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Stipe-n Cap


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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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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


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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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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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Stipe-n Cap


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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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Stipe-n Cap


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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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rockyfungus
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Speaking from a human physiology point of view. Scalloped edges, uneven borders, thicker borders, raised areas, change in colors, etc are not NORMAL. Basically things that human pattern recognition software picks up. Are these abnormalities idiopathic (genetics for simple sake) or true pathology?
Something is either impeding growth, taking up mass, stealing nutrients or blood (let's call it water?), or hacking cellular mechanisms. Obviously these mycoparasites like most human pathogens don't want to kill the host outright. They need to parasitize a host but not disrupt it's lifeblood.
We are only talking about "biotic" life here aren't we. What about the billions of viruses that most likely causing issues as well.
Wouldn't a germination plate have the highest concentration of microbes if we are working in true aseptic fashion?
We try and grab way from those notches as best chances are far away from the "odd" part. Did those clean plates come from anywhere near the notch?
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Kizzle
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I'm pretty sure mycogone is strictly a fruit body pathogen. It's not something that could hide away in mycelium like Trichoderma can. I seem to recall they did some experiments with adding it to the compost and it had no effect. The spores had to be in the casing layer where they would come in contact with the fruit bodies, same with verticillium. Presumably the mycelium can produce metabolites that inhibits in growth and fruit bodies can't.
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sandman420
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Re: Myco Parasites [Re: Kizzle]
#27526109 - 11/01/21 08:15 AM (2 years, 2 months ago) |
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Quote:
Kizzle said: I'm pretty sure mycogone is strictly a fruit body pathogen. It's not something that could hide away in mycelium like Trichoderma can. I seem to recall they did some experiments with adding it to the compost and it had no effect. The spores had to be in the casing layer where they would come in contact with the fruit bodies, same with verticillium. Presumably the mycelium can produce metabolites that inhibits in growth and fruit bodies can't.
I know what you are talking about, here is my explanation of these studies:
Big commercial growers do not grow their own spawn, and if they do it is in a totally separate facility with that goal in mind. They use elite worldwide banked monocultures and they know what they are doing. They are not running foil prints to agar made in some hippies closet.
Clearly they can grow while not on a mushroom fruit body, see the pics of it growing on agar as proof, it goes without saying that is not a fruit body.
There are studies I have seen that show it can germinate in sterile water.
So it would make sense that their studies are mainly in line with after spawning because that's all they do is spawn the bulk and case and assume that they spawn is clean because that is a separate professional thing with it's own concerns. Does that make any sense? Mushroom growing isn't the same thing as spawn making or culture making, commercially/professionally that is.
Verticillium in particular is interesting because it sporulates on the caps of mushrooms only, which would be a big ass problem for a print...
PLEASE NOTE I DO NOT HAVE A FULL UNDERSTANDING OF THIS JUST AS ANYONE ELSE COMMENTING JUST CONTRIBUTING THE BEST I CAN, I'M NO AUTHORITY
edit: I think that was a bad explanation let me try again.
Commercial growers of the type in these studies don't use spore prints, spores at all, agar, or probably even have anything to do with their grain spawn. The just spawn bulk substrate from grain spawn that is provided to them professionally. And when grain spawn and cultures ARE made they aren't made from spore prints, only elite cultures. So therefore this is not a concern of these studies, they all seem to only ever discuss spawning and casing not the in between print life cycle that we do.
Those studies mainly are saying that for them, they get it at casing and the stuff wont really grow in the casing until it touches primordia. But it germinates and waits. I think.
Edited by sandman420 (11/01/21 08:44 AM)
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Stipe-n Cap


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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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sandman420
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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.
if my above post could be distilled this is it:
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.
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Stipe-n Cap


Registered: 08/04/12
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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.
Quote:
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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rockyfungus
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I could be completely off base. For commercial growers they just transfer these known commercial strains? Are they reaching senescence and weakening the "immune system" of the myc.
Like my 3015 is god knows how far out it is? It came from agar, I've received LCs for gourmet. I've tried to go back to spores from own grows but at that point it's not 3015 I'd imagine.
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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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sandman420
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Re: Myco Parasites [Re: Kizzle]
#27526299 - 11/01/21 11:40 AM (2 years, 2 months ago) |
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It's also been my experience with "wet bubble looking" infections appearing early and leading to a normal second flush. But probably related to severity/time of infection just like aspects with button mush. Maybe that's a sign of infection after spawning for example (out of my ass)
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Stipe-n Cap


