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 flavoraid
now with twicethe ketamine andopiates!
Registered: 12/05/07
Posts: 1,678
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 hydroponic scletoria production 
#8461810 - 05/29/08 09:01 PM (3 years, 11 months ago) |
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so I was reading about ergot production and it's produced in an airated liquid solution
so a culture is made, put into a 1gal jar, has a air filter/gas exchange lid
a bubbler airates the liquid solution.
how would this work?
ergot is a scletoria producing strain but I've never cultivated it before.
this is what has inspired me to make this thread.
http://www.shroomery.org/forums/showflat.php/Number/6225737#6225737
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coda said:
imachavel, Man you really need to do some reading, the amount of bullshit you put into almost every single one of your posts is absolutely astounding.
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 solumvita
Q.B.E.
Registered: 02/12/08
Posts: 1,828
Loc: South Africa
Last seen: 1 hour, 50 minutes
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Re: hydroponic scletoria production [Re: flavoraid]
#8479519 - 06/03/08 12:51 PM (3 years, 11 months ago) |
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The article does not specifically state that sclerotia are produced in the liquid culture rather only mycelium or "ergot alkaloids". I do not think you will find sclerotia forming in a LC.
But i like the tek to use it to produce an aerator.
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One of these days all the answers will be revealed until then we learn from each other!
www.mushrush.co.za
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flavoraid
now with twicethe ketamine andopiates!
Registered: 12/05/07
Posts: 1,678
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Re: hydroponic scletoria production [Re: solumvita]
#8480287 - 06/03/08 04:04 PM (3 years, 11 months ago) |
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oh
it seemed sort of confusing to me heh. I was unsure of how the sclertoria was harvested or grown
:O
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coda said:
imachavel, Man you really need to do some reading, the amount of bullshit you put into almost every single one of your posts is absolutely astounding.
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waixingren
Registered: 03/14/05
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Loc: SW Florida
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Re: hydroponic scletoria production [Re: flavoraid]
#8494721 - 06/06/08 10:48 PM (3 years, 11 months ago) |
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this is a little different but i had a p. atlantis sclerotia form in a liquid culture once.
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flavoraid
now with twicethe ketamine andopiates!
Registered: 12/05/07
Posts: 1,678
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Re: hydroponic scletoria production [Re: waixingren]
#8497066 - 06/07/08 05:41 PM (3 years, 11 months ago) |
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nznz
the intention of the lsd sytnh tek thing is mycelium production.
I've been doing way too much drugs... decided yesterday 100days of complete sobriety living the spartan ideal 99 more days to go!
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coda said:
imachavel, Man you really need to do some reading, the amount of bullshit you put into almost every single one of your posts is absolutely astounding.
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truth seeker
Stranger
Registered: 12/20/07
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Re: hydroponic scletoria production [Re: flavoraid]
#8522033 - 06/13/08 10:22 PM (3 years, 11 months ago) |
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i have been contemplating cunsuming spawn i have added oyster spawn to stir fry when very hungery and think a jar would work in off season or as tea
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Plasmid
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Re: hydroponic scletoria production [Re: truth seeker]
#8522257 - 06/13/08 11:50 PM (3 years, 11 months ago) |
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If you are interested in cultivating claviceps, the best sources for information are patents or the book Ergot: The Genus Claviceps by Vladimir Kren and Ladislav Cvak. Cvak holds a number of patents for ergot cultivation on unheard of scales (10s of thousands of kg) with the specific aim of extracting various ergot alkaloids. The earliest patents go back to Hofmann and Stoll and the like.
It seems to me that the biggest difficulty is in isolating a strain appropriate for alkaloid production. Many of the patents refer to specific strains, which are available from various stockpiles, but it's not the kind of thing you'd probably be able to order.
I believe the method you're referring to is called a saprophytic culture.
Edited by Plasmid (06/14/08 12:20 AM)
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fastfred
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Registered: 05/17/04
Posts: 6,242
Loc: Dark side of the moon
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Re: hydroponic sclerotia production [Re: Plasmid]
#8524918 - 06/14/08 07:57 PM (3 years, 11 months ago) |
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You're right. Every strain I've looked up on ATCC is listed as "unavailable" and a search on "claviceps" turns up nothing.
I'm curious if anyone actually has a culture or access to wild strains? I've been thinking about going out hunting, but I don't think there is anything very close to me.
Most of the high producing strains are the result of lab work. Attaining a heterokaryotic state has been reported as important and several papers detail mutating strains for higher production. Some even report getting higher yields with multiple rounds of mutation.
One of the problems with these strains is senescence. The claviceps lifecycle is not easy to complete as it requires a host and an overwintering stage.
JLF is possibly still selling ergot samples, but I've only seen one thread here where it seems someone has tried culturing it. If anyone has any experience in the area it would be nice to hear about it...
-FF
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happyday
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Registered: 06/22/04
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Re: hydroponic scletoria production [Re: waixingren]
#8526437 - 06/15/08 10:04 AM (3 years, 11 months ago) |
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Did the sclerotia form within the liquid itself or on the side of the jar? I like the pics.
