CASIROZ - 1st meeting: Ectomycorrhizal studies in WP1 & WP2

CASIROZ - 1st meeting: Ectomycorrhizal studies in WP1 & WP2

Hojka KRAIGHERSlovenian Forestry Institute, Ljubljana, Slovenia Slovenian Forestry Institute, Ljubljana, Slovenia

Nutrient inputs, immobilization & mineralization in a forest ecosystem & common mycorrhizal networks

Nutrient inputs, immobilization & mineralization in a forest ecosystem & common mycorrhizal networks

Rain (stemflow, throughfall)

Gaseous

loss gain

FrassLeaves, buds, flowers

Twigs, branches

Litter inputSpatial and temporal redistribution

CWD

Immobilization

Mineralization

Mineral weathering

Uptake

Modified after: DIGHTON & BODDY 1988

COMMON MYCORRHIZAL NETWORKS

Mycorrhizal mycelium affects field performance of forest trees

through:•capture & uptake of nutrients•protection against pathogens & toxic elements•extending feeder root longevity•spatial & temporal linkages between sinks & sources of nutrients - common mycorrhizal networks

& depends on: •functional compatibility of species & strain of the fungus & the plant•therefore identification of the fungal partner is important

The importance of the diversity & function of mycorrhizae:

The importance of the diversity & function of mycorrhizae:

Neto transfer of asimilates from a birch to a shaded Douglas fir (13C, 14C; by SIMARD & al., Nature, 1997).

Interactions between ectomycorrhizal fungi and wood decomposing fungi (32P; by LINDAHL & al, 1998)

Weathering of minerals (rock-eating fungi; by JONGMANS & al, Nature, 1997).

Wood-wide-web supporting the productivity & biodiversity (READ, 1998 & VAN DER HEIJDEN et al, Nature 1998).

Ectomycorrhizae alter quality & quantity of carbon allocated belowground (RYGIEWICZ & ANDERSEN, Nature 1994 and subsequent publications).

Cantharellus tubaeformis linking

spruce & nutrients in OH

Mycorrhizae alter quality & quantity of carbon allocated belowground

Mycorrhizae alter quality & quantity of carbon allocated belowground

In forest ecosystems 2/3 of C is contained in soils & peat deposits.

ECM reduces the overall retention of C in the symbiosis by increasing C in roots & below-ground respiration.

ECM shifts C to pools that are rapidly turned over (fine roots, hyphae, fungal respiration).

O3 can reduce belowground respiration

O3 can reduce C allocation to the mycorrhizal fungus.

Right: Lactarius theiogalus x Picea abies

(ANDERSON & RYGIEWICZ, 1994 & 1995 etc.)

Standardized sampling core (270 ml, 3,5 cm, 0-18 cm deep), cleaning, separation & storage.

Counting & image analyses of all root tips (non-mycorrhizal, old non-identifiable types, ECM morphotypes).

Anatomical & molecular identification or characterisation of the separated morphotypes, comparisons to the reference material, PCR-RFLP library or GenBank.

Presentation of lists, tables, pies & biodiversity indices. Check for physiological data on a single sp.(?) Statistics & modelling with environmental & physiol. data.

Identification & quantification of ECMIdentification & quantification of ECM

©M.BRUNDRETT, CSIRO 1999

Identification of types of ectomycorrhizae are done by:

Identification of types of ectomycorrhizae are done by:

anatomical characteristics (AGERER 1987-2000) molecular methods (PCR-ITS-RFLP & sequencing) (GARDES & BRUNS 1993,

as described in KRAIGHER & al. 1995; seq.: MARTĺN 2000)

•Several spp. disappear, others proliferate in polluted sites.•Occurrence of ECM types superior to fruitbody mapping, since:---ECM types are present throughout the year,---hypogeus fungi, corticiaceous fungi, ex Deuteromycotina - are difficult to observe, find or identify,---some ECM types have been found as restricted to polluted or unpolluted plots, while the identity of the fungus was not yet determined (Piceirrhiza terraphila & Piceirrhiza inflata).•Selective sensitive or unsensitive spp: Hydnum rufescens & Paxillus involutus.

STUDY 1: Mycobioindication by mapping of types of ECM

By KRAIGHER & al. 1996

STUDY 2: NATURAL SPRUCE REGENERATION, LIGHT, SOILS & MYCORRHIZAE

STUDY 2: NATURAL SPRUCE REGENERATION, LIGHT, SOILS & MYCORRHIZAE

ECM were differentiated into three groups, providing a tentative interpretation of their ecology: •Selected genera of ectomycorrhizae were correlated positively with:-- E horizon (podzolization) (Lactarius sp., Q-type), -- humus (Cenococcum geophilum) or -- fermentation layer (OLOF, Elaphomyces sp.),

•contributing several new physiological & ecological informations to the limited data on the selected genera known so far (CAIRNEY & CHAMBERS (Eds) 1999).

