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OfflineUzimyco
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Lurker finally posting question on “inconsistency of cultivation” * 1
    #27038563 - 11/14/20 10:35 AM (3 years, 2 months ago)

Hi All

I already know that my question is vague to the point it will be hard to answer but figured I’d take a shot as the frustration is a bit more than I can tolerate.

I am a highly experienced horticulturalist in a wide array of complex genera however new to mycology.

I have had a circumstance that simply doesn’t compute to me.  On multiple occasions I have had situations where (in a truly sterile environment) I have created spore syringes, cake media, sealing process and inoculation procedures however invariably some of my jars will roar out of the gate with mycelium development. Others almost weak and unimpressive and others virtually no development at all.

Now since media is all together.  Spores are dispersed in a communal beaker prior to being up drawn by syringe and all are inoculated and incubated together, how on earth can there be so much disparity in result. Again as an experienced horticulturalist I do not understand. This is all in controlled environments and purposely the variables have been excluded as to avoid such circumstances.

Just wanted to throw it out there if there were any opinions?  It is massively frustrating to me. Note:  I have no signs of contamination in any way shape or form. Again by using protocols that are on par with laboratory settings that is not an issue I have dealt with. It is the variability of the result that I cannot fathom.

I will admit that the only thing I can think of is the “amount of solution” introduced during inoculation however I am trying to use an equivalent amount of agitated solution which may be 1/10th of a cc diffferent here or there. It doesn’t seem likely that would be enough to be so pronounced.

Oh lastly. I have had the experience with a variety of cubensis strains so it isn’t limited to one type.

Thanks for reading. Please let me know if you have any thoughts.


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OfflineCorundum
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco] * 1
    #27038592 - 11/14/20 10:54 AM (3 years, 2 months ago)

Each pair of spores that come together to form a colony is like 2 people having a kid. The genes are a random combination of the genes from the gametes. This is why multispore is so random. Consider looking into mushroom cloning techniques


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Edited by Corundum (11/14/20 10:55 AM)


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InvisibleWall.E
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Corundum] * 1
    #27038616 - 11/14/20 11:11 AM (3 years, 2 months ago)

Because that's how fungi work. They're not seeds, they're hyphae. To increase their chances for success they have a huge genetic load that they blow containing insane amounts of DNA.

Some hyphae are great at what they do and forge forward, some suck and grow slowly.

That's why we do agar work first instead of ms syringes. Ms syringes are a crapshoot.


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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Wall.E]
    #27038994 - 11/14/20 03:19 PM (3 years, 2 months ago)

Many thanks for your replies.

They are genuinely appreciated. To go a step further I would very much like to take your advice and move to agar as this is wildly frustrating.

I realize that a quick search of the forums along with the internet will provide countless versions of material explaining the process. I would however much prefer to take some direction from folks like yourself as you’re experienced where I am not.

Would you be so kind as to post up a link of an agar process that is suitable however not overly complicated?  I just prefer to obtain information that has been vetted by experienced people vs stuff floating around. If you’d be willing to post a link I would be very appreciative.

Many thanks for taking the time to reply. Lurking only goes so far and I’m already glad I posted based on what you’ve provided.  I did not realize the potential benefits of agar vs a brf style tek. I can see it is obviously VERY worthwhile as going through the process for this level of hit and miss isn’t very fun.


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InvisibleWall.E
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27039006 - 11/14/20 03:23 PM (3 years, 2 months ago)

500mL water + 7.5g agar + 7.5g LME + Pressure Cooker + 50 Petri Dishes = Agar work

Boil water, add agar and LME, blend/mix/whisk it together. Pour into jar with vent hole, cover with foil PC for 20 minutes after venting for 10. Take it out, let it cool down a bit. Move everything to SAB, pour plates.

When working in SAB use small, slow movements, don't bump the box and keep everything sterile.

Or just follow D3monic's tek, he's good at agar.

https://www.shroomery.org/forums/showflat.php/Number/26628234#26628234


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OnlineLogicaL ChaosM
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27039041 - 11/14/20 03:42 PM (3 years, 2 months ago)

What kind of spore syringe did u have? Was it mostly clear?

