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Tsjerk
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Don't transfer sterile broth to the incubator, sterilize the broth in the incubator. If you can't do 120 degrees (which kills anything including
spores), just heat the container including the broth too 100 degrees, wait for 24 hours for any surviving spores to germinate, and heat too 100 again
(one hour orso).
This way you only have to sterile transfer the small amount of inoculation culture, not the whole lot of broth, which would be pretty tedious without
a clean bench.
Edit: one hour at 100 already kills almost anything, I don't know if the two times boiling for an hour with 24 in between is really different from
just boiling once for two hours, but it works. Bacillus subtilis spores are quite hardy and don't really survive more than ten minutes boiling, after
twenty they are gone.
[Edited on 6-8-2018 by Tsjerk]
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JJay
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That seems reasonable.
I tend to think that if a lot of culture can be used for inoculation, the ergot would have a big head start and would hopefully manage to defeat minor
bacterial infections (it's antibacterial, right?), but that intuition may well be wrong, and of course it would be best to utterly eliminate all
bacteria. Since ergot is a parasite, it stands to reason that it won't compete very well with saprophytes like yeast and trichoderma (or even common
mushrooms like agaricus).
I've boiled water in a 5-gallon bucket before... the plastic degrades after days of boiling, but it would be fine for an hour or two.
Apparently, ergot is pretty common on Festuca grasses: https://dl.sciencesocieties.org/publications/cs/abstracts/34...
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S.C. Wack
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I didn't read the article but BTW it looks like that's not ergot they're writing about. So, tall fescue may already have alkaloids detectable without
any Claviceps. Infection with the common endophyte is sold as a superior lawn product; such infected seed is reported to have only 1% of the alkaloid
there would be if it was ergot. Obviously not all fescue ergot is the same and different seed heads could have different strains and have more or less
or different alkaloids.
BTW in some places those with some interest in ergot and an abundance of time and laziness can deposit rye on neglected tall weed type areas in late
summer into fall before a little rain. Rye will establish itself in fall while everything not a grass dies. Some years the ergot will be just
ridiculous. It's hard to imagine that even a thousand years ago rye bread countries didn't bother screening out the ergot, which is bigger than the
grain...it says something about us fundamentally...none of the human condition problems involved seem to be going away any time soon.
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JJay
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That's interesting... about half of the ergots I have found so far were growing on fescues. This article suggests that the ergolines in fescues are
caused by Claviceps: www.ncbi.nlm.nih.gov/m/pubmed
Wild rye is pretty common here, but the small variety that is maturing seems to be immune to ergot, and I'm not sure about the larger wild varieties.
I just might give that a shot with a commercial variety.
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S.C. Wack
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BTW this shows helpful hints like ergot growing on different species.
PS speaking of sacks of grass seed I'd really like to have one of Achnatherum inebrians...if anyone in China is listening...My lawn needs to be
replaced and it's supposed to be tough.
[Edited on 14-8-2018 by S.C. Wack]
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andy1988
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Quote: Originally posted by Tsjerk  | | , just heat the container including the broth too 100 degrees, wait for 24 hours for any surviving spores to germinate, and heat too 100 again (one
hour orso). |
This is called the tyndallization process, also called fractional sterilization. A good write up here. I don't have a pressure cooker... this is what I do.
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JJay
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I think I have all the materials I need to grow small quantities of ergot in petri dishes on agar. I've done some reading on the subject, and a few
amateurs have tried it before. Apparently, there are some factors that keep the ergot colonies from growing beyond a certain point. In industry, they
do something akin to giving the ergot cancer with UV radiation to create a mutant strain that can multiply infinitely.
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Tsjerk
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Ethyl methanesulfonate is also used to induce mutations, the procedure is straight forward and mutants are easy to select as they keep growing. There
is a small chance you also mutate the ergoline synthesis pathway though.
[Edited on 3-9-2018 by Tsjerk]
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JJay
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Ethyl methanesulfonate is probably something that could be produced at home... (perhaps reflux sodium sulfite in molten potassium methyl sulfate,
purify and esterify). I'm not really equipped to mess around with it, though... at this point, I'll be happy if I get a single ergot culture growing.
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