3 2 4 
Journal of Agricultural Research 
Vol. I, No. 4 
shoots treated in the same way but unpunctured. Groups A and B were held as checks. 
Group C consisted of 8 shoots bearing io nuts the hulls of which were punctured with 
a sterile needle and sprayed with a sterile distilled-water suspension of the conidia. 
Group D consisted of 6 shoots bearing 8 pecans which were inoculated like group C, 
except that the hulls were not punctured. Group E consisted of io nuts removed 
from the shoots, their hulls punctured, and inoculated with a suspension of conidia 
as in groups C and D, and then placed in damp chambers. 
At the end of three days distinct infection had occurred on all the nuts with punc¬ 
ture inoculations, the tissue of the hulls being blackened for a radius of 2 to 5 mm. 
around the needle punctures. The first checks had the tissue blackened for a radius 
of about 0.5 mm. around the needle punctures, while the nonpunctured check and 
inoculated nuts at this time showed no evidence of infection. Many of the leaves 
on the inoculated shoots were developing small, irregular brownish areas, while the 
uninoculated leaves were all green and healthy. 
After nine days groups A and B appeared as on the third day. The very narrow 
margin of blackened tissue in the punctured checks was due merely to the mechanical 
injury to immediately surrounding cells, and no further injury occurred throughout 
the experiment. (PI. XXXIII, fig. 1, A.) All the pecans in group C (PI. XXXIII, 
fig. 1, C ) were blackened over half to the whole of their surface, and acervuli were 
beginning to develop over the dead parts, with an occasional exudation of the pink 
spore masses. In group D half of the eight nuts had blackened, and acervuli had 
begun to develop, but the others gave no evidence of infection. (PI. XXXIII, fig. 
1, B.) In group E all the nuts were blackened, and very numerous acervuli with 
exuding spore masses had developed. Reisolations of this fungus were made as strain 
144. Plate XXXIII, figure 2, shows three of the inoculated nuts after further develop¬ 
ment of acervuli. 
apples 
Experiment No. i (Dec. 30, 1911).—Five sound Jonathan apples direct from cold 
storage were placed in damp chambers and inoculated by needle punctures, two of 
them with conidia and three with ascospores inserted directly into the punctures. 
Three sound apples were punctured with sterile needles and also placed in damp 
chambers. All were kept overnight at 35 0 C., and subsequently throughout the 
experiment at laboratory temperatures. Examination after seven days showed a 
decay very similar in appearance to bitter-rot around all the inoculation punctures. 
The check apples were perfectly sound. These cultures were kept for 10 days, with 
a gradual increase in the size of the decayed areas and formation of incipient fruiting 
bodies but no spore production. 
Experiment No. 2 (Mar. 5, 1912).—Twelve Yellow Newtown apples were similarly 
inoculated and placed in damp chambers, one half being inoculated with conidia 
and the other half with ascospores from an 8-weeks-old com-meal-agar culture (strain 
17). Three apples from each set were held at 28° to 30° C. and three from each set 
at laboratory temperature. Six apples punctured with sterile needles and placed in 
damp chambers were held as checks, one half at 28° to 3o° and the other half at labo¬ 
ratory temperature. At the end of a week the cultures were examined, and all the 
inoculated apples had developed a decay apparently identical with bitter-rot, but 
the brownish and somewhat sunken spots had increased in size much more rapidly 
at the higher temperature. Two weeks later perithecia were forming and the conidia 
were developing in considerable numbers, but not in such amount as to give the pink 
spore masses characteristic of bitter-rot. The check apples at both temperatures 
remained sound to the end of the experiment. 
Experiment No. 3 (Nov. 30, 1912).—Sound Jonathan and Yellow Newtown apples 
direct from cold storage were inoculated with three strains of Glomerella obtained 
from the pecan and with one strain obtained from the apple. The cultures used. 
