s - Mycological Society of America
s - Mycological Society of America
s - Mycological Society of America
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G.W. Hesseltine. 5407 N. Isabell, Peoria, IL<br />
61614<br />
New food ,fermentations: ragi and related<br />
products.<br />
In the Orient there are several different<br />
classes <strong>of</strong> food fermentations. One class<br />
including ragi, bubod, murcha, Chinese yeast,<br />
and look-pang are based upon solid dry inoculum<br />
used to start a variety 03 beverages and<br />
food fermentations. They are mixed cultures<br />
containing bacteria, yeast and fungi. A<br />
study <strong>of</strong> 114 strains <strong>of</strong> Mucor from 45 <strong>of</strong><br />
these starters showed that they belong to<br />
Mucor circinelloides (82) and g. indicus (32)<br />
with 6 not identified. Identification was<br />
based, in part, upon sexual reactions with<br />
known tester strains. None <strong>of</strong> the 56 Mucor<br />
strains tested utilized starch. fifty-three<br />
isolates from the amylase starters from 7<br />
regions showed that picros~orus (B.<br />
chinensis) and B. orvzae were the species<br />
present, and all <strong>of</strong> them caused hydrolysis <strong>of</strong><br />
starch. A third group <strong>of</strong> RhiZoDuS strains<br />
(7) were intermediate between B. orvzae and<br />
- R. micros~orus. Fourteen strains <strong>of</strong> RhiZoDuS<br />
tested for growth under anaerobic conditions<br />
grew bud did not sporulate. The third mold<br />
present was Amvlomvces rouxii, found in 7<br />
countries; 70 strains were isolated: and<br />
these produced amylases. When various combinations<br />
<strong>of</strong> microorganisms, isolated from<br />
amylase starters, were tested as mixed-culture<br />
fermentations, the only combinations<br />
that produced a good lao-chao product were<br />
the combinations <strong>of</strong> Saccharomvces fibuliaera<br />
with Mucor circinelloides, M. indicus, Rhizo~us<br />
micros~orus and Amvlomvces rouxii.<br />
is more closely related to the Polyporaceae<br />
than to the Tricholocataceae. Morphological<br />
characters (dimitic hyphae and hyphal pegs)<br />
are consistent with this hypothesis. The<br />
results also suggest that neither Lentinus or<br />
the Polyporaceae is monophyletic.<br />
T.*,<br />
H. C. HOCH, and B. TERHUNE. '~ept. <strong>of</strong> Biology,<br />
Rhodes College, Memphis, TN 38112, and Dept. <strong>of</strong> Plant<br />
Pathology, N. Y. State Agric. Exp. Sta., Cornen Univ., Geneva NY<br />
14456. lmmunolocalization <strong>of</strong> Coiled Cytoplasmic Filaments in<br />
Uromyces appendiculatus uredospore germlings.<br />
Uredospore germlings <strong>of</strong> Uromyces appendiculatus were homo-<br />
genized with glass beads, and a microparticulate fraction was<br />
produced by differential ultracentrifugation. The material was<br />
further fractionated on a column <strong>of</strong> Sephacryl S-1000, and a<br />
peak containing mainly flexuous coiled filaments was obtained.<br />
Their diameter was about 13 nm, and their typical maximum<br />
length was 500 - 540 nm. These appear similar to the "WFR"<br />
class <strong>of</strong> "virus-like particle" described by McDonald and Heath<br />
(1978. Can. J. Bot. 56:963-975). The major polypeptide <strong>of</strong><br />
the filament fraction has a molecular weight <strong>of</strong> about 36 kD, as<br />
measured by SDS-PAGE, with weaker bands between 29 and 34<br />
kD. Monoclonal antibodies were produced that were shown by<br />
immunoblotting to recognize an antigen that co-migrates with a<br />
34 kD protein <strong>of</strong> the filament fraction. With indirect immuno-<br />
fluorescence microscopy, the antibody labels fusiform struc-<br />
tures distributed throughout the cytoplasm. With EM immuno-<br />
gold labeling, the antibody binds to bundles <strong>of</strong> cytoplasmic<br />
filaments most closely resembling structures previously identi-<br />
fied as "filamentous distalsomes" (Hoch and Staples. 1983.<br />
Mycologia 75:795-824).<br />
David S. Hibbett and Rytas Vilgalys.<br />
Department <strong>of</strong> Botany, Duke University,<br />
~ukham, NC 27706. -<br />
Taxonomic relationships <strong>of</strong> Lentinus to the W. E. HIhTZ, J. C. ROYER, M. HUBBES and P. A. HORGEN.<br />
Polyporaceae: evidence from restriction<br />
Center for Plant Biotechnology, University <strong>of</strong> Toronto.<br />
analysis <strong>of</strong> enzymatically amplified<br />
Transformation-mediated insemonal mutagenesis <strong>of</strong> the dimorphic<br />
DNA. pathogen Ophoswma ulmi.<br />
Evolutionary relationships <strong>of</strong> Jentinus Fr. to<br />
the Polyporaceae were elucidated using<br />
restriction analysis <strong>of</strong> ribosomal DNA (rDNA).<br />
Five species in Mntinus sensu Pegler, three<br />
species in the Polyporaceae and two species<br />
in the Tricholomataceae were examined. RDNA<br />
phenotypes were determined by restriction<br />
analysis <strong>of</strong> enzymatically amplified rDNA (PCR<br />
Fingerprinting). This method generates<br />
Restriction Fragment Length Polymorphism<br />
(RFLP) data without Southern hybridization or<br />
autoradiography. With five different four-<br />
base restriction enzymes, one hundred unique<br />
restriction fragments were resolved. Among<br />
the 16 individuals examined, there were<br />
twelve unique rDNA phenotypes. A similarity<br />
matrix based on presence or absence <strong>of</strong><br />
comigrating restriction fragments was<br />
analyzed by clustering (UPGMA) and distance<br />
algorithms (Fitch-Margoliash analysis).<br />
Results <strong>of</strong> all analyses were highly<br />
consistent and strongly suggest that Lentinus<br />
Rotoplasts <strong>of</strong> Ophiostoma ulmi, the causal agent <strong>of</strong> Dutch Elm<br />
disease, were aansfmed to hygromycin resistance using a DNA<br />
vector (vPS57) containing a promoter for i~openicillin N synthetase<br />
from ~enicillibn chryso@& fused to a bac'terial gene f&<br />
hygromycin phosphotransferase. The transformation efficiency<br />
was 5 X 103 transformants per ug DNA per 10' protoplasts. The<br />
transforming DNA was stably integrated (mitotically) in single or<br />
tandem copies at unique or several &spersed sites within the<br />
transformant genornes. There appeared to be no targetting <strong>of</strong><br />
specific integration sites within the genome for the transforming<br />
DNA. This provided a useful approach for generating mutations by<br />
the insemonal disruption <strong>of</strong> functional genes. The crippled genes<br />
were conveniently tagged by the transforming DNA and could be<br />
recovered from genomic libraries. We have generated a<br />
developmental mutant (U9) in which the dimorphic phenotype has<br />
been altered. Unlike the untransformed parental strain the U9<br />
mutant fails to switch from yeast-like growth to mycelial pwth when transferred to selective medium. The transforming DNA<br />
integrated at a single locus and we hypothesize that the<br />
transforming DNA interupted one <strong>of</strong> the genes or regulatory<br />
regions essential to phenotype transition. We are currently<br />
analysing the site <strong>of</strong> inteption.