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How to perpetuate an Aloha slant?
    #14219506 -

So I've read that a slant should last a lifetime. I plan on purchasing a Shiitake 75 slant from Aloha Medicinals. They are expensive so I want to make sure I get the most out of it. Further down in this post are some of RR's notes on senescense. I wanted to compile a step-by-step list of how to perpetuate a slant while maintaining its maximum fruiting punch/power. I started the following list and was hoping you guys could help me complete it and correct it:
  • 1. Purchase one 75mm X 250mm slant.
  • 2. Inoculate one agar dish.
  • 3. Use agar dish to inoculate twelve or so fresh master culture slants (mcs).
  • 4. Use one of the mcs to inoculate two agar dishes.
  • 5. Use the two agar dishes to inoculate four to five grain jars (quart).
  • 6. Use the grain jars to inoculate four to five large filter patch bags full of grain?
  • 7. Use the filter patch bags to inoculate many supplemented sawdust bags?
  • 8. Repeat steps 4-7 until the first mcs has expired, then move to the next mcs.
  • 9. When all twelve mcs have expired, start from step 2 again.

Are steps 6 and 7 correct? I'm confused. How do you inoculate sawdust bags using colonized grain bags? With a scoop or spoon? I used RR's 8/15/10 note below to create steps 6 and 7. Please help.

------------- RR's notes below ------------------------------------

I'm still using ten year old cultures, except my shiitake 75 which was received six years ago.

Considering one can stretch out an agar wedge to fill an 18 wheel truck or two, a properly maintained culture slant should last several lifetimes.

I use wood in my slants, and just recently got my original shiitake 75 from six years ago out, and made a fresh batch of culture slants.  These are now only P2 from the original after all this time. I'm sure I'll grow old and die before my master cultures have grown even as much as a single spawn bag of rye, which is still good for 4 to 5 additional grain to grain transfers.

Remember, senescence is related to both the actual age of the culture and the number of cell divisions, both of which are limited during refrigeration, and nearly stop when the culture is refrigerated or frozen while colonized in wood.  Even my cultures which die on agar in the slant, are still good once I pull the wood out and place a piece of it on a petri dish.
RR 9/23/10
-------------
When growth slows to a crawl, you'll know senescence has occurred. It's not so much about how many grain to grain transfers you've done but about how many cell divisions the mycelium has undergone.  Note that mycelium which has had 4 G2G with pint jars will have undergone far less cell divisions than mycelium which has had 4 G2G with large spawn bags.

My favorite shiitake strain was received by me in 2001 as a P5, and is now a P8.  I generally expand two petri dishes with it, and then inoculate grains. After that, I do 4 to 5 G2G transfers with large spawn bags before using the grains to spawn to supplemented sawdust for fruiting.  This turns each petri dish wedge into about 10,000 quarts of rye, and I do it without senescence. However, I wouldn't recommend anyone buy my shiitake in a store and try to clone it to start over.  Performance would be poor at best.
RR 8/15/10
-------------
Actually, yes.  I just got it out last year and it was still viable, so I used it to inoculate a petri dish, which in turn inoculated a dozen or so fresh master culture slants.  The original slant was labeled P3 from aloha medicinals, so counting the petri dish in the middle, I labeled all the new slants P5.

This was one of the 75mm X 250mm slants that aloha sends cultures in with a hardwood stick inserted.  You won't get 9 years from a smaller slant.  It also benefited from being opened a few times over the years, allowing fresh filtered air to enter.

However, what I meant by 'have been using it ever since' is that I've been using dishes and slants from that original culture ever since.  Some are already at P11, but still perform as well as the original.  Considering you can take a sliver of mycelium from a slant and grow enough grain spawn to fill a tractor trailer in 8 weeks or so, 11 petri dish transfers hardly has exercised the culture at all.  I also only allow the cultures to grow to about the size of a US quarter before making the next transfer, so that's not too many cell divisions.
RR 7/6/10

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Re: How to perpetuate an Aloha slant? [Re: RiverFish]
    #14219755 -

simplify your process. Your slant is the best those genetics are going to get when you get them in the tube.

1. subculture form slant onto plate. Label plate F2
2. From F2 plate subculture into many master slants, and several more plates. these are all F3
3. every generation of plates from f2 are labeled subsequent generation, F4, F5, F6, etc.

each plate can be used to make a jar G1, G2, G3 etc.

each jar can be used to make 6 to 10 bags

Each bag can be fruited or spawned or whatever

If you had generation 150, or 150 subcultures from you master slant, senescence might set in. Preserving the integrity of the slants and referring back to a slant when you generations exceed 10 to 20 subcultures is good practice.