Ive noticed with spore syringes that have really high concentrations of spores, for example one spore print for one 12cc spore syringe, u see explosive, consistent growth as the spore concentration on the substrate is so high.

I would say spore concentration is the biggest reason for varied germination times as well as inconsistencies in the PF material (too wet, too dry, wrong ratios, etc).


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Offlinetravels_slightly
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: LogicaL Chaos] * 1
    #27039058 - 11/14/20 03:55 PM (3 years, 2 months ago)

Quote:

LogicaL Chaos said:
What kind of spore syringe did u have? Was it mostly clear?

Ive noticed with spore syringes that have really high concentrations of spores, for example one spore print for one 12cc spore syringe, u see explosive, consistent growth as the spore concentration on the substrate is so high.

I would say spore concentration is the biggest reason for varied germination times as well as inconsistencies in the PF material (too wet, too dry, wrong ratios, etc).





Wait, so by this logic it's ideal to apply as much spore as possible to induce quicker colonization? Am I missing something or is that the sum of it? I only ask because I feel like I see the same reoccurring information being shared here: smaller the amount of MS inoculate, the better. Am I mistaken?


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InvisibleWall.E
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: travels_slightly] * 1
    #27039155 - 11/14/20 04:57 PM (3 years, 2 months ago)

The larger the spore density the more opportunities for mating which leads to more dikaryon which leads to more potential fruiting bodies.


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OnlineLogicaL ChaosM
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: travels_slightly]
    #27039206 - 11/14/20 05:36 PM (3 years, 2 months ago)

I think so.

My logic is that by using more spores, you are getting more genetic matches competing for each other. Its like survival of the fittest at the microscopic mycelium level.


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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: LogicaL Chaos]
    #27040030 - 11/15/20 07:21 AM (3 years, 2 months ago)

Thank you for that information.

Without question the syringes were quite dilute. Not necessarily on purpose but my prior spore experience relates to rare tropical ferns. VERY different situation. Too many spore in the fern world leads to over crowding which becomes a competitive mess.

But no doubt I “could” have used much denser concentration. As a matter of fact I believe I poured like 1 cup of spore solution down the drain as I had made many syringes and figured that it was just overkill.

Again as a new person to the hobby there are some things that aren’t obvious. For example. I would think that any activity even being it a bit would just go the distance and fully colonize anything. Maybe that’s true but it would take an amazingly long time. I have some jars that have “hard core” mycelium in them but it’s the size of a quarter and doesn’t seem to keep spreading out. Don’t know why but I’m learning very fast that what seems logical to me through my experiences are highly not applicable here. This is why I am thankful for such a forum as obtaining the info is slow even when speaking with highly experienced people. I don’t want I think what it would be relying on purely books and the like.

It’s an interesting thought however would likely require a significant number of prints if you’re suggesting using a print per 10-12cc. I’ve been innoculating these half pint jars with about 3-4cc. Maybe I could barely get 3 out of 1 syringe and this process isn’t exactly overnight so it seems counterproductive unless I had like a whole bunch of prints.

I like the idea of cloning or using agar to determine the higher quality of growth “before” the process really gets underway” as it makes much more sense.

I freely admit I will have to study the agar methods repeatedly for a bit.  I know for those experienced it looks easy. I’m a bit of an expert in some areas so things as such are simple to me whereas highly complex to others. It’s a kind of understanding that develops over time. So I do need to read, re-read, re-read, etc until I really understand the whole thing.

Again many thanks to all who’ve contributed. I attest a great deal of knowledge on quite a few subjects to forums such as this.  And this one is no different.  It’s likely one of the few things that I can categorically say is “good” about the internet.

Many thanks!


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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27040048 - 11/15/20 07:38 AM (3 years, 2 months ago)

I would cut out the middle man and just make prints, why bother going through the trouble and resources for a syringe?
Print to foil, use inoculation loop to collect spores from foil, streak agar plate.


