Spagnum Jones, Erasmus Boomfork and Günther Schlonk
Introduction
Some time ago, our colleague and friend Dr Spagnum Jones did what he had been threatening to do for years: throw his laptop in woodchipper, move to Sardinia and open a bakery. The catalyst for this drastic action was provided by Reviewer 2’s suggestion that Jones “have you’re manuscript poofread by a neutral english speeker”. This irksome comment was the tip of a bureaucratic iceberg that had ground an outstanding mycologist down into a shrivelled husk.
During cleanup-week last month, we discovered a Tupperware container in the tea-room fridge belonging to Dr Spagnum. The original nature of its contents was not apparent, for they had been obscured by a spectacular grown of myriad fungi. A cursory visual survey indicated the presence of Cladosporium, Penicillium, Rhizopus and Kleptomyces colonies. Evidently, Spahgnum had been developing a novel medium for fungiculture, and we had stumbled upon an experiment left unfinished. To honour his memory, we therefore took it upon ourselves to complete what he had begun.
Results and Discussion
Our first task was to determine the original nature of Spagnum’s growth medium. We began by carefully removing the fungal surface coating with a teaspoon, and subjecting the malodourous goop beneath to a number of chemical and spectroscopic tests. Olifactometry indicated the presence of piperine, lactic and butyric acids, alkyl amines and short-chain aldehydes, which were likely decomp products of lipids and amino acids. HPLC-MS confirmed this hypothesis, with the detection of β-carotene, lactose, assorted lipids and peptides, vitamins A, C, and B1–12. A Holtzwerfer test indicated the presence of cellulose and hemicellulose, and the Fußkitzler assay gave a positive result for lignin.1,2 These observations suggested that the material in Spagnum’s container was biological in origin. As such, we performed a Hinternküsser test, which gave a positive result for the presence of DNA.3
To determine the origin of this DNA, we performed shotgun sequencing using the Clyde-Bonnie method.4 The first step in this procedure was to shoot an aliquot of the sample with some No. 112 (10 µm) birdshot, fired from a sawn-off 12-gague at a range of 0.15 m. This served to lyse the cell walls, and also to physically fragment the DNA into segments of ~100 base-pairs (Figure 2).
The DNA was then extracted, filtered and precipitated with Jack Daniel’s Absolute Whiskey. Deep Southern Blots (BSB’s) were then performed, to match the isolated DNA fragments with a library of common plant and animal genetic markers.5 We identified genetic material from Allium sepa, Cucurbita moschata, Allium sativum, Daucus carota, Solanum tuberosum, Ipomoea batatas and Petroselinum crispum. These findings, in combination with the olifactrometric and HPLC data unequivocally confirmed the identity of Dr. Jones’ fungal growth medium as a creamy pumpkin soup.
Pumpkin soup has many attractive features for mycoculture. It is cheap and environmentally friendly, non-toxic and biodegradable. It can be prepared in large batches, is easily poured, and can be frozen for later use. Pumpkin soup is rich in fungal macronutrients (starch, cellulose and lignin), while also containing all the minerals, vitamins and other micronutrients required by fungi. This makes it superior to conventional media such as agar gel, as it does not require additional nutrients.
To demonstrate pumpkin soup’s superiority as a growth medium, we prepared a batch following a procedure by Nagi and coworkers (with 1% added gelatine as a gelling agent), and plated out a range of fungal spore (Figure 3).6 Agar plates were prepared and treated with the same spores in a control experiment. We selected three fungi for our assay: Flatularium acridencia, Trichophallus lotsococcus and Yukistufus caerulus.
F. acridensia is a mould that grows on beans of the Fabaceae family, and can cause Rectal Turbulence Syndrome in humans. This causative link was disputed by the medical establishment, until the maverick physician Harry Marshall chugged a broth of the stuff and blew a centrifuge off a bench with one blast.7 While it grew slightly more vigorously on agar than on soup, we still obtained a sufficient mass of fungus to get an emu airborne.
Yukistufus caerulus is the active agent in the psychedelic French cheese “frére puant”. It grew far more rapidly on soup than on agar, probably due to the presence of fats and lactose. The growth of T. lotsococcus or “echidna crotch-fungus” was comparable in both media.
Conclusion
Our findings indicate that pumpkin soup is an effective medium for fungal culture, and may be superior to agar for certain species. It also tastes significantly better. Future work in our laboratory will explore the applications of other soups in microbiology. In particular, we have anecdotal evidence that a lobster bisque can function as a potent culture medium for Escherichia coli, Salmonella bongori, and Listeria monocytogenes.
Notes and references
1. “Ist diese Zellulose, die ich vor mir sehe?” C. F. Holzwerfer, 1954, Angewanker Chemie, 2, 31–33.
2. “A novel test for lignins: turn the substrate into paper and see if it goes yellow after a few years” H. Fußkitzler, 1927, Helvetica, 8, 176–187.
3. “A novel test for DNA: splice it into drosophila and see what happens” K. J. Hinternküsser, 1989, Frenetica Chimia, 9, 111–222.
4. “Strapping DNA to a chair and making it talk: shotgun genomics ” B. E. Parker, C. C. Barrow, 1934, Journal of Literal Genetics, 2, 18457–28499.
5. “How to make DNA Square-Dance: the Deep Southern Blot” W. Melone, 1989, Choreochem, 1(2), 12–34
6. https://www.recipetineats.com/classic-pumpkin-soup/
7. “Well Blow Me Down, Fungi Does Cause Rectal Turbulence Syndrome” H. Marshall, 2011, Australian Journal of Chemistry, 193, 1770–1778.