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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
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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?
Quote:
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
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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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Stipe-n Cap


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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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sandman420
Saint PP



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Re: Mycoparasite/Mycovirus Discussion [Re: Stipe-n Cap]
#27526727 - 11/01/21 06:28 PM (2 years, 2 months ago) |
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Some young lad in a mycology PhD trajectory totally should write your thesis on The Viral Infections of Clandestine Psiolcybin Mushroom Cultivation.
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Stipe-n Cap


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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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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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Yeetusdeetus



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sandman420
Saint PP



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Alright scooters thanks to good ol p9s library link I found some new interesting shits.
From this study
Basically they find that mycogone will germinate on nearly anything from the order Agaricale (20 of 28 tested showed favorable results)
Cubensis and most of our mushrooms we grow are of the fungal order Agaricales.
Quote:
Results are shown in Tables 1 and 2. Unfortunately, because of the time-consuming nature of bioassay procedures and the varying availability of material, complete data relating to tissues, extracts and mycelia (set out in Table 1) were obtained for only 10 species from 9 families. Germination was induced by contact with basidiome tissues of most fungi used here, although germination levels were usually significantly below those found for Agaricw brunnescens. The single exception was a species of Cortinarius on which germination was comparable with that on A. brunnescens. Basidiomes of some species failed to activate conidia and there was additionally a wide variation in effects within most families. Extracts of Agaricw brunnescens basidiomes induced germination, as did those from several other species, although to a lesser extent, and again some extracts failed to activate conidial cells. With the exception of Agaricus species and Coprinus cornatus, germination levels on extracts were below those occurring on corresponding whole tissues. Most mycelia tested induced germination, levels for Agaricus brunnescens being similar to those occurring on its basidiome tissues and extracts. For other species the levels were also relatively high. For instance, in C. comafw, hccaria laccafa (Scop.: Fr.) Cooke, Armillaria bulbosa (Barla) Kile & Watling and Phallus impudicus Pers. they either approached or exceeded those for basidiome tissues. Germination occurred on mycelia of Amanita nrbescens (Pers.: Fr.) S. F. Gray and Sclerodema citrinum Pers., yet not on their basidiomes or basidiome extracts.
Quote:
DISCUSSION Sometimes severe technical difficulties attended preparation of tissues and extracts for bioassay and attempts to obtain mycelial cultures from basidiomes. Therefore the data presented here are incomplete. Nevertheless, the breadth of the survey allows some firm conclusions to be drawn concerning induction of germination in thick-walled conidial cells of M. pemiciosa and permits speculation on the ecology of this mycopathogen. Turning first to Agaricales, basidiomes of 28 wild species were bioassayed to 20 of which M. perniciosa conidia responded positively, with two further species inducing just below 1 % germination. Germination occurred on basidiomes of fungi typical of a variety of habitats and on mycorrhizal, leaf-litter decomposing and lignicolous species. Although germination levels were usually below those for Agaricus bnmnescens, they were commonly above 20% and ranged as high as 66%, which gives