Spatial associations between actin filaments, endoplasmic reticula, mitochondria and fungal hyphae in symbiotic cells of orchid protocorms

Abstract

The relationships between endoplasmic reticula (ER), mitochondria, and actin filaments (Afs) were observed in uncolonized and colonized cells of symbiotic protocorms ofSpiranthes sinensis (Orchidaceae) germinated in the presence of the fungus,Ceratobasidium cornigerum. Mitochondria and ER were observed by transmission electron microscopy, and with the fluorescent probe DiOC6 (3) (3,3′-dihexyloxacarbocyanine) combined with confocal laser scanning microscopy (CLSM). An indirect immunofluorescence method using CLSM and an indirect, pre-embedding immunogold method at the ultrastructural level were used for observation of Afs. In uncolonized cells, cortical ER showed a polygonal pattern and ER formed a network throughout the cytoplasm. In the cortex, a smooth face of ER contacted the plasma membrane. Mitochondria were associated with ER. Afs were in close proximity to ER, mitochondria and amyloplasts. Colonized cells retained cortical ER, and a smooth face of ER was also closely associated with the perifungal membrane. ER and mitochondria were present in the cytoplasmic channels bridging between the central peloton and the peripheral cytoplasm. This distribution of ER and mitochondria during fungal colonization and senescence coincided with that of Afs. The changes in the arrays of Afs accompanying symbiotic fungal colonization and senescence occurred concomitantly with the changes in ER.