LEVINE: CyTOLOGY OF HYMENOMYCETES 145 
MATERIALS AND METHODS 
Spores of Pholiota praecox and numerous Boleti were collected 
by placing the pilei on circular sheets of filter paper and covering 
the whole with a bell jar. The spore prints were kept in sterilized 
Petri dishes. Miss Ferguson’s (1902) method of collecting spores 
was also used. The Petri dishes with the spore prints were kept 
in an ordinary refrigerator until needed. A great number of cul- 
ture media were tried; among those which gave the best results 
were string beans, and fresh horse manure prepared in a manner 
similar to that described by Duggar (1901). Cherry agar was 
prepared by boiling 250 gms. of fresh cherries in a liter of water. 
The cherries on becoming soft were strained through a cheese 
cloth and the skins, pits, and stalks were discarded. The juice of 
the cherries was again boiled with 15-20 gms. of agar agar and the 
decoction was then filtered and sterilized in an autoclave. Malt- 
beef agar was made by adding 25 gms. of Loeflund’s malt and 
25 gms. of Liebig’s beef extract toa liter of water. This was then 
mixed with 15-20 gms. of dissolved agar agar and boiled and 
neutralized with n/1o00 NaOH. The solution was then filtered 
and sterilized. 
The spores germinated best in the malt-beef medium. In the 
case of Pholiota praecox 65 per cent of the spores germinated in 
this medium. Cherry and malt-beef agars were used for the 
propagation of the mycelia of several Polypores. The cultures 
were kept in dark boxes at temperatures varying from 23°—30° C. 
The early stages of spore germination from cultures in Van 
Tieghem’s cells were killed and fixed to slides by a modification of 
Harper’s (1899) and Lutman’s (1910) methods. The cover glass 
on which the culture is grown is removed from the ring and a few 
drops of the fixing solution are added. The cover glass with the 
culture is then inverted over slides covered with a film of albumen 
fixative and gently tapped till part of the drop falls on it. The 
spores in the drop become fixed to the slide by the coagulation of 
the albumen. The slide is then set aside till the liquid partly 
evaporates. The preparation is hardened by pouring graded 
alcohol over it, and it is then ready for staining. More fixing 
solution may be added to the culture drop and the process repeated 
till all the spores have been removed. A minimum of disturbance 
of the delicate germ tubes is achieved by this method. 
