Journal of Agricultural Research 
Vol. VIII, No. 5 
170 
liberates a small amount of carbon dioxid (4, p. 21-22). These sub¬ 
stances, therefore, must be the first things tested out experimentally on 
plants subject to crowngall. On the start it seemed to me probable that 
ammonia, given off in small quantities within the cell by the multiplying 
bacteria, must be the sole determining factor in the abnormal cell pro¬ 
liferation, since as a diffusible stimulant it would enter cells readily, would 
increase the osmotic pressure, and would tend to enter into soapy com¬ 
binations with the lipoids of the cell surface, thus changing the surface 
tension beyond the point of physiological cell restraint, whereupon cell 
division would take place. At the same time that ammonia and other 
concentrated bacterial products move outward, toward the surface of the 
tumor, where usually growth is most abundant, water and dissolved 
foods would move inward into the tumor, thus supplying copiously, 
especially at the periphery of the tumor, both the necessary stimulus and 
the substances needed for the constantly increasing abnormal growth. 
Minute continuous doses of other alkalies might act on the cells in the 
same way, but, as already intimated, it is here necessary to consider only 
such substances as are likely to be liberated within the cells of the tumor 
by the metabolism of the crowngall organism. If the substance which 
removes the growth inhibition is ammonia or any other compound pro¬ 
duced by the crowngall bacterium as the result of its growth within the 
cell then the rate of cell division in the tumor would depend on the rate 
of bacterial multiplication and metabolism within the cells and on the 
relative juiciness of the tissues, which together would determine the 
rate of osmotic movement, while the rate of the bacterial multiplication 
were added to the distillate. That acetone was present was confirmed by adding to a portion of the dis¬ 
tillate an equal portion of strong potassium hydroxid solution and then a few drops of io per cent alcoholic 
solution of salicyl aldehyd. An orange red color is developed on warming to 4o°-so° C. (Csonka, Jour. 
Biol. Chem, 1916, 27, p. 209). 
“ The acetone was removed from a separate portion of the distillate by aerating a few minutes at 40° C., 
and when there was no further positive test for acetone, alcohol was detected by means of the KOH-iodine 
reaction when iodoform was produced abundantly on warming to 6o° C. 
“The presence of aldehyd was detected by the Tollens’s silver reduction and the fuchsin tests. 
“Acetone, alcohol, and aldehyd in varying amounts were found in the distillates from each of the nine 
inoculated flasks. None of these was found in the controls. 
“The residue in the distilling flask, representing about one-half the original volume, was diluted to 
original volume with distilled water, 25 cc 5 N sulphuric acid were added and the mixture submitted to 
steam distillation. The distillate was neutralized with barium hydroxid solution, evaporated in vacuum 
to small volume, and the barium salts obtained by further concentration on the steam bath. There 
appeared to be a mixture of barium salts present, but as the quantities were small only qualitative tests 
were made. Acetic and small amounts of formic acids were detected in flasks A, B, C (Flats Poplar), 
and E, F, G (Rose). Flasks I, K, and I, (Daisy) gave no indication of acetic acid; formic acid only was 
detected (silver nitrate reduction). 
“No nonvolatile acids were found in the acid residue on extracting with ether and evaporating. No 
fixed acids were found on acidifying the calcium carbonate residue in the culture flask and extracting 
with ether. 
"(Signed) J. F. Brewster. 
“January 5, 1917-” 
REPORT ON VOLATILE ALKALI 
“In the alkaline distillates from flasks A, B, C, E, and F the following tests were made: 
Ammonia.: ..Present. 
Amines.. Do. 
Trimethylamine.Absent. 
“(Signed) H. E. Woodward.’" 
