phalisshould be isolated and grown below
20 C (18C works well). Members of Kickxel-
lales and some members of the genusMortier-
ella (Mortierellales) can be hosts for
zygomycete mycoparasites, especially species
of Piptocephalis and Syncephalis (Cuthbert
and Jeffries 1984 ; Jeffries and Young 1978 ).
Benjamin (1985b) discussed and illustrated
the ontogeny of both the anamorph and tele-
omorph of Syncephalis hypogenaR.K. Benj.
Ultrastructural studies on merosporangiospore
ontogeny inPiptocephalis indicaB.S. Mehrotra
& Baijal and Syncephalis sphaerica Tiegh.
(Baker 1979 ; Baker et al. 1977 ) indicate that
they are similar (invagination of plasmalemma)
but differ from that ofSyncephalastrum race-
mosumCohn ex J. Schro ̈t. (cytoplasmic cleav-
age) (Fletcher 1972 ).
Syncephalisoften grows best on the original
host, but some taxa may not survive more than
one or two transfers. Many species ofSyncepha-
lis, however, can be transferred to another host,
such asZygorhynchusorCokeromyces. Other
taxa ofSyncephalisare restricted to a species
ofMortierella. Some members of Zoopagales
can be found in an aquatic environment includ-
ingZoophagusand some species of the Cochlo-
nemataceae.
AenigmatomycesR.F. Castan ̃eda & W.B.
Kendr. is an ascomycete according to Myco-
bank (http://www.mycobank.org/) and Species
Fungorum (http://www.speciesfungorum.org/).
Aenigmatomyces, however, is probably a mem-
ber of Zoopagomycotina (Cochlonemataceae,
Zoopagales) according to Degawa (2002a) and
Seifert et al. ( 2011 ).Aenigmatomyces zygos-
pores were illustrated by Degawa (2002b).Basi-
diolum fimbriatum Cienk. (White 2003 ), a
possible member of Zoopagales, resembles a
small, aquaticSyncephalis.
LecophagusandZoophagus(Dick 1990 )were
both treated as members of Zoopagales. Recent
morphological (light microscopy and transmis-
sion electron microscopy) and molecular (small
subunit sequences) studies demonstrate that
members ofLecophagusare ascomycete ana-
morphs (Morikawa et al. 1993 ; Tanabe et al.
1999 ).Zoophaguswas retained in the zygomy-
cota based on the comparative analysis of 18S
rDNA sequences (Tanabe et al. 1999 ).
VII. Maintenance and Culture
Accounts of the collection and incubation of
substrates and the isolation and cultivation of
most zygomycetes are discussed in detail by
Benjamin ( 1959 ), O’Donnell ( 1979 ), Krug et al.
( 2004 ), and Benny ( 2008 ). Similar information
can be found for Entomophthoromycota (Ento-
mophthorales) (Fuller 1978 ). Ellis ( 1963 , 1966 )
described methods for the culture ofRhopalo-
myces elegansCorda and species ofSyncephalis
without a host.
Tansey ( 1984 ) recommended incubating
samples at 45C on media emended with anti-
biotics and benomyl when isolating thermo-
philic and thermotolerant Mucorales. Isolation
of both psychrophilic and psychrotolerant
zygomycetes is best done on media incubated
at 5C (Botha et al. 1999 ). The addition of
antibiotics and benomyl to the media might
inhance isolation, but it could eliminate some
Mortierellaspecies (Strauss et al. 2000 ).
Barron ( 2004 ) described techniques useful
for studying nematode parasites. Benjamin
( 1959 ,1985b), Jeffries and Kirk ( 1976 ), and
Benny ( 2008 ) include several techniques that
are valuable for making microscope slide pre-
parations and for isolating, growing, and cul-
turing zygomycetous fungi.
Fungi in Mucorales, Dimargaritales, Kick-
xellales, some Zoopagales (Helicocephalida-
ceae, Piptocephalidaceae), and many species
of Entomophthorales can be grown on ordinary
laboratory culture media (Benny 2008 ; Humber
2008 ). Potentially, these fungi may be isolated
from any organic substrate (e.g., dung, humus,
leaf mold, bark).The addition of the antibiotics streptomycin sulfate,
0.1 g–1l (filter sterilized), and chlortetracycline HCl,
0.05 g–1l (in 95 % ethanol), and the fungicide benomyl,
10–20 ppm (aqueous), to the isolation medium
increases the chances of isolatingCoemansiaspp. and
the recovery of other zygomycotan taxa from soil; see
Strauss et al. ( 2000 ) for other formulations.
The majority of members of Mucorales and Kick-
xellales grow best at pH 6 and require an exogenous
thiamine source.Pilobolusis an obligate coprophile but
will grow and sporulate only on slightly alkaline culture
media containing hemin or dung extract (Levetin and
Caroselli 1976 ).Zygomycetous Fungi: Phylum Entomophthoromycota and Subphyla Kickxellomycotina,... 237