Amyloid (mycology)

In mycology a tissue or feature is said to be amyloid if it has a positive amyloid reaction when subjected to a crude chemical test using iodine as an ingredient of either Melzer's reagent or Lugol's solution, producing a blue to blue-black staining. The term "amyloid" is derived from the Latin amyloideus ("starch-like"). It refers to the fact that starch gives a similar reaction, also called an amyloid reaction. The test can be on microscopic features, such as spore walls or hyphal walls, or the apical apparatus or entire ascus wall of an ascus, or be a macroscopic reaction on tissue where a drop of the reagent is applied. Negative reactions, called inamyloid or nonamyloid, are for structures that remain pale yellow-brown or clear. A reaction producing a deep reddish to reddish-brown staining is either termed a dextrinoid reaction (pseudoamyloid is a synonym) or a hemiamyloid reaction.

Melzer's is used by exposing fungal tissue or cells to the reagent, typically in a microscope slide preparation, and looking for any of three color reactions:

Among the amyloid reaction, two types can be distinguished:

Melzer's reactions are typically almost immediate, though in some cases the reaction may take up to 20 minutes to develop.

The function of the chemicals that make up Melzer's reagent are several. The chloral hydrate is a clearing agent, bleaching and improving the transparency of various dark-colored microscopic materials. The potassium iodide is used to improve the solubility of the iodine, which is otherwise only semi-soluble in water. Iodine is thought to be the main active staining agent in Melzer's; it is thought to react with starch-like polysaccharides in the cell walls of amyloid material, however, its mechanism of action is not entirely understood. It has been observed that hemiamyloid material reacts differently when exposed to Melzer's than it does when exposed to other IKI solutions such as Lugol's, and that in some cases an amyloid reaction is shown in material that had prior exposure to KOH, but an inamyloid reaction without such pretreatment.

An experiment in which spores from 35 species of basidiomycetes were tested for reactions to both Melzer's and Lugol's showed that spores in a large percentage of the species tested display very different reactions between the two reagents. These varied from being weakly or non-reactive in Lugols, to giving iodine-positive reactions in Lugol's but not in Melzer's, to even giving dextrinoid reactions in Lugol's while giving amyloid reactions in Melzer's.

Melzer's degrades into a cloudy precipitate when combined with alkaline solutions, hence it cannot be used in combination or in direct series with such common mycological reagents such as potassium hydroxide or ammonium hydroxide solutions. When potassium hydroxide is used as a pretreatment, the alkalinity must be first neutralized before adding Melzer's.

Hemiamyloidity in mycology refers to a special case of cell wall amyloidity where the blue staining by iodine only occurs when the tissue was pretreated with potassium hydroxide solution (KOH) or other strong bases, whereas direct application of iodine causes a red reaction when using Lugol's solution, but no reaction when using Melzer's reagent. Hemiamyloidity is so far only known in Ascomycota, but here widespread and an important taxonomic distinction criterion. If cell walls stain blue by iodine reagents without pretreatment with KOH, this is called euamyloid. The term amyloid comprises both variants.