Correlations (SKUPAJ4.STA 56v*35c)

OLOF AH E H PH_CACL2 C_N DIR5

SM

CEN_GEO

TOM_SPP

TYL_SPP

ELA_SP

LAC_P

LAC_Q

STARO

VIT_MIK

SK

Factor 1

Fa

cto

r 2

DER_CIN

PIC_COR

PIN_EPI

PIN_STE

CEN_GEO

COR_SP

TOM_SP

TYL_SP

TT724

-1.2

-1

-0.8

-0.6

-0.4

-0.2

0

0.2

0.4

-1.2 -1 -0.8 -0.6 -0.4 -0.2 0 0.2 0.4

STUDY 3: BIODIVERSITYSTUDY 3: BIODIVERSITY

Research plot Pokljuka

Stand phase Adult (a) Young (a) Clear-cut (b)

H index (ECM) 2.23 1.52 1.48H index (vegetation) 1.08 1.71 1.17

Below-ground diversity can counterpart the poor plant community in a natural stand.

(After KRAIGHER 2000, based on data by TROŠT & al. 1999 (a), SIMONČIČ & al. 1998 (b), H index for ground vegetation

by URBANČIČ & KUTNAR 1998).

Share (%) of types of ectomycorrhizae (ECM) on Norway spruce and indices of biodiversity

Some general conclusionsSome general conclusions

fungal fruitbodies occurrence does not correspond to the abundance of ECM types (the active mycelia in the soils),

in almost 3/4 million root tips:-1% were non-mycorrhizal,-40-60% of types per plot were old unidentifiable types,-36 out of ca 70 types have been identified to species or group level on spruce & 25 out of 65 types on beech, -a big number needs comprehensive characterisation,

the contemporary characterisation methods are a combination of anatomical & molecular techniques,

correlations need to be done with physiological & env. Data.

CASIROZ - Ectomycorrhizal studies (CR 6)CASIROZ - Ectomycorrhizal studies (CR 6)

WP1: ECOPHYsiology & Modeling

Y1 9 Y2 14 Y3 13 TOTAL MM 36

SS 8PhD 22T 13

WP2: Biochemistry & MOLecular Biology

Y1 2 Y2 3 Y3 4 TOTAL MM 9

SS 5PhD 4T 0

CASIROZ - Ectomycorrhizal studies WP1: ECOPHYSIOLOGY & MODELING

Y1 - 3: •Mapping of fruitbodies & ECM on the whole plot•Identification & characterisation of types of ECM•Collection of herbarium material•Regular analyses, preparation of discussions & reports, presentations & inputs to WP2 & for other partnersY1: •Pilot study of roots & ECM of beech seedlings in containersY2:•ECM identification & quantification on the beech O3 & C plot

Y3:•ECM identification & quantification on the beech O3 & C plot•Study of roots & ECM of beech seedlings in containers

CASIROZ - Ectomycorrhizal studies WP2: BIOCHEMISTRY & MOLECULAR BIOLOGY

CASIROZ - Ectomycorrhizal studies WP2: BIOCHEMISTRY & MOLECULAR BIOLOGY

Y1 - 3: Preparation of PCR-ITS-RFLP database from fungal exiccates & ECM

from the whole plot Regular analyses, preparation of discussions & reports, presentations &

inputs to WP1 & for other partnersY1: Build-up of the molecular databases (RFLP & sequencies)Y2: Molecular identifications of ECM Preparation of quantitative PCR for selected sp. / spp. (optional)Y3: Study of ECM of beech seedlings in containers

Contribution to CASIROZ & interfaces with other participants & studies

Contribution to CASIROZ & interfaces with other participants & studies

New ECM characterisations & molecular databases Diversity of ECM & differences among plots &

containers / ozone impacts & sensitivities of whole mycorrhizal trees & seedllings

A contribution to studies of above & below-ground interactions / hormonal relationships, roots & ECM in beech seedlings

A contribution to simulations of responses of the entire stand to ozone impact

Anatomical & molecular identifications of ECM:

Tine Grebenc, SFI, Ljubljana

Prof. Dr. Reinhard Agerer & Dr.Stefan Raidl, LMU Munich, Germany

Helmut Blaschke & al, TUM, Germany

Samar Al Sayegh Petkovšek, MSc, ERICo Velenje

Technicians & students: Jana Janša, Irena Tavčar & Peter Železnik

CASIROZ (2002-2005) & other EU & bilateral projects:

NAT-MAN (2000-2003), BIODIBERIA (2002) &

bilateral project Molecular ecology of ectomycorrhizae intemperate forests

(with Dr. M.P.Martín, Real Jardín Botanico, Madrid, Spain

Co-financed by:

Ministry of Education, Science & Sport (MESS),

Ministry of Agriculture, Forestry & Food (MAFF),

Ministry of Environment: Bureau for Nature

Present collaborators: