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CaptainAhab


Registered: 12/25/09
Posts: 1,875
Last seen: 6 years, 1 month
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Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ????
#14351526 - 04/25/11 08:38 PM (12 years, 10 months ago) |
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Two different species of Panaeolus, cambodginiensis and suphanburi (goliath) were transferred to agar in a flow hood, and took over 2 weeks to show signs of life. After sitting out for about 1.5 to 2 months, they had grown out to about the size of a quarter. This was transferred to pc'd grain (rye berries), and nothing has happened in a month.
What can explain this? Anyone have this happen to them, before? According to my research, pans are supposed to be as quick as cubensis on agar, LC, grain, etc. I'm trying to do a pictorial/TEK for my thread http://www.shroomery.org/forums/showflat.php/Number/13249652 but now am in need of some prints.
Check out my marketplace post if you can help out, that would be great. Thanks http://www.shroomery.org/forums/showflat.php/Number/14351397
Here's my research on the issue: From the Panaeolus FAQ: http://www.shroomery.org/8421/Panaeolus-cyanescens-FAQ#agar (just scroll up a little bit) "How fast do Panaeolus cyanescens grow? I found the Panaeolus cyanescens to be very fast compared to P. cubensis. I germinate spores on agar and transfer to Manure:rye seed:perlite in quart jars. These run through in a max of 14 days, but usually in under 10 days. Once laid out in tray and cased with a very thin layer of Peat:perlite, the pin in about 7 days, and mature by the 13 day max. Everything is fast."
I've seen lots of LC's people say only take a week to fully colonize, too.
Here's a quote from Blue Helix at http://www.shroomery.org/forums/showflat.php/Number/4872154#4872154 "Liquid Culture - Days 3, 5, and 7 (completion). Starting with a few mls of live culture rather than few spores can shorten this stage to around only 48 hours rather than 7 days... I have since seen Goliath Pan Cyans develop from 3ccs of liquid starter culture in only 48 hours, making them faster than cubensis."
The only difference between Blue Helix's LC and malt extract agar is the dextrose. Still, if LC's and quart jars of spawn colonize as fast or faster than cubensis, then it would seem that it would be the same with agar. The agar I used is also recommended for pan species (see link in panaeolus faq above).
Here's a thread on Panaeolus on agar: http://www.shroomery.org/forums/showflat.php/Number/10992165#10992165 "Started off with eight petri dishes of light malt extract agar, using one drop of spores from a multispore syringe. Out of the eight the following three are all that show growth after nine days. the ones that don't show any growth have excess moisture on the surface of the agar that I can see run when I tilt the petri dish. I believe I poured the agar a little too thick in the dish, and the surface I pored them on wasn't level. So, one side of the dish is thicker than the other."
Here's a test of different agars by a trusted cultivator I've never heard of: http://www.shroomery.org/forums/showflat.php/Number/5673196#5673196 "I wanted to know on which agar panaeolus mycelium grows faster. PDYA or MEYA. So I made a MEYA Agar plate and cut the half out. I filled it with PDYA. The I cut 3 pieces out and filled this "holes" with YGC Agar (Yeast Glucose Chloramphenicol) to look if the mycelium with grow on it too. And the winner is PDYA by now"
An interesting read on Panaeolus bisporus where it was recommended to hot-pour the agar onto spores. This is from StoneSun and someone named Lipa: http://www.shroomery.org/forums/showflat.php/Number/14116777#14116777 "As per lipa's advice spores were germinated on MEA in a fashion of hot pouring the agar on the spores. Germination occurred in just 48 hours after the petri has been poured. Sterilized organic rye berries (1hr @20psi) were inoculated with agar wedges. The mycelium is very wispy and kinda grayish, but colonizes very fast. Took less than a week for the quart of grains."
I'm getting tired of dredging up threads to post on here, but it looks like there's conflicting info about which agar formula is best, and then there's the hot pouring agar onto spores bit in the bisporus notes. Sometimes I wish that stonesun would magically appear and answer some of these questions.
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Edited by CaptainAhab (04/26/11 01:58 AM)
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anonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: CaptainAhab]
#14351647 - 04/25/11 08:55 PM (12 years, 10 months ago) |
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i'm not clear on what agar work u did exactly
-------------------- The above post is fictional, hypothetical, or downright nonsensical.
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higgledy-piggledy
Registered: 08/24/10
Posts: 966
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: anonjon]
#14351819 - 04/25/11 09:22 PM (12 years, 10 months ago) |
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maybe its due to the age of the spores?
too old = no es bueno
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afrosheen
9Lives the cat



Registered: 03/06/10
Posts: 1,878
Last seen: 2 years, 5 months
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: higgledy-piggledy]
#14351923 - 04/25/11 09:39 PM (12 years, 10 months ago) |
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If they took that long to grow out, conditions weren't right for them. Was it 50 degrees in the room? Was it a standard MEA or PDA agar recipe?
I would have at least tried a few generations of isolation to pick up the growth speed before putting it on grain.

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CaptainAhab


Registered: 12/25/09
Posts: 1,875
Last seen: 6 years, 1 month
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: anonjon]
#14351987 - 04/25/11 09:46 PM (12 years, 10 months ago) |
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Quote:
anonjon said:

i'm not clear on what agar work u did exactly
Malt extract agar, two different kinds, same result. Prepoured and self poured.
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higgledy-piggledy
Registered: 08/24/10
Posts: 966
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: afrosheen]
#14352018 - 04/25/11 09:49 PM (12 years, 10 months ago) |
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Quote:
afrosheen said:
 
what does that mean?
too much for you afrosheen?
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CaptainAhab


Registered: 12/25/09
Posts: 1,875
Last seen: 6 years, 1 month
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: afrosheen]
#14352128 - 04/25/11 10:02 PM (12 years, 10 months ago) |
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Quote:
afrosheen said: If they took that long to grow out, conditions weren't right for them. Was it 50 degrees in the room? Was it a standard MEA or PDA agar recipe?
I would have at least tried a few generations of isolation to pick up the growth speed before putting it on grain.
 
I see that you still haven't changed your overly offensive cat in the microwave picture, afrosheen. They were grown in the right conditions. As far as isolations go, I tried a few, but they were as slow as the original. I thought that I'd start fresh and got the same result. That's what was transferred to the grain.
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Edited by CaptainAhab (04/25/11 10:02 PM)
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afrosheen
9Lives the cat



Registered: 03/06/10
Posts: 1,878
Last seen: 2 years, 5 months
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: CaptainAhab]
#14352179 - 04/25/11 10:08 PM (12 years, 10 months ago) |
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Actually I thought pans and other similar strains were much slower than cubensis. I could be wrong since I haven't touched them before, just read some threads here and there, but it seems to me the more potent strains just grow more slowly.
Regardless, what was your source for these? Prints?
Oh and about my icon...yeah...took a poll, nobody cared. Sorry bro.
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Primal Call
Earth Mage



Registered: 09/05/10
Posts: 2,766
Loc: Here until here
Last seen: 4 years, 4 months
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: afrosheen]
#14353039 - 04/26/11 12:50 AM (12 years, 10 months ago) |
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Quote:
afrosheen said: Actually I thought pans and other similar strains were much slower than cubensis. I could be wrong since I haven't touched them before, just read some threads here and there, but it seems to me the more potent strains just grow more slowly. 
I thought the same, but also have yet to work with these.
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CaptainAhab


Registered: 12/25/09
Posts: 1,875
Last seen: 6 years, 1 month
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: Primal Call]
#14353161 - 04/26/11 01:28 AM (12 years, 10 months ago) |
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From the Panaeolus FAQ: http://www.shroomery.org/8421/Panaeolus-cyanescens-FAQ#agar (just scroll up a little bit) "How fast do Panaeolus cyanescens grow? I found the Panaeolus cyanescens to be very fast compared to P. cubensis. I germinate spores on agar and transfer to Manure:rye seed:perlite in quart jars. These run through in a max of 14 days, but usually in under 10 days. Once laid out in tray and cased with a very thin layer of Peat:perlite, the pin in about 7 days, and mature by the 13 day max. Everything is fast."
I've seen lots of LC's people say only take a week to fully colonize, too.
Here's a quote from Blue Helix at http://www.shroomery.org/forums/showflat.php/Number/4872154#4872154 "Liquid Culture - Days 3, 5, and 7 (completion). Starting with a few mls of live culture rather than few spores can shorten this stage to around only 48 hours rather than 7 days... I have since seen Goliath Pan Cyans develop from 3ccs of liquid starter culture in only 48 hours, making them faster than cubensis."
The only difference between Blue Helix's LC and malt extract agar is the dextrose. Still, if LC's and quart jars of spawn colonize as fast or faster than cubensis, then it would seem that it would be the same with agar. The agar I used is also recommended for pan species (see link in panaeolus faq above).
Here's a thread on Panaeolus on agar: http://www.shroomery.org/forums/showflat.php/Number/10992165#10992165 "Started off with eight petri dishes of light malt extract agar, using one drop of spores from a multispore syringe. Out of the eight the following three are all that show growth after nine days. the ones that don't show any growth have excess moisture on the surface of the agar that I can see run when I tilt the petri dish. I believe I poured the agar a little too thick in the dish, and the surface I pored them on wasn't level. So, one side of the dish is thicker than the other."
Here's a test of different agars by a trusted cultivator I've never heard of: http://www.shroomery.org/forums/showflat.php/Number/5673196#5673196 "I wanted to know on which agar panaeolus mycelium grows faster. PDYA or MEYA. So I made a MEYA Agar plate and cut the half out. I filled it with PDYA. The I cut 3 pieces out and filled this "holes" with YGC Agar (Yeast Glucose Chloramphenicol) to look if the mycelium with grow on it too. And the winner is PDYA by now"
An interesting read on Panaeolus bisporus where it was recommended to hot-pour the agar onto spores. This is from StoneSun and someone named Lipa: http://www.shroomery.org/forums/showflat.php/Number/14116777#14116777 "As per lipa's advice spores were germinated on MEA in a fashion of hot pouring the agar on the spores. Germination occurred in just 48 hours after the petri has been poured. Sterilized organic rye berries (1hr @20psi) were inoculated with agar wedges. The mycelium is very wispy and kinda grayish, but colonizes very fast. Took less than a week for the quart of grains."
I'm getting tired of dredging up threads to post on here, but it looks like there's conflicting info about which agar formula is best, and then there's the hot pouring agar onto spores bit in the bisporus notes. Sometimes I wish that stonesun would magically appear and answer some of these questions.
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Edited by CaptainAhab (04/26/11 01:57 AM)
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ManicMongrel
Paragorn of pointless comments


