Brier ley. — Spore Germination in Onygena equina , Willd. 129 
in water, but here no further growth was obtained. Hanging drops of cow- 
dung extract, 1 glue, gelatine, and water were then prepared, fresh mature 
spores were sown, and the cells were left at room temperature and in the 
incubator at 23° C. These cells were carefully preserved and when necessary 
the medium was renewed. From September 18 until February 2 no change 
was observed, but on the latter date the contents of many of the spores in 
the dung extract in the incubator appeared to become slightly clearer. On 
February 8 germination was noted, and two days later this was fairly 
abundant, about 40 per cent, of the spores putting out germ tubes. In the 
glue germination occurred on February 11, in the gelatine on February 19, 
whilst no germination occurred in the hanging drops of water. The spores 
which had been kept at room temperature germinated about the same time, 
but more irregularly. In this case also no germination occurred in water. 
The minimum resting period under these conditions was therefore 143 days. 
On February 2, 191 5, sowings were again made of spores from the original 
specimens in the tin box, which had thus been in a state of desiccation for 
twelve months. On February 14 germination occurred in cow-dung extract, 
on February 16 in glue, and the following day in hanging drops of gelatine. 
No germination occurred in water. In the nutrient media only about 20 to 
25 per cent, of the spores germinated. 
On August 15, 1915, hanging drops containing spores from the original 
material, after eighteen months’ desiccation, were again prepared, but no 
germination could be obtained. 
The ripe ascospores are brown in colour with a slightly thickened wall, 
and if their formation be carefully traced it will be noted that this thickening 
and coloration occur subsequent to the attainment of their full dimensions. 
In the unripe condition they are clear and hyaline, and usually lying free 
within the peridium. By therefore carefully splitting away the peripheral 
chlamydospore crust of a nearly mature fructification and breaking open the 
latter, almost pure sowings of the full-grown but unripe spores may be 
obtained. On March 5, 1914, such spores were placed in hanging drops of 
cow-dung extract, glue, gelatine, and water, and the cells left at room tem- 
perature, 23° C. and 37 0 C. No germination occurred at the latter temperature, 
but on March 8 vigorous growths began from about 70 per cent, of the 
spores at 23 0 C. and on March 9 from those at room temperature. In water 
many of the germ tubes were abnormal, and growth ceased after about the 
third day. These spores had received no preliminary treatment of any kind. 
The observations so far described would appear therefore to show : — 
(a) That the resistance which the mature ascospores offer to direct and 
immediate germination is correlated with changes in the thickening and 
coloration of the spore wall. 
1 50 grm. of air-dried fresh cow-dting well shaken in 1,000 c.c. tap- water, filtered and heated 
in Koch steamer on three successive days. 
K 
