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anarchOi
Ellenalien is fat.



Registered: 08/06/07
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degradation - senescence
#7929026 - 01/25/08 09:26 AM (16 years, 7 days ago) |
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i've been trying to get this sorted out in my head for awhile now and i've been able to find anything conclusive everytime i find something on the forums i think is fact, i find something else saying it's wrong and most people just say "it'll be ok to do this, but not this" etc
so my question is, at what point do you experience negative effects of genetic degradation in cloning?
if you're growing an isolate on agar and you take a little peice of it and put it on another piece of agar, is this cloning since this would be 2nd generation agar?
would putting a slab of agar into a jar of grain count? would the grain jar be 2nd generation? then the jars you innoc with it using g2g be third generation? or would they also be 2nd generation?
is it a bad idea to use a fruit grown from an isolate to clone? (isolate via MS fruit biopsy)
it's all so confusing
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Edited by anarchOi (01/27/08 10:58 PM)
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RogerRabbit
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Re: degradation [Re: anarchOi]
#7929286 - 01/25/08 11:02 AM (16 years, 7 days ago) |
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Quote:
everytime i find something on the forums i think is fact, i find something else saying it's wrong and most people just say "it'll be ok to do this, but not this" etc
Welcome to the world of noobs giving other noobs advice when they don't have a clue.
Quote:
so my question is, at what point do you experience negative effects of genetic degradation in cloning?
Probably never. Senescence causes slow growth and over all weak performance, not necessarily genetic mutations.
Quote:
if you're growing an isolate on agar and you take a little peice of it and put it on another piece of agar, is this cloning since this would be 2nd generation agar?
No. That isn't cloning, it's transferring. It also isn't second generation, because it's the same organism.
Quote:
would putting a slab of agar into a jar of grain count? would the grain jar be 2nd generation? then the jars you innoc with it using g2g be third generation? or would they also be 2nd generation?
None of the above. You would still be at first generation after ten, or even fifty grain to grain transfers, but senescence would definitely be a factor.
A clone keeps mycelium from a particular cell line growing. After about three months of rapid cell division, most mycelium will give up the ghost and performance will be poor. This is why we recommend no more than three grain to grain transfers. The reason is to make sure that once the mycelium from the 3rd G2G colonizes the bulk substrate, it still has the energy to produce a nice flush.
A generation is just like with humans or other animals. If you do a grow from spores, it's first generation, no matter how long you grow it or how many transfers you make, just like you are your mom and dad's son, no matter how long you live. Once you fruit it, take the spores and germinate them, this new cell line is what would be 2nd generation, and so on. Mycelium can go for many, many generations. The PE strain of cubensis is living proof of this. It's degrading, but still viable after all these years. My PE6 and Workman's work with PE albino, etc., are efforts to infuse fresh genetics into the old strain. 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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anarchOi
Ellenalien is fat.



Registered: 08/06/07
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thanks a thousand RR
hopefully others will find this
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Culland
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Registered: 09/12/07
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Great post, I too have been wondering the same thing and would love to toss in my own selection of questions.
Is there a way to slow down senescencing? So for example I have my block or grain jar 3/4 colonized if I put it in cold storage, will that keep it from aging as fast because it slows its cell divisions from taking place?
Does the number of agar transfers you do not matter then? The amount of time its growing is more important?
If I flush a block that is senescencing, its tired, produces poorly and I get just a couple fruits. If I clone one of those fruits to agar, does it start all over, or will it carry on the senescencing of the mushroom you cloned from?
If you have something senescencing can you revitalize it by changing its food source? So for example if it was growing on straw and you move it to sawdust will it give it a burst of energy? Something I read, but wasnt sure about.
Thanks!
Cul
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fastfred
Old Hand



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Re: degradation [Re: Culland]
#7935769 - 01/26/08 03:32 PM (16 years, 6 days ago) |
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It's related to the number of cell divisions, so cold storage will nearly stop it.
Quote:
If I flush a block that is senescencing, its tired, produces poorly and I get just a couple fruits. If I clone one of those fruits to agar, does it start all over, or will it carry on the senescencing of the mushroom you cloned from?
It will still consist of the same tired, old cells so that won't help.
> If you have something senescencing can you revitalize it by changing its food source?
That is only done to prevent the myc from becoming to accustomed to one food source and thus generally weak. It probably won't help much for senescence, but it's still good practice.
-FF
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anarchOi
Ellenalien is fat.



