casiroz - 1 st meeting: ectomycorrhizal studies in wp1 & wp2 hojka kraigher slovenian forestry...
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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
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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
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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
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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
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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.)
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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
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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)
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•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
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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
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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
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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.
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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
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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
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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
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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
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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: