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Offlinethisone
the one theydon't see

Registered: 11/06/02
Posts: 818
Last seen: 19 years, 1 day
cloning, sectors, isolates?
    #2078856 - 11/06/03 10:04 PM (20 years, 4 months ago)

Having trouble finding information on more detail in regards to some of these issues. Would like to know the benefits or cloning live tissue over isolating a strain on agar or vice versa, would also like more specifics on choosing which rhizomorphic growth on the agar dish, until a single strain is isolated. Does each one of those strains need to be singally grown out and tested as a fertile fruiting strain? How do you know which is a sector? Just starting out with agar and curious as to all that can be accomplished and the benefits of each. Thanks! :grin:


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Everything written above this line is completely false. I am terribly bored and write here to pass time. I do not participate or support any illegal activity.

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InvisibleMycomancer
Psi Cubed
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Registered: 09/28/03
Posts: 586
Loc: United States
Re: cloning, sectors, isolates? [Re: thisone]
    #2078926 - 11/06/03 10:31 PM (20 years, 4 months ago)

ok...now there's a tall order...alright, the major benefits of cloning live tissure over isolating a strain on agar is that by using a living tissue sample from a donor shroom you have observable genotypes. Take a tissue sample from your largest healthiest shroom and you get a clone of that shroom and it's inherent genotypes. Plus it's alot less indepth and less time consuming than isolating a single great strain on agar.

For rhizomorphic growth on agar, let mycelium colonize agar dish, then cut it up like a pizza and use the wedges to colonize individual agar dishes. Pay attention to the healthiest growers, repropogate those to new agar, take the best of those and innoculate to fruiting medium. Test for the best fruiting and fertile strain (desired genotypes). In order to be considered a viable strain the mushroom must be capable of reproduction (drop spores).

This is about as much as I can muster as it's getting late and I need to wake up at 5:30am. Anyway Hope this helps.

mycomancer

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Offlinethisone
the one theydon't see

Registered: 11/06/02
Posts: 818
Last seen: 19 years, 1 day
Re: cloning, sectors, isolates? [Re: Mycomancer]
    #2078976 - 11/06/03 10:44 PM (20 years, 4 months ago)

cool, well if you're asleep and anyone else knows, is it probably just as good to just clone live tissue, If so should the tissue culture be placed on the agar dish and then take the same pizza like slices to innoc. grain? Is every piece of growth equal since they all carry the same genotypes, or should rhizomorphic growth still be isolated and grown out?


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Everything written above this line is completely false. I am terribly bored and write here to pass time. I do not participate or support any illegal activity.

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Offlinethisone
the one theydon't see

Registered: 11/06/02
Posts: 818
Last seen: 19 years, 1 day
Re: cloning, sectors, isolates? [Re: thisone]
    #2079445 - 11/07/03 01:06 AM (20 years, 4 months ago)

bump.


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Everything written above this line is completely false. I am terribly bored and write here to pass time. I do not participate or support any illegal activity.

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Offlinemickywilliams2
journeyman
Registered: 05/27/03
Posts: 136
Last seen: 5 months, 17 hours
Re: cloning, sectors, isolates? [Re: thisone]
    #2079585 - 11/07/03 02:09 AM (20 years, 4 months ago)

Ive been wondering the same things.
Ive read of people cloning a good fruit and then getting poor results when they fruit it again.


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InvisibleMycomancer
Psi Cubed
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Registered: 09/28/03
Posts: 586
Loc: United States
Re: cloning, sectors, isolates? [Re: mickywilliams2]
    #2080703 - 11/07/03 11:04 AM (20 years, 4 months ago)



That reminds me of one of the disadvantages of cloning live tissue, cell senescence. It's the point when the cells have almost no telomeres left and are no longer reproducing themselves, So without an influx of healthy young cells, the organism just starts to age and become ederly and proabaly senile.

Basically when using live tissue your not starting a brand new young organism, but instead using already aged cells that have lived through a cycle. If someone takes a tissue sample from a 5th flusher, then they've got some old degenerating mycelium that probably didn't have much life left in them anyway. Best bet is from a very young mushroom(primordia), 1st flush. Mycelium will still be somewhat adolescent and filled with piss and vinegar.

