TY  - JOUR
AB  - Like other plant-microbe symbioses, the establishment of orchid mycorrhiza (ORM) is likely to require specific communication and metabolic adjustments between the two partners. However, while modulation of plant and fungal metabolism has been investigated in fully established mycorrhizal tissues, the molecular changes occurring during the pre-symbiotic stages of the interaction remain largely unexplored in ORM. In this study, we investigated the pre-symbiotic responses of the ORM fungus Tulasnella sp. SV6 to plantlets of the orchid host Serapias vomeracea in a dual in vitro cultivation system. The fungal mycelium was harvested prior to physical contact with the orchid roots and the fungal transcriptome and metabolome were analyzed using RNA-seq and untargeted metabolomics approaches. The results revealed distinct transcriptomic and metabolomic remodelling of the ORM fungus in the presence of orchid plantlets, as compared to the free-living condition. The ORM fungus responds to the presence of the host plant with a significant up-regulation of genes associated with protein synthesis, amino acid and lipid biosynthesis, indicating increased metabolic activity. Metabolomic analysis supported the RNA-seq data, showing increased levels of amino acids and phospholipids, suggesting a remodelling of cell structure and signalling during the pre-symbiotic interaction. In addition, we identified an increase of transcripts of a small secreted protein that may play a role in early symbiotic signalling. Taken together, our results suggest that Tulasnella sp. SV6 may perceive information from orchid roots, leading to a readjustment of its transcriptomic and metabolomic profiles.
AU  - De Rose, S.*
AU  - Sillo, F.*
AU  - Ghirardo, A.
AU  - Schnitzler, J.-P.
AU  - Balestrini, R.*
AU  - Perotto, S.*
C1  - 73971
C2  - 57252
CY  - One New York Plaza, Suite 4600, New York, Ny, United States
TI  - Omics approaches to investigate pre-symbiotic responses of the mycorrhizal fungus Tulasnella sp. SV6 to the orchid host Serapias vomeracea.
JO  - Mycorrhiza
VL  - 35
IS  - 2
PB  - Springer
PY  - 2025
SN  - 0940-6360
ER  - 

TY  - JOUR
AB  - The ecological and biogeochemical relevance of hydrolytic enzymes associated with the fungal cell wall has been poorly studied in ectomycorrhizal (ECM) fungi. We used a modified sequential extraction procedure to investigate the activity of various hydrolytic enzymes (β-glucosidase, acid-phosphatase, leucine-aminopeptidase, chitinase, xylanase and glucuronidase) and their association with the cell wall of three ECM fungi (Rhizopogon roseolus, Paxillus involutus and Piloderma croceum). Fungi were grown on C-rich solid medium under three different P concentrations (3.7, 0.37 and 0.037 mM). The sequential extraction procedure classifies enzymes as: (a) cytosolic, (b) loosely bound, (c) hydrophobically bound, (d) ionically bound and (e) covalently bound. Results showed that for the same fungus absolute enzymatic activity was affected by P concentration, whilst enzymatic compartmentalization among the cytosol and the cell wall fractions was not. The association of enzymes with the cell wall was fungus- and enzyme-specific. Our data indicate also that enzymes best known for being either extracellular or cytosolic or both, do act in muro as well. The ecological implications of cell wall-bound enzymes and the potential applications and limitations of sequential extractions are further discussed.
AU  - Pérez-de-Mora, A.
AU  - Reuter, B.
AU  - Lucio, M.
AU  - Ahne, A.
AU  - Schloter, M.
AU  - Pritsch, K.
C1  - 10696
C2  - 30555
SP  - 185-197
TI  - Activity of native hydrolytic enzymes and their association with the cell wall of three ectomycorrhizal fungi.
JO  - Mycorrhiza
VL  - 23
IS  - 3
PB  - Springer
PY  - 2013
SN  - 0940-6360
ER  - 