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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Stipe-n Cap]
    #27040053 - 11/15/20 07:42 AM (3 years, 2 months ago)

:whathesaid:


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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Wall.E]
    #27046410 - 11/18/20 08:57 PM (3 years, 2 months ago)

Hi all. I’m convinced that I had already posted this but somehow I guess maybe I never submitted it.

Based on the information that you’ve all provided I think I have a pretty solid understanding of the agar process. Where my confusion comes in now is that after seeing the process of spawning on agar  And then further seeing the replating of quality growth on subsequent dishes, I’ve watched videos of the whole agar clump in essence dumped into a container that looks like a growing substrate such as brf? Or perhaps something else?

Is my impression correct? Once the agar spawn has been done and perhaps pieces transferred to other agar dishes to further grow out quality mycelium, is that what I am looking at? Where the entire agar disk with mycelium growth is placed into a container with some kind of nutrition such as brf to take it further?  I swear I know I posted this. I wonder if I did in in a different thread by accident?

Any of your advice again is most genuinely appreciated!


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OfflineGrinchGrower
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27046467 - 11/18/20 09:18 PM (3 years, 2 months ago)

You can put the *clean* colonized agar to BRF cakes (myc slurry) OR do what most people prefer and go to bulk by dropping little pieces in sterilized grain jars/bags (spawn) then put that to a substrate (coir, hpoo etc.) once it is fully colonized.

You can also make LC (liquid culture), but this is a little more advanced.


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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: GrinchGrower]
    #27046474 - 11/18/20 09:27 PM (3 years, 2 months ago)

Once the mycelium looks healthy on agar you would transfer a piece of it to a vessel of hydrated, sterilized whole grain such as rye, oats, wheat, barley, etc.

Brf is usually used in conjunction with inoculation methods involving liquids such as with a spore syringe, liquid culture, or liquid inoculant

Once the mycelium has colonized your sterilized grain you mix it with your hydrated bulk substrate such as coir, in a fruiting vessel (shoebox, monotub)


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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Yeetusdeetus]
    #27046503 - 11/18/20 09:44 PM (3 years, 2 months ago)

Many thanks to your replies. I think that’s what I meant in essence. I will have to just find a reliable easy bulk media to use as this all should eliminate the touch And go aspect of spore syringe to brf tek.

Any recommendations as to the best formulation (general consensus)) as to the grain mix or is it simply sterilized grain as you’ve described?  I assume it is grain in its original form vs “flour” structure or at least it looks as such in videos?


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OfflineGrinchGrower
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27046527 - 11/18/20 09:58 PM (3 years, 2 months ago)

The search function on this site has literal DECADES of information including grain prep TEKs, bulk substrate recipes, etc.

I suggest finding one of the Trusted Cultivator members here you think sound like they know what they are doing (just kidding, they all do) and simply following one of their TEKs step-by-step.


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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27046548 - 11/18/20 10:21 PM (3 years, 2 months ago)

Quote:

Uzimyco said:
Any recommendations as to the best formulation (general consensus)) as to the grain mix or is it simply sterilized grain as you’ve described?  I assume it is grain in its original form vs “flour” structure or at least it looks as such in videos?




I believe most people have been so patient and willing to help you because of how well spoken and intellectual you seem. However, that could fade very fast here for asking a bunch of questions that could very easily be answered if you did a little bit of exploring and research on the forums.

Here are a few tips:
Sticky posts at the top of the forum usually contain compiled lists of teks from many Trusted Cultivators.

Check the agar portal.
Check the Noob Forum.

Search for people like bodhisatta, pastywhyte, eatyualive, D3onic, a.k.a., verum, mushboy, mushroomnate and read through their posts and teks. You could literally spend hundreds of thousands of hours researching this website.

Finally, find a few good teks AND STICK TO THEM. Don't use a bunch of info from 20 different teks and make your own plan. That almost never works.

Good luck


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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: tiptrippy]
    #27046564 - 11/18/20 10:50 PM (3 years, 2 months ago)

I can appreciate your reposnse and in fact do appreciate it.

