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OfflineSandala
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Registered: 02/20/20
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Last seen: 1 year, 9 months
Is this 'tufty' growth normal?
    #26789249 - 06/28/20 09:01 AM (3 years, 6 months ago)

Does this 'tufty' growth look Healthy? The lack of rhizomorphic growth has me questioning everything. 

Conditions have not been ideal, the jars were done too early and stored for a few weeks before colonisation (4 weeks ago), then after seeding from agar temps got up to 85F for a couple of weeks and even tho it's now cooled down a bit everything seems to have stalled.

I tried to get a pic of the clone plates showing how 'tufty' the growth looks, it doesn't show too well but that fine 'cotton wooly' looking growth is 1/4" thick off the plate.

I've kept all my plates from spore, transfers up to T4, and clone transfers to T3, although all the plates are now fully colonised to the edge, they all seem to have some of this 'tufty' growth.

Is it worth starting again from spore, or is this 'tufty' growth normal for cubes in which case I can knock up some more grain and seed straight away.



got no black spores or discolour on anything, all samples looks white pretty much the same as in these pics

I did 4 jars at the same time, 3 out of 4 didn't seem to bounce back after shaking here's one of them...




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Edited by Sandala (06/28/20 09:26 AM)


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Invisiblesandman420
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Re: Is this 'tufty' growth normal? [Re: Sandala]
    #26789317 - 06/28/20 09:28 AM (3 years, 6 months ago)

It's not normal. I think you have a ride-a-long contamination that is intertwined and living in symbiosis with the mushroom mycelium. Probably white mold AKA mycogone aka wet bubble. Also could be verticillium aka dry bubble which is also white mycelium. Notice on agar there is no group effort at a defining edge. Each individual infected sub strain is afraid of each other and will not consolidate because there is a sickness in it. For fun you could make a selective agar for verticillium that allows it to outgrow the cubesnsis.

Quote:


FROM https://extension.psu.edu/verticillium-dry-bubble about vert

The easiest method to identify this pathogen is to culture it on selective media. This selective media was developed to allow Verticillium to outgrow any competitors. Details on how to prepare the media are Rinker et al., 1993. When Verticillium is grown on potato-dextrose agar (PDA), at room temperature, the colonies are white and the underside of the plates is colorless to yellow. Some cultures may have a low profile with rounded, scalloped edges. If one can measure the spores under a microscope, they would be about 8-10 mm long by 1-3 mm wide and tapering to 0.5-1.0 mm at the tip. The important morphological characteristic from a dissemination and disease control perspective is that the spores are produced in a gelatinous matrix. This gummy material will collect and hold many spores together.




I'm not sure on the exact makeup of a selective media I will look into it. PDA based though.

EDIT: Found one but it's like a damn 5 star chef recipe including 2 fungicides which I was under the impression kills vert but anyway here it is

https://www.researchgate.net/publication/240511870_A_selective_medium_for_Verticillium_fungicola

edit: that last jar pic you added is bacterial


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Edited by sandman420 (06/28/20 09:46 AM)


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Invisiblemaxmush
Always learning...
Registered: 06/13/20
Posts: 440
Re: Is this 'tufty' growth normal? [Re: Sandala]
    #26789346 - 06/28/20 09:40 AM (3 years, 6 months ago)

It looks to me that your agar transfers where not sectored in the "optimal" spots to start off. Although most will prefer rhizo vs tomentose either will likely produce the results you are looking for. I personally dont see any negative signs of contam in there, but i think there are better experts at eying this out.

It is very possible the temp swings had something to do with it as i noticed more  tomantose on tubs that had inconsistent temp vs constant temp which usually shows more rhizo.

How long after the shake up was that pic taken?


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Disclaimer: all information presented is intended for educational purposes only. All photos are only representations and not directly from the user.


Edited by maxmush (06/28/20 09:42 AM)


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OfflineSandala
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Registered: 02/20/20
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Re: Is this 'tufty' growth normal? [Re: maxmush]
    #26789395 - 06/28/20 10:01 AM (3 years, 6 months ago)

Thanks for replies chaps.

Quote:

sandman420 said:
It's not normal. I think you have a ride-a-long contamination that is intertwined and living in symbiosis with the mushroom mycelium. Probably white mold AKA mycogone aka wet bubble. Also could be verticillium aka dry bubble which is also white mycelium. Notice on agar there is no group effort at a defining edge




Bugger.

Read Rinker et al 1993, no way I'm going to be able to source those chemicals to make selective agar.

I've got access to a decent microscope but even if I sample up some slides I'm not sure I could tell the difference between Verticillium & Cube myc :justdontknow: and doubt I'd be able to get pics to share either

since my spores were taken from the same grow as the clone was taken from, is it likely they will also have this contam?



