Home | Community | Message Board


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: North Spore Cultivation Supplies   Original Sensible Seeds High THC Strains   OlympusMyco.com Olympus Myco Bulk Substrate   Left Coast Kratom Buy Kratom Extract   MagicBag.co All-In-One Bags That Don't Suck   Kraken Kratom Red Vein Kratom   Mushroom-Hut Liquid Cultures   Myyco.com Golden Teacher Liquid Culture For Sale   Unfolding Nature Unfolding Nature: Being in the Implicate Order

Jump to first unread post Pages: 1
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
OfflineBajaManitoba
Oysters, mostly

Registered: 09/16/12
Posts: 31
Loc: Baja Manitoba (think abou...
Last seen: 1 month, 1 day
One step...spore print to grain spawn? No agar or plates? * 1
    #17731205 - 02/02/13 06:53 AM (11 years, 3 months ago)

I searched the archives, and don't think that this paper has been discussed.

In 1984, J.P. San Antonio and P.K Hanners of the USDA published a paper on how to take a spore print of an oyster mushroom, and make grain spawn.  No petri plates, agar, etc.

In the method, they made a spore print on wax paper, took the spore print, cut a piece of the wax paper, and suspended the spores in sterile water in a test tube.  They diluted the suspension to a specified turbidity (judged by holding the test tube against a book, and judging how obscured the text is by viewing through the test tube).  Then, they did a dilution of the suspension (something like 0.3 mL in 500 mL of sterile water), and used the diluted spore suspension to inoculate the grain.  They found that by diluting the spore suspension, that contaminants were diluted to extinction, being present in far smaller numbers than the spores. 

I give the citation and quote the abstract below. 

Is this a workable procedure?


San Antonio, J. P. and P. K. Hanners, P. K. 1984.  Using basidiospores of the oyster mushroom to prepare grain spawn for mushroom cultivation.  HortScience  19 (5): 684-686 

Abstract:
    Serial dilution of basidiospores of a spore print to prepare grain spawn of Pleurotus ostreatus is described. The method, which has a contamination rate of about 5%, requires sterilization of a grain medium and the use of some simple clean procedures (described). The method is especially advantageous for small scale and part-time cultivation of oyster mushrooms on logs. P. ostreatus mycelium grew rapidly into the wood of freshly cut logs inoculated with discs of grain spawn prepared using spore inoculum. Unike pure-culture spawn discs, however, discs prepared from spore-inoculated grain did not become completely tightly bonded to the wood surface.

Extras: Filter Print Post Top
Offlinemenlee
Old but New
 User Gallery

Folding@home Statistics
Registered: 11/10/12
Posts: 76
Loc: Flag
Last seen: 11 years, 21 days
Re: One step...spore print to grain spawn? No agar or plates? [Re: BajaManitoba]
    #17731232 - 02/02/13 07:10 AM (11 years, 3 months ago)

:popcorn:

Extras: Filter Print Post Top
Offlinechaka333
mountain grunt.
Male User Gallery

Registered: 07/16/12
Posts: 1,097
Loc: on a hill in a pasture.
Last seen: 8 months, 6 days
Re: One step...spore print to grain spawn? No agar or plates? [Re: menlee]
    #17731253 - 02/02/13 07:23 AM (11 years, 3 months ago)

Interesting. So how did they measure that small of an amount of spores per water? Sorry I just got a little confused by the way it was worded.


--------------------
If you want to achieve greatness
Stop asking for permission.

Extras: Filter Print Post Top
OfflineTerry M
Stranger in a Strange Land
Male


Registered: 06/18/10
Posts: 1,502
Loc: Rhode Island
Last seen: 9 years, 3 months
Re: One step...spore print to grain spawn? No agar or plates? [Re: chaka333]
    #17731285 - 02/02/13 07:37 AM (11 years, 3 months ago)

I don't see why this wouldn't work. Nature does it all the time! The disadvantage is that mating spores will generate a huge variety of different child strains. There's no opportunity to directly obtain a monoculture, as there would be for agar isolation or cloning. So you pretty much take pot luck as to the dominant strain or strains. Propagating a strain with particular desirable traits, like high yield or large fruit size, isn't an option unless you subsequently do fruit cloning.