Check out Growing gourmet and medicinal mushrooms by paul stametes for further information. Make sure to use RR's "little peice of wood " tec in you slant making process, and just for grins try to keep a couple of spore prints on file,. You can also clone any of your mushrooms from this strain to revitalize in case you loose your masters


--------------------
Mycoelf

Sterility is a process that can be likened unto infinity, which is a long walk, the closer to the end you start before beginning, the more achievable  the goal of infinity becomes.  Remember, cleanliness in next to goddessness

:aliendance::aliendance::wicca::aliendance::aliendance::pipesmoke:

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Re: How to perpetuate an Aloha slant? [Re: mycoelf]
    #14219790 -

I forgot the most important part, distribute your quality genetics to your friends on shroomery so that we can do the same and only loose a generation or so in our preservation process. That way if you totally loose your stock, goddess forbid, you can get a bump back from your friends you invested postage in.:goodluck:


--------------------
Mycoelf

Sterility is a process that can be likened unto infinity, which is a long walk, the closer to the end you start before beginning, the more achievable  the goal of infinity becomes.  Remember, cleanliness in next to goddessness

:aliendance::aliendance::wicca::aliendance::aliendance::pipesmoke:

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Re: How to perpetuate an Aloha slant? [Re: mycoelf]
    #14219837 -

mycoelf said:
each jar can be used to make 6 to 10 bags


In this step you are using one quart jar to inoculate 6 to 10 bags of grain?

mycoelf said:
Each bag can be fruited or spawned or whatever


In this step you are scooping the grain out and inoculating supplemented sawdust bags? You wouldn't put a colonized grain bag into fruiting would you?

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Re: How to perpetuate an Aloha slant? [Re: mycoelf]
    #14219852 -

mycoelf said:
distribute your quality genetics to your friends on shroomery


Heck ya! I would love to be able to give back to the shroomery.

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Re: How to perpetuate an Aloha slant? [Re: RiverFish]
    #14222395 -

RiverFish said:
mycoelf said:
each jar can be used to make 6 to 10 bags


In this step you are using one quart jar to inoculate 6 to 10 bags of grain?



or wood, or coir, or whatever the sub of the day is

mycoelf said:
Each bag can be fruited or spawned or whatever


In this step you are scooping the grain out and inoculating supplemented sawdust bags? You wouldn't put a colonized grain bag into fruiting would you?


I am transferring colonized grain from a jar into a bag. Then the bag colonizes and can then be either fruited or move into the next sub, like pasteurized straw out doors. There are about one million variations on how to design a spawn run. Every myc is a little different and is going to want different things. The game is to figure out not only the subs, but the fruiting environment also.

IN your second question you are asking a species specific question. Cubes will fruit straight from grain. That is the exception not the rule. Most mushies use grain as an intermediate source of transfer and nutrition along the way. Most mushies work on shit, straw or wood. cubies ae possibly the easiest mushroom in the world to grow, because you just have to cook some (almost anything) and BAM you got mushies.

GGMM is the best 35 dollars you will ever spend in this hobby. You wont be sorry:goodluck:


--------------------
Mycoelf

Sterility is a process that can be likened unto infinity, which is a long walk, the closer to the end you start before beginning, the more achievable  the goal of infinity becomes.  Remember, cleanliness in next to goddessness

:aliendance::aliendance::wicca::aliendance::aliendance::pipesmoke:

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Re: How to perpetuate an Aloha slant? [Re: mycoelf]
    #14222437 -

mycoelf said:
GGMM is the best 35 dollars you will ever spend in this hobby. You wont be sorry:goodluck:


I second that. The next best $35 you spend would be on Mycelium Running
IMO.


--------------------
Click Below To Enjoy
EvilMushroom666's Take on BRF Cakes
How EvilMushroom666 Prepares Grain Spawn (Jars & Bags)
Proper Pasteurization of Substrates
EvilMushroom's Goodies
Trade List

June 1st 2010

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Re: How to perpetuate an Aloha slant? [Re: mycoelf]
    #14222610 -

Thanks for the clarification. Buying the book now.

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Re: How to perpetuate an Aloha slant? [Re: RiverFish]
    #14242056 -

Ive been wondering the SAME thing for the SAME reason - that GOD AWFUL expensive S75.