Quote:

maxmush said:

How long after the shake up was that pic taken?




2-3 weeks


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InvisibleRoger Clemency
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Re: Is this 'tufty' growth normal? [Re: Sandala]
    #26789422 - 06/28/20 10:14 AM (3 years, 6 months ago)

The jars don't look good. Neither that first one or the one that never recovered from shake. The plate looks like a germ plate with some crazy looks going on. Did you innoc from that plate or from plates that had been transferred until clean?


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Invisiblemaxmush
Always learning...
Registered: 06/13/20
Posts: 440
Re: Is this 'tufty' growth normal? [Re: Sandala]
    #26789435 - 06/28/20 10:18 AM (3 years, 6 months ago)

wow 2-3 weeks. there is definite contam then :frown: jars may have had too high moisture content and/or grains burst with contam in them. sorry


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OfflineSandala
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Registered: 02/20/20
Posts: 197
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Last seen: 1 year, 9 months
Re: Is this 'tufty' growth normal? [Re: Roger Clemency]
    #26789496 - 06/28/20 10:42 AM (3 years, 6 months ago)

Quote:

Roger Clemency said:
The jars don't look good. Neither that first one or the one that never recovered from shake. The plate looks like a germ plate with some crazy looks going on. Did you innoc from that plate or from plates that had been transferred until clean?





Nah the innoc was from a T4 that looked clean, originally from MSspore.

downside of using sauce pots is it's real hard to get a decent pic, this transfer was taken from the same sample went into jars (best pic out of 10 attemps  :whiteflag: )



since my spores were taken from the same grow as the clone was taken from, is it likely they will also have this contam?


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Invisiblesandman420
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Re: Is this 'tufty' growth normal? [Re: Sandala]
    #26789546 - 06/28/20 11:02 AM (3 years, 6 months ago)

It is probably that all the prints from that grow are contaminated if that was verticillium. The way that verticillium normally works is it attaches itself to the cap of the mushroom at the primordia stage by lying dormant on the surface of the mycelium only activating when rhyzomorphs appear. Then after the mushroom matures the vert makes a grey or brown slimy sticky colony on the top middle of the cap which is where it is sporulating. So naturally when you go to take a print you get sticky vert spores all over that and then the mycoparasite is alive and well in your culture through it's entire life.

Verticillium is one of if not the biggest problem in commercial mushroom growing and it has devastated multi million dollar edible operations. Some times you gotta move states and change religions to get away. Even fungicides are not effective anymore I am reading as it developed resistance to all they were using.


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Marijuanaut escapes earth to cultivate - Grow-room is church temple of the new stoner breed


Edited by sandman420 (06/28/20 11:09 AM)


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OfflineSandala
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Registered: 02/20/20
Posts: 197
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Re: Is this 'tufty' growth normal? [Re: sandman420]
    #26789580 - 06/28/20 11:15 AM (3 years, 6 months ago)

Best options for going forward?

I have 2x cube spore syringes from same vendor each with about 1ml left, one is the line all these pics are from, that grew out a kit grow that I collected spores from, same grow as my avatar. 2nd syringe came as a free sample, I tried to start some on agar but only contam showed up (mite have been my SAB technique tho)

Although I got a few shrooms from the kit (and a spore print) I fucked it up keeping it too wet with not enough FAE so it contamed out

So bin all my agar samples, and the prints I took from the kit grow, and start again onto agar from these two spore syringes hoping they weren't the source of the contam?

I suppose I could also put some syringe spore on a microscope plate to see if there's any contam, I've not got a calibaration slide but surely I would see a difference in spore size?


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Edited by Sandala (06/28/20 11:31 AM)


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Invisiblesandman420
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Re: Is this 'tufty' growth normal? [Re: Sandala] * 1
    #26789713 - 06/28/20 12:30 PM (3 years, 6 months ago)

I would order a brand new print from a brand new supplier and start new agar plates with no pour style filtered lid agar.


--------------------
- Sandbag Tek - How To Sterilize Spawn Bags - All About Static Pressure / Pressure Drop for DIY Flow Hoods - Sandman's LC Tek-

Marijuanaut escapes earth to cultivate - Grow-room is church temple of the new stoner breed


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OfflineSandala
Noob Shroomer

Registered: 02/20/20
Posts: 197
Loc: UK Flag
Last seen: 1 year, 9 months
Re: Is this 'tufty' growth normal? [Re: sandman420]
    #26790077 - 06/28/20 02:58 PM (3 years, 6 months ago)

Thanks Sandman, toally get what you're saying, thanks for all your help.

Ordering new print.

in the meantime I got my scope out of storage, cleaned & set her up, see if I can't learn something useful from this disaster while I wait for the print to arrive.



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