I'd love to see if you are successful at this, though! It could do an end run around having to mate on agar, skipping purification from contaminants entirely, and then isolating many different monocultures and fruiting them separately.


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

Extras: Filter Print Post Top
OfflineBajaManitoba
Oysters, mostly

Registered: 09/16/12
Posts: 31
Loc: Baja Manitoba (think abou...
Last seen: 1 month, 1 day
Re: One step...spore print to grain spawn? No agar or plates? [Re: Terry M]
    #17731462 - 02/02/13 08:55 AM (11 years, 3 months ago)

They took 1-2 cm2 of spore print and suspended the spores in 5 mL of sterile tap water in a test tube (18 mm inner diameter).  The suspension was too turbid to be able to read the letters of a book, when the test tube was held against the book, so they diluted the suspension with sterile water until they could barely make out the letters.  That level of turbidity was associated with a spore concentration of about 6 million spores per milliliter.  They then took 0.3 mL of this suspension, and injected it into 500 mL of sterile water.  Then they took 0.2 mL of this final, very dilute, suspension to inoculate jars of grain spawn.

So, 6,000,000 x (0.3) x (0.2/500) = about 700 spores per jar of grain spawn.

They looked at many spore prints from oyster mushrooms, and found 12 or fewer contaminants per million spores, so it was pretty easy to get rid of the contams by dilution.

I think the dilution concept has merit for obtaining agar cultures from spore prints.  Make a spore suspension, and then make 10-fold serial dilutions, like you did in Micro 101.  Take a loopfull of each dilution, and place a spot on an agar plate.  In theory, you will eventually have plates where the contaminants in the spores have been diluted to extinction.  Perhaps this is already a common practice.

I can e-mail a scan of this paper to anyone who requests by PM.  It is a government document (USDA work product) and the journal has been open access since 1991, so I think that it would be OK.

Extras: Filter Print Post Top
OfflineKing Reishi
Ninja
Male


Registered: 01/11/13
Posts: 396
Loc: Nibiru
Last seen: 11 years, 1 month
Re: One step...spore print to grain spawn? No agar or plates? [Re: BajaManitoba]
    #17732571 - 02/02/13 01:11 PM (11 years, 3 months ago)

someone should make a nice video or TEK of this.


--------------------
Reishi (fruit) Double Extraction
http://www.shroomery.org/forums/showflat.php/Number/17728431

drying Reishi mycelium
http://www.shroomery.org/forums/showflat.php/Number/17727919

how to take Reishi Mycelium
http://www.shroomery.org/forums/showflat.php/Number/17727387

"THE PSYCHEDELIC EXPERIENCE IS THE TRUE RELIGION"

Extras: Filter Print Post Top
InvisibleArmFromTheAbyss
Old Hand

Registered: 10/09/02
Posts: 1,368
Loc: Down here in Babylon
Re: One step...spore print to grain spawn? No agar or plates? [Re: BajaManitoba]
    #17732661 - 02/02/13 01:23 PM (11 years, 3 months ago)

It might work but it's still MS. I don't see any advantage to this idea. Professional growers use monocultures so they could just scrape some spores on an agar dish and take it from there. The amateurs who use MS can either order a syringe or make one so what's the point?


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

Extras: Filter Print Post Top
OfflineTerry M
Stranger in a Strange Land
Male


Registered: 06/18/10
Posts: 1,502
Loc: Rhode Island
Last seen: 9 years, 3 months
Re: One step...spore print to grain spawn? No agar or plates? [Re: ArmFromTheAbyss]
    #17732708 - 02/02/13 01:31 PM (11 years, 3 months ago)

Quote:

ArmFromTheAbyss said:
It might work but it's still MS. I don't see any advantage to this idea. Professional growers use monocultures so they could just scrape some spores on an agar dish and take it from there. The amateurs who use MS can either order a syringe or make one so what's the point?



I don't see any advantage for ordinary oysters, as the experimenters used. But given that you have just a spore print of some rare species, this would be a great deal less trouble than growing it out into many sectors on agar, then isolating, growing, and fruiting each strain separately.


