Home | Community | Message Board

MushroomMan Mycology
This site includes paid links. Please support our sponsors.


Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Shop: Original Sensible Seeds Autoflowering Cannabis Seeds   Kraken Kratom Red Vein Kratom   Left Coast Kratom Buy Kratom Extract   Myyco.com APE Liquid Culture For Sale   Mushroom-Hut Liquid Cultures   MagicBag.co All-In-One Bags That Don't Suck   PhytoExtractum Buy Bali Kratom Powder   North Spore Bulk Substrate

Jump to first unread post Pages: 1 | 2 | 3 | 4  [ show all ]
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
OfflineFGL
Stranger

Registered: 06/25/05
Posts: 572
Last seen: 9 years, 10 months
differences betwen strain isolation by transfers and by cloning?
    #5831095 - 07/07/06 11:13 AM (17 years, 8 months ago)

If I take a clone from a multispore growing am I getting the same strain qualities than doing isolations on petri dishes? :rolleyes:

Extras: Filter Print Post Top
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery

Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 29 days
Trusted Cultivator
OG Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: FGL]
    #5831103 - 07/07/06 11:15 AM (17 years, 8 months ago)

No.


--------------------
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

Extras: Filter Print Post Top
OfflineFGL
Stranger

Registered: 06/25/05
Posts: 572
Last seen: 9 years, 10 months
Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #5831131 - 07/07/06 11:24 AM (17 years, 8 months ago)

Quote:

RogerRabbit said:
No.




So for getting the best of a strain is it better to do transfers?

Extras: Filter Print Post Top
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery

Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 29 days
Trusted Cultivator
OG Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: FGL] * 1
    #5831155 - 07/07/06 11:30 AM (17 years, 8 months ago)

One swipe of spores on agar will yield hundreds of strains. By selecting a dozen or so of the best rhizomorphic strains and fruiting each one separately, you can find the super performer that will cover every spot of your casing layer with healthy pins. It might take hundreds of multispore grows to find that strain, if ever, because multispore inoculated substrates usually end up with only one or two strains by the time they fruit because they've all combined. (anastomosis)

This means the good fruiting and potent strains combine with the poor fruiting and bunk strains. You never know what you're going to get. Isolate on agar, then fruit separately and test each strain. When you find the best one, you can keep master slants in the refrigerator and grow that isolate forever.
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

Extras: Filter Print Post Top
OfflineFGL
Stranger

Registered: 06/25/05
Posts: 572
Last seen: 9 years, 10 months
Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #5831356 - 07/07/06 12:20 PM (17 years, 8 months ago)

Got it! :thumbup:
Many thanks for your help

Extras: Filter Print Post Top
InvisiblePsychoslut
The Mother Fucking Bear-o-dactyl
 User Gallery
Registered: 12/10/02
Posts: 20,917
Loc: all up in ya
Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #5831383 - 07/07/06 12:25 PM (17 years, 8 months ago)

How do you avoid not running out of original master slants? How do you keep growing out the same isolate and avoid senescence and all that?


Ive read that you cant do too many transfers before the isolate goes bunk. So how do you not run out of "P1"??


--------------------



[quote]KristiMidocean said:
Good now thats clear.WHO FUCKING CARES. If I am fat u all keep pointing it out like its suppose to be a secret.LIke u really have nothing better to do then make fat jokes. If o know its like I do I know yall can come up with NEW AND BETTER SHIT . This shit is old and boring . I left in the first place cause this shit got boring not because of the fat jokes . Fat jokes dont bother me but seriously its old[/quote]

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: differences betwen strain isolation by transfers and by cloning? [Re: Psychoslut]
    #5832740 - 07/07/06 05:53 PM (17 years, 8 months ago)

> One swipe of spores on agar will yield hundreds of strains.

Come on RR, stop calling each individual a "strain". They're sibling individuals and share 50% of their DNA, that hardly qualifies calling each of them a strain.

> How do you avoid not running out of original master slants?

You only take a speck from your master slant and then put it back in the fridge. You only take a tiny bit each time and you grow it out to however much you need. You also keep active cultures going if you need to frequently access that sample.


-FF

Extras: Filter Print Post Top
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery

Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 29 days
Trusted Cultivator
OG Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: fastfred] * 1
    #5833313 - 07/07/06 08:50 PM (17 years, 8 months ago)

Two compatible hyphae pairing up and producing a dykaryon is the mycological definition of a strain. The OMC use of the word strain to denote the geographical area the parent stock came from is marketing driven.
RR

Extras: Filter Print Post Top
Offlinehyphae
born to grow
 User Gallery

Registered: 12/13/02
Posts: 6,228
Loc: the rain forests
Last seen: 12 years, 10 months
Trusted Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: fastfred]
    #5833318 - 07/07/06 08:52 PM (17 years, 8 months ago)

Quote:

fastfred said:
> One swipe of spores on agar will yield hundreds of strains.

Come on RR, stop calling each individual a "strain". They're sibling individuals and share 50% of their DNA, that hardly qualifies calling each of them a strain.

> How do you avoid not running out of original master slants?

You only take a speck from your master slant and then put it back in the fridge. You only take a tiny bit each time and you grow it out to however much you need. You also keep active cultures going if you need to frequently access that sample.


-FF



True fastfreddy they are indeed all substrains, I believe thats what RR means but I'm glad you caught that shows good solid mycological understanding.


--------------------
Getting the most out of your casings!, A pinning strategy.
Oyster Shell "Flour" $2 for 1lb. a hell of a deal :wink:
Not what is overlay but rather what overlay is
Gas Exchange vs. FAE

"We all have priorities. I used a closet once setup a nice little lab trouble was all the shit that was in there ended up in the bedroom that pissed off the GF then I ended up dumping her as she was getting in the way of my sterile culture technique! Ya I got priorities too!!!"

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: differences betwen strain isolation by transfers and by cloning? [Re: hyphae]
    #5833581 - 07/07/06 10:21 PM (17 years, 8 months ago)

Strain: Biology A group of organisms of the same species, having distinctive characteristics but not usually considered a separate breed or variety.

Strain: (biology) a group of organisms within a species that differ in trivial ways from similar groups.

Strain: A strain is sometimes used to indicate a group of plants with similar (but not identical) appearance and/or properties. The term has no official status.


> Two compatible hyphae pairing up and producing a dykaryon is the mycological definition of a strain.

References?

The mating of two monokaryons from the same parent, as is referred to in this post, represents the absolute minimum of genetic variation resulting from the completion of the sexual cycle. It isn't a "strain" and even calling it a "substrain" is being VERY generous.


