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this can be done, in the dark, by bacteria to which only pure mineral salts—e.g. carbonates, sulphates and chlorides of ammonium, sodium and magnesium—were added. In other words these bacteria can build up organic matter from purely mineral sources by assimilating carbon from carbon dioxide in the dark and by obtaining their nitrogen from ammonia. The energy liberated during the oxidation of the nitrogen is regarded as splitting the carbon dioxide molecule,—in green plants it is the energy of the solar rays which does this. Since the supply of free oxygen is dependent on the activity of green plants the process is indirectly dependent on energy derived from the sun, but it is none the less an astounding one and outside the limits of our previous generalizations. It has been suggested that urea is formed by polymerization of ammonium carbonate, and formic aldehyde is synthesized from CO2 and OH2. The Nitro-bacteria are smaller, finer and quite different from the nitroso-bacteria, and are incapable of attacking and utilizing ammonium carbonate. When the latter have oxidized ammonia to nitrite, however, the former step in and oxidize it still further to nitric acid. It is probable that important consequences of these actions result from the presence of nitrifying bacteria in rotten stone, decaying bricks, &c., where all the conditions are realized for preparing primitive soil, the breaking up of the mineral constituents being a secondary matter. That “soil” is thus prepared on barren rocks and mountain peaks may be concluded with some certainty.

Bacteriology 14.png
Fig. 14.—Stages in the formation of a colony of a variety of Bacillus (Proteus)
vulgaris (Hauser), observed in a hanging drop. At 11 a.m. a rodlet appeared (A); at 4
p.m. it had grown and divided and broken up into eight rodlets (B); C shows further
development at 8 p.m., D at 9.30 p.m.—all under a high power. At E, F, and G further
stages are drawn, as seen under much lower power.  (H. M. W.) 

In addition to the bacterial actions which result in the oxidization of ammonia to nitrous acid, and of the latter to nitric acid, the reversal of such processes is also brought about by numerous bacteria in the soil, rivers, &c. Warington showed some time ago that many species are able to reduce nitrates to nitrites, and such reduction is now known to occur very widely in nature. The researches of Gayon and Dupetit, Giltay and Aberson and others have shown, moreover, that bacteria exist which carry such reduction still further, so that ammonia or even free nitrogen may escape. The importance of these results is evident in explaining an old puzzle in agriculture, viz. that it is a wasteful process to put nitrates and manure together on the land. Fresh manure abounds in de-nitrifying bacteria, and these organisms not only reduce the nitrates to nitrites, even setting free nitrogen and ammonia, but their effect extends to the undoing of the work of what nitrifying bacteria may be present also, with great loss. The combined nitrogen of dead organisms, broken down to ammonia by putrefactive bacteria, the ammonia of urea and the results of the fixation of free nitrogen, together with traces of nitrogen salts due to meteoric activity, are thus seen to undergo various vicissitudes in the soil, rivers and surface of the globe generally. The ammonia may be oxidized to nitrites and nitrates, and then pass into the higher plants and be worked up into proteids, and so be handed on to animals, eventually to be broken down by bacterial action again to ammonia; or the nitrates may be degraded to nitrites and even to free nitrogen or ammonia, which escapes.

That the Leguminosae (a group of plants including peas, beans, vetches, lupins, &c.) play a special part in agriculture was known even to the ancients and was mentioned by Pliny (Historia Naturalis, viii). These plants will not only grow on poor sandy Bacteria
and Legumin-
soil without any addition of nitrogenous manure, but they actually enrich the soil on which they are grown. Hence leguminous plants are essential in all rotation of crops. By analysis it was shown by Schulz-Lupitz in 1881 that the way in which these plants enrich the soil is by increasing the nitrogen-content. Soil which had been cultivated for many years as pasture was sown with lupins for fifteen years in succession; an analysis then showed that the soil contained more than three times as much nitrogen as at the beginning of the experiment. The only possible source for this increase was the atmospheric nitrogen. It had been, however, an axiom with botanists that the green plants were unable to use the nitrogen of the air. The apparent contradiction was explained by the experiments of H. Hellriegel and Wilfarth in 1888. They showed that, when grown on sterilized sand with the addition of mineral salts, the Leguminosae were no more able to use the atmospheric nitrogen than other plants such as oats and barley. Both kinds of plants required the addition of nitrates to the soil. But if a little water in which arable soil had been shaken up was added to the sand, then the leguminous plants flourished in the absence of nitrates and showed an increase in nitrogenous material. They had clearly made use of the nitrogen of the air. When these plants were examined they had small swellings or nodules on their roots, while those grown in sterile sand without soil-extract had no nodules. Now these peculiar nodules are a normal characteristic of the roots of leguminous plants grown in ordinary soil. The experiments above mentioned made clear for the first time the nature and activity of these nodules. They are clearly the result of infection (if the soil extract was boiled before addition to the sand no nodules were produced), and their presence enabled the plant to absorb the free nitrogen of the air.

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Fig. 15.—Invasion of leguminous roots by bacteria.
a, cell from the epidermis of root of Pea with
“infection thread” (zoogloea) pushing its way
through the cell-walls. (After Prazmowski.)
b, free end of a root-hair of Pea; at the right are
particles of earth and on the left a mass of bacteria.
Inside the hair the bacteria are pushing their way up
in a thin stream.
(From Fischer's Vorlesungen über Bakterien.)
Bacteriology 16.png
Fig. 16.
a, root nodule of the lupin, nat. size. (From Woromv.)
b, longitudinal section through root and nodule.
g, fibro-vascular bundle.
w, bacterial tissue. (After Woromv.)
c, cell from bacterial tissues showing nucleus and
protoplasm filled with bacteria.
d, bacteria from nodule of lupin, normal
undegenerate form.
e and f, bacteroids from Vicia villosa and
Lupinus albus. (After Morck.)

(From Fischer's Vorlesungen über Bakterien.)

The work of recent investigators has made clear the whole process. In ordinary arable soil there exist motile rod-like bacteria, Bacterium radicicola. These enter the root-hairs of leguminous plants, and passing down the hair in the form of a long, slimy (zoogloea) thread, penetrate the tissues of the root. As a result the tissues become hypertrophied, producing the well-known nodule. In the cells of the nodule the bacteria multiply and develop, drawing material from their host. Many of the bacteria exhibit curious involution forms (“bacteroids”), which are finally broken down and their products absorbed by the plant. The nitrogen of the air is absorbed by the nodules, being built up into the bacterial cell and later handed on to the host-plant. It appears from the observations of Mazé that the bacterium can even absorb free nitrogen when grown in cultures