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Anonymous

Will this work? (grain to grain and senescence)
    #1258046 - 01/29/03 09:56 AM (14 years, 2 months ago)

I want to do a multispore inoculation of grain with P. Mexicana spores. I'll be using a high-sclerotia, low-fruiting strain. I don't intend on ever fruiting it, just collecting sclerotia. For this reason, for subsequent batches, I intend on doing grain to grain transfers.

I've read a little about senescence, and my understanding of it is that it is when a pure strain (a genetically unique individual) gets old and eventually degenerates. Is this correct? Does senescence occur in a multi-strain culture? Or is it possible to transfer from grain to grain to grain ad infinitum?

It would seem that within a multi-strain culture, sexual reproduction is constantly occuring and new individuals are always being formed, and so senescence wouldn't be a problem. Are there any problems with numerous grain to grain transfers? Fruiting these mushrooms is not something I'm interested in trying to undertake, but I would like to be independant of having to order spores. Will the grain transfer idea work?


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OfflineRaadt
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Re: Will this work? (grain to grain and senescence) [Re: ]
    #1258094 - 01/29/03 10:11 AM (14 years, 2 months ago)

from a multispore I would imagine the grain to grains could go much further, without senescense - however, no it will not go ad infinium.

If you want to save some, innoculate a petri.


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Raadt

-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--


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Anonymous

Re: Will this work? (grain to grain and senescence) [Re: Raadt]
    #1258101 - 01/29/03 10:14 AM (14 years, 2 months ago)

I'm really just looking for a way around having to order spores or fruit the cultures. Is this possible?


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OfflineRaadt
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Re: Will this work? (grain to grain and senescence) [Re: ]
    #1258118 - 01/29/03 10:21 AM (14 years, 2 months ago)

definantly, clone a piece of sclerotia on agar - then keep the culture refrigerated once it grows about 3/4 the dish.

Or you can multispore onto a dish, and just keep that - or isolate a strain, and keep that isolate, then you can transfer dish to dish for quite some time. Because 1 dish will innoculate hundreds more.

Agar cultures are the way most mushroom companies do their work, I doubt they ever find need to reisolate from spores.


--------------------
Raadt

-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--


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Anonymous

Re: Will this work? (grain to grain and senescence) [Re: Raadt]
    #1258204 - 01/29/03 10:44 AM (14 years, 2 months ago)

could i just stick a colonized jar in the fridge and use that to start all my new batches?


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Anonymous

Re: Will this work? (grain to grain and senescence) [Re: Raadt]
    #1258210 - 01/29/03 10:46 AM (14 years, 2 months ago)

how come going from dish to dish is ok, but not jar to jar?


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Re: Will this work? (grain to grain and senescence) [Re: ]
    #1258314 - 01/29/03 11:20 AM (14 years, 2 months ago)

because the dish goes into like 1000 dishes not from one dish to the next, etc. also the mycelium doesn't "age" whole lot in the process of colonizing a petri as opposed to a whole jar of substrate.


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OfflineRaadt
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Re: Will this work? (grain to grain and senescence) [Re: ]
    #1258349 - 01/29/03 11:29 AM (14 years, 2 months ago)

because you can't innoculate 1000 jars from 1 jar, not to mention the mycellium doesn't have nearly as much surface area to cover, therefore doesn't tire as quickly.

From 1 dish, the original multispore - assuming you never wanted to isolate it, you can innoculate like 100 dishes, by little pieces. 1000 was an exaggeration. That has only gone down generation, wheras your jar would innoculate maybe 10 jars before you would have to step down another generation.

Don't get me wrong, you can probably go on a long time via multispore, but for say a commercial endeavour - that sort of quantity - you wouldn't be able to. You would need to save a culture.


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-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--


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Anonymous

Re: Will this work? (grain to grain and senescence) [Re: Raadt]
    #1258388 - 01/29/03 11:45 AM (14 years, 2 months ago)

So you can go from one culture directly to 100 cultures, but not from one culture to ten cultures, then pick one of them and do ten more, then pick one of them and do ten more... ?


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OfflineAzmodeus
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Re: Will this work? (grain to grain and senescence) [Re: ]
    #1258433 - 01/29/03 12:04 PM (14 years, 2 months ago)

I think what was implied was that the 100 agar you would get from the first could be used to innoculat thousands of jars that will still only be 2nd generation.
(correct me if im wrong)


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Lest we forget. "


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OfflineRaadt
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Re: Will this work? (grain to grain and senescence) [Re: Azmodeus]
    #1258507 - 01/29/03 12:31 PM (14 years, 2 months ago)

That is the basic gist of it.


the multispore on a plate would be generation one. And you have 100 sources from that, that would all be gen 2. The multispore into a jar will then have to be transferred to another jar (which is much more surface area for the mycellium to run, and the better mating pairs selectively reproduce) which can probably innoculate ~10 jars, which are gen 2. However, as stamets states in TMC, you can safely go 3 generations in grain to grains. So jar 1 is innoculated and will be referred to as Gen 1, and further jars Gen 2 and so forth.

so

MS->Gen1
x10
so have 10 Gen2 jars
then those will each innoculate successfully 10gen3 jars.
so you'll have 100 jars at gen3, which is what stamets suggests staying at. I imagine you could go to 1000 jars at gen4, but it's not suggested by stamets.