-FF

Extras: Filter Print Post Top
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery

Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 29 days
Trusted Cultivator
OG Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: fastfred] * 1
    #5834378 - 07/08/06 05:43 AM (17 years, 8 months ago)

I've given the mycological definition of a strain above. I won't argue semantics with you because your old biology book says something different. What I gave is the accepted mycological definition of a strain. Perhaps you can write letters to Stamets, Holliday, Volk, etc., and tell them they're all wrong.
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

Extras: Filter Print Post Top
Offlinehyphae
born to grow
 User Gallery

Registered: 12/13/02
Posts: 6,228
Loc: the rain forests
Last seen: 12 years, 10 months
Trusted Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: fastfred]
    #5834432 - 07/08/06 07:34 AM (17 years, 8 months ago)

Quote:

fastfred said:
Strain: Biology A group of organisms of the same species, having distinctive characteristics but not usually considered a separate breed or variety.

Strain: (biology) a group of organisms within a species that differ in trivial ways from similar groups.

Strain: A strain is sometimes used to indicate a group of plants with similar (but not identical) appearance and/or properties. The term has no official status.


> Two compatible hyphae pairing up and producing a dykaryon is the mycological definition of a strain.

References?

The mating of two monokaryons from the same parent, as is referred to in this post, represents the absolute minimum of genetic variation resulting from the completion of the sexual cycle. It isn't a "strain" and even calling it a "substrain" is being VERY generous.


-FF




fastfreddy we work with different strains ie; PESA, PESH, Amazonians, B+, etc; they all have different charateristics that remain consistant. Now when we take one of thoses strains and do a multispore we create substrains I don't care how you interpret what you read this is simple and easy for anyone/everyone to understand. Were not trying to get too technical here, substrain is a well understood term used in mycology that at least gets us all on the same page. TC bro


--------------------
Getting the most out of your casings!, A pinning strategy.
Oyster Shell "Flour" $2 for 1lb. a hell of a deal :wink:
Not what is overlay but rather what overlay is
Gas Exchange vs. FAE

"We all have priorities. I used a closet once setup a nice little lab trouble was all the shit that was in there ended up in the bedroom that pissed off the GF then I ended up dumping her as she was getting in the way of my sterile culture technique! Ya I got priorities too!!!"

Extras: Filter Print Post Top
Offlinezeegos
Shroomagator
 User Gallery

Registered: 04/03/06
Posts: 827
Loc: bat country
Last seen: 3 years, 2 months
Re: differences betwen strain isolation by transfers and by cloning? [Re: hyphae]
    #6293261 - 11/17/06 08:50 AM (17 years, 4 months ago)

I think theres something i dont quite understand. RR, iv read your write-up on strain isolation on mycotopia and understand that you select different strains from multispore on agar to find the best performing strain. first off your looking for a fruiting strain (of course  :rolleyes:), one which produces large fruits and good pinsets. but i dont understand how this cant be achieved at the same standard as cloning? i mean, isnt cloning different healthy shrooms the same as isolating different strains on an agar dish?

woops, i forgot that only 1 or 2 dominant strains make up the fruits, nevermind  :tongue:

Edited by zeegos (11/17/06 08:53 AM)

Extras: Filter Print Post Top
OfflineBluemoondreamer
Stranger
Registered: 02/11/05
Posts: 107
Last seen: 17 years, 3 months
Re: differences betwen strain isolation by transfers and by cloning? [Re: zeegos]
    #6293375 - 11/17/06 09:28 AM (17 years, 4 months ago)

And strains also share more than 50% of their genes too.

"We found the average nucleotide identity (ANI) of the shared genes between two strains to be a robust means to compare genetic relatedness among strains, and that ANI values of ≈94% corresponded to the traditional 70% DNA–DNA reassociation standard of the current species definition. At the 94% ANI cutoff, current species includes only moderately homogeneous strains, e.g., most of the >4-Mb genomes share only 65–90% of their genes, apparently as a result of the strains having evolved in different ecological settings."

Edited by Bluemoondreamer (11/17/06 09:54 AM)

Extras: Filter Print Post Top
OfflineBluemoondreamer
Stranger
Registered: 02/11/05
Posts: 107
Last seen: 17 years, 3 months
Re: differences betwen strain isolation by transfers and by cloning? [Re: Bluemoondreamer]
    #6293400 - 11/17/06 09:41 AM (17 years, 4 months ago)

To be clearer - when multispore wipes from the same parents are used, they typically retain high 90's % of the same genes. Very little if at all genetic drift.

Extras: Filter Print Post Top
OfflineBluemoondreamer
Stranger
Registered: 02/11/05
Posts: 107
Last seen: 17 years, 3 months
Re: differences betwen strain isolation by transfers and by cloning? [Re: Bluemoondreamer]
    #6293417 - 11/17/06 09:49 AM (17 years, 4 months ago)

strain -

1. The collective descendants of a common ancestor; a race, stock, line, or breed.
2. Any of the various lines of ancestry united in an individual or a family; ancestry or lineage.
3. Biology A group of organisms of the same species, having distinctive characteristics but not usually considered a separate breed or variety: a superior strain of wheat; a smooth strain of bacteria.

There is RR's reference if u really need one.. I think I learned it back in HS.

*edit a perfect example would be compatable hyphae from PF albino and PE pairing - another example would be compatible hyphae from spores within the same PE syringe pairing up. Regardless of the level of DNA similarities - inbreeding or outbreading results in new strains. 1% of genetic drift may not look like worlds apart on paper, but it is worlds apart on the genome level.

Edited by Bluemoondreamer (11/17/06 10:13 AM)

Extras: Filter Print Post Top
OfflineMycoMaster83
FlowHood Builder& HPoo Wrangler
 User Gallery

Registered: 10/28/06
Posts: 46
Last seen: 17 years, 3 months
Re: differences betwen strain isolation by transfers and by cloning? [Re: Bluemoondreamer]
    #6293624 - 11/17/06 11:18 AM (17 years, 4 months ago)

^Interesting

Extras: Filter Print Post Top
Invisiblecreamcorn
mad scientist
Male User Gallery

Registered: 03/13/06
Posts: 2,962
Trusted Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #6293684 - 11/17/06 11:41 AM (17 years, 4 months ago)

Quote:

RogerRabbit said:
One swipe of spores on agar will yield hundreds of strains.  By selecting a dozen or so of the best rhizomorphic strains and fruiting each one separately, you can find the super performer that will cover every spot of your casing layer with healthy pins.  It might take hundreds of multispore grows to find that strain, if ever, because multispore inoculated substrates usually end up with only one or two strains by the time they fruit because they've all combined. (anastomosis)

This means the good fruiting and potent strains combine with the poor fruiting and bunk strains.  You never know what you're going to get.  Isolate on agar, then fruit separately and test each strain.  When you find the best one, you can keep master slants in the refrigerator and grow that isolate forever.
RR




My apologies to responding to part of this thread from months ago, but somebody else bumped it and has me thinking again, I still don't understand this... and I remember talking with RR about it a while back.