I will be preparing petris with PDA, colonizing and tossing in a toothpick.  Once said toothpick colonizes, I'll be dropping it in a culture tube filled to cap with sterile, distilled water.

Sterile, inter, nutrient-free environments cause myc to shift into dormancy - suspended animation.  The reintroduction of nutrients and oxygen reanimates the suspended culture.

When I use it again, I'll just tweezer it out onto a fresh plate or shove it into a grain jar.

As far as I've understood it, sterile water preservation is by far the best catch-all (ease, cost, longevity) way to preserve mushroom cultures in general.  While each strain / species has it's own "best method" sterile water preservation has been proven to maintain efficacy, generally, for up to 20 years.

-SJ


--------------------
Shiitake BRF Growlog - Picture Heavy
1st Shiitake Grow /w Filter Bag
Long Term Preservation TEK

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Re: How to perpetuate an Aloha slant? [Re: shroom-jitsu]
    #14242225 -

Thanks shroom-jitsu. I will definitely try that. Just keep the culture tube at room temp after dropping the toothpick in?

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Re: How to perpetuate an Aloha slant? [Re: RiverFish]
    #14242396 -

Yessir.  Keeping it cool might help, but the write-up I saw said clearly that cultures can be kept in sterile distilled h2o at room temp.


--------------------
Shiitake BRF Growlog - Picture Heavy
1st Shiitake Grow /w Filter Bag
Long Term Preservation TEK

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Re: How to perpetuate an Aloha slant? [Re: shroom-jitsu]
    #14242420 -

Enjoy!

General:
http://www.hardydiagnostics.com/articles/Saving-fungal-Cultures-Sterile-Water-By-Harris.pdf

Efficacy:
http://www.springerlink.com/content/x6m13523648x6357/

Abstract
Five-hundred ninety-four strains of fungi were studied. They were found being preserved with Castellani's method with distilled water during 1 to 20 years. 62% of the strains (n = 368) did grow when subcultured and maintained their main morphological features. 90% of the 20 years old strains of different species were viable. It is argued that the technique of introduction of the strains into the water and their optimal condition will determine survival. The Castellani's method is recommended as easy, cheap and satisfactory for preservation of most species of fungi.


As many seem not to know, please share widely! :smile:


--------------------
Shiitake BRF Growlog - Picture Heavy
1st Shiitake Grow /w Filter Bag
Long Term Preservation TEK

Edited by shroom-jitsu (04/05/11 03:14 PM)

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Re: How to perpetuate an Aloha slant? [Re: mycoelf]
    #14242471 -

mycoelf said:
each jar can be used to make 6 to 10 bags ...

I am transferring colonized grain from a jar into a bag. Then the bag colonizes and can then be either fruited or move into the next sub, like pasteurized straw out doors.


I've been using the RR tek where one jar goes into one bag. As I understand it, the higher your spawn to substrate ratio (actually lower, which is confusing! The more substrate vs. spawn.), the cheaper your cost is, but at the risk of more frequent contamination.

RR's spawn ratio is about 30%. You are using a 2% to 4% ratio. I know this is in the normal range for commercial growers. Did it take any significant time or special effort to achieve this spawn ratio?


--------------------
Liberté, égalité, humidité.

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Re: How to perpetuate an Aloha slant? [Re: Terry M]
    #14247057 -

Terry M said:
mycoelf said:
each jar can be used to make 6 to 10 bags ...

I am transferring colonized grain from a jar into a bag. Then the bag colonizes and can then be either fruited or move into the next sub, like pasteurized straw out doors.


I've been using the RR tek where one jar goes into one bag. As I understand it, the higher your spawn to substrate ratio (actually lower, which is confusing! The more substrate vs. spawn.), the cheaper your cost is, but at the risk of more frequent contamination.

RR's spawn ratio is about 30%. You are using a 2% to 4% ratio. I know this is in the normal range for commercial growers. Did it take any significant time or special effort to achieve this spawn ratio?


I really thought long and hard about this.  My brain divided by zero and I ended up staring blankly at my LCD for an unknown span of time.  I need to understand this better.


--------------------
Shiitake BRF Growlog - Picture Heavy
1st Shiitake Grow /w Filter Bag
Long Term Preservation TEK

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Re: How to perpetuate an Aloha slant? [Re: shroom-jitsu]
    #14247223 -

shroom-jitsu said:
Terry M said:
mycoelf said:
each jar can be used to make 6 to 10 bags ...

