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5.Cell Surface Characterization of Ectomycorrhizal Hyphae Using Fluorochrome Labeled Lectins and Its Application as a Probe to Detect Ectomycorrhizal Hyphae in Soils of Forest Ecosystems

Takahiro Tateishi and Ayato Kohzu(Center for Ecological Research, Kyoto University)

Surface carbohydrates of the vegetative mycelium of ectomycorrhizal fungi were analyzed using fluorochrome labeled lectins and two fluorochromes in order to investigate the feasiability on the selective detection of their hyphae in forest soils. All of the hyphea in cultured aerial mycelium of ectomycorrizae were mainly composed of β-1,4-glucan, β-1,3-glucan and side residues of oligomer of N-acetyl-glucosamine by binding assay with calcofluor white M2R (CW), aniline blue (AB) and FITC labeled wheat germ agglutinin (WGA). Binding assay with FITC labeled peanut agglutinin (PNA) showed that side residues of β-D-galactosamine and its derivatives were existed on the cell walls of both vegetative and aggregated mycelia of Sect. Amanita in Amanitaceae, but of other ectomycorrhizal fungi except some strains. This suggested that PNA was available to detect the external hyphae and mycorrhizae of Sect. Amanita in Amanitaceae in soils. Binding assay with FITC-WGA and FITC-PNA was made for the ectomycorrhizae and hyphae collected from the soils where fruiting bodies of Amanita pantherina appeared. FITC-WGA could bind the hyphae on the mantle surface of many ectomycorrhizae examined, but some did not. FITC-PNA did not bind hyphae on the mantle surface of ectomycorrhizae. Hyphal length bound by FITC-PNA was 12 m/g dry soil, which was considered to be the external hyphae of A. pantherina in soils. This corresponded to 0.3% of the total haphal length in the soils examined.

外生菌根/ectomycorrhizae、FITC結合レクチン/FITC labeled lectin、菌糸/hyphae、表面糖鎖/surface carbohydrates、細胞壁構造/cell wall structure