I still don't understand what the fundamental difference in the "amount" of varied genes would be between a pure isolate, and a clone.  It still seems to me that both (assuming proper procedure) are isolated strains, one single unified organism, with one single set of genetic material.  So at least it SEEMS to me that both are a different means to the same end.

The difference is that multispore on agar yields many strains (substrains, however you want to word it), so you have many to pursue for isolation, and several to choose from.  By looking for rhizomorphic growth, you have a pretty good idea you'll have a fruiter, but its not a gauruntee.  You have no idea how good of a fruiter it is, until you have fruited it.  You may need to try several, but again you have several to choose from, so one is bound to stand out above the others.  Still, you're not garunteed what you have is near the "best"... you may have been unlucky and got a bunch of crappy strains to choose from - but you don't know until a considerable investment of time, materials, and effort has taken place and you've fruited each and compared.

On the other hand, cloning doesn't leave you with that choice.  On a single grow, that's it.. through anastamosis you've likely narrowed your substrate down to a few (sub)strains, and it still has to do with chance, but your choices are now limited.  So you clone a fruitbody... at least you *know* you're taking genetic material from something capable of fruiting, because the proof is in hand already.  If you happen to get lucky and not only grew a good performer by chance, but chose a good performer, then haven't you reached the game goal? 

Seems to me you're tackling the same problem, only starting at the beginning (agar) versus starting at the end (cloning) and there's pros and cons to each. 

I guess I still don't understand or know for sure that a clone is truly "isolated"... you might have a mix of things that made it up due to the anastamosis... but couldn't that also occur right from inoculation on a petri?  I know you can observe two sectors coming together and a third one emerging, I've seen it.  Isn't the same thing that happens as your particular grow's substrate "joins up" into a single body happening there, just at an earlier stage?  Are we concerned that the third sector came from two others, and didn't purely emerge from spores?  (If so, that's news to me about how you should choose sectors to pursue on agar!) 

At any rate, my 2 cents is I've done it both ways, and cloning really works for me.  (In fact I'm gonna get some plates going next week, I've had a sleeve of petris left over untouched for over a year now and enough agar around to make a thanksgiving jell-o mold with haha, might as well play around :wink:)  But from my personal experiences, while cloning isn't necessarily as reliable to get off the ground with, and the choices are limited to basically the "result" of any given fruiting tray, it has produced the best results for me in the long run, with less effort and specialty materials involved I might add.  The LC cloning I describe in my other thread is basically the conclusion I came to after doing things both ways, its a method that requires little skill, few materials, and even with the "chance" involved in getting a quality specimen to clone, and found that supposed chance at least for me, to *always* be in my favor, with noticably more abundant and potent crops coming from each and any clone I've ever taken, versus starting from multispore.  Maybe I'm alone on this and just lucky with some good mushroom karma? :smile:

Extras: Filter Print Post Top
OfflineEraserhead
Lost Soul
Male User Gallery

Registered: 05/26/06
Posts: 1,363
Loc: Earth
Last seen: 14 years, 4 months
Re: differences betwen strain isolation by transfers and by cloning? [Re: MycoMaster83]
    #6293685 - 11/17/06 11:41 AM (17 years, 4 months ago)

Why not call the seperate isolates "phenotypes" ? like with MJ, each strain has many variant phenotypes.


--------------------

Extras: Filter Print Post Top
Invisiblecreamcorn
mad scientist
Male User Gallery

Registered: 03/13/06
Posts: 2,962
Trusted Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: Eraserhead]
    #6293696 - 11/17/06 11:46 AM (17 years, 4 months ago)

Quote:

Eraserhead said:
Why not call the seperate isolates "phenotypes" ? like with MJ, each strain has many variant phenotypes.




I believe "phenotype" would be reserved for describing characteristics of an actual fruitbody.  For example, I have a culture that produces mushrooms with dimples in their caps (like the pic in my avatar)... I'd consider the "dimple cap" culture I have, a phenotype.  It all points back to my confusion on my long rant above, is a phenotype then, a single set of genetics, or is it a description of a single set of characteristics?  Is it truly an isolated organism?  Would that explain why we have all these words to describe it? :wink:

Extras: Filter Print Post Top
OfflineEraserhead
Lost Soul
Male User Gallery

Registered: 05/26/06
Posts: 1,363
Loc: Earth
Last seen: 14 years, 4 months
Re: differences betwen strain isolation by transfers and by cloning? [Re: creamcorn]
    #6293705 - 11/17/06 11:49 AM (17 years, 4 months ago)

Phenotype
Quote:

1.
1. The observable physical or biochemical characteristics of an organism, as determined by both genetic makeup and environmental influences.
2. The expression of a specific trait, such as stature or blood type, based on genetic and environmental influences.
2. An individual or group of organisms exhibiting a particular phenotype.

phenotypic phe'no·typ'ic (-tĭp'ĭk) or phe'no·typ'i·cal (-ĭ-kəl) adj.
phenotypically phe'no




It goes on if you click the link, I would think isolates would be phenotypes.


--------------------

Extras: Filter Print Post Top
Invisiblecreamcorn
mad scientist
Male User Gallery

Registered: 03/13/06
Posts: 2,962
Trusted Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: Eraserhead]
    #6293743 - 11/17/06 11:59 AM (17 years, 4 months ago)

Yes, isolates would be phenotypes.  But is the reverse true?  Are phenotypes *always* necessarily isolates?  I'm not just playing on semantics I promise, its a legitimate question if you follow me :wink:

If two mushrooms grow side by side and have some defining characteristic that makes them unmistakably the same phenotype (i.e., the dimple spots in the caps would be an example), is it safe to assume they have the same exact genetic make-up?  If I took a clone from each, then fruited them out under the same exact conditions, would I find undistinguisable results in separate grows from either sample?  Or can other characteristics come about that makes the two differ, because they weren't in fact the same to begin with?

To sum up all my babbling thus far, all I'm getting at is this: is there something about a proper agar isolate that makes it more "pure" in a genetic sense than the resulting culture from a cloned tissue sample?  (and if so, how  :confused:)

Extras: Filter Print Post Top
OfflineEraserhead
Lost Soul
Male User Gallery

Registered: 05/26/06
Posts: 1,363
Loc: Earth
Last seen: 14 years, 4 months
Re: differences betwen strain isolation by transfers and by cloning? [Re: creamcorn]
    #6293759 - 11/17/06 12:05 PM (17 years, 4 months ago)

I would think other charateristics would come out, if it was from a multi spore, but if you just had 2 mated spores, I would think they would both produce identically.

Like MJ, has strains, northern lights, kush, blah blah, but each seed is it's own phenotype, but if you take cutings from that plant, all the cuttings will produce _virtually_ the same, if the conditions they are grown under are the same, and they are the same size, and have the same nodes, ect.


--------------------

Extras: Filter Print Post Top
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery

Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 29 days
Trusted Cultivator
OG Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: Eraserhead] * 1
    #6293765 - 11/17/06 12:08 PM (17 years, 4 months ago)

Why not call them what they are? Strains. The definition of a strain is not the geographical area the original print was taken from. That's marketing.

As for cloning vs strain isolation, they're not related. By the time a substrate fruits, hundreds or perhaps thousands of strains have exchanged DNA, either weakening or strengthening the mass. What you get is a 'heinz 57' that may or may not be that great because the weaker genes and the stronger genes(mycelium) have all combined. An example would be mixed breed dogs. We've all seen good examples and others that are dumber then hell.

Strain isolation on agar begins when the spores first start to germinate. I make the first transfers as soon as I can see mycelium growing from the point of inoculation, long before sectoring can be detected. By doing this, and by continuing to separate each individual growth, you can isolate mycelium prior to the process of anastomosis combining dikaryons into a single mass.

You don't isolate looking for one super rhizomorphic strain. You isolate down to single sectors and then fruit out each one to determine the best performer. When you transfer mycelium to a grain master, the original petri dish the mycelium was taken from is placed into a clean refrigerator. By doing this, when you find the best performing strain, you then go back to your well marked petri dishes, thus your original P1 culture. This petri dish can be used to inoculate a few test tube slants that can be incubated for a week, then placed in cold storage. Whenever you need mycelium, a tiny piece the size of a grain of rice can be taken from the test tube and put on agar to grow out, while the test tube is placed back into the refrigerator. These stored test tube cultures preserve the low P value of your isolated strain for years.

I have a complete video tek on strain isolation and master slant preparation and use already filmed. I'll release it when I get the rest of the teks filmed, and editing completed. Hopefully soon.
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

Extras: Filter Print Post Top
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery

Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 29 days
Trusted Cultivator
OG Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #6293777 - 11/17/06 12:11 PM (17 years, 4 months ago)

"To sum up all my babbling thus far, all I'm getting at is this: is there something about a proper agar isolate that makes it more "pure" in a genetic sense than the resulting culture from a cloned tissue sample? (and if so, how )"

Yes. More than one strain can be present in an individual fruit. Thus, a clone is not an isolated strain, unless it was grown from an isolated line of mycelium.
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

Extras: Filter Print Post Top
Invisiblecreamcorn
mad scientist
Male User Gallery

Registered: 03/13/06
Posts: 2,962
Trusted Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #6293792 - 11/17/06 12:15 PM (17 years, 4 months ago)

Quote:

RogerRabbit said:
Strain isolation on agar begins when the spores first start to germinate.  I make the first transfers as soon as I can see mycelium growing from the point of inoculation, long before sectoring can be detected.  By doing this, and by continuing to separate each individual growth, you can isolate mycelium prior to the process of anastomosis combining dikaryons into a single mass.




Ok, well this clears it up mostly for me. :smile:  There was in fact problems with that minor detail there in my procedure... like I mentioned you can see anastamosis take place right in front of you sometimes, when new sectors emerge from where two meet.  I had never considered that as something to avoid transferring (or transfer early to prevent happening), just thought of it as another choice to choose from. :wink:

So essentially you have more choices from an agar isolation process, but those choices are likely going to exhibit a wider range of characteristics in the end, because we (hopefully) captured a single dikaryotic strain from two and only two spores.  Seems reasonable then that the more mating that goes on, the more things tend to become "average" on you... and that's truly the pro then to isolation... you're starting at the top of the family tree and can travel any of those paths, rather than end up with some kid way down at the bottom. :wink:

To use your analogy, cloning leaves us with a "mutt" :smile:  But there still is that element of chance that says this mutt is actually a fantastic performer.  So again, maybe luck, but growing my mutts out has gotten the job done  :thumbup:  :smile:

Extras: Filter Print Post Top
Invisiblecreamcorn
mad scientist
Male User Gallery

Registered: 03/13/06
Posts: 2,962
Trusted Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #6293827 - 11/17/06 12:29 PM (17 years, 4 months ago)

Quote:

RogerRabbit said:
Yes.  More than one strain can be present in an individual fruit.  Thus, a clone is not an isolated strain, unless it was grown from an isolated line of mycelium.
RR




Ok, I guess this is where I need to do some research and make myself understand that... as in on a cellular level I guess.  I remember seeing pictures of yours a while back that was actually of tri-karyotic mycelium, and if i recall right the cells had 3 nucleii or something to that effect?  (and i realize that was to show trikaryotic mycelium happens, but isn't necessarily the norm.. besides the point really)... but i'm supposing there's some mixing and matching of those nuclei which is what allows for more than one strain to be present in the fruit body?

i know that's probably grossly over-simplified and i guess i don't really expect an answer, just mulling it over... have some more advanced reading ready to go when i find the time on the subject :smile:

Extras: Filter Print Post Top
Offlinezeegos
Shroomagator
 User Gallery

Registered: 04/03/06
Posts: 827
Loc: bat country
Last seen: 3 years, 2 months
Re: differences betwen strain isolation by transfers and by cloning? [Re: Eraserhead]
    #6294293 - 11/17/06 03:33 PM (17 years, 4 months ago)

Q - I understand that spores can germinate on agar with peroxide if enough spores are present on the one spot. i read that colonisation speeds of the agar are reduced but would it still work for isolation? or would the dominant substrain take over before the mycelium has time to stretch over the agar and for a workable sized piece to be transfered?

Extras: Filter Print Post Top
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery

Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 29 days
Trusted Cultivator
OG Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: zeegos]
    #6294328 - 11/17/06 03:49 PM (17 years, 4 months ago)

Peroxide should never be used when germinating spores. It's self defeating. If you use a massive amount of spores to find some that will survive the peroxide, it makes sense that you'll also be placing a massive amount of contaminants, some of which also will survive.