I do know a few things:

Yes I can look on forums. I can internet search like few can. What I can tell you is I did significant reading and research and ended up deriving information that was incomplete snd misled me causing wasted time and resources.

My request was simply that all information of forums and the internet are not created equal. It is for that reason that I requested the information from you all as you were kind enough to provide valid and succinct answers to questions which have eluded me.

An example is the commentary above having to do with spore density and or spore load having a. material effect on syringe inoculation success. It is the single first thing I’ve read detailing such snd I have read more material than I care to admit.  So I urge you not to mistakenly interpret my questions as either laziness or stupidity regarding searching through the internet. Your interpretation is erroneous. It is an intellectual mind that focuses on information and the source.vs information with no inherent validity.

I don’t care to waste effort and resources on making mistakes anymore. That is what has occurred and that is what prompted my post. All is not what it appears on the surface.

Again however I can appreciate your feedback.


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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27046568 - 11/18/20 10:58 PM (3 years, 2 months ago)

You seem like a thoughtful, intelligent human. Stop making spore syringes, they're for noobs.

This is how to succeed:

1. Acquire spore print; (if you have a spore syringe just Streak the solution)

2. Streak spores to agar using an inoculation loop;

3. Clean culture through isolation;

4. Use clean agar to inoculate grains or liquid culture;

5. Use grains to spawn tubs, or use as masters for gtg. If you made LC with your agar then use it to inoculate the grains that will be used for tubs.

Fuck spore syringes, fuckin forget that they even exist. Spore syringes can eat a dick....also, spores to grains or LC is fucking retarded, don't do that. If you become a mushroom growing God and feel like fucking around with any of that shit after you've learned how not to fuck everything up, feel free....until then, fuck all of that.

Any issues that you've previously experienced are directly correlated to your failure to follow that protocol. 

This is the way.


Edited by Stipe-n Cap (11/18/20 11:38 PM)


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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Stipe-n Cap]
    #27046729 - 11/19/20 03:49 AM (3 years, 2 months ago)



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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Wall.E]
    #27046851 - 11/19/20 07:56 AM (3 years, 2 months ago)

Thank you both. I will take your direction without question. That was always the intent.


For the sake of fun, I did want to use an illustration of what random info on the net provides, even so from this forum of course. (I am not referencing all the information you’ve all provided).

The attached image is the result of reading lots of tecs and doing things with precision.  You will see no such contamination as I rely heavily on sterilization with 257nm and 222nm radiation and do not waste much time with things such as bleach (I’m referencing inorganic items). Such radio sterilization virtually ensures the destruction of any contaminant. Even ones you cannot access or areas that cannot be seen.

This just 6 of 18 jars (remaining ones illustrate similar disparity) and this is my 4th unique run. Everything was done simultaneously. All variables were fixed. Things we’re introduced inside of a laminar flow hood. Spore was used from a communal vessel. Media was prepared all together and yet I followed instructions beyond the precision of the advice.  The photo illustrates what you already know. It however illustrates what was not discussed in stacks of literature.  You have all kindly filled in the blanks. I am only shocked at how significant time and effort in learning was stifled by incomplete information floating around.  Had I known before what I now know due to all of your patience, I would have avoided such mistakes.  This is the nature of learning. True it forced me to dig deeper and as a result I have moved to a greater level of understanding.  As long as I ultimately achieve the desired result, I’m ok with failure along the way. 

And if you’re wondering, yes I am well aware of the fact that I am absurdly verbose and blather incessantly. It is simply how I’m wired up when in a self education mode.  Such qualities have helped me in life equal to the damage they’ve done. I have accepted the benefits along with price one must pay.  Again although I’m sure you all do not require the reiteration: your patience and the information you’ve given is genuinely appreciated- it gives me comfort knowing a world dominated by selfish and unyielding people is offset by those who chose another path.  I’m actually quite typical in “real life”. Even “goofy” by the assessment of friends and contemporaries. But God did bestow an ability to write in the form you’re reading and allow me to articulate my thoughts in a professional manner.  You wouldn’t believe what contracts and legal documents look like when written by me...... they are quite ridiculous :wink:



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Offlinetravels_slightly
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Wall.E]
    #27046853 - 11/19/20 07:57 AM (3 years, 2 months ago)

I'm super grateful for threads like this. I also learned something, and I feel like p9hu7 just answered in a single post many of the things I was too shy to ask about out of fear I'd be punished for not UingTFSE my damn self, subsequently spending a whole lot of hours compiling conflicting information from which to draw my own (often incorrect) assumptions.

