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OfflinerenagaderX
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Registered: 08/13/18
Posts: 103
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Newbie's first time with Agar. Need help differentiating between cobweb vs tomentose?
    #25471665 - 09/19/18 01:57 AM (2 years, 7 months ago)

Hey everyone !

I've had 2 successful grows so far, mini monotubs, and was fortunate enough to have purchased good MS syringes from a local supplier that were contaminant free.

I've started getting into agar, and I find I'm having trouble telling between cobweb mold vs tomentose growth on my agar plates.

I'm lead to believe it is tomentose, and some of the strains haven't mated to produce the rhizomorphic growth that is desired, but here are some pictures of some of my agar plates.

Guidance, criticism, and comments are needed.

Please and thanks

Agar plates I need help distinguishing between:

#1

#2

#3

#4

#5

#6

#7



Some rhizomorphic growth :



This guy looking pretty good to me !



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OfflineDcevLearn
Team Tomentose

Registered: 07/07/18
Posts: 289
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Re: Newbie's first time with Agar. Need help differentiating between cobweb vs tomentose? [Re: renagaderX]
    #25471954 - 09/19/18 07:13 AM (2 years, 7 months ago)

I don't have a ton of experience but I can tell you that I've put 7 different varieties from print or syringe to agar and they all look like your examples.  I still don't have any that look close to your final good looking ones even after many transfers. 

A lot of people say it comes down to how much nutrition is in your agar mix.  You should reply back with what recipe you are using.

I have heard of an experiment you can try.  Take a wet piece of cardboard and pressure cook it in a plastic container.  Do an agar transfer to the cardboard and the culture should really extend it's legs looking for nutrients.


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Current Project: LAGM: Stones From Scratch in 2020



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Offlineteladi
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Re: Newbie's first time with Agar. Need help differentiating between cobweb vs tomentose? [Re: DcevLearn]
    #25471998 - 09/19/18 07:51 AM (2 years, 7 months ago)

What happened with 4-6? 4 looks to have a gigantic agar wedge and 5-6 looks like their transfer wedges rolled around.

The look fine for now, even though your camera focused on the lid. Don't worry too much about rhizo vs tomentose growth, especially with MS cultures.


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OfflinerenagaderX
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Re: Newbie's first time with Agar. Need help differentiating between cobweb vs tomentose? [Re: DcevLearn]
    #25472323 - 09/19/18 11:15 AM (2 years, 7 months ago)

It was all transferred from 2 different agar plates, T-2 (Transfer 2) from Z strain.

Those good looking ones are clones, which are demonstrating nice rhizo growth imo.

10g Light Malt Extract
10g Agar
500g Water


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OfflinerenagaderX
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Re: Newbie's first time with Agar. Need help differentiating between cobweb vs tomentose? [Re: teladi]
    #25472328 - 09/19/18 11:17 AM (2 years, 7 months ago)

4 is a gigantic agar wedge ( Still working on my transferring tek, getting as little as possible / only what is needed to transfer )

5-6 I tried to center it a bit more I think, when I accidentally missed the center when dropping.



Just gonna wait it out and hope I'll be able to use a few of these plates into some jars


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OfflineSpikeSpiegel
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I'm a teapot


Registered: 07/30/18
Posts: 162
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Re: Newbie's first time with Agar. Need help differentiating between cobweb vs tomentose? [Re: renagaderX]
    #25472564 - 09/19/18 01:45 PM (2 years, 7 months ago)

those all look like cube mycelium to me.


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Offlineteladi
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Re: Newbie's first time with Agar. Need help differentiating between cobweb vs tomentose? [Re: renagaderX]
    #25472570 - 09/19/18 01:47 PM (2 years, 7 months ago)

Ha! You don't need that much, except when you're dong A2G (Agar to grain). Less is more: You'll get fewer strains and less of a chance of transferring hidden contams cruising along for the ride. My transfers are tiny, just a little spec of tissue.

Here are some pics for you to examine: https://www.shroomery.org/forums/showflat.php/Number/22020260. You'll notice it's often easier to spot hidden contamination when you let your plates grow out; easier to identify uniformity or the lack thereof.

Overall, it looks like you're on a good track.


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OfflineSolipsis
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Re: Newbie's first time with Agar. Need help differentiating between cobweb vs tomentose? [Re: teladi]
    #25472586 - 09/19/18 01:54 PM (2 years, 7 months ago)

cobweb is more hairy and faint looking..

tomentose growth can be a sign of having just a bit much macronutrients IIRC, or a humidity thing although the latter would result rather in aerial growth.

Although you do wanna pick rhizo sectors I wouldn't worry about it too much.

But also I would not try to fully isolate starting from MS without testing unless you are going to save, store and test a shitload of isolates.


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OfflinerenagaderX
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Re: Newbie's first time with Agar. Need help differentiating between cobweb vs tomentose? [Re: teladi]
    #25472678 - 09/19/18 02:29 PM (2 years, 7 months ago)

Thanks for the link and the help from everybody!

So with that reference link to hidden contams teladi posted, can we refer to the sneaky bacteria pic vs pic 3 of my Z strain?

Not sure if that bit of slight white film can be considered hidden contam then? If you look at #4 its much more clear.

Thoughts and opinions?


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Invisiblemary fairchild
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Registered: 12/06/14
Posts: 772
Trusted Cultivator
Re: Newbie's first time with Agar. Need help differentiating between cobweb vs tomentose? [Re: renagaderX]
    #25472858 - 09/19/18 03:42 PM (2 years, 7 months ago)

From what I can see your plates all look ok. Cobweb looks more like loosely organized filaments, nothing at all like tomentose form which is much denser.

Here's a pic of a cobweb- its at the edge of plate at the 10 oclock position:


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OfflineSolipsis
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Re: Newbie's first time with Agar. Need help differentiating between cobweb vs tomentose? [Re: renagaderX]
    #25472868 - 09/19/18 03:45 PM (2 years, 7 months ago)

Take #6:

things that aren't a good sign are a region of wetness (although i have on occasion seen certain agar wedges respond to certain media by "melting" a little, it seems), and also myc growing only in one direction from a wedge.

That suggests your myc is having a standoff with bacteria which were already there on your previous plate or got on the wedge during transfer..

I think cobweb mold is unlikely to just take the place of where your own myc should be in a stealthy way.. If you take a piece of agar which has a reasonable amount of good myc on it, in case of contam you should expect to see some kind of "fighting" going on with visible effects. Regions of inhibited growth between colonies and stuff like that.

Just seeing fuzzy growth is no reason to get suspicious or paranoid IMO..

Seeing a satelite colony on your plate gets exceedingly suspicious the further away it is from your wedge/point of inoculation.

Close to it is definitely possible cause the myc does not just grow on top of the agar but into and through it, so it can just dig and pop up for whatever reason although normally it wouldn't do that.. but funky things can happen with unresolved MS, but also just if a bit of myc touched or landed there during transfer of course.

Seems like you have enough plates right? just give the dubious ones a little more time, or make better sharp pics without reflection.


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OfflineGonzo the Eternal
In Sterquiliniis Invenitur


Registered: 05/09/18
Posts: 480
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Re: Newbie's first time with Agar. Need help differentiating between cobweb vs tomentose? [Re: Solipsis]
    #25524392 - 10/09/18 03:05 PM (2 years, 6 months ago)

Kind of an old thread but something you said Solipsis is intriguing. Why is it suspicious when a satellite colony appears far away from the inoculation point? I've had this problem recently. Both with a couple satellite colonies at the very edge of the plate and also with distinguishing cobweb from tomentous.


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