Characterization of a Lectin from Lactarius deterrimus (Research on the Possible Involvement of the Fungal Lectin in Recognition between Mushroom and Spruce during the Early Stages of Mycorrhizae Formation).

摘要

A lectin (LDetL) was isolated from carpophores of the mushroom Lactarius deterrimus, a specific symbiont of the spruce, by a combination of affinity, hydroxylapatite, and gel-filtration chromatography. Its molecular mass, as determined by gel filtration, is about 37,000 D, and its structure is dimeric, with two identical subunits assembled by noncovalent bonds. It appeared homogeneous on high-performance liquid chromatography gel filtration, but isoelectric focusing revealed microheterogeneity, with a main band in the pH zone near 6.5. Amino acid analysis showed that LDetL contains a large proportion of glycine and especially methionine. Hapten inhibition assay indicated that LDetL is most specific for [beta]-D-galactosyl(1->3)-D-N-acetyl galactosamine residues. The lectin was formed in the in vitro-cultivated mycelium, and anti-lectin antibodies revealed by indirect immunofluorescence the presence of lectin in the cell wall. Receptor sites for LDetL were found on the roots, especially on the root hairs, of axenically grown spruce seedlings. The lectin LDL previously isolated by us from the taxonomically related mushroom Lactarius deliciosus, a symbiont of the pine, does not bind to the spruce radicle. This suggests a role of the fungal lectin in recognition and specificity during the early stages of mycorrhizae formation.