52 BULLETIN 727, U. S. DEPARTMENT OF AGRICULTURE. 
In a total of more than 4,000 seeds tested no anthracnose was 
found. From the 3,100 seeds tested in sterile soil no diseased seed- 
lings developed. Where culture media were used the profuse devel- 
opment of bacteria and fungi upon and around each seed proved 
that the latter bore an abundant and varied surface flora. On 
old seed the flora was less varied. The rapid development of these 
forms rendered anthracnose isolation by cultural methods a matter 
of great difficulty. 
To determine whether or not the methods employed above were 
reliable, tests were made with seed previously dipped in a spore 
suspension. The results of these tests warrant a more detailed 
account. On March 13 about 500 seeds were immersed for 50 minutes 
in a heavy suspension of spores of the anthracnose fungus and then 
dried on filter paper. The next day 50 of these seeds were planted 
in sterile moist sand in a damp chamber. The seedlings were sprayed 
from time to time with sterile water. Sixteen days after planting, 
one seedling, which had carried its seed coat up with the cotyledons, 
showed an anthracnose lesion on the stem near the ground line and 
another on a cotyledon. Owing to the stem lesion the seedling had 
already fallen over, asin damping-off. On thestem lesion were numer- 
ous sporulating acervuli. In 22 days after planting, three more 
seedlings had damped-off with anthracnose and four others bore 
lesions at the ground line. In 29 days after planting, 16 out of the 
45 seedlings had succumbed to anthracnose. 
These results corroborate those secured by Carsner,! who obtained 
damping-off of cucumber and muskmelon seedlings when seeds pre- 
viously dipped in a spore suspension were planted im sterile soil. 
Such results suggest that perhaps in the field the originally diseased 
plant may damp-off and disappear, so that the first lesions dis- 
covered are the result of spores splashed from this plant upon those 
adjacent to it. 
To test the filter-paper culture method, 28 of these inoculated 
seeds were planted in four sterile Petri dishes on filter paper mois- 
tened with 2 per cent dextrose solution. Six days later, acervuli of 
anthracnose were found on two seed coats, and by nine days one 
more seed bore acervuli and its seedling was diseased. Two weeks 
after planting, two more seedlings were diseased. Mold and bacte- 
rial growth was as usual abundant on these seeds. The results of 
these and further tests with these seeds are presented in Table VI. 
While it is evident that diseased seedlings may result from mocu- 
lated seed, it is noteworthy that not all inoculated seeds yield diseased 
seedlings in soil. Owing largely to the profusion of other organisms 
on the seeds, the methods of anthracnose detection used with the 
commercial seed, as summarized in Table V, show a low efficiency 
Ww Hen appa to seed known to be heavily inoculated with ee fungus. 
1Carsner, E, Op. cit, 
