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Kmacmo
The aborted pin



Registered: 08/14/19
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Cultivating mushroom generations/agar work/clones. Need help with few questions, please.
#26346709 - 11/26/19 12:54 PM (4 years, 2 months ago) |
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Gday everyone, I just have few questions about mushroom genetics and generations, I spent time looking for the specific answers but couldn't find any unfortunately.
Q1. When you grow a tub from multispore and clone the best fruit does that make your clone you put on the agar plate generation 2?
Q2. If you use that clone plate to inoculate loads of jars say, could you transfer some of the new mycelium growth from that plate onto a new plate, that's how you keep the same clone going forever and ever?
I have 6 b+ plates all transferred from two T1 plates (3 from both plates) they went to T2 on Sunday. Hoping to use one of these plates to knock up a couple jars for a test run and leave the other 5 to go into transfer 3.
My mycelium is 'cold' growing so don't see any rhizmorphic growth but I can tell which plates are more aggressive and just look like no weird shit is going on. I could take different sectors of growth from the mycelium plate to inoculate the jars? (Just want to get some jars colonising, the aim to so just get an actual clone and use that from here on)
Any advice or critique on my method is appreciated
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Phycus
Blister Lips



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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: Kmacmo]
#26346778 - 11/26/19 01:23 PM (4 years, 2 months ago) |
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A1) yes, that first fruit is your master in a sense. Your isolate. every subsequent fruit would be a clone if i am not mistaken. A2) yes. that is called a master plate. you can use it to inoculate whatever you want (even make LC syringes with it) you can do the same with grains.
-------------------- disclaimer - nothing i post is real. this account is for fictitious purposes and posts should not be taken literally
Edited by Phycus (11/26/19 01:26 PM)
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AyePlus
Stony Danza



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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: Phycus]
#26346797 - 11/26/19 01:34 PM (4 years, 2 months ago) |
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I mean technically yeah its a later generation but for all practical intents and purposes its the beginning of a new line,
Ideal circumstance Master plate gets made into master slant, at the same time 2nd gen master plate is made from the master plate to be used for inoculating and the master plate and slant go into cold storage. Once the 2nd gen master is gone a new plate can be made from the og master until that one is gone, then at that point you make a new master plate from the slant and continue on like that till the slant is almost gone then you make a new slant from the last master plate.
Keep the genetics as young as possible. Cold storage slows the aging process almost to a stop but after ~ 5 years slants should be plated and grown out and re-slanted, plates redone every 4 months or so.
-------------------- Learn about breeding
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Kmacmo
The aborted pin



Registered: 08/14/19
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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: AyePlus]
#26346876 - 11/26/19 02:12 PM (4 years, 2 months ago) |
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Thanks for clearing that up guys, makes sense.
So only fruiting the mycelium changes the generation I guess? Doing what you said, my mycelium will be the same generation forever aslong as i inoculate jars from the same master clone plate or the slant?
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Edited by Kmacmo (11/26/19 02:15 PM)
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AyePlus
Stony Danza



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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: Kmacmo]
#26346977 - 11/26/19 03:25 PM (4 years, 2 months ago) |
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well not literally forever but yeah effectively. Long enough anyways.
-------------------- Learn about breeding
  C10ās agar guide Good surface conditions = Good pinsets Read more, post less. š
š° š¼ š“ š
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š“ š° š¼ š² š» šø š½ š¶ š
š
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curious.psychonaut
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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: Kmacmo]
#26347006 - 11/26/19 03:42 PM (4 years, 2 months ago) |
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I've noticed and accepted that generation gets used in all sorts of ways here (transfers, clones, etc.) but, technically, it's all just growth of the same culture/DNA/"individual". Though, ofc, you will reduce the number of strains ("individuals") present with each step. Come to think of it, "individual" is not a very good term, because you effectively divide them but still get to keep the "individual" in both containers, very much unlike what it's like when you do that to a cat.
A 2nd generation requires sexy times (spores*), and if that doesn't happen, it's just the same old "individual" (just a little older and more tired in fungi that display senescence).
*Interestingly (if you find such stuff interesting), it's not very clear cut where exactly sex occurs, but it probably makes most sense to say it's when spores are created in the basidia (at gills on cubes and other gilled shrooms), because that is when the two haploid nuclei of the dikaryon actually fuse into a diploid nucleus, actually combining genetics. (The diploid nucleus soon divides again in order to form haploid spores.) You can also look at monokaryons mating from a sexy angle, as there are selection mechanics and compatibility, but nothing genetically new is created at this stage**, and in fact one or both of the nuclei may yet mate with others: lookup "di-mon pairing". Here's a fun article on the subject.
**Except the unique phenotype created, since both nuclei in the dikaryon influence metabolism & development.
Edit: clarification b/c I forgot to answer directly: this all means "no", a clone of the best fruit is not a second generation, since there were no spores involved.
-------------------- My LAGM2020 grow log
Edited by curious.psychonaut (11/26/19 03:53 PM)
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Kmacmo
The aborted pin