Registered: 03/14/11
Posts: 253
Loc: Across the Atlantic
Last seen: 5 months, 18 days
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: CaptainAhab]
#14353658 - 04/26/11 05:54 AM (12 years, 10 months ago) |
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Panaeolus are not really a big fan of grains. You can grow on it for sure, but they prefer dung and compost as their main habitat.
I have the impression that they are very symbiotic, I mean they seem to need bacterias from dung to fight off contams like other fungus. If you dont believe it just germinate fresh unpasteurized poop and look how easily they colonize it.
You have healthy mycelia so its all good so far. Are you sure the temperature is right? Try tweaking it.
When I work with new pan strains from the wild, I usually transplant small pieces of mycelia to a pre-sterilized jar filled with carefully pasteurized horse dung, then tweak the temperature until I find where they take off.
-------------------- Rules and laws are guidelines, to follow them by definition is equivalent to ignoring responsibility - Let me know if anything in my Trade List got your attention! New asian pan varieties up for trade [10.july.2013] "Familiarity breeds contempt"
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Javadog
Continuing along



Registered: 05/03/10
Posts: 7,385
Loc: USA
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: ManicMongrel]
#14354237 - 04/26/11 09:42 AM (12 years, 10 months ago) |
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I have read that they can be slower at times. They rip on manure though.
The course seems to be: hpoo+verm+gypsum+??? for a thinner than usual substrate, and cased with pH balanced 50/50.
Shaking can stall growth, use it carefully.
I will be following this. Along with Jalisco, I have a bone to pick with Pans.
JD
-------------------- Boyd Rice told my brother that life is a corny pack of freesakes Myco-tek.org
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thesoundd
Candy Coater



Registered: 04/14/10
Posts: 178
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: Javadog]
#14354411 - 04/26/11 10:29 AM (12 years, 10 months ago) |
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Eh, I just obtained some pan cam 'goliath'...didn't realize their growing prefs might be that drastically different from cubes. So...are you guys saying that we can throw some pan cam mycelia into pasteurized quart jars of manure/verm/gypsum and then go bulk from there? Not starting off with sterile growth conditions sounds very scary.
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anonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: thesoundd]
#14354522 - 04/26/11 10:58 AM (12 years, 10 months ago) |
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Quote:
thesoundd said: Eh, I just obtained some pan cam 'goliath'...didn't realize their growing prefs might be that drastically different from cubes. So...are you guys saying that we can throw some pan cam mycelia into pasteurized quart jars of manure/verm/gypsum and then go bulk from there? Not starting off with sterile growth conditions sounds very scary.
You sterilize it, inoculate it, and fruit it. It's not spawn.
-------------------- The above post is fictional, hypothetical, or downright nonsensical.
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Boozie
I like beer.



Registered: 02/18/10
Posts: 1,226
Loc: :ↄo⅃
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: thesoundd]
#14354537 - 04/26/11 11:00 AM (12 years, 10 months ago) |
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Quote:
didn't realize their growing prefs might be that drastically different from cubes.
They're not. Besides wanting a minor increase in temperature for fruiting (upper 70s, low 80s), a bit more FAE, and the fact they require a manure-based substrate, they'll grow in the exact same conditions as cubes.
In my experience, panaeolus mycelium grows at just about the same speed as cube mycelium on grain. Maybe even a tad quicker. But, like Javadog said, it really rips through the manure after you spawn it. Maybe 4-5 days to fully colonize a 2 quart (2 inch depth) tray of substrate.
If it took over a month to get to the size of quarter on agar, then nothing after you transferred to grain, i'd imagine some sort of contamination was present as well. Maybe bacteria? Though you should be able to notice that on that agar to begin with.
-------------------- "After silence, that which comes nearest to expressing the inexpressible is music." - Aldous Huxley
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ManicMongrel
Paragorn of pointless comments


