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Communities of
ectomycorrhizal fungi in a Pb
contaminated pine forest site
Nan Hui, Rauni Strömmer, Teija Koivula,
Martin Romantschuk,
Dept. of Ecological and Environmental Sciences, University of
Helsinki, Niemenkatu 73, FI-15140 Lahti, Finland
Introduction
 Hälvälä an old shotgun shooting range located at a pine
forest site, located in southern Finland, is used as the
study area.
Study areas
Contamination Level
mg/kg
Clean
Polluted
Mycorrhiza
 A mycorrhiza is a symbiotic association between a fungus
and the roots of a plant.
root
Types of Mycorrhizas
 The two most common types
 ectomycorrhizas
Basidiomycota and Ascomycota
 endomycorrhizas (Arbuscular mycorrhizas)
associated with the roots of herbaceous plants
other mycorrhizas
Pine root tips and Ectomycorrhizal fungi
1mm
The main point of the study
 Find the Ectomycorrhizal fungi species
difference between Clean and Lead Polluted
area.
 Find the species which are particularly Lead
tolerable.
 Find the species which are Lead sensitive.
Method and material
extraction
Amplifying certain area
of Fungal Ribosomal
DNA by PCR
Aligning the
sequence with the
genebank database
sequence the PCR
product
Total DNA
DNA Extraction from pine root tips
 Root tips isolation
 FastDNA SPIN Kit for soil
modified the original protocol
PCR amplification
 Fungal ribosomal RNA gene
 Primer selection
Fun18f
ITS1
18S
ITS2
5.8S
28S
ITS4
Sequencing and Bioinformation
 In Biocenter Viikki, Helsinki
 Staden Package
 EMBL
A Nucleotide Sequence Database
http://www.ebi.ac.uk/embl/
 UNITE
A molecular database for the identification of
ectomycorrhizal fungi
http://unite.ut.ee/
Result
 Altogether 351 sequences were distributed into 37
different groups. These groups were aligned to EMBL-EBI
database and UNITE database.
The phylogenetic trees are made by
neighborjoining method (PHYLIP).
Dermocystidium salmonis is used as an
outgroup.
The tree colored according to the distribution of
species in the three environmental conditions.
Clean: gray; Medium: magenta; Polluted(III): red;
Both in Medium and polluted:Yellow
The uncolored four species could be found in all the
three environmental conditions.
Fig.1 Species tree in the clean area
Frequency of the species in
different study areas
Clean
Medium
Polluted
Lactarius rufus
Piloderma fallax
Common species in all areas
Piloderma reticulatum
Piloderma croceum
Tylospora asterophora
Tylospora fibrillosa
Species found only in clean area
Russula decolorans
Mortierella verticillata*
Russula betularum
Tomentella stuposa
Suillus luteus
Cortinarius armeniacus
Species found only in polluted area
Nolanea conferenda
Thelephora terrestris
Amanita spissa
Conclusion
 Clear difference is found between clean and polluted
area.
 In clean area, all the species are recognizable by
genebank database.
 In polluted area, some unknown novel species are found
and given a number.
Thanks for your attention