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OfflineJustAHobby
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Making a Good Substrain (Agar, etc.)
    #7089799 - 06/25/07 07:12 AM (14 years, 7 months ago)

All,

*Note to mods: if this is in the wrong forum, let me know. I thought that I would post this agar/substrain isolation question in the Advanced Mycology section so that more experienced eyes would see it.*

Quote:

RogerRabbit said:
They're fine for multispore inoculated fruits.  A few aborts and otherwise deformed mushrooms are normal.  People like to post their killer flushes with wall to wall fruits, but those are the exception, not something you should hold yourself to until you learn strain isolation.  Once you get a great fruiting strain isolated, you can save it in master culture slants to use from now on.  As long as you're using spores to grow, your flush is what to expect.  That's nothing to be ashamed of by the way.
RR




So here's my question. I want wall to wall fruits (who doesn't want to perfect their hobby, as RR said somewhere else :wink: ). Essentially, how do I go about doing this?

Here are some ideas:

1. Multispore innoc to agar. Continually isolate the fastest growing myc and then use this to fruit. Will this lead to a great pinset? I don't see why.
2. Take samples from the stems of the biggest fruits. Clone to agar, then use this to make new fruits. Repeat. Won't this just give big fruits?
3. Multispore innoc to agar. Isolate several sections on their own, and basically "branch" them in different directions (by this I mean now you have three different "types" of myc.). Now fruit, see which fruits the best?
4. Some combination of the above?

I don't want to do 3...it seems like it will take forever and that it's essentially just trial and error to find a good fruiting substrain. But it seems like 1. will just isolate genetics that give great mycelium growth and 2. will just isolate genetics to make big shrooms.

How do you all do this? I can't imagine you do #3, which appears to be guesswork, but I can't come up with another way.

Also, maybe I'm thinking about this the wrong way. If you see any major flaws with my thought process, please let me know. I have heard that by isolating a single piece of mycelium, you get a substrain that is a single genetic crossing. This will make it all fruit at the same time....thus leading to a "near perfect" pinset. Is this true?

Also, please keep your responses on the subject of genetics. I know that the environment plays a huge role in the whole song and dance, but let's pretend for a moment that I have the environment thing worked out to perfection and I just need genetics.

Hopefully, I can get some Old Hands' responses (Agar, RR, hyphae?)

Thanks,

JaH


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Invisiblefastfred
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Re: Making a Good Substrain (Agar, etc.) [Re: JustAHobby]
    #7090538 - 06/25/07 01:30 PM (14 years, 7 months ago)

> thus leading to a "near perfect" pinset. Is this true?

No. Gene by environment interaction is very significant. Simply having a monoculture is no guarantee of good results. It helps, but it's no guarantee that you'll get uniform results.

As far as getting the best genetics, I suggest going with direct multispore inoculation for awhile. It's very easy to clone fruits and switch over to monoculture.

Multispore gets dissed way too much here. Some say it's a crapshoot, but what is really a crapshoot is just picking a random strain from a petri dish. If you do multispore then you are assured of having a good selection of substrains to choose from.


-FF


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OfflineRogerRabbitM
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Re: Making a Good Substrain (Agar, etc.) [Re: fastfred]
    #7092575 - 06/25/07 11:42 PM (14 years, 7 months ago)

You need to do number 3. However, a single swipe of spores on a petri dish will yield at least 50 substrains. I'm doing a series of isolations now that started with two swipes, each on its own dish. I've only been transferring the best looking mycelium, and am now up to over 30 substrains of pure rhizomorphic mycelium, and this is from just two swipes of spores onto agar.

I plan to fruit out each and every one of these strains separately after placing the individually well marked dishes in the lab refrigerator. After fruiting, I will determine the best strains that I want to keep. I will then go back to the individual petri dishes that are in the fridge, allow them to warm back to room temp and start growing again, and then transfer a small piece of mycelium from each into master culture slants for long term storage.

I will then be able to grow those same strains for 20 to 30 years, without having to go back to spores. A lot of work? Perhaps, but once the strains are isolated and stored in culture slants, it's a breeze after that and the results are dependable.

Multispore inoculation certainly works, and it's how nature works. However, by isolating strains, you can get those killer flushes of wall to wall fruits with whatever 'other' qualities you seek. Those 'other' qualities might not come from the fastest growing or largest strain, and might not even come from the best flushing strain. That's why each strain needs to be individually tested.

By the way, there is a full strain isolation tek and also a master culture slant tek on my DVD. You can see some short low res previews here: http://www.mushroomvideos.com/1809563.html
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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Offlinedeformedreality
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Re: Making a Good Substrain (Agar, etc.) [Re: JustAHobby]
    #7093129 - 06/26/07 02:04 AM (14 years, 7 months ago)

Are the video's actually made and ready to buy? I thought you said something about being released in fall? I wonder if i wait long enough if hollywood video would get some copies in and and i can rent them ;-) lol. If they are ready to buy ill probally look into with the next pay check.


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OfflineRogerRabbitM
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Re: Making a Good Substrain (Agar, etc.) [Re: deformedreality]
    #7095826 - 06/26/07 07:15 PM (14 years, 7 months ago)

They've been out for a few months now. Click the link above and visit the site. A sequel will be out this fall, but will deal with using mushrooms to recycle. The one that's out now is a two DVD set on cultivation.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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InvisibleLiquidkick
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Re: Making a Good Substrain (Agar, etc.) [Re: deformedreality]
    #7095875 - 06/26/07 07:32 PM (14 years, 7 months ago)

They are ready to purchase.


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OfflineJustAHobby
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Re: Making a Good Substrain (Agar, etc.) [Re: RogerRabbit]
    #7096117 - 06/26/07 08:26 PM (14 years, 7 months ago)

RR et al,

I was afraid you'd say that. Now what would you say if I were to ask

1. Can I isolate a bunch of strains and do micro grows of them (i.e. small casings so I can do them all at once and see how each one does?)
2. What if I clone from a good looking and potent boomer, then substrain isolate? In theory, I will probably have less strains to have to fruit out, and likely that more of them will have good characteristics, right?
3. If you had to pick one strain of cubies to do all the substraining with, what would you choose?

Thanks for all the advice.

-JaH


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OfflineRogerRabbitM
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Re: Making a Good Substrain (Agar, etc.) [Re: JustAHobby]
    #7096273 - 06/26/07 09:03 PM (14 years, 7 months ago)

The 'named' strains are irrelevant. Just do strain isolation and find one you like that way. They're all the same species.

Yes, you can isolate a bunch of strains and grow a pf cake or two with each one, and then clone from the cakes you like best. However, you'll have younger cell lines if you'll isolate and then save back a bit in the refrigerator while you grow each one out. That way, you can eat the fruits, and then go back to the corresponding petri dish to make your master culture slants from. It's all explained in the DVD on disk 2.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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InvisibleLiquidkick
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Re: Making a Good Substrain (Agar, etc.) [Re: RogerRabbit]
    #7171155 - 07/13/07 03:09 PM (14 years, 6 months ago)

is the PF cake the most effecient way to test an isolate or a grain jar, etc...


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Invisiblefastfred
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Re: Making a Good Substrain (Agar, etc.) [Re: JustAHobby]
    #7171344 - 07/13/07 03:56 PM (14 years, 6 months ago)

> is the PF cake the most efficient way to test an isolate or a grain jar, etc...

If you are looking for a substrain that grows well on PF cakes, then yes. Otherwise it might be fairly important to assess their performance with casing.


-FF


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