That's my best explanation, where as agar dishes aren't considered generations.

You can take generation 1 (multispore), and transfer that to 10 plates, which you can refrigerate 2-3 of, and use the rest to each innoculate about 4 jars (give or take).

SO basically, generation 1, can be a. saved, for further transfers to many more plates, and b. can be used to innoculate upwards of 40 jars, which is even more space for grain to grains.

If I sound wrong to anyone, please let me know. This is my best understanding of it.



--------------------
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-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--


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Re: Will this work? (grain to grain and senescence) [Re: Raadt]
    #1258522 - 01/29/03 12:34 PM (14 years, 2 months ago)

I cant wait till i get my petri dishes and agar!! :grin:


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Lest we forget. "


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Re: Will this work? (grain to grain and senescence) [Re: ]
    #1258529 - 01/29/03 12:35 PM (14 years, 2 months ago)

i have transferred agar wedges up to 5 and 6 times and the isolates are still perfect fruiters, so I do not know that agar is considered the same, as g2g's.


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-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--


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Re: Will this work? (grain to grain and senescence) [Re: Raadt]
    #1258569 - 01/29/03 12:45 PM (14 years, 2 months ago)

perfect explanations. I get the whole process perfectly but have very limited agar experience as my teknique utilizes different methods. i do not think that the agar really counts as a generation either. if i had to put a number on it... i'd say a petri generation equals roughly 1/3 of a grain generation. age is solely based on the # of cell divisions the mycelium has undergone.


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Re: Will this work? (grain to grain and senescence) [Re: debianlinux]
    #1258710 - 01/29/03 01:16 PM (14 years, 2 months ago)

That's correct, stamets measures this with his P value system. You always want to stay as close to the original as possible, more is worse, generally speaking.


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-- The information I provide is only information from readings, growing of gourmet mushrooms, and second hand stories--


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Re: Will this work? (grain to grain and senescence) [Re: Raadt]
    #1261984 - 01/30/03 01:58 PM (14 years, 2 months ago)

Right. And stamets also mentions cell divisions, which is the best way to understand the mechanics. See, senescence is due to randomn mutations. These mutations occur rarely in an organisms lifetime, and increase only very slightly everytime the cell has to divide. Over millions of cell divisions from sexual reproduction (spores mating), mutations build up and eventually render the organism (strain) useless.

Following this, keep in mind that you always want to use cultures which have the fewest number of cell divsions since the original culture. These "younger" (in #s of cell divisions) cultures will have fewer mutations and will therefore be more vigorous.

Stamets even gives estimated #'s of cell divisions required to colonize a petri if you're interested. Since this is a relatively low number of divisions, and is relatively standard for each species, he measures the age of his cultures according to the P-value. The P value is how many petris have been colonized since the original culture. Knowing the P value gives an indication of how many cell divisions the culture has gone through.

And obviously, it takes far fewer cell divisions to colonize a petri than it does to colonize a jar/bag of grain. That's why you can have many more petri generations (p-value) than you can have grain generations.

That said, most people don't have problems with the G3 jars, so. You could start say 10 jars from a syringe. Use one to make 10 G2's, and use those to make 100 G3's. The other masters are stored in the fridge until you've used the G3 jars. Then you can repeat this process for 9 more cycles, effectively producing 1000 jars from your original syringe. The practical limitiation in this method though is that you can't really store grain jars in the fridge indefinitely, especially a home fridge. But it illustrates that if you manage your "young" cultures well, you will rarely run into senescence problems and you won't have to buy spores very often. If you do, you are growing on a much too large a scale to be talking about it on the internet.


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Re: Will this work? (grain to grain and senescence) [Re: ]
    #1263779 - 01/31/03 12:54 AM (14 years, 2 months ago)

Greetings all,

I received my first gift of a living carph in the middle 70's. Rye in tapered pints and/or shook out in glass cake pans was the way then. Grain transfers were done with small spoons, individually wrapped and sterilized. One spoon per seed jar was used to inoculate about ? doz fruit jars. The fastest jars were used for the next round of transfers.

Largest and blueist fruits were tissue cultured to agar plates. 2 oz. French Square bottles were used for backups. Regrown out on a 2 to 3 month rotation with different agars used. Over time a half dozen stable phenotypes were established. No spores were ever used for culturing. Some were collected for fun and just in case.

That one single gift was in my care for a decade without senescence before circumstance changed.

Peace

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Giant King - GULLIVER'S TRAVELS


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Re: Will this work? (grain to grain and senescence) [Re: JaComet]
    #1264407 - 01/31/03 06:46 AM (14 years, 2 months ago)

A mushrooms history... :grin:


--------------------
"Know your Body - Know your Mind - Know your Substance - Know your Source.

Lest we forget. "


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