhere together with this for the OMC.
Quote:
Viruses Associated with A Die-Back Disease of Cultivated Mushroom M. HOLLINGS
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Kim, Y.-J., S. Park, S.W. Yie, and K.H. Kim. 2005. RT-PCR detection of dsRNA mycoviruses infecting Pleurotus ostreatus and Agaricus blazei Murrill. Plant Pathol. J. 21, 343–348.
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Ihrmark, K., H. Johannesson, E. Stenstron, and J. Stenlid. 2002. Transmission of double-stranded RNA in Heterobasidium annosum. Fungal Genet. Biol. 36, 147–154.
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Morris, T.J. and J.A. Dodds. 1979. Isolation and analysis of double-stranded RNA from virus-infected plant and fungal tissue. Phytophalology 69, 854–858.
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Preisig, O., B.D. Wingfield, and M.J. Wingfield. 1998. Coinfection of fungal pathogen by two distinct double-stranded RNA viruses. Virology 252, 399–406.
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Rao, J.R., D.W.A. Nelson, and S. McClean. 2007. The enigma of double-stranded RNA (dsRNA) associated with mushroom virus X(MVX). Curr. Issues Mol. Biol. 9, 103–122.
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Raper, J.R. and C.A. Raper. 1972. Genetic analysis of the life cycle of Agaricus bisporus. Mycologia 64, 1088–1117.
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Revill, P.A. and P.J. Wright. 1997. RT-PCR detection of dsRNAs associated with La France disease of the cultivated mushroom Agaricus bisporus (Lange) Imbach. J. Virol. Methods 63,
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Romaine, C.P. and M.M. Goodin. 2002. Unravelling the viral complex associated with La France disease of the cultivated mushroom Agaricus bisporus, 237–257. In S.M. Tavantzis (ed.), dsRNA Genetic elements: Concepts and applications in agriculture, forestry and medicine. CRC Press, USA.
Quote:
Rong, R., S. Rao, S.W. Scott, and F.H. Tainer. 2001. Common multiple dsRNAs are present in populations of fungus Discula destructiva originating from widely separated geographic locations. Curr. Microbiol. 42, 144–148.
Quote:
Seo, J.J., W.-S. Lim, J.H. Jeong, Y.B. Yoo, S.W. Yie, and K.-H. Kim. 2004. Characterization and RT-PCR detection of dsRNA mycovirus from Oyster Mushroom, Pleurotus ostriatus.
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Van Diepeningen, A.D., A.J.M. Debets, and R.F. Hoekstra. 2006. Dynamics of dsRNA mycovirus in black Aspergillus populations. Fungal Gen. Biol. 43, 446–452.
Quote:
Yu, H.J., D. Lim, and H.S. Lee. 2003. Characterization of a novel single-stranded RNA mycovirus in Pleurotus ostreatus. Virology 314, 9–15.
Edited by sandman420 (11/01/21 01:45 PM)
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rockyfungus
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Didn't read the whole thing just what you got. Viruses are in you, part of your DNA. Probably along those lines.
All this genetic material is prayed on by viruses. So spores are a honeypot I guess.
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rockyfungus
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Re: Mycoparasite/Mycovirus Discussion [Re: Stipe-n Cap] 1
#27526825 - 11/01/21 07:36 PM (2 years, 2 months ago) |
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QM33
(NOT A PUPPET!) ❤❤❤❤❤



Registered: 04/09/20
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So you guys figure anything out ha?
Sound like me a couple months ago.
Glad someone else decided to start a mycogone thread as well.
But don't forget about mine ha!
https://www.shroomery.org/forums/showflat.php/Number/27450000/page/3
-------------------- OmManiPadmeHum,OmManiPadmeHum, OmManiPadMeHum... There are known knowns, there are known unknowns, there are also unknown unknowns. With great privilege comes great responsibility.
  Quantom Qups PROOF AND Soft Drops Turn your Swab to a Syringe and Syringe to Multiple Syringes! No Pours (QuantomStyal)Magic Fruit Leather DMT for IandI
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Forrester
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Re: Myco Parasites [Re: Kizzle] 1
#27560425 - 11/28/21 07:57 AM (2 years, 1 month ago) |
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Quote:
Kizzle said: Also in all my encounters with mycogone-like fruit body deformations I've never seen any indication of a disease spreading. They usually appear in late colonization or early pinning and then normal fruit bodies follow.
FWIW, I've only run into mycogone infections a couple times, both with edible or medicinal species.
I think once was king oysters, and that time the substrate only produced one, giant MP bubble. No other fruit bodies ever formed before or after that. I left it to sit quite a while with the giant bubble because I didn't know at the time what it was. Never fruited.
I think the other time may have been with reishi. Again, IIRC no reishi fruits ever were produced, the MP seemed to entirely prevent it from doing so.
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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Stipe-n Cap


Registered: 08/04/12
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This is my experience as well, nof fruits, no pins.
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rockyfungus
dirty


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I'll keep it short as I'm not 100% sure on the language and only read your excerpts. Also nothing is a "virus" in the study. It's a parasite they took genetic info from and inserted into a bacteria (plasmid)??
Mycoviruses don't traditionally hijack a cell extracellularly. Viruses typically find a receptor attach and inject genetic material.
Fungal viruses are active in hyphael networks and spores? How do they get in?
The 2nd blurb is how they got the genetic material in. They had to destroy the cell walls of a parasite so it could take up genetic info and form spheroblasts. Sphereoblasts could only enter a (damaged?) hyphal network. They destroyed the hyphaes and rebuilt them? Which I suppose this parasite is resembling the mycovirus to prove they can only infect through those processes?
So where in growth does hyphal colony have shedded cell walls that can be infected. Or how do spores get infected? Basically is there a place in fungal reproduction where it's hyphael network is bare and exposed?
Rambled if you have a direct question I can try and break it down. Or if you want a term broken down. I hate explaining stuff to other mycologist like they don't understand science. I've done a fuck ton of biology so some words are standard. The more educated people get in a singular field the less they understand what normal people know. So if you need it explained like your 5. No worries I'll try.
Edited by rockyfungus (02/23/22 08:28 AM)
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QM33
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By asexual spores do they mean chlamydospores?
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QM33
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Forrester
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Wow, that's insane what you've got going on there. Eesh...
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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rockyfungus
dirty


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Someone should send you a known culture already colonized, see what happens? Or flip. Work somewhere else with fresh clothes, tools, print. Fruit at normal place and elsewhere and compare?
Or you just picked a turd or need to dial those conditions in
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sandman420
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Mycogone Wet Bubble Disease:


Cobweb: Please note I have confused cobweb with pin molds in the past. Real cobweb can look nearly the same as cubensis mycelium.






Some pic dumps
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Stipe-n Cap


Registered: 08/04/12
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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
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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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multifractal
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Never said it was the end of a conversation. I just don't think it's so black and white that if a plate has these features then it is necessarily being parasitized and thought that those plates might not be good examples of the topic that you are discussing due to their apparent cleanliness in producing healthy and copious mushrooms. I understand using those plates to add nuance to the discussion, offering a counter example, but not using them to exemplify contamination. As you mentioned, I looked at some plates exhibiting similar patterns under scope and saw nothing to indicate the presence of any organism besides cubensis.
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