1. Nan Hui, Rauni Strömmer, Teija Koivula, Martin Romantschuk,
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 Lahti, Finland

2. Introduction
Hälvälä an old shotgun shooting range located at a pine forest site, located in southern Finland, is used as the study area.

3. Study areas

4. Contamination Level
mg/kg

5. Clean Polluted

6. Mycorrhiza
A mycorrhiza is a symbiotic association between a fungus and the roots of a plant.
root

7. Types of Mycorrhizas The two most common types ectomycorrhizas
Basidiomycota and Ascomycota
endomycorrhizas (Arbuscular mycorrhizas)
associated with the roots of herbaceous plants
other mycorrhizas

8. Pine root tips and Ectomycorrhizal fungi
1mm

9. 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.

10. Method and material Amplifying certain area Total DNA
extraction
Amplifying certain area
of Fungal Ribosomal DNA by PCR
Aligning the sequence with the genebank database
sequence the PCR product

11. DNA Extraction from pine root tips
Root tips isolation
FastDNA SPIN Kit for soil
modified the original protocol

12. PCR amplification Fungal ribosomal RNA gene Primer selection ITS1 ITS2
Fun18f
ITS1
ITS2
18S
5.8S
28S
ITS4

13. Sequencing and Bioinformation
In Biocenter Viikki, Helsinki
Staden Package
EMBL
A Nucleotide Sequence Database
UNITE
A molecular database for the identification of ectomycorrhizal fungi

14. Result
Altogether 351 sequences were distributed into 37 different groups. These groups were aligned to EMBL-EBI database and UNITE database.

15. The phylogenetic trees are made by neighborjoining method (PHYLIP)
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.

16. Fig.1 Species tree in the clean area

17. Frequency of the species in different study areas
Clean
Medium
Polluted

18. Common species in all areas
Lactarius rufus
Piloderma fallax
Piloderma reticulatum
Piloderma croceum
Species found only in clean area
Tylospora asterophora
Tylospora fibrillosa
Russula decolorans
Mortierella verticillata*
Russula betularum
Species found only in polluted area
Tomentella stuposa
Suillus luteus
Cortinarius armeniacus
Nolanea conferenda
Thelephora terrestris
Amanita spissa

19. 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.

20. Thanks for your attention