Registered: 03/14/11
Posts: 253
Loc: Across the Atlantic
Last seen: 5 months, 18 days
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: thesoundd]
#14354672 - 04/26/11 11:29 AM (12 years, 10 months ago) |
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From my experience you can, and your odds are great, as long as temperature is ideal few other fungi can compete with pans in terms of growth speed, if they're in their right environment.
Pans are decomposers and the competition for nutrients in manure and compost should be pretty insane. They are ment to thrive in company of certain types bacteria, that you typically find in compost and manure. You can sterilize for sure but, at best it will only save you a day or two in terms of colonization, the drawback is that it will be prone to mold infection.
Sterile growth conditions are tricky if you work with pans, fermented substrate is just a lot easier. In theory you should get a better harvest with sterile substrate, but people say they contaminate a lot easier than cubes.
-------------------- Rules and laws are guidelines, to follow them by definition is equivalent to ignoring responsibility - Let me know if anything in my Trade List got your attention! New asian pan varieties up for trade [10.july.2013] "Familiarity breeds contempt"
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4204ME
eatingtheagareater



Registered: 12/11/10
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Loc: Australia
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: afrosheen]
#14354698 - 04/26/11 11:33 AM (12 years, 10 months ago) |
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Quote:
afrosheen said: Actually I thought pans and other similar strains were much slower than cubensis. I could be wrong since I haven't touched them before, just read some threads here and there, but it seems to me the more potent strains just grow more slowly.
Regardless, what was your source for these? Prints?
Oh and about my icon...yeah...took a poll, nobody cared. Sorry bro. 
I may be missing something, I cannot see a cat in the microwave...
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thesoundd
Candy Coater



Registered: 04/14/10
Posts: 178
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: ManicMongrel]
#14355028 - 04/26/11 12:34 PM (12 years, 10 months ago) |
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Quote:
ManicMongrel said: Pans are decomposers and the competition for nutrients in manure and compost should be pretty insane. They are ment to thrive in company of certain types bacteria, that you typically find in compost and manure. You can sterilize for sure but, at best it will only save you a day or two in terms of colonization, the drawback is that it will be prone to mold infection.
Sterile growth conditions are tricky if you work with pans, fermented substrate is just a lot easier. In theory you should get a better harvest with sterile substrate, but people say they contaminate a lot easier than cubes.
Ok, cool. So it seems like a good method is to use liquid mycelium to inoculate pasteurized spawn bags that are heavy on the manure, and after colonization spread the substrate out and throw on a thin casing layer, say, in a monotub? I don't have spawn bags, but I wouldn't mind making my own with this tek...if it isn't outdated.
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ManicMongrel
Paragorn of pointless comments


Registered: 03/14/11
Posts: 253
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: thesoundd]
#14355369 - 04/26/11 01:54 PM (12 years, 10 months ago) |
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That should work pretty smooth :-)
I do it the lazy way and spray the spores directly on the manure mix. When the jars are done I just make plenty of holes in the layer of bulk substrate and place pieces from the jars in the holes. I can usually see that the mycelium webs has started to stretch outwards the next morning, takes little over a week before its fully colonized. Its all a little sloppy but, I have done it 4 times now and everything went fine.
-------------------- Rules and laws are guidelines, to follow them by definition is equivalent to ignoring responsibility - Let me know if anything in my Trade List got your attention! New asian pan varieties up for trade [10.july.2013] "Familiarity breeds contempt"
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Javadog
Continuing along



Registered: 05/03/10
Posts: 7,385
Loc: USA
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Re: Panaeolus Q: Spores took 2 weeks to show signs of life on agar, won't grow on grain. ???? [Re: ManicMongrel]
#14355717 - 04/26/11 03:06 PM (12 years, 10 months ago) |
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That's a wild approach MM. Interesting.
The standard thinking is that one cannot directly inoculate bulk substrates with spore solution, or even LCs, without having to deal with competitors and contamination. (I know, there are exceptions).
One either uses a pasteurized sub, or a sterilized sub kept in sterile conditions. The cleanliness of the inoculant is critical when doing a sterilized substrate.
Stonesun is probably our best example of doing this well.
As I struggle to grow Gymnopilus I may see where a sterilized sub might work where initial attempts lost to a mold. --> Using a sterilized substrate gives a perfect advantage to one species of fungus that you are not able to give perfect conditions to otherwise.
Anyhow, just stopping in from hell....work, gotta run.
JD
-------------------- Boyd Rice told my brother that life is a corny pack of freesakes Myco-tek.org
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