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

Hälvälä an old shotgun shooting range located at a pine

forest site, located in southern Finland, is used as the

study area.

Introduction

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

Total DNA

extraction

Amplifying certain area

of Fungal Ribosomal DNA by PCR

sequence the PCR product

Aligning the sequence with the

genebank database

DNA Extraction from pine root tips

Root tips isolation

FastDNA SPIN Kit for soil

modified the original protocol

PCR amplification

18S 5.8S 28SITS1 ITS2

Fun18f

ITS4

Fungal ribosomal RNA gene

Primer selection

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

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

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