reason to suppose that many wild species could act as hosts for M. perniciosa. In this regard it is of interest to note that over 50% germination was recorded on basidiomes of Agaricw campestris L.: Fr., the close relative of Agaricw brunnescens. This view is reinforced by the generally high germination levels produced by contact with mycelia, since it is probable that association of M. perniciosa with undifferentiated mycelium precedes the appearance of disease on an epidemic scale during mushroom cultivation. However, there is little or no direct evidence for the existence of such disease reservoirs outside the mushroom production unit, and so far it has proved to be unfeasible to carry out pathogenicity tests on basidiomes of wild species either in the field or laboratory. A notable feature of the bioassays was the wide variation in response even to basidiomes from fungi within a single genus. Thus Panaeolw semiovatus (Sow.: Fr.) Lund stimulated over 50% germination, but for P. sphinctrinus (Fr.) Quel. this was zero. Similarly Amanita vaginafa (Bull.: Fr.) Vitt. promoted 23 % germination and A. rubescens (Pers.: Fr.) S. F. Gray zero; Lactarius rufus (Scop.: Fr.) Fr. 34% and L. furpis (Weinm.) Fr. zero. Curiously, whilst basidiomes of Hypholoma fasciculare and their extracts induced germination, contact with mycelium did not. Three of the six non-agaric Basidiomycotina investigated gave positive results; basidiomes, extracts and mycelium of C. sfercorew all inducing low levels of germination, as did mycelium of Scleroderma citrinum Pers. Results using P. impudicus were of particular interest in that contact with its exoperidium brought about 25 % germination but whole egg extracts had little effect. By contrast, contact with its mycelium induced nearly 50% germination. This survey clearly demonstrates that germination-inducing factors are widespread within Agaricales but that they are by no means confined to them. In addition, it appears that whilst these factors can sometimes be extracted in aqueous solution from basidiomes, this may not always be the case; as, for example, for L. laccafa and P. impudicus. Similar factors are present in mycelia but, by contrast, cannot be extracted from them; nor, generally, are they found in media within which mycelia have been grown (Holland, 1988). Work is now in progress to identify the compounds involved in triggering germination of dormant, thick-walled conidial cells of M. perniciosa and some preliminary findings might be mentioned here. They are ethanol-soluble and are to some extent wallbound (Holland, 1988). Furthermore, two or more groups of factors are implicated which act either synergistically or in sequence (Rawlins, 1990). It is hoped to publish details in due course.
Some other important information, mycogone makes 2 types of spores. 1 one which germinates easily on anything and one of which survives a long time but only germinates in the presence of mushroom tissue/mycelium.
Quote:
Mycogone pemiciosa sporulates heavily on Agaricus basidiomes, producing small thin-walled phialospores together with large bicellular conidia each consisting of a dark, spherical, thick-walled, vermcose apical cell and a thin-walled basal cell. After secession the latter dies, leaving the thickwalled component as the major survival propagule (Holland et al., 1985). By contrast with phialospores, which germinate freely on a variety of substrata, thick-walled conidial cells are dormant and will germinate only when activated by uncharacterized factors emanating from vegetative mycelia and basidiome tissues. As well as being present in Agaricus bmnnescens Peck, such activators have also been detected in expressed juices from Lepiofa procera QuPl., Schizophyllum commune Fr., and in single, unidentified species of Collybia, Coprinus and Hygrophorw (Vincent-Davies, 1973). This suggests at least the possibility of a potentially wide host range for M. perniciosa within Agaricales and the existence of disease foci in the field.
So it is NOT a wild theory that mycogone can infect cubensis, the study would definitely support this.
Edited by sandman420 (11/05/21 05:15 PM)
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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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rockyfungus
dirty


