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Invisiblesandman420
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Re: Myco Parasites [Re: rockyfungus]
    #27525251 - 10/31/21 11:26 AM (2 years, 2 months ago)



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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Re: Myco Parasites [Re: Kizzle]
    #27526109 - 11/01/21 08:15 AM (2 years, 2 months ago)

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.


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Edited by sandman420 (11/01/21 08:44 AM)


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Invisiblesandman420
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Re: Myco Parasites [Re: Stipe-n Cap]
    #27526132 - 11/01/21 08:55 AM (2 years, 2 months ago)

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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Re: Myco Parasites [Re: Kizzle]
    #27526299 - 11/01/21 11:40 AM (2 years, 2 months ago)

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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Re: Myco Parasites [Re: Stipe-n Cap] * 1
    #27526330 - 11/01/21 12:10 PM (2 years, 2 months ago)

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

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




Quote:

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.





Quote:

Ihrmark, K., H. Johannesson, E. Stenstron, and J. Stenlid. 2002. Transmission of double-stranded RNA in Heterobasidium annosum. Fungal Genet. Biol. 36, 147–154.




Quote:

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.




Quote:

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.




Quote:

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.




Quote:

Raper, J.R. and C.A. Raper. 1972. Genetic analysis of the life cycle of Agaricus bisporus. Mycologia 64, 1088–1117.




Quote:

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,




Quote:

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.




Quote:

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.




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Edited by sandman420 (11/01/21 01:45 PM)


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Invisiblesandman420
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Re: Mycoparasite/Mycovirus Discussion [Re: Stipe-n Cap]
    #27526727 - 11/01/21 06:28 PM (2 years, 2 months ago)

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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Invisiblesandman420
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Re: Myco Parasites [Re: Yeetusdeetus]
    #27531650 - 11/05/21 05:13 PM (2 years, 2 months ago)

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.


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Edited by sandman420 (11/05/21 05:15 PM)


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Invisiblesandman420
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Re: Myco Parasites [Re: Stipe-n Cap]
    #27531854 - 11/05/21 08:15 PM (2 years, 2 months ago)

I read in another one of these studies they were using 30 watt phillips TUV lights for mutagenesis. But now were off topic again!


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Edited by sandman420 (11/05/21 08:16 PM)


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Invisiblesandman420
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Re: Myco Parasites [Re: Stipe-n Cap]
    #27532146 - 11/06/21 06:09 AM (2 years, 2 months ago)

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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Re: Myco Parasites [Re: Kizzle]
    #27532242 - 11/06/21 08:37 AM (2 years, 2 months ago)

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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Re: Myco Parasites [Re: rockyfungus]
    #27560350 - 11/28/21 06:09 AM (2 years, 1 month ago)

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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Re: Myco Parasites [Re: QM33]
    #27560364 - 11/28/21 06:35 AM (2 years, 1 month ago)

Not yet but I will be ordering some Mycogone perniciosa culture soon from Germany after Christmas.


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Re: Myco Parasites [Re: xVadisx]
    #27670149 - 02/23/22 07:31 AM (1 year, 10 months ago)

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.


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Edited by sandman420 (02/23/22 07:36 AM)


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Re: Myco Parasites [Re: QM33]
    #27670251 - 02/23/22 09:43 AM (1 year, 10 months ago)

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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Re: Myco Parasites [Re: QM33]
    #27670296 - 02/23/22 10:37 AM (1 year, 10 months ago)

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.


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Edited by sandman420 (02/23/22 10:49 AM)


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Re: Myco Parasites [Re: sandman420]
    #27670318 - 02/23/22 11:13 AM (1 year, 10 months ago)

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 :crazy2: :rocket: :evil:...

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.


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Edited by sandman420 (02/23/22 03:18 PM)


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Re: Myco Parasites [Re: Forrester]
    #27670330 - 02/23/22 11:29 AM (1 year, 10 months ago)

:fuck:

I wanted to move anyway.:feelsbatman:

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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Re: Myco Parasites [Re: Dendrocopos]
    #27675630 - 02/27/22 02:27 PM (1 year, 10 months ago)

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 :tomatoface:


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