I am transferring colonized grain from a jar into a bag. Then the bag colonizes and can then be either fruited or move into the next sub, like pasteurized straw out doors.


I've been using the RR tek where one jar goes into one bag. As I understand it, the higher your spawn to substrate ratio (actually lower, which is confusing! The more substrate vs. spawn.), the cheaper your cost is, but at the risk of more frequent contamination.

RR's spawn ratio is about 30%. You are using a 2% to 4% ratio. I know this is in the normal range for commercial growers. Did it take any significant time or special effort to achieve this spawn ratio?


I really thought long and hard about this.  My brain divided by zero and I ended up staring blankly at my LCD for an unknown span of time.  I need to understand this better.


:grin:

I made a tiny bit of an Excel spreadsheet to compute this. The trick is that you have to subtract out the spawn from the full bag volume to get the volume of substrate. Then you just divide the spawn volume by the substrate volume.


--------------------
Liberté, égalité, humidité.

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Re: How to perpetuate an Aloha slant? [Re: Terry M]
    #14274125 -

I did not mean to throw a wrench in to the works, In GGMM I beleive stametes defines a G1 as 220 grams in a 1 quart jar. I sometimes push this depending on the spawn run to 240 or 260g. I easily spawn 6 5lb'ers from 1 jar, reishi shitake, cubensis or whatever. If I am in a big hurry for the spawn I have gone as high as 2 g1's per bag, but the colonization is so quick you risk experiencing thermal death from the resulting heat created from the metabolism building to critical levels in the sub and killing the myc. I am always careful to maintain a lower incubation temp and proper bag spacing in that scenario.

I have text book colonization times for every strain I spawn at that ratio. I work diligently to maintain my cultures with the youngest possible cell lines, resulting in the most vigorous capture times.when I really want to maximize a spawn ratio, I use a mycelial fragmentation/fermentation technique, through which I can create 60 1 quart G1's from one 100mm petri, that in turn will colonize and be usable in most cases anywhere from 4 to 10 days for the slower growing species. I do not want to spend a ton of time transferring wedges into jars, using the LC technique mentioned I can noc those 60 jars in 20 minutes of session, including pre and post cleaning. All that results in more time for the :bigblunt:


--------------------
Mycoelf

Sterility is a process that can be likened unto infinity, which is a long walk, the closer to the end you start before beginning, the more achievable  the goal of infinity becomes.  Remember, cleanliness in next to goddessness

:aliendance::aliendance::wicca::aliendance::aliendance::pipesmoke:

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Re: How to perpetuate an Aloha slant? [Re: mycoelf]
    #14343468 -

mycoelf said:
GGMM is the best 35 dollars you will ever spend in this hobby. You wont be sorry:goodluck:


Got my copy of GGMM! Ebay for $24. You are right man, this book tells you how to do everything!

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Re: How to perpetuate an Aloha slant? [Re: RiverFish]
    #14344327 -

once you get some time in with GGMM the next book I recommend is it's older companion "the mushroom cultivator" it dates to like 81 and has some old info in there, but there is some things in MC that are not in GGMM, like specific spawn runs for different species, and the section on compost and steam is more comprehensive then the new book

Those two book comprise my "Bible" there is MC wich I refer to as "the old testament" and GGMM that I call "the book of paul":laugh2:

Psilocybes of the world is the book of "revelations" :lighter:


--------------------
Mycoelf

Sterility is a process that can be likened unto infinity, which is a long walk, the closer to the end you start before beginning, the more achievable  the goal of infinity becomes.  Remember, cleanliness in next to goddessness

:aliendance::aliendance::wicca::aliendance::aliendance::pipesmoke:

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Re: How to perpetuate an Aloha slant? [Re: mycoelf]
    #14344634 -

Sorry, I'm just jumping in here to remember this thread.  Good Stuff!


--------------------
I need tropical cultures, ABM, V.v....!!!!

Well things don't always look as they are and things can be misread and mistaken for what they realy are so don't read too much into what I say since I might be mistaken myself. And remember I rarely ever give a definate answer.

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Re: How to perpetuate an Aloha slant? [Re: trout]
    #14356293 -

Much thanks for that Jitsu, it's nice to see someone else mentioning the sterile water method.

I hope once I have sufficient time, I can transfer my cultures into duplicate sterile water storage to double up my library and hopefully have a "non-refridgerated" library set aside to test with years later. :thumbup:


--------------------
Master Slants Made Simple

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Mushrooms, Mycology and Psychedelics > Gourmet and Medicinal Mushrooms
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