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What's the NM on that thing? UV leads to reactive oxygen and nitrogen that damages major cellular pathways. DNA and lipid membranes mainly. This leads to cytotoxicity (cell death), mutations, and alterations in cell signaling.
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Stipe-n Cap


Registered: 08/04/12
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Loc: Canada
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No idea, I don't know much about the application of UV.
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sandman420
Saint PP



Registered: 06/17/04
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I read in another one of these studies they were using 30 watt phillips TUV lights for mutagenesis. But now were off topic again!
Edited by sandman420 (11/05/21 08:16 PM)
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Stipe-n Cap


Registered: 08/04/12
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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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sandman420
Saint PP



Registered: 06/17/04
Posts: 5,384
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No, the only way this is getting solved is as follows
Quote:
4.1. Fungal Isolates and Morphological Characteristics Mycogone perniciosa isolates (Table 1) were obtained from the fruiting bodies of A. bisporus showing typical symptoms of wet bubbled disease in mushroom farms located in Fujian, Gansu, Hubei, and Shandong provinces of China. The disease survey was carried out in 2014–2015. The sampled infected tissues were sterilized in 2% sodium hypochlorite (NaClO) solution for 60 secs and washed three times with sterilized deionized (DI) water, then plated on Petri dishes containing PDA (potatoes 200 g/L, glucose 20 g/L, agar 15 g/L) amended with 100mg/L Kanamycin and incubated at 25 C for 5 days in darkness. Pure cultures were subsequently obtained through single spore isolation from all colonies showing the morphological characteristic of a typical M. perniciosa and five representative purified isolates sub-cultured on PDA without antibiotics. The growth characteristics, colony morphology, and conidial characteristics—such as shape, length, width—were examined for a total of 18 representative isolates. Colony color was assessed 7–10 days after single spores were transferred to PDA. A minimum of 30 conidial characters was observed under a Leica DMR HC microscope (Leica Microsystems Imaging Solutions Ltd., Cambridge, UK) fitted with Leica DFC320. The sporulation of the isolates was estimated as described by Santos et al. [27]. Briefly, 100 mg of the fungal mycelium of each isolate was collected and transferred to an Eppendorf tube, in which it was homogenized with 1 mL of Tween 80 solution (0.05%). The conidia count of each such suspension was then determined using a Neubauer chamber. Conidia counts were performed in triplicate for each isolate. All cultures were conserved on PDA in slant tubes and deposited in the Engineering Research Center of the Chinese Ministry of Education for Edible and Medicinal Fungi of Jilin Agricultural University (HMJAU) in China.
4.2. DNA Extraction and Molecular Identification Total genomic DNA was extracted from 7-day old mycelia mat growing on PDA plates with cellophane sheets using Nuclear Plant Genomic DNA Kit of CWBIO (CWBIOTECH, Beijing) following the manufacturer’s protocol. The DNA quality and quantity were measured using a BioSpec-nano spectrophotometer (Shimadzu Biotech, Tokyo, Japan) at a wavelength of 260 and 280 nm, respectively. The DNA was stored at 80 C until required for further use. PCR amplification and sequencing of the internal transcribed spacer regions of the rDNA was performed for each isolate utilizing the primer set ITS4 and ITS5 [28]. The obtained sequences were individually checked by BLAST analysis against the NCBI GenBank (http://www.ncbi.nlm.nih.gov/) database and highly corresponding sequences were retrieved, aligned, and the phylogenetic tree constructed with the maximum likelihood method using the Tamura and Nei substitution method [29] in MEGAX [30].
Quote:
4.3.2. Inoculum Preparation and Disease Assessment The inoculum for each M. perniciosa isolates were prepared from 7-day old cultures on PDA, by washing down the pathogen conidia with sterile distilled water and sieving the deferment via six layers of sterile cheesecloth. The spore/conidial concentration was estimated using a hemocytometer. The optimal spore/conidial concentration of the suspension for the isolates to cause disease was determined by inoculating 104, 105, 5 105, and 106 conidia/ml suspension on A. bisporus strains CCMJ1020 and CCMJ1036. A spore/conidial concentration of 1 x 105 was standardized and used for disease inoculation for all the A. bisporus strains. Three days after application of casing soil and regulation of temperature and relative humidity, approximately 50 ml of inoculum (spore/ conidial concentration105/ml) were sprayed into each basket containing the cultivated button mushroom. The controls were sprayed with 50 ml of sterile distilled water. Three replications were evaluated per isolate per mushroom strain. Disease assessment was recorded for the first flush. Pathogenicity was determined by the number of A. bisporus strains on which an isolate caused wet bubble disease and the order of susceptibility of strains to individual isolates. Pathogenicity tests to confirm Koch’s postulate was assessed on A. bisporus strain CCMJ1020. Disease severity was rated on sporocarp of individual mushroom strains for 30 days after inoculation using a 0–5 visual rating scale, where 0 = no symptom; 1 = 1–10%; 2 = 11–25%; 3 = 26–50%; 4 = 51–75%; and 5 = >75% based on the number of sporocarp showing disease against the total mushrooms harvested from the baskets. Based on the rating scale, the A. bisporus strains were classified as either resistant or susceptible ( 3 = resistance (R) and >3 = susceptible (S)). The severity indexes were subjected to one-way analysis of variance, and significant mean dierences (P = 0.05) were determined with Duncan’s multiple range test using GenStat 12th Edition version 12.0.0.3033 (VSNI, Hemel Hempstead, England). The experiment was repeated three times in a completely randomized design with three replicates per M. perniciosa isolate. The same batch of compost was used for each experimental trial. Also prior to each trial, the pathogenicity of each isolate was tested on the A. bisporus caps to confirm their pathogenicity before the trial. All trials were subjected to the same environmental conditions (temperature and relative humidity) and routine rigid management was maintained in a clean environment to prevent contamination from other pathogens. M. perniciosa isolate WH001 inoculated on A. bisporus strain CCMJ1020 was used as a standard for each trial to detect the eect of variation in growth room conditions on symptom expression.
4.4. AFLP Analysis The AFLP reactions were carried out as described by Vos et al. (1995) [31] with modifications. The adapters and primers used in this study are shown in Table 3 and were purchased from Genset Oligos, France and IBB PAN, Poland. Restriction digestion and adapter ligation were performed simultaneously in a 20 L reaction volume made of 4 L (50 ng/L genomic DNA, 1 L Adapter, 2 L (5 units (U)) HindIII/MseI (New England Biolabs Inc., Ipswich, MA, USA), 2.5 L 10X Reaction buer, 2.5 L 10 mM ATP, 1 L (1 unit) T4 DNA Ligase (New England Biolabs Inc., Ipswich, MA, USA) and 7 L H2O. The reaction mixture was centrifuged for 15 s, incubated at 37 C for 5 h, held at 8 C for 4 h and stored overnight at 4 C. The quantity and quality of the digested products were observed using electrophoresis on 1.5% agarose gels stained with GelRed, visualized and photographed using Bio-Rad Gel Doc XR+ system (Bio-Rad Laboratories Inc., Hercules, CA, USA). Nonselective PCR pre-amplification was performed on the digested and ligated template DNA using non-selective primer pair HindIII/MseI in a total volume of 25 L. The PCR was performed in a T-Personal thermal cycler (Biometra, Göttingen, Germany) with the following settings: 94 C for 2 min followed by 30 cycles of 30 s at 94 C, 30 s at 56 C, and 80 s at 72 C. The final thermal cycle was followed by a 5 min extension at 72 C and (hold temperature conserved at 4 C for the moment) and stored at 20 C before gel electrophoresis. The PCR products were diluted in 20-fold with TE buer. The selective PCR amplification was performed in 25 L total volume containing eight dierent primer pairs consisting of HindIII combined with MseI (Table 3). All reactions were carried out in a T-Personal thermal cycler (Biometra, Göttingen, Germany) with the following settings; first-round amplification, 94 C for 2 min followed by 12 cycles of amplification, with a decreasing annealing temperature of 0.7 C/cycle: 94 C for 30 s, first annealing for 30 s at 65 C (the annealing temperature was influenced by primers Tm), 72 C for 80 s, and next 23 amplification cycles of 94 C for 30 s, 55 C (the annealing temperature was influenced by primers Tm) for 30 s, and 72 C for 80 s. The final thermal cycle was followed by the extension of 5 min at 72 C. The PCR yields were stored at 4 C till subsequent analysis. Five L of loading buer (GelTM Vilber Lourmat, Collégien, France) were added to 25 L of the PCR products. The mixture was loaded on 1% polyacrylamide gel in 1 TBE buer (89 mM boric acid, 89 mM Tris base, 2 mM EDTA, pH8.0) and run in the Agagel Mini, Biometra electrophoresis system (Biometra, Göttingen, Germany) was run at 200 V in TBE buer for 20 min. The gels were stained with GelRed (Biotium, Inc., Fremont, CA, USA), visualized and imaged on a UV transilluminator (Vilber Lourmat FLX-20M, Collégien, France). The DNA samples of each isolate were extracted three times from fresh fungal cultures and fingerprinted twice to estimate the reproducibility of the AFLP band patterns. The electrophoretograms were examined using GeneScan® Analysis Software (Applied Biosystems, Inc., Foster City, CA, USA). AFLP markers were physically scored as binary data for the existence or nonexistence of fragments between 70 and 500 bp. This binary data obtained was later used to estimate the Jaccard’s pairwise similarity coecients as applied in FreeTree version 0.9.1.50 program [32]. The unweighted-pair-grouping method with arithmetic average (UPGMA) dendrogram was produced from DNA band patterns using the Nei and Li correlation coecient [14]. The phylogenetic tree was viewed and edited using NTSYSpc version 2.02 (Exeter Software, Setauket, New York, USA).
From the single ultimate study I have found so far, Genetic and Pathogenic Variability of Mycogone perniciosa Isolates Causing Wet Bubble Disease on Agaricus bisporus in China
so it would seem that this needs big daddy money lab and that bodhs previous post disputing the existence of mycogone p. in cubensis is... uhh... insufficient. This requires DNA analysis not 30 minutes peeping at a few slides.
Of great note is this
Quote:
On PDA chlamydospores were not observed on Hp2 and Hp9, similarly, conidia were not observed for H2, Hp8, and Hp9 (Supplementary Table S1). The isolates with brown colony morphology produced more conidia compared to the isolates with white colony morphology.
errr duh why didn't I think if this. I'm sure it would be a lot easier for one of us to get ahold of this "M. perniciosa isolate WH001"
or any other mycogone p. isolate
and we should be able to just fuck with some healthy fruits/tubs/plates and observe.
Obs we aint doing DNA but if any readers have access...
wait..is that what you just said lmao
well I'm not allowed within 1000 feet of a school by national treaty, so one of you fuckers will have to get access to some mycogone perniciosa isolates please and thank you
Edited by sandman420 (11/06/21 06:34 AM)
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Kizzle
Misanthrope


