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Offlineloggrower
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The whole P-Value/Generation thing needs to be rethought...
    #19026953 - 10/24/13 05:33 PM (10 years, 3 months ago)

I was thinking about the P-Value/Generation thing for cloning and it is extremely inaccurate. We are trying to approximately measure the number of cellular divisions from the master/original, correct? Then distance is the key.

If someone clones a mushroom on a plate and then takes a slice from the edge of a plate it will have grown twice the distance of a slice taken from halfway between the middle and the edge of the plate. Different samples taken from different locations on a plate can have three times or more the divisions of others on the plate.

Wouldn't it make much more sense just to record growth distance? It wouldn't be complicated and it would be much more accurate... I propose a D-Value measured in cm!

The only way P-Value/Generations would make any sense is if each cutting and transfer is the main thing damaging the genetics of the strain...


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Invisiblelaughingsol
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Re: The whole P-Value/Generation thing needs to be rethought... [Re: loggrower]
    #19027068 - 10/24/13 05:59 PM (10 years, 3 months ago)

Quote:

loggrower said:
I was thinking about the P-Value/Generation thing for cloning and it is extremely inaccurate. We are trying to approximately measure the number of cellular divisions from the master/original, correct? Then distance is the key.

If someone clones a mushroom on a plate and then takes a slice from the edge of a plate it will have grown twice the distance of a slice taken from halfway between the middle and the edge of the plate. Different samples taken from different locations on a plate can have three times or more the divisions of others on the plate.

Wouldn't it make much more sense just to record growth distance? It wouldn't be complicated and it would be much more accurate... I propose a D-Value measured in cm!

The only way P-Value/Generations would make any sense is if each cutting and transfer is the main thing damaging the genetics of the strain...





... or if you take each transfer at the same point under very tightly controlled conditions... or if you're trying to advertise a marketing gimmick to separate your company from others... 

The p-value system, although it may have its virtues, has never been subjected to a peer reviewed double blind study proving its efficacy.  I postulate that most mycologists would agree that there are more factors involved in senescence than just the number of cell divisions a culture has undergone.


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Offlineloggrower
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Re: The whole P-Value/Generation thing needs to be rethought... [Re: laughingsol]
    #19027483 - 10/24/13 06:53 PM (10 years, 3 months ago)

Yes, sugars are also important... Agar plates with simple sugars will drive a strain into senescence real fast, while complex sugars maintain it better.

Has there been any thorough research into which agar's sustain Oyster and Shiitake the best?


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Invisiblelaughingsol
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Re: The whole P-Value/Generation thing needs to be rethought... [Re: loggrower]
    #19027904 - 10/24/13 07:53 PM (10 years, 3 months ago)

Quote:

loggrower said:
Yes, sugars are also important... Agar plates with simple sugars will drive a strain into senescence real fast, while complex sugars maintain it better.

Has there been any thorough research into which agar's sustain Oyster and Shiitake the best?




My limited understanding suggests that a rotation of agar recipes is preferable to a single repeated recipe.  Also, I believe the agar recipe used is secondary to general good culture preservation practices.  For example, a long term preservation slant should be less nutritious than an agar plate poured to produce vigorous mycelium for grain spawn inoculation.  Other factors like temperature fluctuation/maintenance in your culture fridge will have a much larger effect on long term culture health than the agar recipe(s) that you use.  The practices in place between taking your slants out of long term storage and returning them to long term storage will also have a greater effect than the agar recipe used. 

Heres a recently posted and interesting article about long-term storage in sterile distilled water.


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Offlinewood chip
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Re: The whole P-Value/Generation thing needs to be rethought... [Re: laughingsol]
    #19029386 - 10/25/13 02:10 AM (10 years, 3 months ago)

Loggrower said  "If someone clones a mushroom on a plate and then takes a slice from the edge of a plate it will have grown twice the distance of a slice taken from halfway between the middle and the edge of the plate"

The mycelium at the edge is excreting more extracellular enzymes into the substrate and is not differentiating.  It is acquiring the material to assemble itself. The vegetative growth is occurring faster.  Older differentiating cells are growing slower through the substrate and seem to have obtained the material to assemble and manifest themselves ultimately as the reproductive fruit body or mutation of one. 

The tips cells have not grown twice the distance.  They did not exist when the clone hit the plate.  They are younger cells. even then the master.  younger is not better. It is the same but has come to be alive in the present unless a mutation occurred due to the environment.  Time does not exist for fungi.


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Offlineloggrower
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Re: The whole P-Value/Generation thing needs to be rethought... [Re: wood chip]
    #19029515 - 10/25/13 03:22 AM (10 years, 3 months ago)

You mean that the cells at the edge of the plate have undergone the same number of cellular divisions as cells near the middle where the mycelium grew from?

I cannot see how that is possible...

Should be less cellular divisions and a closer age to the original nearer the center of the plate... When I refer to age, I just mean cellular divisions. It's the same thing really... Humans even get old from too many cellular divisions.


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OfflineRogerRabbitM
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Re: The whole P-Value/Generation thing needs to be rethought... [Re: loggrower]
    #19029765 - 10/25/13 06:45 AM (10 years, 3 months ago)

Quote:

The whole P-Value/Generation thing needs to be rethought...




Nobody uses that system but Paul.  It's inaccurate.

Here's what I do(4 year old thread)

Furthermore, generations have nothing to do with senescence since every generation starts over with fresh cell lines.
RR


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Offlinewood chip
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Re: The whole P-Value/Generation thing needs to be rethought... [Re: RogerRabbit]
    #19030964 - 10/25/13 12:51 PM (10 years, 3 months ago)

"You mean that the cells at the edge of the plate have undergone the same number of cellular divisions as cells near the middle where the mycelium grew from?"

No. Generally, a single cell divides. Then there are two, (half the material is from the original cell) they divide then there are four lateral branching occurs causing exponential growth, The cell that gave life to number two is older.  The two cells that gave life to four may or may no be the the same age.  However, overtime the division of cells makes many different ages simultaneously.

To me and older cell is one that has differentiated usually as a colony. 

A a single individual cell at the tip or edge of the plate could be thought of as a stem cell in the case of a pure strain with predictable growth patterns. 

All the DNA is there to differentiate into any possible outcome depending on the environment and interconnectedness of all the cells as a unit, but the cell has not yet done so.  This material at the edge.  It is closest to the master for this reason.

This is my theory, 
I am sure there are other themes depending on individual fungi that contradict the idea.

I looked at RR thread he posted a link to which uses of the number of days out of refrigeration.
 
I think this if useful, but have stored slants at room temperature for years with no degradation of culture. (I do not what the how number of days at room temperature effect the culture once it has colonized the slant matter for shiitake) I think it would be different in you ran the live mcyleium across the petri plate for years by transferring every five days due to fluctuation and non exact replication of the DNA. But if the cells breed true the culture should be the same age from the fungi perspective, even you if are 10 years closer to death.  Some strains may be effectively immortal.


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