. Bulletin. 1901-13. Agriculture; Agriculture. 108 CROWN-GALL OP PLANTS. It does not stain by Gram. It stains readily a uniform deep blue if amyl alcohol be substituted for ethyl alcohol in the washing after exposure to the anilin gentian violet and the iodine-potassium iodid of Gram's stain. Taken from beef bouillon 7 weeks old it did not show glycogen stain when exposed to iodine water; i. e., there was only a uniform yellow color. It is not acid-fast. Brizi's method was employed on daisy galls without success. By the use of methylene green (not methyl green, but that was tried also), withou
. Bulletin. 1901-13. Agriculture; Agriculture. 108 CROWN-GALL OP PLANTS. It does not stain by Gram. It stains readily a uniform deep blue if amyl alcohol be substituted for ethyl alcohol in the washing after exposure to the anilin gentian violet and the iodine-potassium iodid of Gram's stain. Taken from beef bouillon 7 weeks old it did not show glycogen stain when exposed to iodine water; i. e., there was only a uniform yellow color. It is not acid-fast. Brizi's method was employed on daisy galls without success. By the use of methylene green (not methyl green, but that was tried also), without sub- sequent exposure to acid, numerous cell inclusions, con- sisting of bacteria-like gran- ules, were demonstrated, but whether really bacteria re- mained undetermined. The bacteria Brizi suc- ceeded in stainmg readily in poplar tumor tissues by an acid-fast method were probably not-this organism. In our hands the gall-producing organism in American poplar galls stains like the daisy. It is not an acid-fast organism. In sections it is often stained with difficulty, and wte have seldom been able to differentiate it well from the surrounding tissue. It seems to us to occur, for the most part, at least, in the interior of the parenchyma cells rather than in the intercellular spaces or vessels. Repeated efforts to stain in situ have not yielded, as a rule, well- stained, sharpl}' defined rods such as one would expect, but occa- sionally stained and unstained we have seen rods inside the cells which we believe to be the Fig. 3.—Daisy organism. Slime from pellicle on beef- bouillon culture three weeks old. Stained with carbol fuchsin, and camera-drawn by Miss Brown. CULTURAL CHARACTERS. NUTRIENT AGAR.'' Colonies.— When the organisms are obtained from a crushed TcTiot the colonies come up in from 3 to 12 days (usually 4 to 6) at a room temperature of about 25° C. on poured agar plates. They come up very much slower when taken from knots than when taken from young c
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