Identification of endophytic hyphae of <i>Lophodermium piceae</i> in tissues of green, symptomless Norway spruce needles by immunoelectron microscopy

Abstract

Antiserum specific for Lophodermium piceae hyphae was obtained by absorbing a rabbit L. piceae antiserum with hyphal material of different fungal isolates. The specificity of this absorbed antiserum was tested with hyphae of endophytic fungi isolated from green, asymptomatic needles of Norway spruce (Picea abies Karst.) using the on-section immunogold labeling technique. With this specific, absorbed antiserum, a homogenous labeling was obtained with cultured hyphae of L. piceae, whereas all other spruce needle endophytes examined remained practically unlabeled. Insignificant cross-reactivity was also observed with needle tissues. The absorbed antiserum was then applied for the immunoelectron microscopical identification of L. piceae hyphae in situ in infected tissues of green, asymptomatic Norway spruce needles. The function of papillalike structures observed in infected needle mesophyll as well as the role of the hyphal sheath surrounding endophytic L. piceae hyphae are discussed in connection with the interaction between L. piceae and asymptomatic Norway spruce needles.