TY  - JOUR
AB  - The aim of a joint effort by different research teams was to provide an improved procedure for enzyme activity profiling of field-sampled ectomycorrhizae, including recommendations on the best conditions and maximum duration for storage of ectomycorrhizal samples. A more simplified and efficient protocol compared to formerly published procedures was achieved by using manufactured 96-filter plates in combination with a vacuum manifold and by optimizing incubation times. Major improvements were achieved by performing the series of eight enzyme assays with a single series of root samples instead of two series, reducing the time needed for sample preparation, minimizing error-prone steps such as pipetting and morphotyping, and facilitating subsequent DNA analyses due to the reduced sequencing effort. The best preservation of samples proved to be storage in soil at 4-6 °C in the form of undisturbed soil cores containing roots. Enzyme activities were maintained for up to 4 weeks under these conditions. Short-term storage of washed roots and ectomycorrhizal tips overnight in water did not cause substantial changes in enzyme activity profiles. No optimal means for longer-term storage by freezing at -20 °C or storage in 100% ethanol were recommended.
AU  - Pritsch, K.
AU  - Courty, .P.E.*
AU  - Churin, J.L.*
AU  - Cloutier-Hurteau, B.*
AU  - Ali, MA.*
AU  - Damon, C.*
AU  - Duchemin, M.*
AU  - Egli, S.*
AU  - Ernst, J.
AU  - Fraissinet-Tachet, L.*
AU  - Kuhar, F.
AU  - Legname, E.*
AU  - Marmeisse, R.*
AU  - Müller, A.*
AU  - Nikolova, P.
AU  - Peter, M.*
AU  - Plassard, C.*
AU  - Richard, F.*
AU  - Schloter, M.
AU  - Selosse, M.A.*
AU  - Franc, A.*
AU  - Garbaye, J.*
C1  - 5793
C2  - 29033
CY  - Berlin
SP  - 589-600
TI  - Optimized assay and storage conditions for enzyme activity profiling of ectomycorrhizae.
JO  - Mycorrhiza
VL  - 21
IS  - 7
PB  - Springer
PY  - 2011
SN  - 0940-6360
ER  - 

TY  - JOUR
AU  - Schubert, R.*
AU  - Raidl, S.*
AU  - Funk, R.*
AU  - Bahnweg, G.
AU  - Müller-Starck, G.*
AU  - Agerer, R.*
C1  - 9979
C2  - 21066
SP  - 159-165
TI  - Quantitative detection of agar-cultivated and rhizotron-grown Piloderma croceum Erikss. & Hjortst. by ITS1-based fluorescent PCR. 
JO  - Mycorrhiza
VL  - 13
PY  - 2003
SN  - 0940-6360
ER  - 

TY  - JOUR
AB  - The bacterial community structure of ecto-mycorrhizospheres on beech (Fagus sylvatica) grown in natural forest soil in southern Germany was examined by fluorescence in situ hybridization (FISH) using fluorescent oligonucleotide probes, targeting phylogenetic relevant sequences of the 16S and 23S rRNA. Lactarius subdulcis, L. vellereus, L. rubrocinctus and Laccaria amethystina were found to be the prevalent fungi forming ectomycorrhizae with F. sylvatica. For FISH studies using confocal laser scanning microscopy, oligonucleotide probes labeled with carboxymethylindocyanine-succinimidyl ester allowed detection of associated bacteria, because the autofluorescence of ectomycorrhiza samples could be overcome in the infrared. Bacteria of the alpha-, beta and gamma-subclasses of the proteobacteria were detected in high numbers on mantle surfaces, while members of other phylogenetically defined groups were found in smaller numbers. This contrasts with previous published results on the cultivation of mycorrhiza-associated bacteria. Hybridizing bacteria were also found within damaged cells of the hyphal mantle of L. rubrocinctus, as well as on emanating hyphae of L. amethystina. Using a newly developed extraction protocol for bacteria associated with ectomycorhizas, the two most common fungi on F. sylvatica, L. vellereus and L. subdulcis, were mostly associated with members of the alpha- and beta-subclasses of the proteobacteria. The proportion of hybridizing bacteria varied between the two ectomycorrhizae, which were thus host to distinct populations of bacteria.
AU  - Mogge, B.
AU  - Loferer, C.*
AU  - Agerer, R.*
AU  - Hutzler, P.
AU  - Hartmann, A.
C1  - 23018
C2  - 31141
SP  - 271-278
TI  - Bacterial community structure and colonization patterns of Fagus sylvatica L. ectomycorrhizospheres as determined by fluorescence in situ hybridization and confocal laser scanning microscopy.
JO  - Mycorrhiza
VL  - 9
IS  - 5
PB  - Springer
PY  - 2000
SN  - 0940-6360
ER  - 

TY  - JOUR
AU  - Pritsch, K.
AU  - Munch, J.-C.
AU  - Buscot, F.*
C1  - 21671
C2  - 19828
SP  - 87-93
TI  - Identification and differentiation of mycorrhizal isolates of black alder by sequence analysis of the ITS region.
JO  - Mycorrhiza
VL  - 10
PY  - 2000
SN  - 0940-6360
ER  -