692 
Journal of Agricultural Research 
Vol. XXHI, No. 9 
Observations made frequently thereafter failed to show any signs of 
the development of a perfect stage. 
Experiment II.—February 22. Sclerotia collected from castor-bean plants under 
natural conditions were sterilized for two minutes in 1 to 1,000 mercuric chlorid, 
washed in sterile water three changes, and placed partly on plates of agar, partly on 
plates of moist sterilized sand. The agar plates developed a typical culture of the 
fungus (B 39, isolation record, p. 688). On the sand, the mature sclerotia produced an 
abundance of normal conidiophores and conidia. 
FIRST APPEARANCE OF APOTHECIA 
The first appearance of the perfect stage of this organism was more or 
less unexpected; consequently it might be of interest to record its history 
in detail. On one of the cultures sent to Prof. Whetzel at Cornell Univers¬ 
ity, No. B21, collected November 25, 1918, he noticed an apothecial stalk 
arising from one of the sclerotia. Prof. Whetzel immediately notified 
the writer at his temporary station, Orlando, Fla., and he found (Mar. 8) 
in his culture of the same strain (B 21) and also in one other strain (B 1), a 
similar condition. The cultures had remained on the shelf and had not 
been examined for some time. These cultures were then watched with 
jealous care at both places. In the culture at Cornell the stalk developed 
a little more, then shriveled and dried up. Under what were undoubtedly 
the more favorable conditions at Orlando, Fla., the culture continued to 
develop until the cup opened out and reached maturity. Thus there was 
secured a perfect specimen of the apothecial stage of this fungus which 
until then had appeared to be merely a new species of Botrytis. 
Over anxiety to permit this specimen to reach full maturity resulted in 
its becoming slightly overmature, as was evidenced by the development 
of a mealy condition of the disk. Other apothecia were meanwhile 
rapidly developing, however, and it was clear that working material was 
to be had in sufficient abundance for immediate needs. Extraordinary 
precautions were taken with this culture tube to obtain absolutely 
authentic single ascospore cultures, for it was not known at this time 
whether or not further specimens could readily be obtained. From the 
first specimen to mature, mounts were made and drawings and measure¬ 
ments secured. The second was used to start cultures, but these were 
discarded in favor of the third, a more perfect specimen, from which 
more clearly reliable results were secured. This apothecium, taken in 
situ, is shown in Plate 11 A. For the sake of subsequent references it 
will be designated apothecium C. Its maturity was indicated on March 
27 when a cloudiness different from that due to moisture was observed on 
the inside of the tube near the disk of the apothecium. A scraping was 
made from this deposit with a sterile platinum needle and examination 
showed it to consist of long fusoid spores, obviously ascospores that had 
been ejected from the mature apothecium. 
In preparation for the actual work of securing single ascospore cul¬ 
tures, several Van Tieghem cells, slides, and thin cover glasses were 
sterilized in the hot air oven, within Petri dishes. In addition, tubes of 
sterile water and sterile vaseline were made ready. Then some of the 
cover glasses were covered within a small circular area on one side with 
a thin, smooth film of clear corn meal agar, applied directly from a tube 
of melted agar with a platinum loop. Apothecium C was then removed 
from the tube and placed in a sterile Petri dish. 
Isolations were made from ascospores derived in three different ways, 
as follows: (a) Spores ejected directly from the apothecium to an agar- 
