Pine microsatellite markers allow roots and ectomycorrhizas to be linked to individual trees

Linking roots and ectomycorrhizas (EcM) to individual host trees in the field is required to test whether individual trees support different ectomycorrhizal communities. Here we describe a method that identifies the source of EcM roots by PCR of polymorphic pine nuclear microsatellite loci using fluorescently labelled primers and high-throughput fragment analysis. ITS-PCR can also be performed on the same EcM DNA extract for fungal identification. The method was tested on five neighbouring Scots pine (Pinus sylvestris var scotica) trees in native woodland. Successful host tree identification from DNA extracts of EcM root tips was achieved for 93% of all root fragments recovered from soil cores. It was estimated that each individual mature pine sampled was colonised by between 15 and 19 EcM fungi. The most abundant fungal species were found on all five trees, and within the constraints of the sampling scheme, no differences between trees in EcM fungal community structure or composition were detected.     

Morphological and anatomical characterisation of black alder Alnus glutinosa (L.) Gaertn. ectomycorrhizas

 Ectomycorrhizal types of black alder [Alnus glutinosa (L.) Gaertn.] collected over a 3-year period within an alder forest were characterised by morphological and anatomical features. Of the total of 16 types, 14 are described for the first time in this paper. Eight identified types belong to the genera Russula, Lactarius, Naucoria, and Cortinarius, while eight further types remained unidentified. In some cases, similarities of mantle features indicate relationships to identified mycorrhizas. Mycorrhizas of Naucoria escharoides and N. subconspersa were not distinguished. Two unidentified mycorrhizal types exhibited hyphal mantle structures very similar to these Naucoria species. Within the genus Cortinarius, mycorrhizas of C. cf. helvelloides were easily distinguished from all other Cortinarius-like mycorrhizas described on Alnus, which in general showed little anatomical variation. Two further unidentified mycorrhizas, "Alnirhiza lilacina" and "A. violacea", probably also belong to Cortinarius. The ectomycorrhiza of Russula pumila was the only identified type within the genus Russula, but the unidentified type "Alnirhiza cremicolor" also likely belongs to this genus. Three Lactarius species were present in the experimental plot. Two species (L. obscuratus and L. omphaliformis) had indistinguishable mycorrhizal types, but were easily differentiated from the mycorrhizas of L. lilacinus, which caused intracellular penetration of Hartig net hyphae into epidermal and cortical cells. All other mycorrhizal types of black alder exhibited a paraepidermal Hartig net without penetration of root cells. Two unidentified mycorrhizal types "Alnirhiza atroverrucosa" and "A. cystidiobrunnea", already described from North American Alnus rubra as unnamed morphotypes, showed no similarity to identified mycorrhizas. All 16 mycorrhizal types appeared to be specific or at least typical for alders, since they have not yet been reported from other tree species. Accepted: 29 August 1997     

The response of seedlings of two dipterocarp species to nutrient additions and ectomycorrhizal infection

An experiment was conducted to investigate the effect of ectomycorrhizal infection on growth and nutrient uptake, especially of P and K of dipterocarp seedlings.Hopea helferi (Dyer) Blanco andHopea odorata Roxb. seedlings were grown in a sandy loam soil given a basal dressing. Nutrient treatments were unamended soil (NIL), amended soil with the addition of P and K (F), amended soil without P but with K (-P), amended soil without K but with P (-K), amended soil without P or K addition (-PK). Seedlings grown in the amended soil treatments showed foliar symptoms suggestive of calcium deficiency. Ectomycorrhizal infection appeared to improve shoot Ca concentration and relieved the foliar symptoms. Ectomycorrhizal infection inH. odorata plants increased shoot P concentration and increased shoot and total dry weight to the same or greater extent than those of uninfected plants growing on P amended soil.H. helferi showed a positive response to ectomycorrhizal infection in shoot, root and total dry weight in all nutrient treatments but no response to the nutrient treatments themselves.     

Ericoid mycorrhizal fungi are common root associates of a Mediterranean ectomycorrhizal plant (Quercus ilex)

Mycorrhiza samples of neighbouring Quercus ilex and Erica arborea plants collected in a postcutting habitat were processed to see whether plants differing in mycorrhizal status harbour the same root endophytes. Three experiments were performed in parallel: (i) isolation, identification and molecular characterization of fungi from surface-sterilized roots of both plant species; (ii) re-inoculation of fungal isolates on axenic E. arborea and Q. ilex seedlings; (iii) direct inoculation of field-collected Q. ilex ectomycorrhizas onto E. arborea seedlings. About 70 and 150 fungal isolates were obtained from roots of Q. ilex and E. arborea, respectively. Among them, Oidiodendron species and five cultural morphotypes of sterile isolates formed typical ericoid mycorrhizas on E. arborea in vitro. Fungi with such mycorrhizal ability were derived from both host plants. Isolates belonging to one of these morphotypes (sd9) also exhibited an unusual pattern of colonization, with an additional extracellular hyphal net. Ericoid mycorrhizas were also readily obtained by direct inoculation of E. arborea seedlings with Q. ilex ectomycorrhizal tips. Polymerase chain-restriction fragment length polymorphism and random amplified polymorphic DNA analyses of the shared sterile morphotypes demonstrate, in the case of sd9, the occurrence of the same genet on the two host plants. These results indicate that ericoid mycorrhizal fungi associate with ectomycorrhizal roots, and the ecological significance of this finding is discussed.     

Quantities and types of ectomycorrhizal and endophytic fungi associated with Betula platyphylla var. japonica seedlings during the initial stage of establishment of vegetation after disturbance

Ectomycorrhizal and endophytic fungi of Betula platyphylla Sukatchev var. japonica Hara seedlings were investigated by bioassay using soils from sites where the surface layer had been removed by destructive disturbances. Soil samples were taken from sites A, B, C and D, where 1, 2–3, 4–5, and 7–8 years, respectively had passed since disturbance. Naturally regenerated B. platyphylla var. japonica seedlings grew at sites C and D, but not at sites A or B. The percentages of ectomycorrhizal formation in seedlings were significantly lower in the soils from site A (4%) and site B (13%), compared to those in the soils from site C (53%) and site D (37%). The numbers of ectomycorrhizal morphologic types in sites A, B, C, and D were eight, five, one, and seven, respectively. The same dominant type of ectomycorrhiza was found in sites C and D, and this type was different from those in sites A and B. The frequencies of colonization of seedling roots by endophytic fungi, especially Mycelium radicis atrovirens Melin (MRA) in soils from sites A and B were 31 and 33%, respectively; these frequencies were significantly higher than those for site C (0%) and site D (2%). During the initial stage of establishment of vegetation following disturbance, the quantities and types of ectomycorrhizal fungi in the field that have the potential to associate with B. platyphylla var. japonica might rapidly change after invasion of the host plant. Ectomycorrhizal fungi seemed to compete with endophytic MRA fungi for colonization of the roots of B. platyphylla var. japonica seedlings.