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Reagent matrix

Reagent against taxon, for the records published so far. A negative result is recorded rather than left blank, because a negative result is evidence: it tells you what the specimen is not.

MELZ Melzer's reagentWeight 3.00

Preparation
0.5 g iodine + 1.5 g potassium iodide + 20 g chloral hydrate in 20 ml distilled water
What it tests
Spore wall and spore ornamentation, examined at 1000x in a water mount
Verdict vocabulary
Amyloid — blue-black to grey-blue · Dextrinoid — reddish-brown to rusty · Inamyloid — pale yellow to unchanged, the colour of the reagent itself

Chloral hydrate softens the spore wall so the iodine can reach the inner layers; without it (Lugol's) only the outer wall reacts and amyloid spores read as a much weaker blue. Read the result within 2 minutes — chloral hydrate slowly bleaches the preparation.

RecordTierVerdictReaction
Agaricus xanthodermusT1inamyloidSpore wall stays yellow-brown; no blue-black development.
Amanita phalloidesT3inamyloidSpores stay pale; a faint grey wash on some spores is not a blue-black amyloid reaction and should not be read as one.
Boletus edulisT0inamyloidSpore wall remains olive-yellow; no blue or red-brown development.
Cantharellus cibariusT0inamyloidSpore wall stays pale yellow; no blue or red-brown development over 5 minutes.
Chlorophyllum molybditesT1dextrinoidSpore wall turns a deep reddish-brown to rusty within 2 minutes — a strong pseudoamyloid reaction that looks amyloid at low magnification and must be read at 1000x.
Clitocybe dealbataT3inamyloidSpore wall stays pale; no blue-black development at any magnification.
Coprinus comatusT0inamyloidSpore wall stays dark yellow-brown; no blue-black amyloid development.
Cortinarius orellanusT3inamyloidThe spore wall does not turn blue-black, but the coarse warts are strongly outlined and the ornamentation is far easier to count than in water.
Cortinarius rubellusT3inamyloidNo blue-black reaction; the warts outline sharply and are easier to count than in a water mount.
Entoloma sinuatumT2inamyloidSpore wall stays pinkish; no blue-black development, and the facets do not darken.
Galerina marginataT3inamyloidSpore wall does not turn blue-black, but the ornamentation is thrown into sharp relief and the smooth plage becomes obvious.
Gyromitra esculentaT3inamyloidSpore wall and the ascus tips stay pale yellow; no blue-black amyloid reaction in either.
Hypholoma fasciculareT1inamyloidSpore wall stays yellow-brown and the germ pore does not darken; no blue-black development.
Lepiota brunneoincarnataT3dextrinoidSpore wall turns reddish-brown within 2 minutes; the reaction is uniform and easy to read at 400x.
Macrolepiota proceraT0dextrinoidSpore wall and the inner wall layers turn reddish-brown to rusty within 2 minutes — the classic dextrinoid reading.
Marasmius oreadesT0inamyloidSpore wall stays pale yellow; no blue-black or rusty development.
Paxillus involutusT2inamyloidSpore wall stays ochre-brown; no blue, no red-brown intensification.
Pleurocybella porrigensT2inamyloidSpore wall stays pale; no blue-black development.
Rubroboletus satanasT1inamyloidSpore wall stays olive; no blue-black or rusty development.
Scleroderma citrinumT1inamyloidSpore wall and spines do not turn blue-black; the ornamentation is however outlined more sharply than in water.
Tricholoma equestreT2inamyloidSpore wall stays pale yellow; no blue-black development at any concentration.

KOH5 Potassium hydroxide 5%Weight 2.00

Preparation
5 g KOH pellets in 95 ml distilled water
What it tests
Cap cuticle, flesh in the cap centre, and the stem base
Verdict vocabulary
Chrome yellow — immediately, then fading to brown · Olive to greenish-grey · Reddish-brown to vinaceous · No change

The reaction is a surface phenomenon: wipe the cap cuticle with the drop rather than flooding a cut section. Chrome yellow on a yellowing agaric is the single most useful bedside test in this bench, because the colour is strong enough to read in daylight and survives the specimen drying out.