This may be what accounts for (at least in part) to the poor results some people get from cloning.

mycomancer

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OfflineVertigo
Mycovoire
Registered: 03/05/03
Posts: 289
Loc: Panhandle
Last seen: 5 years, 7 months
Re: cloning, sectors, isolates? [Re: Mycomancer]
    #2081133 - 11/07/03 01:28 PM (20 years, 4 months ago)

What are these telomeres that you speak of?


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"One side will make you grow taller, and the other side will make you grow shorter."

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Offlineshirley knott
not my real name
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Registered: 11/11/02
Posts: 9,105
Loc: London Flag
Last seen: 7 years, 2 months
Trusted Cultivator
Re: cloning, sectors, isolates? [Re: Vertigo]
    #2081201 - 11/07/03 01:50 PM (20 years, 4 months ago)

long dangly bits on the ends of chromasomes that get shorter with successive child-bearing. kinda like a woman's breast muscles


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buh

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InvisibleMycomancer
Psi Cubed
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Registered: 09/28/03
Posts: 586
Loc: United States
Re: cloning, sectors, isolates? [Re: Vertigo]
    #2081514 - 11/07/03 04:03 PM (20 years, 4 months ago)

Think of telomeres like they are a bomb fuse or something. Each time a cell divides the length of telomeres at the end of the chromosone shorten. After x divisions, when telomeres are virtually gone, from that point on the cell will no longer divide. The gradual halting of cell reproduction is basically the aging process.

Remember when they cloned the sheep Dolly? Well, they ran into a simmilar problem. Since she was cloned from a donor cell, she didn't have the full regimine (fuse length) of telomeres as normal sheep. Now she is getting old quite early and has lost something like half her life span!

Kids with Progeria are born with extremely short telomeres and have ederly bodies by the time they are 10. Not to long later they die of old age, which is in early adolescents, if that.

The human body has a built in system which scientist have taken advantage of. Each cell has the ability to produce an enzyme called Telomerase. Telomerase actually rebuilds the shortened telomeres after each division, but it's only active a relatively short time. This system is necessary durring foetal development, because in this stage the foetus is going through many many cell divisions as a whole body/person is being constructed, without it we would be born ederly babies. Soon after we're are born, and our bodies are pretty much developed, then Telomerase production is shut down, allowing the aging process.

Scientist have found a way to reactivate production of Telomerase in cells. I believe it was a team of Swedish scientist, but they were testing forskin samples that normally went sescent after approx. 60 divisions. By the time they made their offical report to the medical journals, the skin cells had already divided at least 300 times, with no sign of slowing down, aging, or mutations. They discovered cellular immortality. Just sit for a second and think of the implications of that!

This research was quickly quieted as you would expect, and very little has been heard about it since, but you can do a search for telomeres and run into an article about it.

Death was built into living things to aid in natural selection. The obsolete models needed to perish so that the new improvements could flood the market, so to speak. Then ponder the concept of when and how we evolved a death mechanism. Long long long ago in early life, the aging mechanism might not have been built into the DNA.

Not to end in a bummer or anything, but this also is the same principle that cancer cells work on. Cancerous cells have the Telomerase production going all the time, which is why they are so hard to kill because the are immortal on the cellular level.


mycomancer

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Offlinethisone
the one theydon't see

Registered: 11/06/02
Posts: 818
Last seen: 19 years, 1 day
Re: cloning, sectors, isolates? [Re: Mycomancer]
    #2082599 - 11/07/03 10:37 PM (20 years, 4 months ago)

does the substrate make a difference when selecting a fruit (primordia) to clone? How many generations before it degenerates in comparison to the speed or isolate agar strains (I think you said maybe up to ten but some only four)?


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Everything written above this line is completely false. I am terribly bored and write here to pass time. I do not participate or support any illegal activity.

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OfflineMycoCat
a.k.a. ShroomCat

Registered: 10/09/03
Posts: 1,042
Last seen: 10 years, 3 months
Re: cloning, sectors, isolates? [Re: thisone]
    #2082625 - 11/07/03 10:53 PM (20 years, 4 months ago)

That's pretty cool Mycomancer.


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No question is so difficult to answer as that to which the answer is obvious.

Meow.

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