The primary materials needed for the process of humification are plant detritus and dead animals and microbes, excreta of all soil-dwelling organisms, and also black carbon resulting from past fires. The composition of humus varies with that of primary (plant) materials and secondary microbial and animal products. The decomposition rate of the different compounds will affect the composition of the humus.
It is difficult to define humus precisely because it is a very complex substance which is still not fully understood. Humus is different from decomposing soil organic matter. The latter looks rough and has visible remains of the original plant or animal matter. Fully humified humus, on the contrary, has a uniformly dark, spongy, and jelly-like appearance, and is amorphous; it may gradually decay over several years or persist for millennia. It has no determinate shape, structure, or quality. However, when examined under a microscope, humus may reveal tiny plant, animal, or microbial remains that have been mechanically, but not chemically, degraded. This suggests an ambiguous boundary between humus and soil organic matter, leading some authors to contest the use of the term humus and derived terms such as humic substances or humification, proposing the Soil Continuum Model (SCM). However, humus can be considered as having distinct properties, mostly linked to its richness in functional groups, justifying its maintenance as a specific term.
Fully formed humus is essentially a collection of very large and complex molecules formed in part from lignin and other polyphenolic molecules of the original plant material (foliage, wood, bark), in part from similar molecules that have been produced by microbes. During decomposition processes these polyphenols are modified chemically so that they are able to join up with one another to form very large molecules. Some parts of these molecules are modified in such a way that protein molecules, amino acids, and amino sugars are able to attach themselves to the polyphenol "base" molecule. As protein contains both nitrogen and sulfur, this attachment gives humus a moderate content of these two important plant nutrients.
Radiocarbon and other dating techniques have shown that the polyphenolic base of humus (mostly lignin and black carbon) can be very old, but the protein and carbohydrate attachments much younger, while to the light of modern concepts and methods the situation appears much more complex and unpredictable than previously thought. It seems that microbes are able to pull protein off humus molecules rather more readily than they are able to break the polyphenolic base molecule itself. As protein is removed its place may be taken by younger protein, or this younger protein may attach itself to another part of the humus molecule.
There is little data available on the composition of humus because it is a complex mixture that is challenging for researchers to analyze. Researchers in the 1940s and 1960s tried using chemical separation to analyze plant and humic compounds in forest and agricultural soils, but this proved impossible because extractants interacted with the analysed organic matter and created many artefacts. Further research has been done in more recent years, though it remains an active field of study.
Much of the humus in most soils has persisted for more than 100 years, rather than having been decomposed into CO2, and can be regarded as stable; this organic matter has been protected from decomposition by microbial or enzyme action because it is hidden (occluded) inside small aggregates of soil particles, or tightly sorbed or complexed to clays. Most humus that is not protected in this way is decomposed within 10 years and can be regarded as less stable or more labile. The mixing activity of soil-consuming invertebrates (e.g. earthworms, termites, some millipedes) contribute to the stability of humus by favouring the formation of organo-mineral complexes with clay at the inside of their guts, hence more carbon sequestration in humus forms such as mull and amphi, with well-developed mineral-organic horizons, when compared with moder where most organic matter accumulates at the soil surface.
Stable humus contributes few plant-available nutrients in soil, but it helps maintain its physical structure. A very stable form of humus is formed from the slow oxidation (redox) of soil carbon after the incorporation of finely powdered charcoal into the topsoil, suggested to result from the grinding and mixing activity of a tropical earthworm. This process is speculated to have been important in the formation of the unusually fertile Amazonian terra preta do Indio. However, some authors suggest that complex soil organic molecules may be much less stable than previously thought: "the available evidence does not support the formation of large-molecular-size and persistent 'humic substances' in soils. Instead, soil organic matter is a continuum of progressively decomposing organic compounds.″
Humus has a characteristic black or dark brown color and is organic due to an accumulation of organic carbon. Soil scientists use the capital letters O, A, B, C, and E to identify the master soil horizons, and lowercase letters for distinctions of these horizons. Most soils have three major horizons: the surface horizon (A), the subsoil (B), and the substratum (C). Some soils have an organic horizon (O) on the surface, but this horizon can also be buried. The master horizon (E) is used for subsurface horizons that have significantly lost minerals (eluviation). Bedrock, which is not soil, uses the letter R. The richness of soil horizons in humus determines their more or less dark color, generally decreasing from O to E, to the exception of deep horizons of podzolic soils enriched with colloidal humic substances which have been leached down the soil profile.
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