Harmful and parasitic unicellular eukaryotes persist in a shallow lake under
reconstruction (L. Karla, Greece)
Eleni Nikouli1, Konstantinos Ar. Kormas1*, Panagiotis Berillis1, Hera Karayanni2, Maria
Moustaka-Gouni3
1
Department of Ichthyology and Aquatic Environment, School of Agricultural Sciences,
University of Thessaly, 384 46 Volos, Greece
2
Department of Biological Applications and Technology, University of Ioannina, 451 10
Ioannina, Greece
3
Department of Botany, School of Biology, Aristotle University of Thessaloniki, 541 24
Thessaloniki, Greece
*
Corresponding author: Tel.: +30-242-109-3082, Fax: +30-242-109-3157, Email:
kkormas@uth.gr
Supplementary material
Submitted to “Hydrobiologia”
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Figure S1. 18S rRNA gene clone library coverage based on Good’s C estimator of the
unicellular eukaryotes Lake Karla, Greece. O/P = ratio of observed-to-predicted number of
phylotypes based on SChao1.
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Figure S1a. The phylogenetic tree of the Fungi 18S rDNA relationships (ca. 1600 bp) of the
representative unique for each sample (grouped on ≥98% similarity) phylotypes found in
the Lake Karla water column in May, August and November 2010. The tree is based on the
neighbour-joining method as determined by distance Jukes-Cantor analysis. One thousand
bootstrap analyses (distance) were conducted. Scale bar represents 2% estimated distance.
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Figure S1b. The phylogenetic tree of the rest of the Fungi 18S rDNA relationships (ca. 1600
bp) of the representative unique for each sample (grouped on ≥98% similarity) phylotypes
found in the Lake Karla water column in May, August and November 2010. The tree is based
on the neighbour-joining method as determined by distance Jukes-Cantor analysis. One
thousand bootstrap analyses (distance) were conducted. Scale bar represents 2% estimated
distance.
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Figure S2. The phylogenetic tree of the Mesomycetazoa 18S rDNA relationships (ca. 1600
bp) of the representative unique for each sample (grouped on ≥98% similarity) phylotypes
found in the Lake Karla water column in May, August and November 2010. The tree is based
on the neighbour-joining method as determined by distance Jukes-Cantor analysis. One
thousand bootstrap analyses (distance) were conducted. Scale bar represents 2% estimated
distance.
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Figure S3. The phylogenetic tree of the Copepoda 18S rDNA relationships (ca. 1600 bp) of
the representative unique for each sample (grouped on ≥98% similarity) phylotypes found
in the Lake Karla water column in May, August and November 2010. The tree is based on
the neighbour-joining method as determined by distance Jukes-Cantor analysis. One
thousand bootstrap analyses (distance) were conducted. Scale bar represents 2% estimated
distance.
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Figure S4. The phylogenetic tree of the Cryptophyta 18S rDNA relationships (ca. 1600 bp)
of the representative unique for each sample (grouped on ≥98% similarity) phylotypes
found in the Lake Karla water column in May, August and November 2010. The tree is based
on the neighbour-joining method as determined by distance Jukes-Cantor analysis. One
thousand bootstrap analyses (distance) were conducted. Scale bar represents 2% estimated
distance.
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Figure S5. The phylogenetic tree of the Chlorophyta 18S rDNA relationships (ca. 1600 bp)
of the representative unique for each sample (grouped on ≥98% similarity) phylotypes
found in the Lake Karla water column in May, August and November 2010. The tree is based
on the neighbour-joining method as determined by distance Jukes-Cantor analysis. One
thousand bootstrap analyses (distance) were conducted. Scale bar represents 2% estimated
distance.
8
Figure S6. The phylogenetic tree of the Cercozoa 18S rDNA relationships (ca. 1600 bp) of
the representative unique for each sample (grouped on ≥98% similarity) phylotypes found
in the Lake Karla water column in May, August and November 2010. The tree is based on
the neighbour-joining method as determined by distance Jukes-Cantor analysis. One
thousand bootstrap analyses (distance) were conducted. Scale bar represents 2% estimated
distance.
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Figure S7. The phylogenetic tree of the Stramenopiles 18S rDNA relationships (ca. 1600
bp) of the representative unique for each sample (grouped on ≥98% similarity) phylotypes
found in the Lake Karla water column in May, August and November 2010. The tree is based
on the neighbour-joining method as determined by distance Jukes-Cantor analysis. One
thousand bootstrap analyses (distance) were conducted. Scale bar represents 2% estimated
distance.
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Figure S8. The phylogenetic tree of the Alveolata 18S rDNA relationships (ca. 1600 bp) of
the representative unique for each sample (grouped on ≥98% similarity) phylotypes found
in the Lake Karla water column in May, August and November 2010. The tree is based on
the neighbour-joining method as determined by distance Jukes-Cantor analysis. One
thousand bootstrap analyses (distance) were conducted. Scale bar represents 2% estimated
distance.
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Scanning Electron Microscopy (SEM) protocol
Water samples were fixed with glutaraldehyde (final concentration 2.5%) overnight at 4°C
and then filtered on an isopore polycarbonate membrane filter (0.2μm). The filters
were dehydrated with a graded alcohol series (50%, 70%, 80%, 90%, 95%, 100%) and then
were air-dried and covered with a thin layer of gold using a sputter coater (Bal-tec SCD
004), before their examination under a scanning electron microscope (Cambridge
Stereoscan 240).
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Figures S9. Scanning electron microscopy photographs of unicellular eukaryotes identified
in the Lake Karla water column during the first year of the lake’s water refilling.
a, b: Euglena sp.; c: Peridiniopsis sp.; d Pfiesteria sp. (arrow); e: Cyclotella sp.; f: Tetraedron
minimum; g: Coleps sp.
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Figure S10. DAPI prokaryotic cell counts in Lake Karla, Greece.
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