Save peroxide for control of cobweb mold. It has little other use in mycology besides possibly helping to control mold in humidifiers.
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

Extras: Filter Print Post Top
Offlinezeegos
Shroomagator
 User Gallery

Registered: 04/03/06
Posts: 827
Loc: bat country
Last seen: 3 years, 2 months
Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #6294374 - 11/17/06 04:06 PM (17 years, 4 months ago)

hmm, ok. when doing the first transfer of mycelium, you say to transfer the first mycelium you see to a new dish. doesn't that basically mean transfer all of the visible mycelium? seeing as it'll be so minute. im just puzzled how this would isolate away from contams.
thats my last q, i promise  :grin:

Extras: Filter Print Post Top
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery

Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 29 days
Trusted Cultivator
OG Cultivator
Re: differences betwen strain isolation by transfers and by cloning? [Re: zeegos]
    #6294389 - 11/17/06 04:12 PM (17 years, 4 months ago)

No. The mycelium doesn't originate from a tiny dot. You 'swipe' the spores onto the agar so that the mycelium begins to grow from the zig zag pattern you make with your loop.(you can even sign your name in agar with spores) That means mycelium will begin growing in streaks. Transfer tiny pieces the size of grains of rice along these streaks to new dishes to begin the isolation process.
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

Extras: Filter Print Post Top
InvisibleRoYaL_fLuSh
Stranger
Male

Registered: 01/04/08
Posts: 499
Loc: UK
Re: differences between strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #9450232 - 12/17/08 02:02 PM (15 years, 3 months ago)

I'm bumping this up 1 because its a great thread and 2 because i got a question !

would then isolating clone tissue transfers be almost a shortcut to achieve the best possible results rather than doing multiple agar transfers from ms prior to any fruiting at all ? if not a shortcut would it at least be the most surefire way to pin down preferred characteristics already tangible with less work before-hand ? not having to do control grows in essence

Extras: Filter Print Post Top
Invisiblearp180
student of life
 User Gallery

Registered: 10/17/08
Posts: 1,449
Loc: Macondo
Re: differences between strain isolation by transfers and by cloning? [Re: RoYaL_fLuSh]
    #9450295 - 12/17/08 02:14 PM (15 years, 3 months ago)

No, isn't that clearly already explained in this two year old thread?

Although a clone is a genetic match to it's parent, unless that parent was created from an isolated strain, you still don't have an isolate.

You are basically breeding a mut.


--------------------
"Given the choice between the experience of pain and nothing, I would choose pain."  William Faulkner

"That which exists without my knowledge exists without my consent."
-A quote from the Judge in the novel Blood Meridian; or the Evening Redness in the West by Cormac McCarthy

"Let there be light" My Quick Reference Guide to Lighting
My AutoMono (11oz First Flush)
My Monster Mushroom Mono (9.3oz First Flush)


Extras: Filter Print Post Top
Offlinedead
grateful
Male User Gallery


Registered: 08/28/08
Posts: 2,761
Loc: North pole
Last seen: 9 years, 3 months
Re: differences between strain isolation by transfers and by cloning? [Re: arp180]
    #9450306 - 12/17/08 02:16 PM (15 years, 3 months ago)

Well, cloning is still beneficial, because even if you get several strains you will know they will all be efficient fruiters. Or at least capable of fruiting.


--------------------
"The third eye. You spend years doing everything you can to open it and then the damn thing opens and your friends laugh at you when you tell them you can see their souls behind their eyes burning like rainbows."

Links:
:regularshroom:Nibin's Guide for Noobs
:regularshroom:some easy teks on bulk & grain prep. (my journal)

Extras: Filter Print Post Top
Invisiblearp180
student of life
 User Gallery

Registered: 10/17/08
Posts: 1,449
Loc: Macondo
Re: differences between strain isolation by transfers and by cloning? [Re: dead]
    #9450333 - 12/17/08 02:22 PM (15 years, 3 months ago)

By no means am I saying cloning doesn't have it's place in mycology. I just answered his question.


--------------------
"Given the choice between the experience of pain and nothing, I would choose pain."  William Faulkner

"That which exists without my knowledge exists without my consent."
-A quote from the Judge in the novel Blood Meridian; or the Evening Redness in the West by Cormac McCarthy

"Let there be light" My Quick Reference Guide to Lighting
My AutoMono (11oz First Flush)
My Monster Mushroom Mono (9.3oz First Flush)


Extras: Filter Print Post Top
InvisibleRoYaL_fLuSh
Stranger
Male

Registered: 01/04/08
Posts: 499
Loc: UK
Re: differences between strain isolation by transfers and by cloning? [Re: arp180]
    #9450347 - 12/17/08 02:24 PM (15 years, 3 months ago)

Quote:

arp180 said:
No, isn't that clearly already explained in this two year old thread?

Although a clone is a genetic match to it's parent, unless that parent was created from an isolated strain, you still don't have an isolate.

You are basically breeding a mut.




i understand that its not going to become a pure isolate .. what im looking for is the best possible results by getting a great preforming mutt and strengthening it ..

Extras: Filter Print Post Top
InvisibleJ3illy
Trainee

Registered: 10/18/08
Posts: 3,344
Re: differences between strain isolation by transfers and by cloning? [Re: RoYaL_fLuSh]
    #9450811 - 12/17/08 03:50 PM (15 years, 3 months ago)

You will definitely have benefits using a clone that are comparable to using an isolate.  You should get a very even, full pinset, w/ all the shrooms growing together w/ similar potency.  It could very well BE an isolate, or it could still be multiple strains - but all the strains will be productive fruiting strains.

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: differences between strain isolation by transfers and by cloning? [Re: J3illy]
    #9451776 - 12/17/08 07:05 PM (15 years, 3 months ago)

Cloning is the easiest way to isolate from a multispore grow.  It works just fine.  Regardless if it sectors a little or not you at least know you are getting the genetics you're looking for.

There could be di-mon matings, other substrains, or whatever but it's a good first step.


-FF

Extras: Filter Print Post Top
Invisiblearp180
student of life
 User Gallery

Registered: 10/17/08
Posts: 1,449
Loc: Macondo
Re: differences between strain isolation by transfers and by cloning? [Re: fastfred]
    #9453180 - 12/17/08 10:43 PM (15 years, 3 months ago)

agreed, but if you isolate many strains and then fruit, to find what fruits best in your conditions, it's an even better step


--------------------
"Given the choice between the experience of pain and nothing, I would choose pain."  William Faulkner

"That which exists without my knowledge exists without my consent."
-A quote from the Judge in the novel Blood Meridian; or the Evening Redness in the West by Cormac McCarthy

"Let there be light" My Quick Reference Guide to Lighting
My AutoMono (11oz First Flush)
My Monster Mushroom Mono (9.3oz First Flush)


Extras: Filter Print Post Top
Invisibleseven
.
Male

Registered: 06/09/08
Posts: 1,478
Loc: north carolina
Re: differences between strain isolation by transfers and by cloning? [Re: arp180]
    #9453564 - 12/17/08 11:57 PM (15 years, 3 months ago)

you can acheive simular results to isolation by taking clones from a couple diferent MS grows. Lets say you have four ms trays. Take two clones from each tray>grow each clone batch out. Now you have eight clone batches to pick from.choose the best clone culture to keep. This isnt isolation but it is growing out a selection of cultures with different genetics to pick from. Note - an isolate is more dependable and consistant. With cloning: there is a chance that the fruits will not turn out the same as the fruit you cloned from. its rare but it happens. Other substrains in the clone culture can become dominate and express different phenotypes the next clone grow. < its rare. point being an isolate has been choped down to a single mating- thus no room for variation except for growing conditions.


--------------------
grind

Extras: Filter Print Post Top
InvisibleShroominit
Part Time Mycologist


Folding@home Statistics
Registered: 11/29/08
Posts: 4,662
Re: differences between strain isolation by transfers and by cloning? [Re: arp180]
    #9453570 - 12/17/08 11:58 PM (15 years, 3 months ago)

Yes and no to your original question. Yes it's a shortcut if you just want to isolate a single fruitable strain. Not necessarily if you want to produce the strongest/fastest growing mycelium or most potent fruit. This is because the combination of strains may make it possible for a set of DNA with the coding for 'potent' to combine with a fruitable strain that is not potent. While when you separate them out the potent strain may not be fruitable. Does that make sense?

The different characteristics that make up a fruit body may not be presented when separated out.

Extras: Filter Print Post Top
Invisibleseven
.
Male

Registered: 06/09/08
Posts: 1,478
Loc: north carolina
Re: differences between strain isolation by transfers and by cloning? [Re: seven]
    #9453669 - 12/18/08 12:14 AM (15 years, 3 months ago)

i think cloning to agar and transfering each sector that comes from a fruit would be a quicker method of getting some isolates. the isolates are somewhat chosen for you as the results of a MS grow, but you will make less transfers.Same could be done to a handfull of other different ms clones for genetics to choose from. this is sort of a question: I havent done this but it seems logical you can use clones to acheive isolates in less transfers? :shrug:


--------------------
grind

Extras: Filter Print Post Top
InvisibleShroominit
Part Time Mycologist


Folding@home Statistics
Registered: 11/29/08
Posts: 4,662
Re: differences between strain isolation by transfers and by cloning? [Re: seven]
    #9453718 - 12/18/08 12:22 AM (15 years, 3 months ago)

Yes seven, of course. Like I said if you just want something that will fruit and give you an even pinset with hopefully stronger mycelium, that will do it just fine.

When you isolate for a specific property, that is when you run into issues with a clone.

Also clones would need way less dishes/transfers.

Extras: Filter Print Post Top
Invisibleseven
.
Male

Registered: 06/09/08
Posts: 1,478
Loc: north carolina
Re: differences between strain isolation by transfers and by cloning? [Re: Shroominit]
    #9453845 - 12/18/08 12:46 AM (15 years, 3 months ago)

Quote:

Shroominit said:
Yes seven, of course. Like I said if you just want something that will fruit and give you an even pinset with hopefully stronger mycelium, that will do it just fine.

When you isolate for a specific property, that is when you run into issues with a clone.

Also clones would need way less dishes/transfers.


A clone will almost always show phenotypes of the fruit its cloned from. Isolation of a substrain takes the variation of phenotypes from other substrains out of the picture. Only growth conditions will likely change results of an isolate's performance. :thumbup:


--------------------
grind

Extras: Filter Print Post Top
Offlinedead
grateful
Male User Gallery


Registered: 08/28/08
Posts: 2,761
Loc: North pole
Last seen: 9 years, 3 months
Re: differences between strain isolation by transfers and by cloning? [Re: seven]
    #9454594 - 12/18/08 06:57 AM (15 years, 3 months ago)

As I understand it is very rare for a clone to have several substrains.


--------------------
"The third eye. You spend years doing everything you can to open it and then the damn thing opens and your friends laugh at you when you tell them you can see their souls behind their eyes burning like rainbows."

Links:
:regularshroom:Nibin's Guide for Noobs
:regularshroom:some easy teks on bulk & grain prep. (my journal)

Extras: Filter Print Post Top
InvisibleJ3illy
Trainee

Registered: 10/18/08
Posts: 3,344
Re: differences between strain isolation by transfers and by cloning? [Re: dead]
    #9454598 - 12/18/08 06:59 AM (15 years, 3 months ago)

I've wondered how common it would be myself, but I don't think it's that rare.  Someone said they once had a clone sector into like 6 sub-strains.  I don't think it's that uncommon for multiple sub-strains to 'collaborate' on a fruit.

Extras: Filter Print Post Top
Offlinedead
grateful
Male User Gallery


Registered: 08/28/08
Posts: 2,761
Loc: North pole
Last seen: 9 years, 3 months
Re: differences between strain isolation by transfers and by cloning? [Re: J3illy]
    #9454989 - 12/18/08 08:54 AM (15 years, 3 months ago)

On the other hand I've heard several people say they have never seen a clone culture sector at all.


--------------------
"The third eye. You spend years doing everything you can to open it and then the damn thing opens and your friends laugh at you when you tell them you can see their souls behind their eyes burning like rainbows."

Links:
:regularshroom:Nibin's Guide for Noobs
:regularshroom:some easy teks on bulk & grain prep. (my journal)

Extras: Filter Print Post Top
Offlinegrod31
Stranger
Male


Registered: 12/06/07
Posts: 2,077
Loc: New York
Last seen: 15 years, 1 month
Re: differences between strain isolation by transfers and by cloning? [Re: dead]
    #9455086 - 12/18/08 09:26 AM (15 years, 3 months ago)

i have had a clone sector, they defiantly do


--------------------
Back the tape up.  I need it again!
Let it roll!  Just as high as the
fucker can go!  And when it comes
to that fantastic note where the
rabbit bites its own head off, I
want you to  THROW THAT FUCKING
RADIO INTO THE TUB WITH ME!
    Not me.  It would blast you through
the wall stone dead in ten
seconds and they'd make me explain
VVVVV Free Myco Thread

Extras: Filter Print Post Top
Offlinebeandip
mycelium
Male


Registered: 04/20/08
Posts: 425
Last seen: 3 years, 2 months
Re: differences between strain isolation by transfers and by cloning? [Re: grod31]
    #11017830 - 09/07/09 11:01 PM (14 years, 6 months ago)

Has anyone ever measured specific chemical compositions while isolating a strain? Theoretically could you isolate any above average mycelium (that showed excellent fruits with higher biochemical composition) to create some type of super strain?

Extras: Filter Print Post Top
Invisibleratdog
wild capture trader
Male User Gallery


Registered: 08/16/09
Posts: 959
Loc: Colorado mount. top
Re: differences between strain isolation by transfers and by cloning? [Re: Bluemoondreamer]
    #11017945 - 09/07/09 11:26 PM (14 years, 6 months ago)

Quote:

Bluemoondreamer said:
And strains also share more than 50% of their genes too.

"We found the average nucleotide identity (ANI) of the shared genes between two strains to be a robust means to compare genetic relatedness among strains, and that ANI values of ≈94% corresponded to the traditional 70% DNA?DNA reassociation standard of the current species definition. At the 94% ANI cutoff, current species includes only moderately homogeneous strains, e.g., most of the >4-Mb genomes share only 65?90% of their genes, apparently as a result of the strains having evolved in different ecological settings."





i think this is quit clear as to the need to isolate then clone instead of just cloning.

some of the impure genetics from a clone of a multi spore will show up as were a isolate which was isolated for a specific purpose to a point of stable repeatable traits will not.

so the end game is there but you need real good luck to win on the first try. cuz the second or third might loose the game for you.


LMAO!!!


--------------------
some people just don't get it:spank:
http://www.shroomery.org/forums/showflat.php/Number/11241796
so here is a video or two or three for you guys:rolleyes:

Extras: Filter Print Post Top
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery


Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 29 days
Trusted Cultivator
OG Cultivator
Re: differences between strain isolation by transfers and by cloning? [Re: ratdog]
    #11018049 - 09/07/09 11:43 PM (14 years, 6 months ago)

Quote:

i think this is quit clear as to the need to isolate then clone instead of just cloning.




When you isolate, you don't need to clone because you save well-labeled mycelium from each culture under refrigeration.  Later, when you determine the best isolates, you go back to your culture collection and grab that specific one to grow out.  This way, you always have very young cell lines, not months old cell lines from a clone. 

These juvenile cell lines stored in master slants are how commercial mushroom farms stay in business.  If they kept cloning, all their cultures would fade out due to senescence.
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

Extras: Filter Print Post Top
Invisibleratdog
wild capture trader
Male User Gallery


Registered: 08/16/09
Posts: 959
Loc: Colorado mount. top
Re: differences between strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #11018107 - 09/07/09 11:53 PM (14 years, 6 months ago)

Quote:

RogerRabbit said:
Quote:

i think this is quit clear as to the need to isolate then clone instead of just cloning.




When you isolate, you don't need to clone because you save well-labeled mycelium from each culture under refrigeration.  Later, when you determine the best isolates, you go back to your culture collection and grab that specific one to grow out.  This way, you always have very young cell lines, not months old cell lines from a clone. 

These juvenile cell lines stored in master slants are how commercial mushroom farms stay in business.  If they kept cloning, all their cultures would fade out due to senescence.
RR





this is a good deff of senescence too thanks RR


--------------------
some people just don't get it:spank:
http://www.shroomery.org/forums/showflat.php/Number/11241796
so here is a video or two or three for you guys:rolleyes:

Extras: Filter Print Post Top
Invisibleanonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
Re: differences between strain isolation by transfers and by cloning? [Re: ratdog] * 1
    #11018130 - 09/07/09 11:56 PM (14 years, 6 months ago)


SENESCENCE
The End is Near


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

Extras: Filter Print Post Top
Offlinebeandip
mycelium
Male


Registered: 04/20/08
Posts: 425
Last seen: 3 years, 2 months
Re: differences between strain isolation by transfers and by cloning? [Re: anonjon]
    #11024051 - 09/08/09 08:50 PM (14 years, 6 months ago)

I can also see where the cost of materials would come into play. It is cheaper to figure out you have a good strain to grow in bulk on a small amount of agar rather than a whole jar, bag, or tub. I am going to look into doing isolation once I get myself a mini fridge. I would prefer to keep my isolated strains separate from my leftover mexican food.

RR do you perhaps have an answer to my original question?

Extras: Filter Print Post Top
Invisibleanonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
Re: differences between strain isolation by transfers and by cloning? [Re: beandip]
    #11028561 - 09/09/09 04:34 PM (14 years, 6 months ago)

took me about 30 seconds to find this:
'I'm going to throw something out here that contradicts the prevailing beliefs in cloning/isolation. I would like to see some of you in a position to do so, attempt to verify or disprove the following.

I have noticed when cloning fruits from multispore inoculation, that more than one strain is cloned. I have seen obvious sectors on the agar from a single piece of cloned tissue. At first, I thougt I must have gotten some spores that were stuck to the stipe into the mix, but later I was very careful to only select tissue from deep inside the stem from fruits that had not yet opened their caps.

My theory based on these observations is that single fruits on a cake or cased substrate from multispore inoculation, can and are sometimes made up of multiple strains(substrains in shroomy speak).

Multiple strains within the same fruitbody? That is how it appears to me. Before anyone answers "impossible", do a bit of research. If there's another explanation, I'm all ears. I'm not talking about different types of mycelium such as cottony, rhizo, airy, etc., but actually separate sectors that when transferred to different petri dishes will not mix with other sectors from the same frutbody. Sometimes fact is stranger than fiction. I'd love to see a few of you myco-geeks jump on this. I know I am. Have fun.
RR '

originally here: http://www.shroomery.org/forums/showflat.php/Number/4345100#4345100


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: differences between strain isolation by transfers and by cloning? [Re: anonjon]
    #11029016 - 09/09/09 06:00 PM (14 years, 6 months ago)

Nobody has ever shown that sectoring is in fact proof that there are different substrains involved.  There are a lot of different possible explanations.

It could be differing life stages, environmental conditions, differentiation of the tissue, haploidization, etc., etc..

While it's not impossible that different individuals have combined to create a fruit body, it's only one possible explanation.  It's also quite common to have myc colonize the surface of fruit bodies.  So it's quite possible that the fruit body simply had a different substrain grow on the surface of it, contaminating the culture.

Anyhow, more research is needed.  Someone who's isolated and identified mating types could haploidize and type the resulting tissue from these sectors to see if they are different mating types or what exactly is going on.


-FF

Extras: Filter Print Post Top
Invisibleanonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
Re: differences between strain isolation by transfers and by cloning? [Re: fastfred]
    #11029838 - 09/09/09 07:58 PM (14 years, 6 months ago)

Weird. Dikaryotic tissue can 'unmate' back? This can be forced? When the cell nucleus loses chromosomes, does it only lose the dna from one spore or the other? i.e. is the resulting tissue exactly like one of the original mating partners, or can it be lose some chromosomes from each side, resulting in a new unique haploid sequence?