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I think you'd be better off isolating it yourself. A strain used for agriculture may not be a strain that infects mushrooms but if you get it from a mushroom substrate it's more likely it will be.
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sandman420
Saint PP



Registered: 06/17/04
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Re: Myco Parasites [Re: Kizzle]
#27532242 - 11/06/21 08:37 AM (2 years, 2 months ago) |
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Well it would just take the guess out if we take known mycogone p. and can infect and cause the symptoms.
I don't have anything doing the bubbles right now plus we aren't in agreeance that the bubbles are even that in the first place. But we can get known mycogone p.
I'm sure there is availability in academy culture banks and it's not like regulated or anything so someone could easily snatch some worry free.
Oh snap here we go $140 euro and it's ours. This is isolated from infested mushrooms. Should do us what we need.
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Yeetusdeetus



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Re: Myco Parasites [Re: Kizzle]
#27532282 - 11/06/21 09:23 AM (2 years, 2 months ago) |
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Quote:
Kizzle said: I think you'd be better off isolating it yourself. A strain used for agriculture may not be a strain that infects mushrooms but if you get it from a mushroom substrate it's more likely it will be.
I’ve read gathering samples that come directly from the fruiting substrate is fairly important when attempting to grow the white jelly mushrooms from the wild.
https://www.mushroomcompany.com/articles/0005/tremella.pdf
The way I understand it, the mushrooms physical features are caused by a species of yeast parasitizing a dark puffball shaped mushroom.
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rockyfungus
dirty


Registered: 03/01/21
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 Not sure if mycovirus. Hispanica I keep germing and getting a feathery appearance with zones of clearing as it grows. It never forms a nice round colony just feathers.
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sandman420
Saint PP



Registered: 06/17/04
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I'm not familiar with that species on agar but here is a pic from workman. Looks like it may be normal for it to make weak feathery mycelium.
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QM33
(NOT A PUPPET!) ❤❤❤❤❤



Registered: 04/09/20
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Loc: Oregon
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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.
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sandman420
Saint PP



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Re: Myco Parasites [Re: QM33]
#27560364 - 11/28/21 06:35 AM (2 years, 1 month ago) |
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Not yet but I will be ordering some Mycogone perniciosa culture soon from Germany after Christmas.
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QM33
(NOT A PUPPET!) ❤❤❤❤❤



Registered: 04/09/20
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Ya keep is posted.
Funny besides p9 using a link to explain his hypothesis I swear I asked alot of these questions originally and even provided one or two of these links.
Thanks P9!

-------------------- 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
aspiring sociopath


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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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xVadisx
MountainsDogsAndFungi



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Re: Myco Parasites [Re: QM33]
#27580963 - 12/14/21 09:52 AM (2 years, 1 month ago) |
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I've been dealing with a series of fucked over the past few days that I suspect is some real nasty nasty. I've posted in a few places but sandman asked me to pop this over here so here are some quotes:
Quote:
xVadisx said: It was near a few bags that looked like this:

Several rosecomb mushrooms. A few with early broken veils, rosecomb heads, and dark ripped-looking (but not split) stipes. Lots of dry-bubble-looking/yellow/blobby stuff. I saw it in ziplocks and from a cake of the same variety. I've thrown away the worst in fear.
I've isolated everything else outside the grow area and am letting things go and debating harvesting/using. I probably will go ahead and let them grow and harvest at least a first flush...I just don't want to fuck my upcoming grows/plates/work area. I have six PF cakes that I already took some tissue to agar from (a super spikey cake) but am now hesitant to continue with as I don't want to transfer ick.
I despise losing actives, but with so many things coming up that are clean, I want to keep them clean and am not that attached. I still don't want to waste unnecessarily. I have a shoebox and two ziplocks as well that looks much much healthier but were from the same timeframe/CV-batch.
So, the grow room just has jars of oats and plates and has been cleaned. Air filter is running for whatever its worth. Everything else is in a completely other room. I'm not going to mist the cakes any more. They do what they do now. One is doing a pushup.
 
Quote:
xVadisx said: Is it okay to dehydrate bacterial fruit at 165F?
Logically, there is a fan... and it will blow hot air at these things moving air around (and I'm mostly curious if this air will then bring clouds of invisible-to-the-naked-eye-fuckedness to my home, a scientific term).
I haven't flipped the switch yet. I hate tossing actives.
 
