That looks REALLY impractical. Where am I going to get a fungus indigenous to patagonia, requiring association with a tree. This species is an
endophyte. That means difficulty. Ergot is difficult enough to work with, and I've considered attempting to isolate grass endophytes, such as that of
Lolium, which produces ergot alkaloids. But generally endophytic fungi are highly specific to the host, and just won't grow in culture. Such as for
example, the Clavicipitaceous endophyte that produces the ergot alkaloids in some strains of hawaiian baby woodrose, or the morning glory endophytes.
You just can't culture them, they are too fussy and won't grow in culture media.
Even parasitic fungi often aren't particularly compliant. I've been doing a bit of work with ergot (Claviceps purpurea) and things like strain
selection for productivity and in the case of Claviceps spp. actually getting productivity, or if you do, then you need to build a bioreactor, monitor
oxygen content, optimize culture media. And its difficult enough with a really rather well known and thoroughly explored family. I've bought the book
'the genus claviceps' by Kren and Cvak, a couple of decades old by now but still, some fairly up to date how-tos on the likes of submerged culture,
strain selection, host infection equipment.
Yet, still, its an utter shitter, starting from some wild-type sclerotia I managed to obtain growing on wild ryegrasses. Come the climate getting a
bit warmer, I'm going to re-activate that project. And build a dedicated bioreactor, and get to experimenting with encapsulation in calcium
alginate/perfluorocarbon emulsion (the alginate microsphere encapsulation of the fungus, which only produces ergot alkaloids in the sclerotia, but
trapping the sphacelial growth stage, which itself is relatively easy to culture, and after mutagen exposure and cloning from the colonies most
reactive when samples are tested with Van Urk reagent or Erlich's, trying for tryptophan autotrophs and subsequent reversion to prototrophy (this
essentially severs a link in the metabolic chain that assembles the ergoline ring structure that acts as a negative feedback control on
tryptophan-DMAT production whereby tryptophan itself is the signalling agent, too much acts as an inhibitory agent on productivity at an early stage,
DMAT synthase repression I think, certainly one of the very early stages in the biosynthetic path)
The encapsulation 'tricks' sphacelial ergot culture tissue into 'thinking' that it is in fact sclerotial growth, the microcapsules of polymerized
alginate acting as a sort of 'pseudosclerotia' and the perfluorocarbon emulsion is for enhancing O2 transport, since naturally, the microspheres have
to be very small, such as produced by electrostatic spraying through fine needles under pressure of an alginate-ergot culture into CaCl2 solution,
which produces very fine, small beads that polymerize instantaneously as Ca alginate, trapping the ergot culture, but after a couple of mm, the
alginate impairs O2 transport so that the result is essentially an anoxic core, surrounded by an outer layer of growth.. The perfluorocarbon emulsion,
along with added surfectants such as tween to improve O2 permeability of the cell walls allows for deeper penetration of the fungus and improved
alkaloid production.
I don't think COD growth is going to be practical. I don't exactly have an easy way of going out to patagonia, wandering around with my microscope,
sampling potential trees for endophytes under the oil immersion lens, carrying around a reagent kit full of stains and a spade, digging up a tree or
two and flying them back home.
Endophytes are just bitchy. |