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

Extras: Filter Print Post Top
Invisibleratdog
wild capture trader
Male User Gallery


Registered: 08/16/09
Posts: 959
Loc: Colorado mount. top
Re: differences between strain isolation by transfers and by cloning? [Re: anonjon]
    #11029935 - 09/09/09 08:08 PM (14 years, 6 months ago)

DE-evolution???  i think not! try mutation.


--------------------
some people just don't get it:spank:
http://www.shroomery.org/forums/showflat.php/Number/11241796
so here is a video or two or three for you guys:rolleyes:

Extras: Filter Print Post Top
Invisiblefastfred
Old Hand
Male User Gallery

Folding@home Statistics
Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
Re: differences between strain isolation by transfers and by cloning? [Re: ratdog]
    #11035973 - 09/10/09 06:35 PM (14 years, 6 months ago)

> Dikaryotic tissue can 'unmate' back?

Yep.

> This can be forced?

Yep.  Stress or the use of haploidizing agents can cause this.  It can also happen spontaneously.


Quote:

When the cell nucleus loses chromosomes, does it only lose the dna from one spore or the other? i.e. is the resulting tissue exactly like one of the original mating partners, or can it be lose some chromosomes from each side, resulting in a new unique haploid sequence?




Usually the parental types will be recovered.  However, if recombination occurs then the recovered types will not be a parental type.


-FF

Extras: Filter Print Post Top
OfflineRailrider
Chronic contaminator


Registered: 07/10/09
Posts: 598
Loc: a van down by the river.
Last seen: 9 months, 30 days
Re: differences between strain isolation by transfers and by cloning? [Re: fastfred]
    #11036895 - 09/10/09 09:18 PM (14 years, 6 months ago)

So this thread is saying that cloning from a fruit is a waste of time?Using the heinz 57 dog analogy, when the cloned myc. starts to fruit you will not know how its going to react due to all of the different genetic material available to it from the original multispore inoculation and myc. growth resulting from it?

Even when you use the iso method it still may be a crap shoot due to great rhizo. growth but crappy fruiting/potency?

If you happen to get lucky enough to find the "golden ticket"(all the great qualities) of spores have it fruit, the spores that drop from that fruit body is going to be as random as when you started originally from the iso?

Then if you take a chunk of that fruit and grow it in an lc the subsequent quality of the fruits there after will degrade if you repeat the process over and over again.

I would think the biggest challenge would be to have every iso tested for potency.It makes you wonder how everyones crappy multispore shrooms even grow or make you trip:smirk:


--------------------

Any post I post here is pure speculation from other read posts
Best tip for cultivation!

Extras: Filter Print Post Top
Invisibleanonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
Re: differences between strain isolation by transfers and by cloning? [Re: Railrider]
    #11037312 - 09/10/09 10:17 PM (14 years, 6 months ago)

It's not a waste of time, you can make a master slant of your clone tissue and use it to make as many as you want/need.

And multispore is not as much of crapshoot given the way these strains are cultured/cloned. I'd expect the gene-pool to diminish over time. I think this is true of penis envy strain in particular.

It's just that you need to have a cycle of sexual reproduction after a few generations of clones. A cell can only divide so many times before the dna replication starts getting faulty. Sexual reproduction gives it a fresh start.

The variety of possible variations within a strain is generally static. I wouldn't expect several generations of multispore to degrade the strain any.


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

Extras: Filter Print Post Top
Offlinebusyskull
Captain Amigo
Male


Registered: 07/01/09
Posts: 62
Loc: Tincan Mountain
Last seen: 12 years, 6 months
Re: differences between strain isolation by transfers and by cloning? [Re: seven]
    #11196395 - 10/06/09 08:10 PM (14 years, 5 months ago)

I isolated feather type mycelium to new dish, now i transferred from a feqather type from that sample onto a new dish and all myceleum is fuzzy again ,- the whole dish
any advice

Extras: Filter Print Post Top
Invisiblefeelfunny
I am you
Male User Gallery


Folding@home Statistics
Registered: 03/11/09
Posts: 8,747
Loc: South
Re: differences between strain isolation by transfers and by cloning? [Re: busyskull]
    #11196420 - 10/06/09 08:13 PM (14 years, 5 months ago)

transfer till you get this and keep going it will get there


--------------------
IF A CAT AND DOG CAN GET ALONG WHY CANT EVERYONE ELSE?
If the sky is falling, don't look up!  :abduction:

Feel Family Founder. :pm: me if you are tired of hearing, "Use the search function".

Extras: Filter Print Post Top
Jump to top Pages: 1 | 2 | 3 | 4  [ show all ]

Shop: Original Sensible Seeds Autoflowering Cannabis Seeds   Kraken Kratom Red Vein Kratom   Left Coast Kratom Buy Kratom Extract   Myyco.com APE Liquid Culture For Sale   Mushroom-Hut Liquid Cultures   MagicBag.co All-In-One Bags That Don't Suck   PhytoExtractum Buy Bali Kratom Powder   North Spore Bulk Substrate


Similar ThreadsPosterViewsRepliesLast post
* Do different strains = different trips?
( 1 2 3 all )
oO_wombat_Oo 11,750 57 07/14/09 09:21 PM
by rovert
* Isolates vs Clones homebrew 2,453 1 04/27/03 01:26 AM
by zeronio
* Is there a strain isolation tek? thecannuck 3,632 2 12/19/02 07:17 AM
by Raadt
* Re: Whats Tha Difference?? (Strains) Lizard King 1,803 5 04/04/00 03:58 PM
by LillSkit
* What is the point of all these different strains?
( 1 2 all )
EgoMagickian 3,236 22 07/10/03 09:15 AM
by YellowPurpleHills
* mixing different strains of spawn thescientist 2,075 1 03/15/02 04:28 PM
by black_dove
* Can different strains of mushrooms be in the same terrianium The_Greater_God 2,774 19 02/02/03 03:40 PM
by ShroomBoarder
* 2 different strains... Mycelium 688 1 06/24/02 04:38 PM
by bassplayer74

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: Shroomism, george castanza, RogerRabbit, veggie, mushboy, fahtster, LogicaL Chaos, 13shrooms, Stipe-n Cap, Pastywhyte, bodhisatta, Tormato, Land Trout, A.k.a
14,990 topic views. 8 members, 116 guests and 28 web crawlers are browsing this forum.
[ Show Images Only | Sort by Score | Print Topic ]
Search this thread:

Copyright 1997-2024 Mind Media. Some rights reserved.

Generated in 0.039 seconds spending 0.005 seconds on 12 queries.