If it is as nasty as I think I'm paranoid about spores getting everywhere, carried on me, etc. Gah.
-------------------- Thanks to everyone for sharing what they know . 165F for 24hrs. Better Holes Tek (Use a Step Drill Bit) Don't forget your towel: THE HITCHHIKER'S GUIDE TO THE SHROOMERY
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sandman420
Saint PP



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Re: Myco Parasites [Re: xVadisx]
#27670149 - 02/23/22 07:31 AM (1 year, 10 months ago) |
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Quote:
In contrast to viruses of plants and animals, mycoviruses uniformly lack an extracellular phase to their replication cycle. Consequently, they are not infectious in the classical sense. Infections cannot be initiated by exposure of uninfected hyphae to cell extracts prepared from an infected fungal strain. Rather, mycoviruses are transmitted by intracellular mechanisms such as anastomosis (fusion of hyphae) or through asexual spores.
Can anyone help me understand this?
Quote:
Both protocols require the generation of cell-wall-free spheroplasts from virus-free C. parasitica strains. For transformation, a plasmid, that contains a full-length hypovirus cDNA copy and an independent selectable marker gene, is introduced into C. parasitica spheroplasts by DNAmediated transformation. Transformants that contain the chromosomally integrated plasmid and cDNA-derived cytoplasmically replicating hypovirus RNA are selected following cell-wall regeneration and growth in the presence of the appropriate antibiotic (Choi and Nuss, 1992). The hypovirus transfection system uses synthetic transcripts corresponding to the hypovirus coding strand RNA (12.7 kb in the case of hypovirus CHV1/EP713) that are synthesized in a T7-polymerase-dependent cell-free transcription system. The synthetic transcripts are introduced into spheroplasts by electroporation and followed by cell-wall regeneration in the absence of any selection (Chen et al., 1994). Replicating hypoviruses are able to migrate through the cytoplasmic network of the regenerated hyphal colony.
You see, I'm not very bright but I have the spirit! I gots moxy!
Quote:
Mycoviruses are able to readily spread through the hyphal network that comprises a fungal colony.
from here
I think this means that only spores from infected hyphae and the hyphae fusing can spread the virus? How does it start then? I don't fully grasp what this means.
Edited by sandman420 (02/23/22 07:36 AM)
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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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Registered: 04/09/20
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By asexual spores do they mean chlamydospores?
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sandman420
Saint PP



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Re: Myco Parasites [Re: QM33]
#27670251 - 02/23/22 09:43 AM (1 year, 10 months ago) |
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I know that the uhh..spores of infected hyphae carry the virus so it spreads that way. I recall reading one paper that said something to the effect of 80%+ of spores from the infected fungi have the virus.
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QM33
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Registered: 04/09/20
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""Rather, mycoviruses are transmitted by intracellular mechanisms such as anastomosis (fusion of hyphae) or through asexual spores.
I mean basidiospores are not asexual right?
I mean this is what I got for 5 seconds on Google
Are Basidiospores asexual? No. The life cycle of Basidiomycota can be divided into two phases – sexual and asexual. Basidiospores are used in sexual reproduction.Jan 14, 2019 https://www.bustmold.com › resources
Idk this might be worth a read https://www.sciencedirect.com/topics/immunology-and-microbiology/asexual-spore
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sandman420
Saint PP



Registered: 06/17/04
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Re: Myco Parasites [Re: QM33]
#27670296 - 02/23/22 10:37 AM (1 year, 10 months ago) |
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Ok well I think the hangup is on that one paper simply using the word asexual spores there. They may have been specifically referring to the fungi in question on that paper, I think it was a rust blight or something and guessing it makes chlamydospores?
It definitely spreads through the spores of mushrooms, which correct me if I'm wrong would be basidiospores right.
Quote:
While sharing some characteristics with animal and plant viruses, mycoviruses also have the following unique characteristics: (1) most mycoviruses lack an extracellular route for infection; (2) mycoviruses are transmitted intercellularly only through cell division, sporulation, and cell fusion; and (3) mycoviruses apparently lack a movement protein, which is essential for the life cycle of animal and plant viruses.
On this paper they don't specify asexual spores.
Edited by sandman420 (02/23/22 10:49 AM)
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sandman420
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Now I present to you exhibit Q
I believe this sandbag to be infected with a mycovirus or mycoparasite.
This is just an example of my last run of several sandbags, every bag was identical to this pretty much.
The culture was a plate pin clone culture that looked, not to toot my own horn but it looked way better than anything you'll ever grow in your life sucka ...
It's natalensis.
The plates were so clean.
LC clean.
Grain was clean but ~80% of the bags stalled.
The grains that finished looked fine, they all looked fine but they would stall after shaking. No stank. No bahturia even on the stalled ones.
So i spawned my few sandbags with the good grain and they colonized mostly ok. Took a long fucking time to pin.
Heres what we gots homey...
Mostly less than 1" tall. NO SPORES. Very shitty. The plate clone was a huge spaghetti monster and sporulated LIKE CRAZY on the plate
Now you may say just genetics but fuck no man. This same shit has been happening to everything all kinds of cultures of cubensis too, getting worse every month till now I think it's hit max fucky.
I know that mycovirus (and probably mycoparasite too) effect the sporulation. I have been getting so many first flush no spores on so many projects.