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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: curious.psychonaut]
#26348634 - 11/27/19 01:25 PM (4 years, 2 months ago) |
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This stuff is so interesting 
So aslong as I can keep mycelium (transfers/clones) alive and healthly on plates you never should worry about going over too many generations?
I thought spores where the beginning of the line, the mycelium would have to 'give birth' (fruit mushrooms) then all those mushrooms are generation 1 right?
It's alot to take in, and I will read that link looks like it will be helpful thanks alot for the knowledge guys
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AyePlus
Stony Danza



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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: Kmacmo]
#26348642 - 11/27/19 01:29 PM (4 years, 2 months ago) |
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No. You want to Keep them at the lowest/youngest gen possible. Less cell divisions= more vigorous culture.
The idea is to keep young cultures in storage and make cultures to be used from them.
-------------------- Learn about breeding
  C10ās agar guide Good surface conditions = Good pinsets Read more, post less. š
š° š¼ š“ š
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LtLurker
Lost Sailor



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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: curious.psychonaut]
#26348654 - 11/27/19 01:34 PM (4 years, 2 months ago) |
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Quote:
curious.psychonaut said: I've noticed and accepted that generation gets used in all sorts of ways here (transfers, clones, etc.) but, technically, it's all just growth of the same culture/DNA/"individual".
Edit: clarification b/c I forgot to answer directly: this all means "no", a clone of the best fruit is not a second generation, since there were no spores involved.
I don't think this is a common occurrence for regulars, only confused newer growers. I can't recall any regulars here calling transfers, expansions, or clones a new generation at any point(maybe colloquially?). Maybe it's the notation of "T" on plates, which just stands for "transfer #", that throws people off.
Quote:
Phycus said: A1) yes, that first fruit is your master in a sense. Your isolate. every subsequent fruit would be a clone if i am not mistaken.
It's not an isolate, just a clone. Isolate is a single genetic code, only 2 spores mated, a strain. Clones still contains many strains working together.
@op Q2. Yes you can transfer, you should be doing a few transfers to ensure you have a clean culture. Noccing with a germ plate is not a good idea since you'd be putting any ride alongs from the cloning procedure(or spores) right into the jar, making your use of agar moot.
Edited by LtLurker (11/27/19 01:41 PM)
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AyePlus
Stony Danza



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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: LtLurker]
#26348660 - 11/27/19 01:39 PM (4 years, 2 months ago) |
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Great now Iām confused lol. Younger is better and we might be using generation incorrectly.
Generation should mean the next offspring but we were using it to mean transfer. Less transfers is better, number of generations not as important as age of culture. But after alot of selfing your genetic diversity will be lessened, which could be a problem if selection for vigor/potency isnt occuring at the same time
-------------------- Learn about breeding
  C10ās agar guide Good surface conditions = Good pinsets Read more, post less. š
š° š¼ š“ š
š“ š° š¼ š
š“ š° š¼ š² š» šø š½ š¶ š
š
š° šæ
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LtLurker
Lost Sailor



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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: AyePlus]
#26348672 - 11/27/19 01:50 PM (4 years, 2 months ago) |
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yea man, it's just age or number of transfers. Working from a slant is an easy way to keep using the original genetics you cloned instead of constantly lessening the number of strains while taking a long chain of transfers.
Although, i don't have a strong opinion or answer about how much truth there is to an ageing culture dying out because of senescense. Some say yes, some say no it's more likely another problem. Personally i can run a series of 8 g2g transfers and they're all as vigorous as the first round, they all die out from contams in the g2g process instead of age. I think you'd need to keep something going a very long time before senesence is an issue.
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Kmacmo
The aborted pin



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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: LtLurker]
#26350549 - 11/28/19 12:46 PM (4 years, 2 months ago) |
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Quote:
AyePlus said: The idea is to keep young cultures in storage and make cultures to be used from them.
Like make a slant from a clone, then use the slant to inoculate a plate, do 3 transfers and then into grain ... Think I'm getting it now 
Quote:
LtLurker said: @op Q2. Yes you can transfer, you should be doing a few transfers to ensure you have a clean culture. Noccing with a germ plate is not a good idea since you'd be putting any ride alongs from the cloning procedure(or spores) right into the jar, making your use of agar moot.
My plates are from spore print and got a few T2 transfers growing atm then going to T3... How many transfers roughly before you could 'weaken' the mycelium?
Quote:
LtLurker said: yea man, it's just age or number of transfers. Working from a slant is an easy way to keep using the original genetics you cloned instead of constantly lessening the number of strains while taking a long chain of transfers.
Slant it is need to wait till I've got fruits(hopefully my mycelium is a fruiter). Would you put the clone tissue into the slant or put it on a plate first for a few transfers before the slant?
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LtLurker
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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: Kmacmo]
#26350653 - 11/28/19 02:07 PM (4 years, 2 months ago) |
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I call my initial plate 0. I'll take 2 to 4 transfers until I'm happy with it before I slant a wedge and inoculate with the rest of the plate. I want my slant as clean as possible so I'm not trying to pull from a contamed slant..
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Kmacmo
The aborted pin



Registered: 08/14/19
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Re: Cultivating mushroom generations/agar work/clones. Need help with few questions, please. [Re: LtLurker]
#26351832 - 11/29/19 11:21 AM (4 years, 1 month ago) |
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Brilliant no wastage thanks for clearing that up for me
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