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OfflineTreycherry
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Agar/mycelium questions
    #26944617 - 09/19/20 05:35 PM (3 years, 4 months ago)

Hey guys! I’m on my third transfer with my clone sample and I’m curious what I should be looking for as an indicator to stop. This dish is a lot more uniform looking than the previous two so I feel as if I’m on the right path.

Secondly, on my first grow the mycelium in the jar was very puffy looking ( I used a multi spore ).
This time I used my clone that I put to agar and it appears way more stringy. I’m assuming it’s a good sign but I’m still curious as to why it appears different and if it’s actually better or not.



Thanks for any information:)


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InvisibleLenz
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Re: Agar/mycelium questions [Re: Treycherry]
    #26946842 - 09/20/20 08:21 PM (3 years, 4 months ago)

It's my understanding that the myc in your multi spore jar was so puffy because when you inoculate with a MS syringe there are countless different genetic strains growing together and through each other. Essentially, there are a bunch of genetically distinct individuals that have all germinated from their respective spores and are growing hyphae and mycelial networks and competing with each other. Hence, a puffy dense mycelial network.  With a clone however, the mycelium grown from a cloned fruit should technically be genetically identical to the fruit you cloned it from. Therefore, you have a pure culture as opposed to the polyculture clusterfuck that is the result of a MS injection.

It looks like your clone culture is rhizomorphic (stringy) culture rather than tomentose (puffy) (which is dependent mostly on genetics but also nutrient content of your agar and probably some other factors). That's why the myc in your jar looks like that. I'm not sure if rhizo is necessarily better, people seem to like rhizomorphic growth probably because it looks cool/is aggressive/is easily identifiable but again, I don't think that really tells you anything about yields or fruits or other favorable traits.

On the question of when to stop, I think 3-5 transfers is usually fine--theoretically, if your sterile tek is on point though, the initial plate inoculated with the clone tissue should be safe to use without transferring. Spores are inherently not sterile so that's why transfers are taken but I'm not certain if it's really necessary with a well done clone inoculation. Basically though, just transfer until you're sure it's clean.

Hope this helps a bit, I'm definitely still learning though so this might not be the most accurate answer.


--------------------
:chems:


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OfflineAlsetAlokin
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Registered: 07/30/20
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Re: Agar/mycelium questions [Re: Lenz]
    #26946865 - 09/20/20 08:45 PM (3 years, 4 months ago)

I would stop when the plate looks uniform with no visible contamination.

Lots of variables. From what I've read, types of growth (in spawn) have little or no impact on the final product, but can be a good indicator of it fighting contamination, and in/correct conditions.

Stringy or rhizo growth on plates is helpful for noticing contamination that may be tagging along because you can see through the myc to the agar underneath.


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OfflineFlufferNutter
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Re: Agar/mycelium questions [Re: Lenz]
    #26946871 - 09/20/20 08:51 PM (3 years, 4 months ago)

Those look good man! Let them grow out a little bit, watch how it fills out the plate. There's more to those colonies than meets the eye rn, trust me.

@Lenz you're pretty much right on man. Only thing is, by the time you can see any fuzzy growth, the mycelium is already dikaryotic(sp?)and clamps are all established. All the combos possible have been made and are the strands that you are seeing.

  Rhizo and tomentose both serve functions that are not mutually exclusive, you need both, and whether it's obvious or not, in any healthy, mature culture you will have both. I'm not formally trained in mycology by any means. I had the very same hang up with the biology as well. It might not be the best, but my podunk way of looking at it is that the fuzzy myc is happy just eating, the rhizo is looking for dinner. Tomentose myc will spring from rhizo and fill out the area around it, set up shop, and ideally, organize more rhizo. Rhizo is suited for claiming new territory quickly. 



Looking good though Trey...keep it up!


Edited by FlufferNutter (09/20/20 09:01 PM)


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OfflineTreycherry
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Re: Agar/mycelium questions [Re: FlufferNutter]
    #26947520 - 09/21/20 10:13 AM (3 years, 4 months ago)

Thanks for all the information guys! That answered a lot of questions I had


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