130 Brier ley. — Spore Germination in Onygena equina , Willd. 
(b) That the ripe ascospores will germinate after a resting period in 
the absence of digestive treatment. 
(c) That digestive treatment with artificial gastric juice eliminates or 
greatly curtails this resting period. 
Influence of low temperature on ascospore germination . Following 
the successful lead given by Eriksson, 1 with spores of the Uredineae, 
Marshall Ward tried to bring about the germination of the ascospores of 
Onygena by first subjecting them to a temperature below zero. In only 
one case was any germination observed, and Marshall Ward 2 himself terms 
this case ‘ doubtful and suggests ‘ that the successful germination without 
gastric juice was really that of the chlamydospores In my own observa- 
tions this method was given a very thorough trial. Freezing mixtures, 
melting ice, and exposure out of doors to frost were used, and with the 
spores in situ in the fructifications, freshly removed as a loose powder, 
suspended in water, and air-dried from water suspensions upon glass slides. 
Not one case of germination was observed, and it would seem that the 
method of forcing the germination of spores by subjecting them to low 
temperatures is inapplicable to the spores of Onygena equina. 
Germination of the chlamydospores . The media in which Marshall 
Ward obtained the germination of chlamydospores in the absence of 
preliminary digestive treatment were hydrolized horn, and cow-dung and 
gelatine plus hydrolysed horn ; these being maintained at a temperature of 
22° to 23° C. In the first case the spores were fresh and germinated well. 
In the second the spores had been soaked in water for two hours at 35 0 C. 
before sowing and the germination was very poor. 
In the present investigation hanging drops containing spores were 
prepared on March 17. Germination occurred on March 23, about 60 per 
cent, of the spores in hydrolysed horn 3 putting out germ tubes and about 
50 per cent, of those in dung extract and gelatine plus hydrolysed horn (equal 
quantities of dung extract and hydrolysed horn plus 10 per cent, gelatine). 
In addition, hanging drops of Witte’s peptone, 4 glue, water, gelatine, and 
cow-dung extract had been prepared, and, with the exception of the spores 
in water, abundant germination (50 to 70 per cent.) occurred after five or 
six days. In water only a few germ tubes were protruded, and many of 
these were abnormal and gave no further growth. Chlamydospores treated 
with gastric juice before sowing possessed no advantage over those lacking 
1 Eriksson, J. : liber die Forderung der Pilzsporenkeimung durch. Kalte. Centralb. f. Bakt., 
Abt. 2, Bd. i, No. 15/16, pp. 557-65, 1895. 
2 Marshall Ward, loc. cit., pp. 277, 279. 
3 10 grm. of shavings from fresh ram’s horn were dissolved in 100 c.c. of boiling 20 % pure 
concentrated sulphuric acid. This was neutralized with barium carbonate, filtered, and sterilized in 
the autoclave. » 
4 100 c.c. distilled water, 1 grm. peptone, 1 grm. sodium chloride, heated and then filtered. 
Sterilized in Koch steamer on three successive days. 