Thanks gang!


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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco] * 1
    #27046898 - 11/19/20 08:25 AM (3 years, 2 months ago)

Quote:

Uzimyco said:
I am a highly experienced horticulturalist in a wide array of complex genera




If you're already using a flowhood and have the above mentioned experience then I would wager that you have experience with plant tissue culture. Don't waste your time with spore solution and BRF. These have their place, of course,  they often serve as a point of contact for very new growers with limited exposure and experience.

Mush cult often seems very "sciency" and advanced when you're unfamiliar with the terminology.
Agar, petri dishes, sterilization/sterile technique, etc; they cause self doubt and hesitation. New growers often want to postpone their pursuit of such topics when they hear about what appears at first glance to be a shortcut to success with spores injected into a simple BRF recipe.

Cakes are often times not very noob friendly due to the complexity of relationships between competitive contamination found in spore syringes, the mushroom life cycle and it's relationship to it's environment. These subjects generate countless questions about their cakes being often contaminated, too dry/too wet, so on and so forth. These questions can only be answered with an experienced eye. Not very set and forget as a new grower may assume. It's not as easy as "hey, just going to squirt water into my jars and then become a mushroom Pablo Escobar in no time, brah"
New growers often find that once failure with that line of reasoning has forced them out of their comfort zone and into the warm welcoming arms of agar they realize there was nothing holding them back, it's actually super easy. Suddenly the world of mush opens up and success builds upon success. They always wish that they had started sooner.


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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Stipe-n Cap]
    #27049990 - 11/21/20 07:42 AM (3 years, 2 months ago)

:wink: you are correct.

The fact that you’re familiar with TC already defines that your experience extends beyond mycology.

I freely admit that TC is something I had outstanding interest in at one time but that interest has waned. Obtaining quality aperical meristem tissue from rare almost one of a kind plants is a scary endeavor. Since it is “live material” I can’t “cheat” by using radiation sterilization.  It destroys all deoxyribonucleic acid 🧬 therefore rendering the tissue quite dead and no longer viable.  The extraction specifically of meristem tissue runs the heavy risk of killing a plant. When that plant is perhaps 1 of the 2 or 3 in the United States, it creates too much stress on myself and I’d virtually rather propagate my plants vegetatively.  Those kinds of mistakes are truly irreparable. Therefore I’ve chosen to leave TC to the true experts with vast resources of MS types of agar, specifically proven for a given genera. The bigger problem is that when dealing with really rare plants your opportunity to tweak media composition may well be the last mistake you make rendering the plant lost for the long term foreseeable future.  I’m sure you get the gist of what I mean.

In this case however such I’m in uncharted territory. For example - I can sterilize fern spore in a physan solution with quite good results. I have not tired this with fungi spore and I feel very sure it would be destructive. I have learned the hard way - fern spore is likely about as similar to muchroom spore as a car is to a dog. Spore is a term that structurally is dissimilar and fern spore maintains an outer layer that protects the delicate components. Fern spore is also exponentially larger than mushroom spore as fern spore can be individually viewed on paper whereas fungi spore are likely impossible to discern from a “pile”.