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Plasmid
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Re: hydroponic scletoria production [Re: happyday]
#8526862 - 06/15/08 12:40 PM (3 years, 11 months ago) |
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Fastfred,
if I recall correctly, a lot of the early work on mutant strains was just done by screening random mutants. I'm not sure how the mutations were caused, maybe radiation. I'm sure that a competent person could mimic the earlier mutant inducing methods without too many problems, but I think that the mutant screening would really be a lot of work. The upside would be that after doing the screening, you would have developed a procedure for isolating the necessary alkaloids. The patent literature is the best place to look for this kind of information.
The only references I've seen for acquiring specific strains of Claviceps purpurea involve various national fungal collection centers. These all seem to be run by government bodies and I'm sure they don't give out to free samples to anonymous people knocking at their doors.
As for hunting, I have seen some papers discussing various aspects of ergot and listing the sample source as "field", "side of road", "wooded area", etc. and listing the nearest town. It does seem possible to actually just go out looking for ergot and finding it, as long as you generally know where it may grow.
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fastfred
Old Hand
Registered: 05/17/04
Posts: 6,242
Loc: Dark side of the moon
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Re: hydroponic scletoria production [Re: Plasmid]
#8527546 - 06/15/08 04:44 PM (3 years, 11 months ago) |
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> I'm not sure how the mutations were caused,
One paper I remember used a chemical mutagen. Can't remember which, but probably EMS. UV would probably work as well.
> I think that the mutant screening would really be a lot of work.
Actually one paper describes a high volume screening method using Van Urk's reagent in which they simply sprayed colonies on a petri dish with Van Urk's and picked ones with the darkest color reactions. Using this method they screened several hundred mutants in just a few days.
As far as the culture collections that might have strains... I've looked up close to a dozen ATCC accession/voucher numbers and they are all listed as "unavailable" with no details provided on them whatsoever. So if ATCC isn't providing them I doubt anyone else is either. There's probably no chance of getting them at all from them. Usually ATCC would just make you jump through some hoops and sign an MTA, so it's pretty weird that they're simply listed as unavailable.
Even if one of the culture collection would send you out a culture you'd still need approval from the USDA before they would send it. It's a crop damaging pest species and is illegal to import/export from states without proper permits.
Locally collected samples would avoid this problem. It's also good to remember that these strains aren't magic. Someone actually went out and collected them from the wild and worked with them in the lab to achieve high alkaloid productions. Nothing I've read that they've done is all that hard to do in the lab so it would not be hard to do the same thing yourself.
I'm interested in the genetics of the ergot alkaloid pathway. There have been some interesting papers recently detailing the genetic basis for conclusions about the evolutionary history of the alkaloid pathway. The pathway has found it's way into many species and various species have different non-functional genes in the pathway resulting in different products. Such as lysergic and paspalic acid in C. paspali and other earlier precursors in more distantly related species.
I think there is still a lot of work and potential medicines to be found in Claviceps. The nature of ergopeptides leaves a ton of variations to be explored.
Quote:
A new industrial producer strain of the microorganism Claviceps purpurea (Fr.) TuL, CCM 8360, was bred from proprietary strain F-721. Sclerotium of this strain was transferred to saprophytic culture - a surface of sclerotium was sterilized and moved on agar-thickened malt medium. After 28 days at temperature 25° Celsius biomass of conidia covered the surface of the agar. The biomass was subjected to mutation using solution of 0.2M N-methyl-N-nitro-N-nitrosoguanidine for 8 hours. Mutated isolates were inoculated onto rye using a syringe, and grown sclerotia were analyzed for the qualitative and quantitative content of ergot alkaloids. Sclerotium with the highest content again was transferred to saprophytic culture that was subject to mutation using 0.2M ethyl methane sulphonate for 16 hours. Mutated isolates so obtained were inoculated onto rye using a syringe, and grown sclerotia were analyzed. Sclerotium with the highest content was passaged on rye in experiments in the small-plot scale (approx. 1 m2). Sclerotium of this strain was transferred to saprophytic culture that was exposed to Gamma radiation, with radiation dose being 3000 Gy. Irradiated isolates were inoculated onto rye using a syringe, and grown sclerotia were analyzed for the qualitative and quantitative content of ergot alkaloids. Sclerotium with the highest content was passaged on rye in pilot-scale experiments (approx. 1 ha). Sclerotium with the highest content, harvested from these plots of land, was transferred to saprophytic culture and kept in the Czech Collection of Microorganisms (CCM) at the Masaryk University, Tvrdeho 14, Brno, Czech Republic, under identification number CCM 8360. The above mutation procedures connected with a positive selection resulted in an increase in the average content of ergot alkaloids in sclerotia by 146 wt% in strain CCM 8360, compared with initial strain CCM F-721. At the same time, the quality of the spectrum of individual alkaloids was improved by changing the ratio of alpha-ergokryptine to beta-ergokryptine from 1.15:1 to 0.79:1 in favour of desirable beta-ergokryptine. [0035] Mutated isolates were then inoculated onto rye using a syringe.
Looks like they used several rounds of different mutation techniques for this strain. However, claviceps is much easier to get to over produce alkaloids that they usually produce.
It might be somewhat difficult to get the amazing production that scientists with big budgets and lots of time have gotten. But up to 2g/L of culture has been reported and I've seen 1g/L reported in many papers. For most purposes I think a lot of people would be happy with 0.5g/L, which shouldn't be all that hard to attain.
As far as breeding a heterokaryotic strain it would be really helpful to have samples from several locations that are geographically distant.
-FF
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