So I know I have a huge fucking problem. I don't know how far it goes is the problem. I think everything in my culture storage is carrying this. I think all of my spores carry this.
I am shutting down my room, selling out, moving, ordering fresh spores from a virgin monk in Indonesia, and starting fresh. It's the only way to be sure.
Edited by sandman420 (02/23/22 03:18 PM)
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QM33
(NOT A PUPPET!) ❤❤❤❤❤



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Forrester
aspiring sociopath


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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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sandman420
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I wanted to move anyway.
But heres the thing, this is a fresh print that was sent to me from someone and I just worked with it on agar in the flowhood besides open air spawning so how the fucks did I get this fuckin fuck?! Fuck.
I would be lead to believe the mycoviruses are not air transmittable by my new learning. But not mycoparasites to my knowledge.
The fuck is going on.
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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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Dendrocopos
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Repost from agar envy thread. Sorry if that is not allowed. My cultures have all started to look kind off funky ever since i bough some maz spores. I get crazy rosecomb/mutants on everything i grow. Along weird mycogone like blobs of stem tissue oozing metabolites. Mr. Sandman told me i most likelly have some sort of mycovirus/mycoparasite based on my plates. Could someone give me a second opinnion before i toss all my shit ? Or, how does one get rid of something like that without tossing everything ?
Looks good, but rhyzomorphs weirdly lift into the air like they are escaping from something.

weirdly ununiform growht

3D/fuzzy look. Looks like bacterial bubbling.

?clean plate? i am so paranoid, i don't know at this point
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sandman420
Saint PP



Registered: 06/17/04
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Bacteria as a mycoparasite is my main interest right now. We need more information on that. I bet it's a big one.
I think your last plate is looking good.
But your other picture sure do show a lot of intersting stuff that if linked to your problem could be handy for others.
First pic the multiple layers is odd. The tendril like upcropping's in the middle are noteworthy.
2nd picture the petering off and confused unorganized look is noteworthy.
3rd picture I mean dang what's going on with the grainy spots?
Love to hear someone else talk about this for a minute too
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rockyfungus
dirty


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Unrelated but possibly related? Was reading about the parasites I may come in contact with in my line of work. The trophozoites are mobile and consume bacteria (which allows for the diagnosis on E. coli plates). The trophozoites form double walled cysts which are incredibly resistant to methods of eradication (including freezing, heating, and irradiation).
Not going to dig but do they eat fungi?
https://en.wikipedia.org/wiki/Trophozoite
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GandalfTheWhite
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Registered: 05/02/21
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Saw that there was a couple posts in General Disc. in regards to this subject, but then noticed this thread was created. Apologies for resurecting an older thread.
Dealing with something quite similar with my APE culture. Swabbed a few plates, put some cotton fibers on the plate, snapped the tip off on a couple dishes and nothing looks right.
This is one of the T1's from the germ plates that I made early on. As you can see the scalloping and disorganized growth is obvious. Even the germ plates are starting to go south with discoloration and some bruising of the myc.

Think this culture is going to be a wash and will have to start over with another source
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Stipe-n Cap


Registered: 08/04/12
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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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GandalfTheWhite
Newb


Registered: 05/02/21
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Re: Growth Inhibitory Compound Discussion (BLIS-Chitinase) [Re: Stipe-n Cap]
#28007229 - 10/19/22 10:40 PM (1 year, 3 months ago) |
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Thats kind of my plan ever since I noticed all this ragged growth. I've been searching around the past few days trying to find something decent that would work for this type of application. Feel like a stereo scope would be good for looking at plates and early transfers, but for this I would need a compound scope to see whats going on. I'd really like to look into stuff like this even though I could easily toss the plate and start over, but this has me curious.
My current dilemma being I want both.. at some point. Just not sure where to start comparing all the different specs or whats decent/trash
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