RecordTierVerdictReaction
Agaricus xanthodermusT1positiveCap cuticle flashes chrome yellow within seconds, then fades through orange to brown over about a minute — the reaction is fast enough to be visible while you are still holding the pipette.
Amanita phalloidesT3negativeCap cuticle lightens to yellow-brown or, on the pale form, stays almost unchanged; no chrome yellow flash.
Boletus edulisT0variableCap cuticle turns orange-brown to red-brown within 30 s on most collections; the flesh stays unchanged, which is the reading that matters.
Cantharellus cibariusT0negativeCap cuticle dulls to greyish-brown but develops no yellow, green or vinaceous tone.
Chlorophyllum molybditesT1positiveCap scales turn orange-brown; the pale ground between them stays unchanged, producing a strongly mottled result.
Clitocybe dealbataT3negativeCap shows no reaction whatever; the white pigment is stable to alkali, unlike that of some pale Clitocybe species which yellow.
Coprinus comatusT0negativeCap surface unchanged; the reddish-brown tips of the scales are stable to alkali.
Cortinarius orellanusT3negativeCap cuticle darkens to a deeper red-brown without turning yellow or green.
Cortinarius rubellusT3positiveCap cuticle turns a deep blood-red to vinaceous within 30 s, then fades to brown — a reaction several Cortinarius share and which is not diagnostic on its own.
Entoloma sinuatumT2negativeCap surface darkens slightly to grey-brown; no yellow, green or red development.
Galerina marginataT3negativeCap cuticle darkens to a dull brown; no chrome yellow, no red.
Gyromitra esculentaT3negativeHead surface darkens slightly and the crust softens; no yellow, green or red development.
Hypholoma fasciculareT1negativeCap surface darkens to olive-brown; the yellow and green pigments are stable to alkali.
Lepiota brunneoincarnataT3negativeCap squamules darken slightly to a deeper brown; no yellow or green development.
Macrolepiota proceraT0positiveCap squamules turn orange-brown to greenish-brown; the pale ground tissue between them shows no reaction.
Marasmius oreadesT0negativeCap shows no reaction whatever; the buff pigment is stable to alkali, which is not true of the pale Clitocybe look-alikes.
Paxillus involutusT2positiveCap cuticle turns a deep red-brown to mahogany within 30 s, markedly darker than the untreated surface.
Pleurocybella porrigensT2negativeCap and gills show no reaction whatever; the white pigment is stable to alkali.
Rubroboletus satanasT1positiveCap cuticle turns orange-brown; the red pore surface darkens to a deeper blood-red without turning yellow.
Scleroderma citrinumT1negativeThe rind darkens to a deeper brown and softens; the gleba does not change colour.
Tricholoma equestreT2negativeCap cuticle darkens to olive-brown, but the yellow pigments are otherwise stable and produce no red flash.

FESO4 Iron(II) sulfate 10%Weight 3.00

Preparation
10 g FeSO4·7H2O in 90 ml distilled water, used fresh
What it tests
Cap flesh, and the surface of the stem in the upper third
Verdict vocabulary
Olive-green — slow, developing over 30 s · Salmon-pink to flesh-pink · Grey-blue to slate · No change

Salmon-pink flesh salt is the one reaction in this bench that separates otherwise identical pale russulas from the Tricholoma group. Use a fresh solution: an oxidised, rust-coloured stock reads as a false positive on everything.

