152 DopGE: METHODS OF CULTURE OF ASCOBOLACEAE 
Ascobolus Leveillei var. americanus Cooke Ascodesmis nigricans v. Bee (Bou- 
& Ellis diera Claussenti P. Hen 
Ascobolus pusillus Boud. Boudiera sp. 
Ascobolus viridis Currey var. (?) Cubomia sp 
Ascobolus viridulus Phil. & Plow. Thecotheus Pelletieri (Crouan) Boud. 
Ascobolus Wintert Rehm Rhyparobius crustaceus (Fuckel) Rehm 
Ascophanus Aurora (Crouan) Boud. Rhyparobius pachyascus Zukal 
Ascophanus carneus (Pers.) Boud. Rhyparobius niveus (Fuckel) Rehm 
Ascophanus yates Boud. Rhyparobius sexdecimsporus (Crouan) 
Ascophanus glaucellus Rehm Sacc. 
Ascophanus PC (Crouan) Lasiobolus equinus (Miill.) Karst. 
Bo 
Saccobolus depauperatus (B. & Br.) 
Asrophowns Holmskjoldit Hansen Rehm 
Ascophanus lacteus (Cooke & Phil.) Phil. Saccobolus Kerverni (Crouan) Boud. 
) 
Ascophanus corater onto (B. r.) Phil. Saccobolus neglectus Boud. 
Ascophanus minutiss Ss Boud. Saccobolus violaceus Boud 
Ascophanus ochraceus dein Boud. Thelebolus stercoreus Tode 
Of this list fourteen species have been grown more extensively 
and studied from the morphological standpoint as to the characters 
of their ascogonia, methods and conditions of spore germination, 
etc. Further studies are in progress, dealing with nuclear phe- 
nomena and methods of reproduction. The discussion of the 
identification of these species forms a part of another paper, 
which is now being prepared. 
I have found aceto-carmin useful for staining young ascogonia, 
although it is not permanent, and as a temporary stain it can not 
always be depended upon to differentiate between the nuclei and 
the granules of the cells. The nuclei are more easily differen- 
tiated in the young hyphae. A drop of the aceto-carmin mixture 
placed on a piece of agar containing the fungus before it is crushed 
under the cover glass, will stain the hyphae and ascogonia, if the 
slide is left for some time in a damp chamber where the stain does 
not dry out. The ascogonia, especially, swell under the action 
of the stain but this is no disadvantage when it is desired only 
to locate them. 
Material stained in toto with iron-hematoxylin can be crushed 
on the slide, perfectly dehydrated, and mounted in balsam without 
shrinkage. Such mounts are useful for showing germinated spores 
and the young mycelium. A still better method is to allow the 
spores to be shot upon a slide where they will stick securely enough 
to be carried through all the processes of heating, germination, 
fixation, etc. 