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

Extras: Filter Print Post Top
Offlinethiotimoline
Stranger
 User Gallery
Registered: 12/01/12
Posts: 898
Loc: Bay Area
Last seen: 6 years, 3 months
Re: One step...spore print to grain spawn? No agar or plates? [Re: Terry M]
    #17733336 - 02/02/13 03:42 PM (11 years, 3 months ago)

Just because you use agar doesn't mean you have to isolate separate strains. I don't see how this saves much effort unless you don't have a still air box.

Extras: Filter Print Post Top
OfflineBajaManitoba
Oysters, mostly

Registered: 09/16/12
Posts: 31
Loc: Baja Manitoba (think abou...
Last seen: 1 month, 1 day
Re: One step...spore print to grain spawn? No agar or plates? [Re: thiotimoline]
    #17737027 - 02/03/13 06:44 AM (11 years, 3 months ago)

thiotimoline wrote:

>I don't see how this saves much effort unless you don't have a still air box.


I think it's another tool for the tool box.  The limited amount of equipment needed is its main appeal.

I will probably try the technique next year in working with some wild oysters I found growing on a stump.

Extras: Filter Print Post Top
OfflineoONoodlesOo
ShroomSauce


Registered: 04/11/10
Posts: 179
Last seen: 10 years, 2 months
Re: One step...spore print to grain spawn? No agar or plates? [Re: BajaManitoba]
    #17766375 - 02/08/13 08:17 AM (11 years, 3 months ago)

i think this would work, if you have a test tube with 0.5ml of purple water :smile: and dilute it to 500ml there should be more spores than contaminants just as you said, im assuming since there should be more spores it should colonize quicker, but why would ya wanna do that when you can just make a lc or a glc? i read a thread where someone made GLC from 2 different strains and they came out 2 different colors :eek:

Extras: Filter Print Post Top
OfflineZenlykos
Wandering One
Male User Gallery


Registered: 12/25/12
Posts: 234
Last seen: 1 year, 4 months
Re: One step...spore print to grain spawn? No agar or plates? [Re: oONoodlesOo]
    #17768444 - 02/08/13 04:02 PM (11 years, 3 months ago)

Definitely bookmarking this.

This is great for those of us (like me) that don't have access to Agar or just don't feel like isolating...

Thanks!!


--------------------
I worship the tree that is not there.

To readers: all of my posts are of knowledge I have obtained from practice, reputable sources, and common knowledge. If anything I say is incorrect, please feel free to correct me so that everyone can benefit.

Anything I post is purely fictional and is not to be taken seriously. :mushroom2:

Extras: Filter Print Post Top
Jump to top Pages: 1

Shop: North Spore Cultivation Supplies   Original Sensible Seeds High THC Strains   OlympusMyco.com Olympus Myco Bulk Substrate   Left Coast Kratom Buy Kratom Extract   MagicBag.co All-In-One Bags That Don't Suck   Kraken Kratom Red Vein Kratom   Mushroom-Hut Liquid Cultures   Myyco.com Golden Teacher Liquid Culture For Sale   Unfolding Nature Unfolding Nature: Being in the Implicate Order


Similar ThreadsPosterViewsRepliesLast post
* 15 year old spore print, viable?
( 1 2 all )
jdm 10,341 31 09/03/03 11:13 AM
by RogerRabbit
* Post deleted by Moe Howard Macey Howard 1,539 8 06/18/02 08:40 PM
by Anno
* Large casings without cakes or grain spawn...
( 1 2 all )
trendalM 3,553 21 04/24/03 02:13 PM
by trendal
* Spore prints from clones maxxjones 1,452 1 03/12/02 10:30 AM
by CLuB99
* spore print knowledge needed compoman 2,861 14 08/02/01 06:39 AM
by Anonymous
* optimal time for azure spore prints Dobie 1,331 9 11/23/02 02:38 AM
by Dobie
* Agar help homebrew 880 2 01/08/02 12:11 AM
by homebrew
* New potent hybrid (Workman and Spore works company)
( 1 2 all )
theshiftingwalls 9,245 36 06/22/03 08:44 PM
by theshiftingwalls

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: RogerRabbit, Pastywhyte, bodhisatta
6,902 topic views. 0 members, 3 guests and 2 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 0.019 seconds spending 0.004 seconds on 12 queries.