RecordTierVerdictReaction
Agaricus xanthodermusT1negativeFlesh unchanged or very faintly grey after 60 s; no salmon-pink.
Amanita phalloidesT3negativeFlesh shows no reaction within 60 s; occasional faint grey-green over the cuticle is within range.
Boletus edulisT0slow-positiveFlesh develops a grey-green to olive wash over 60-90 s; a fast salmon-pink here indicates a different genus entirely.
Cantharellus cibariusT0negativeFlesh shows no colour change after 60 s; a faint grey wash on the cuticle is within normal variation.
Chlorophyllum molybditesT1negativeFlesh shows no reaction within 60 s or a very faint grey-olive wash.
Clitocybe dealbataT3negativeFlesh unchanged after 60 s.
Coprinus comatusT0negativeFlesh shows no reaction within 60 s.
Cortinarius orellanusT3negativeFlesh unchanged after 60 s; a faint grey wash on the cortina zone is within range.
Cortinarius rubellusT3negativeFlesh unchanged after 60 s.
Entoloma sinuatumT2negativeFlesh unchanged after 60 s; no salmon-pink.
Galerina marginataT3negativeFlesh and cuticle unchanged after 60 s.
Gyromitra esculentaT3negativeStem flesh and head tissue unchanged after 90 s.
Hypholoma fasciculareT1negativeFlesh unchanged after 60 s.
Lepiota brunneoincarnataT3negativeFlesh unchanged after 60 s.
Macrolepiota proceraT0negativeFlesh unchanged after 60 s; a slow grey-green wash occasionally appears on the stem context.
Marasmius oreadesT0negativeFlesh unchanged after 90 s at 10% concentration.
Paxillus involutusT2positiveFlesh develops an olive-green to grey-green wash over 30-60 s, especially in the stem context.
Pleurocybella porrigensT2negativeFlesh unchanged after 60 s.
Rubroboletus satanasT1positiveFlesh develops a grey-green to olive wash within 30 s, faster and stronger in the stem base than in the cap.
Scleroderma citrinumT1negativeRind and young gleba unchanged after 90 s.
Tricholoma equestreT2negativeCap flesh unchanged after 60 s. A salmon-pink reaction here would place the specimen in the russulas instead.

ANIS Anisaldehyde (with sulfuric acid)Weight 2.00

Preparation
0.5 ml anisaldehyde + 1 ml concentrated sulfuric acid in 8.5 ml 96% ethanol
What it tests
Cap cuticle and cap flesh, on a fresh or a well-dried specimen
Verdict vocabulary
Grey-violet to black — the strong reaction · Grey-violet only at the disc · Negative — no change, or a pale yellow that is the reagent itself

Anisaldehyde is the reagent that separates the tricholomas, the lepiotas and the entolomas from the pale species they are confused with, and it is the one bench reagent whose result improves on a dried specimen: a cap dried at 40 C gives a cleaner reading than a rain-soaked fresh one. Read at five minutes and again at thirty; the weak reactions drift in over half an hour and the strong ones start to blacken the slide.

RecordTierVerdictReaction
Agaricus xanthodermusT1not tested—
Amanita phalloidesT3negativeNo change on the cap cuticle or on fresh flesh; the drop stays the colour of the reagent itself.
Boletus edulisT0not tested—
Cantharellus cibariusT0not tested—
Chlorophyllum molybditesT1not tested—
Clitocybe dealbataT3negativeNo change on cap or flesh; the white of the fruit body is unaffected at thirty minutes.
Coprinus comatusT0not tested—
Cortinarius orellanusT3not tested—
Cortinarius rubellusT3not tested—
Entoloma sinuatumT2positiveGrey-violet spreading from the cuticle into the flesh at the disc, reading near-black at five minutes.
Galerina marginataT3not tested—
Gyromitra esculentaT3not tested—
Hypholoma fasciculareT1not tested—
Lepiota brunneoincarnataT3positiveGrey-violet on the cap cuticle within two minutes, deepening towards black over the disc at thirty minutes.
Macrolepiota proceraT0not tested—
Marasmius oreadesT0negativeNo change on the reviving cap; the buff tone reads the same wet and dry.
Paxillus involutusT2negativeNo change on the cap; the brown pigment is not displaced by the reagent at any concentration.
Pleurocybella porrigensT2negativeNo change on the white cap or on the translucent flesh.
Rubroboletus satanasT1not tested—
Scleroderma citrinumT1not tested—
Tricholoma equestreT2negativeNothing beyond a faint grey cast on the cap after five minutes; the flesh never darkens, which is what separates it from the strongly reacting tricholomas.

