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Vol. XXV, No. 3 
(9) The bacteria were observed in unstained free-hand sections and 
also in paraffin sections stained with very dilute carbol-fuchsin and light 
green. Apart from the region immediately around the wound, they were 
found only in an intercellular position. They were seen in gall tissue 
of all ages from the day of inoculation until the gall showed the characters 
of maturity about 24 days after inoculation. 
(10) Isolation and microscopic observations indicated that the bac¬ 
teria are present in larger numbers than was previously supposed. The 
treatment of the tissues with mercuric chlorid before plating appears 
to lower the count so as to give a false impression of the number of 
bacteria present. 
(11) A further control on the location of the crowngall bacteria was 
made by observing their multiplication in free-hand sections cut under 
aseptic conditions and embedded in agar, by subsequently isolating 
them from such preparations, and by establishing their identity with 
the usual cultural and inoculation methods. Dilution plates poured 
from such sections have repeatedly yielded thousands of colonies of the 
crown-gall organism with less than one per cent contaminations. 
LITERATURE CITED 
(1) Conn, H. Joel. 
1918. THE MICROSCOPIC STUDY OR BACTERIA AND FUNGI IN SOIL. N. Y. 
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(2) Jones, Leon K. 
1920. A SURVEY OF DISEASES AND INSECT INJURIES OF CANE FRUITS IN WIS¬ 
CONSIN, 1919. In Ann. Rpt. Wis. State Hort. Soc., v. 50, p. 143-150. 
(3) Jones, L. R. 
1909. PECTINASE, THE CYTOLYTIC ENZYM PRODUCED BY BACILLUS CAROTOVORUS 
and certain other soft-rot organisms. In Vermont Agr. Exp. 
Sta. Bui. 147, Pt. II, p. 281-360, 10 fig. Bibliography, 357-360. 
(4) Pfeffer, W. 
1888. tlBER CHEMOTAKTISCHE BEWEGUNGEN VON BACTERIEN, FLAGELLATEN 
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(5) Smith, Erwin F. 
1911. crown-gall and sarcoma. U. S. Dept. Agr. Bur. Plane Indus. Circ. 
8 5 , 4 P . 
( 6 ) - 
1912. ON SOME RESEMBLANCES OF CROWN-GALL TO HUMAN CANCER. In 
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1912. the staining of bact. tumefaciens in tissue. In Phytopathology, 
v. 2, p. 127-128. 
( 8 ) - 
( 9 ) 
(10) 
(11) 
( 12 ) 
1916. STUDIES ON THE CROWN-GALL OF PLANTS. ITS RELATION TO HUMAN 
cancer. In Jour. Cancer Research, v. 1, p. 231-309, 25 pi. (in text). 
1917. EMBRYOMAS IN PLANTS. (PRODUCED BY BACTERIAL INOCULATIONS.) 
In Bui. Johns Hopkins Hosp., v. 28, no. 319, p. 277-294, 1 fig., pi. 
26-53. 
1920. AN INTRODUCTION TO BACTERIAL DISEASES OF PLANTS. 688 p., 453 fig. 
Philadelphia and London. Literature at end of most of the chapters. 
--, Brown, Nellie A., and McCulloch, Lucia. 
I912. THE STRUCTURE AND DEVELOPMENT OF CROWN GALL: A PLANT CANCER. 
U. S. Dept. Agr. Bur. Plant Indus. Bui. 255, 61 p., 2 fig. 109 pi. 
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