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Re: degradation [Re: fastfred]
#7943215 - 01/27/08 10:57 PM (16 years, 5 days ago) |
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so basically the only way to get around senescence is to refridgerate a master
how is it that patches in the wild can survive for so long? they seem to live forever under the lawn
could there be any benefit from adding fresh spores to a senescent dish? might they work togethor providing new cells to a once great culture?
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fastfred
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Re: degradation [Re: anarchOi]
#7943398 - 01/27/08 11:49 PM (16 years, 5 days ago) |
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Hard to say. Perhaps a complex network of myc of various ages is able to avoid senescence because the younger myc provides whatever it needs. Perhaps younger nuclei migrate throughout the network as old ones become weak.
-FF
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badiggity
yo daddy



Registered: 09/11/07
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Re: degradation [Re: fastfred]
#7957700 - 01/30/08 08:29 PM (16 years, 2 days ago) |
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so spores are nothing like seeds per se? why wouldn't the generations essentially restart after being germinated from the spores of the clone?
-------------------- devils speak of the ways in which she'll manifest angels bleed from the tainted touch of my caress need to contaminate to alleviate this loneliness I now know the depths I reach are limitless
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fastfred
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> why wouldn't the generations essentially restart after being germinated from the spores of the clone?
They do.
-FF
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badiggity
yo daddy



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Re: degradation [Re: fastfred]
#7958501 - 01/30/08 11:16 PM (16 years, 2 days ago) |
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so then it would be possible, through many isolations, to perfect a sub strain down to the point that the majority of the spores would exclusively express the desired characteristics a majority of the time, correct? Lets say one were to isolate a decent spore producing PE, and through many isolations and fruitings, would they be able to produce prints with a rather high chance of yielding printable mushrooms? I realize that spores from prints have huge numbers of different substrains, but i'm wondering is if the genetics carried through prints of proven isolates carry more of the desired characteristics.
-------------------- devils speak of the ways in which she'll manifest angels bleed from the tainted touch of my caress need to contaminate to alleviate this loneliness I now know the depths I reach are limitless
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anarchOi
Ellenalien is fat.



Registered: 08/06/07
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aren't most mushrooms printable? sparing the mutants
i don't see any need to isolate just to get prints and of course, you can create your own isolate culture that does exactly what you want it to over time, but that doesn't make it a substrain correct me if I'm wrong, but i don't think it's a substrain until you get your desired quality 100% of the time i.e. if you have 1 leucistic mushroom (lack of pigment) out of 30, even if you clone it you're likely not going to get all leucistic mushrooms, but then if you print the ones that are and restart the process, you can VERY VERY slowly create a substrain
not to mention, this has nothing to do with senescence
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badiggity
yo daddy



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Re: degradation [Re: anarchOi]
#7959402 - 01/31/08 08:21 AM (16 years, 1 day ago) |
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exactly wut im asking. the reason i used PE as an example is do to its sporadic printing capabilities
i do know that this would be far from a quik process, but am willing to put in the work. If any of you guru's could drop me a conclusive answer, id appreciate your input.
-------------------- devils speak of the ways in which she'll manifest angels bleed from the tainted touch of my caress need to contaminate to alleviate this loneliness I now know the depths I reach are limitless
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fastfred
Old Hand



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Re: degradation [Re: badiggity]
#7970021 - 02/02/08 07:04 PM (15 years, 11 months ago) |
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Yes, if you want to breed for spore production, and your initial germplasm has enough variation or proper genes, it shouldn't be very hard.
The thing is that PE probably doesn't have very good spore production genes present and is already pretty inbred, so there isn't much varitation to work with. You'd probably be best off to cross it with another strain and breed for PE traits with good spore production.
Anarch is right to point out that not all traits you see in a clone are purely genetic. A lot of it is GXE interaction (gene x environment).
-FF
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rungi
journeymana

Registered: 07/11/01
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Re: degradation [Re: fastfred]
#8608475 - 07/08/08 02:33 PM (15 years, 6 months ago) |
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RR said:
"If you do a grow from spores, it's first generation, no matter how long you grow it or how many transfers you make, just like you are your mom and dad's son, no matter how long you live. Once you fruit it, take the spores and germinate them, this new cell line is what would be 2nd generation, and so on."
I do not understand "first generation." Do you mean spores collected from the wild before they are grown under optimal conditions. And that the spores collected after the mass flushes only possible by controlled environment. are second generation? The name "first" and "second generation" are not at all clear. Some strains from the wild seem clearly superior and some inferior by degrees determined experimentally. But weather the wild strain is first or second or third is unknown and makes no sense because a species "is" identification (which implies stagnation) comes into being by saying it does verbally and relating this to the consensual human time scale.
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MycoAu
5thKingdomCome


Registered: 07/18/07
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Re: degradation [Re: rungi]
#8608896 - 07/08/08 04:27 PM (15 years, 6 months ago) |
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Your "first" generation is the first culture/isolation that you perform successfully.
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RogerRabbit
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Re: degradation [Re: rungi]
#8609219 - 07/08/08 05:48 PM (15 years, 6 months ago) |
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Don't confuse transfers and generations. If you're growing mycelium from a wild mushroom that you cloned, it's first generation. The spores it produces will start the second generation, etc.
Wild patches stay productive forever because every year, they and patches all around them(and across the world) produce spores which travel on the winds. Some of these spores land and germinate, and the resulting mycelium with it's genetic characteristics are absorbed into the older mycelium. (anastomosis) This prevents senescence by infusing fresh genes into the pool.
When we say to limit to three grain-to-grain transfers, it's just a rough guide based how many cell divisions have been made. If we're growing grains in 1 pint jars, we could make more than three. If we're doing grains in a 55 gallon bucket, it might well be approaching senescence when the drum is colonized. It's all about how far the individual live mycelium has traveled(cell divisions). 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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MycoAu
5thKingdomCome


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I apologize for not clarifying further.
I only meant to state the "starting point" for assigning a "first generation"- not trying to imply anything with respect to number of cloned transfers versus the production of the next set of genetic recombinations.
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rungi
journeymana

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Re: degradation [Re: MycoAu]
#8609905 - 07/08/08 08:20 PM (15 years, 6 months ago) |
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First generation according to Roger Rabbit is the mycelium or spores propagated from a wild collection (he said both). Numbers are assigned each time the fungi undergoes meiosis. What is the practical use of this information? Nature produces both strains of increasing and decreasing vigor. So if you have a "pure" strong strain no matter what generation (only able to determine by experiments) save the mycelium before it fruits in the frige. No telling what meiosis will do the the next generation.
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RogerRabbit
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Re: degradation [Re: rungi]
#8613952 - 07/09/08 05:15 PM (15 years, 6 months ago) |
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Quote:
What is the practical use of this information?
Very little.
Senescence is far more important and is related to the number of cell divisions the mycelium has undergone. Most questions about 'generations' come from former pot growers looking for comparisons, which really don't exist. Fungi and plants are very dissimilar. 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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happyday
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Registered: 06/22/04
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Re: degradation, senescence and genetic mutation, appropriate use of the term "generation" [Re: RogerRabbit]
#8616066 - 07/10/08 02:39 AM (15 years, 6 months ago) |
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Actually senescence is a slow down of metabolic activities, and in many cases, irreversible. It is not possible to have irreversible manifestation without changes in the operators in the pathways of physiological or metabolic activities. And metabolic and physiological changes are invariably tied with changes in the gene controling such attributes. The short answer is that, in most cases, especially, irreversible metabolic changes involve gene mutation.
The word "generation" is utilized interchangably throughout the mushroom industry to designate culture-to- culture transfers (i.e. second generation grain spawn) as well as when mating the offspring of a mushroom and culturing and then subsequently mating those spores for another isolation. It merely comes down to semantics.
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