NH4OH Ammonium hydroxide 10%Weight 2.00

Preparation
10 ml concentrated NH4OH in 90 ml distilled water
What it tests
Cap cuticle; also useful on the ring surface of Amanita
Verdict vocabulary
Yellow to apricot · Green to blue-green · Purplish then fading · No change

Ammonia and KOH overlap on many taxa; run them on opposite sides of the same cap so a shared result does not masquerade as two independent confirmations. Ammonia is markedly better than KOH at pulling colour out of a dry cap.

RecordTierVerdictReaction
Agaricus xanthodermusT1positiveCap and flesh turn yellow immediately, matching the KOH result; the two should be run on opposite sides of the same cap so that the duplicate is not mistaken for independent confirmation.
Amanita phalloidesT3not tested—
Boletus edulisT0negativeCap cuticle darkens slightly or shows no reaction; no yellow or green flash.
Cantharellus cibariusT0not tested—
Chlorophyllum molybditesT1negativeCap surface darkens slightly; no yellow, green or purple development.
Clitocybe dealbataT3not tested—
Coprinus comatusT0not tested—
Cortinarius orellanusT3not tested—
Cortinarius rubellusT3not tested—
Entoloma sinuatumT2not tested—
Galerina marginataT3not tested—
Gyromitra esculentaT3not tested—
Hypholoma fasciculareT1not tested—
Lepiota brunneoincarnataT3not tested—
Macrolepiota proceraT0not tested—
Marasmius oreadesT0not tested—
Paxillus involutusT2positiveCap surface and flesh turn a brownish-red immediately, then fade over several minutes.
Pleurocybella porrigensT2not tested—
Rubroboletus satanasT1positiveFlesh and pore surface turn greenish-blue immediately, especially at the stem base.
Scleroderma citrinumT1not tested—
Tricholoma equestreT2not tested—

COTTON Cotton blue in lactophenolWeight 2.00

Preparation
0.05 g methyl blue + 20 g phenol + 20 ml lactic acid + 20 ml glycerol per 100 ml
What it tests
Spore ornamentation, cystidia walls, and clamped septa
Verdict vocabulary
Cyanophilic — wall or warts stain deep blue · Weakly cyanophilic — pale blue outline only · Acyanophilic — remains unstained

The stain fills the space between warts, so the pattern reads as a negative image. Photograph within 10 minutes; the phenol fraction clears the preparation and the contrast falls off after that.

RecordTierVerdictReaction
Agaricus xanthodermusT1not tested—
Amanita phalloidesT3acyanophilicSpores remain unstained; the thin wall shows a clear, evenly thick outline against the blue back­ground.
Boletus edulisT0not tested—
Cantharellus cibariusT0acyanophilicSpores remain unstained against a blue background; the outline reads as a clear halo.
Chlorophyllum molybditesT1not tested—
Clitocybe dealbataT3acyanophilicSpores remain unstained and appear as clear outlines.
Coprinus comatusT0cyanophilicSpore wall stains deep blue, the central germ pore remaining as an unstained window.
Cortinarius orellanusT3cyanophilicWarts stain blue against a paler wall, giving a strongly patterned outline.
Cortinarius rubellusT3cyanophilicWarts stain blue against a paler wall.
Entoloma sinuatumT2acyanophilicSpores remain unstained and the angular outline reads clearly against the blue background.
Galerina marginataT3cyanophilicThe warty ornamentation stains blue and the plage stays clear, so the spore reads as an ornamented oval with a bare notch.
Gyromitra esculentaT3acyanophilicSpores remain unstained; the two oil drops show as clear refractile bodies against the blue background.
Hypholoma fasciculareT1not tested—
Lepiota brunneoincarnataT3cyanophilicThe thick wall stains blue and the germ pore remains a clear unstained window.
Macrolepiota proceraT0cyanophilicThe thick spore wall stains blue, leaving the germ pore as a clear unstained patch at one end.
Marasmius oreadesT0acyanophilicSpores remain unstained, appearing as clear outlines.
Paxillus involutusT2not tested—
Pleurocybella porrigensT2not tested—
Rubroboletus satanasT1not tested—
Scleroderma citrinumT1acyanophilicSpores remain unstained, but the spines take up a pale blue edge that makes them easier to count.
Tricholoma equestreT2acyanophilicSpores remain unstained; the thin smooth wall shows as a clear outline.

