Howard . —On Trichosphaeria Sacchari , Massee. 619 
chlamydospores were noted seven days after infection. Similar 
flask-cultures made at the same time gave similar results. 
Evidently under these conditions the Melanconium- mycelium 
does not develop in the manner described by Massee in his 
Kew experiments. 
Meanwhile hanging-drop cultures of Melanconium- spores 
were made according to Professor Marshall Ward’s method 
of drop culture 1 in fifteen per cent, saccharose-gelatine con¬ 
taining Klebs’ solution. Germination took place twenty-two 
hours after sowing, and a colourless branched septate myce¬ 
lium was developed. The day temperature of the laboratory 
during the development of this series of drop-cultures varied 
between 26° C. and 29 0 C. After germination the mycelium 
developed rapidly, and in two days the drops were filled with 
mycelium which in many cases showed fusion of the hyphae 
as described by Marshall Ward 2 . Stages in this process 
were observed later and will be described below. In three 
days the mycelium showed extensive vacuolation, and after 
thirteen days the protoplasm of the mycelium of a single 
spore-culture showed a tendency to aggregate in certain 
parts of the hyphae, leaving the rest empty. This proved 
to be the commencement of chlamydospore formation, stages 
in which were observed. In twenty-eight days in the drop 
in question the hyphae were noted to have knotted up 
together in two areas nearly in contact forming dense stro¬ 
mata. In thirty-four days the edges of the stromata were 
found to contain Melanconium- spores. The stromata were 
roughly elliptical in shape and measured -59 x *24 mm. and 
•3 x -24 mm. respectively. The Melanconium- spores obtained 
therefrom varied from 147 /x to 9-2 /x in length and measured 
37 /x in breadth. It was unfortunate that the early stages 
in the formation of these Melanconium- stromata were not 
observed. It was not until after many failures that a hanging- 
drop culture containing a single Melanconium- spore was 
1 The Ginger Beer Plant and the organisms composing it. Phil. Trans., vol. 
183 (1892), pp. 130-132. 
2 A Lily Disease. Ann. Bot., vol. ii, p. 329 (1888). 
