Book of Abstracts (PDF) - International Mycological Association

IMC7 Main Congress Theme V: CELL BIOLOGY AND PHYSIOLOGY Posters
Chitosan is a biopolymer with unique properties favorable
for a broad variety of industrial, biomedical, and
agricultural application. The chitosan is produced from
crab shell through the chemical procedure which leaves a
lot of industrial waste at present. Chitin is converted to
chitosan by chitin deacetylase. To develop an enzymatic
process for chitosan production that reduces the waste, we
have planned to isolate a clone of chitin deacetylase gene
from Phycomyces blakesleeanus cDNA library which we
made in the _ZAP vector. The 218bp-DNA fragment
amplified from genomic DNA of P. blakesleeanus using
PCR (polymerase chain reaction) primers to two highly
conserved sequences within chitin deacetylase gene of
Mucor rouxii was the probe for isolation the clone from the
library. A positive cDNA clone which is composed of
1,613bp was isolated. The deduced protein encoded by this
clone is composed of 459 amino acids. The protein
sequence comparison revealed that the protein is
significantly similar to chitin deacetylase of M. rouxii.
After ligation of the clone to the expression vector pET32
Ek/LIC, chitin deacetylase activity was detected in the
purified His-tagged protein fraction of the crude extract
from a E. coli transformant. These results show that the
chitin deacetylase gene has been successfully cloned from
P. blakesleeanus. The cloned gene was named PbCD and
the nucleotide sequence of PbCD was registered to
GenBank (accession #AB046690).
1154 - Genome analyses of Micromucor ramanniana and
Micromucor isabellina
Á. Nagy 1* , M. Pesti 1 & C. Vágvölgyi 2
1
University of Pécs, Department of General and
Environmental Microbiology, Pécs, Ifjúság u. 6, H-7624,
Hungary. -
2
University of Szeged, Department of
Microbiology, Szeged, P.O. Box 533, H-6701, Hungary. -
E-mail: nagi@freemail.hu
The Micromucor species (genus Mortierella) belong
among the most common saprophytic soil fungi. Some
members of the genus (Mortierella species) are potent
producers of the long-chain polyunsaturated fatty acids
(e.g. arachidonic acid). These compounds are important
both nutritionally and pharmacologically. These fungi are
biotechnologically important, but the organisation of their
nuclear genome has not been described so far.
Electrophoretic karyotype analysis using different
approaches of pulsed field gel electrophoresis (PFGE) has
led to significant progress in fungal genetics: the physical
karyotypes of numerous previously genetically
uncharacterised fungal species have been established. In
the present study, orthogonal field alternation gel
electrophoresis (OFAGE) and the contour clamped
homogeneous electric field (CHEF) technique were applied
to obtain preliminary information on the organisation and
intrageneric variability of the nuclear genome in three
Mortierella (Micromucor) strains of 2 different species (M.
isabellina and M. ramanniana). Conditions for the
preparation of highly-intact chromosome-size DNA
molecules and for the separation of DNA molecules were
established. Furthermore, through the use of homologous
and heterologous gene probes (e.g. 3-hydroxy-3-
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Book of Abstracts
methylglutatyl coenzyme A reductase) gene assignment
experiments have been performed. This research was
supported financially by the Hungarian Scientific Research
Fund (OTKA) D29113 and T 032738.
1155 - Is the interfacial pH a measure of the efficiency
of nutrient transfer in ectomycorrhizas?
A. Nenninger 1* , G.W. Griffith 1 & W. Heyser 2
1 Institute of Biological Sciences, University of Wales
Aberystwyth, Penglais, Aberystwyth, Ceredigion SY23
3DA, Wales, U.K. - 2 Centre of Environmental Research
and Technology, University of Bremen, D-28359 Bremen,
Germany. - E-mail: ann@aber.ac.uk
Interfacial pH-conditions in Pinus nigra ectomycorrhizas
were determined using the pH-sensitive ratio-dye Cl-NERF
and a confocal laser scanning microscope (CLSM). A
pseudo-ratiometric method is commonly used for
determining apoplastic pH-values. In the present study, a
non-invasive ratiometric method relying on a single
fluorescence dye was devised and tested. Results show
homogeneous pH-values near 3 in non-mycorrhizal roots
from different plants and heterogeneous interfacial pHconditions
between 3.1 and 4 in various pine
ectomycorrhizas. In mycorrhizas, reduced sucrose and
phosphate leads to decreasing interfacial pH-conditions,
whereas external applied glucose induced an increase of
the interhyphal pH-values in the mantle. Based on the
results we developed a model for the interfacial transfer
processes in ectomycorrhizas. Transfer processes on the
ectomycorrhizal interface are not pH-regulated. The
processes generate an interfacial pH-value to maintain the
symbiotic balance. In balance you find optimal pHconditions
for the nutrient transfer. The lack of only one of
the transfer goods induces a decrease of the interfacial pHconditions.
A decreasing pH-value in the interface of
ectomycorrhizae leads to reduce transfer processes and the
mutualistic balance in the symbiosis can be maintained.
1156 - Effect of ferulic acid on guaiacylglycerol-βguaiacyl
ether degradation by extracellular enzymes of
Trametes versicolor
G. Nowak, A. Matuszewska * & M. Nowak
Departament of Biochemistry M. Curie-Sklodowska
University, Pl. M. Curie-Sklodowskiej 3, 20-031 Lublin,
Poland. - E-mail: amatusze@hermes.umcs.lublin.pl
Trametes versicolor, the effective lignin decomposer,
produces a set of enzymes acting on bonds in lignin
polymer, especially on most abundant in lignin ether β-O-4
bond. In our studies the most active enzymes are secreted
into the medium in idiophasic cultures of Trametes as a
result of carbon starvation. During experiments destaining
of Remazol Brilliant Blue and/or carminic acid were used
as an indirect method and degradation of guaiacylglycerol-