SULFOVAN Sulfovanillin (SV)Weight 1.00

Preparation
1 g vanillin in 10 ml concentrated sulfuric acid, diluted 1:1 with distilled water
What it tests
Cystidia contents in the gill trama and on the gill edge
Verdict vocabulary
Blue-black · Violet then grey · Negative — unchanged or yellowish

Applied to a squash mount, not to a whole gill. Mix on a clean slide; a trace of iron from a reused needle turns the whole drop black and voids the test.

RecordTierVerdictReaction
Agaricus xanthodermusT1not tested—
Amanita phalloidesT3not tested—
Boletus edulisT0not tested—
Cantharellus cibariusT0not tested—
Chlorophyllum molybditesT1not tested—
Clitocybe dealbataT3not tested—
Coprinus comatusT0not tested—
Cortinarius orellanusT3not tested—
Cortinarius rubellusT3not tested—
Entoloma sinuatumT2not tested—
Galerina marginataT3not tested—
Gyromitra esculentaT3not tested—
Hypholoma fasciculareT1not tested—
Lepiota brunneoincarnataT3not tested—
Macrolepiota proceraT0not tested—
Marasmius oreadesT0not tested—
Paxillus involutusT2not tested—
Pleurocybella porrigensT2not tested—
Rubroboletus satanasT1not tested—
Scleroderma citrinumT1not tested—
Tricholoma equestreT2not tested—

GUAIAC Guaiac tinctureWeight 1.00

Preparation
1 g gum guaiac dissolved in 10 ml 70% ethanol, kept dark and used within a month
What it tests
Fresh context tissue and the cut surface of the stem
Verdict vocabulary
Blue — rapid, within 30 s · Blue after 2 min · Negative

A positive guaiac reaction indicates an extracellular laccase, which places the specimen among the white-rot wood decayers. It is a metabolic marker, not a toxicity marker: a strong blue says nothing about whether the fruit body is safe to handle or to eat.

RecordTierVerdictReaction
Agaricus xanthodermusT1not tested—
Amanita phalloidesT3not tested—
Boletus edulisT0not tested—
Cantharellus cibariusT0not tested—
Chlorophyllum molybditesT1not tested—
Clitocybe dealbataT3not tested—
Coprinus comatusT0not tested—
Cortinarius orellanusT3not tested—
Cortinarius rubellusT3not tested—
Entoloma sinuatumT2not tested—
Galerina marginataT3not tested—
Gyromitra esculentaT3not tested—
Hypholoma fasciculareT1positiveFresh context turns blue within 30 s — a strong positive, consistent with a white-rot wood decayer rather than a litter saprotroph.
Lepiota brunneoincarnataT3not tested—
Macrolepiota proceraT0not tested—
Marasmius oreadesT0not tested—
Paxillus involutusT2not tested—
Pleurocybella porrigensT2positiveFresh context turns blue within 60 s, consistent with a white-rot wood decayer.
Rubroboletus satanasT1not tested—
Scleroderma citrinumT1not tested—
Tricholoma equestreT2not tested—

Legend