Also I use a wide array of endo and ecto mycorrhizae in horticulture. These are in fact fungi and I use them in tandem with products that introduce sub terranian pathinogenic support. I take it further by leveraging bacterium such as Streptomyces griseoviridis.  This bacterium however acts a fungicidal predator. It is known that it is effective as protection from fungi such as fusarium and it does promise to ignore non pathinogenic fungi. I’ve even considered trying such in mycology to see if it would attack cubensis spore or ignore it, while pedatorizing undesirable contaminants. There are other microorganisms I utilize but again I have relied far too much on horticulture knowledge when becoming interested in mycology and found them to be highly counterproductive. I’ve made cakes with the inclusion of things such as powdered black strap molasses as well as propolis since mycorrhizae flourish with it as a nutrient source. Furthermore using exotic combinations of much higher nutrition grains such as hemp flour yielded zero (and I mean zero results) so I backtracked to more traditional means.

Your posts and the posts of others in this thread have given me the education to stop conflating plants with fungi as the fact that they “grow” is roughly where the similarities end.  So yes. Again I’ve done a lot of messing with the hope that my “related knowledge” would supercharge my results. I have no shame in stating they have in fact, wildly done the opposite. I know when I’ve gone down the wrong road. Using control cultures and non control cultures illustrated that tried and traditional  media types categorically perform whereas exotic mixes with grain based superfoods don’t seem to be of any interest to mushrooms or at a minimum, the genera of mushrooms I’m working with.  As I said- even with great intent I suffered much failure and accepted the concept that I’m not looking to blaze new trail and my horticultural experience was quite worthless here.

Subsequent to the beginning of this thread and resulting from the information within, I have already acquired the necessary materials to make agar based dishes and that will be completed no later than tomorrow. Upon fruiting out the few cultures I have which appear to have hit full colonization, I will begin the agar process due to being able to collect fresh spore. I admit I feel empowered by the information I have been provided as failure is quite bothersome.

I thank you for the advice and I feel like I am on the correct path now. Time will tell but again, I feel a much greater understanding of many, many things that were not obvious to me.


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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Uzimyco]
    #27050010 - 11/21/20 08:02 AM (3 years, 2 months ago)

Quote:

Uzimyco said:
. I admit I feel empowered by the information I have been provided as failure is quite bothersome.





Success is an error laden path. By learning what you shouldn't do you simultaneously learn what you should do. The fool who persists in their folly becomes wise. The only reason that I know not to do it that way is because I've tried it and failed in the exact same way.
Your failures are success because they make you wise.
Good luck.


Edited by Stipe-n Cap (11/21/20 08:11 AM)


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OfflineUzimyco
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Re: Lurker finally posting question on “inconsistency of cultivation” [Re: Stipe-n Cap]
    #27107091 - 12/25/20 08:46 AM (3 years, 1 month ago)

Well I am back again. Ill spare you some of the extraneous pitfalls I ran into, but nevertheless I finally had the ability to go spore straight to agar.

I did a couple of different strains on a couple of plates as a test but then decided to stop so that I could wait and see if I was doing it correctly. (This was recent. As in 2-4 days ago).

Admittedly my technique left much to be desired as the inoculation loops definitely did some cutting into the agar. I assumed that was ok but then again I feel like I can’t assume much of anything since my assumptions have proven wrong over and over.

To further make things exciting (I’m being sarcastic) and without a frame of reference I went a bit crazy with streaking the plates (lots of spore streaks) vs just the Z pattern. Again going back to the concept that more spore load - the better (yet again something I can not attest to being correct?).

So within the last couple of days something is certainly growing but I haven’t a clue if it’s right. It looks more like a translucent growth and not white mycelium at this point. Through radiation sterilizing it is virtually impossible to be just a containment as nothing I’ve ever done has exhibited “contamination type characteristics”.

So we come to the question at hand. Would anyone be able to provide any info as to timeframes of what should be determined by when?  I am incubating the plates at around 82 degrees in darkness 24 hrs a day.  I have read about 100 agar teks and they seem to detail much about prep and concept of spawning to bulk afterwards but not a lot on the process of agar colonization such as timeframes.  I do understand what I believe to be the aspect of selection a portion of healthy growth to replate as an isolate and grow on but that step would be after where I am at currently. Again just curious - what are you all incubating your plates at temp-wise and what is the timeframe that you generally make a determination as to what tissue you would want to isolate for growing on re-plated?

Many thanks to you all!


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