Divergence of Funneliformis mosseae populations over 20 years of laboratory cultivation, as revealed by vegetative incompatibility and molecular analysis

Arbuscular mycorrhizal fungi (AMF) are widespread, important plant symbionts. They absorb and translocate mineral nutrients from the soil to host plants through an extensive extraradical mycelium, consisting of indefinitely large networks of nonseptate, multinucleated hyphae which may be interconnected by hyphal fusions (anastomoses). This work investigated whether different lineages of the same isolate may lose the ability to establish successful anastomoses, becoming vegetatively incompatible, when grown separately. The occurrence of hyphal incompatibility among five lineages of Funneliformis mosseae, originated from the same ancestor isolate and grown in vivo for more than 20 years in different European locations, was assessed by systematic detection of anastomosis frequency and cytological studies. Anastomosis frequencies ranged from 60 to 80% within the same lineage and from 17 to 44% among different lineages. The consistent detection of protoplasm continuity and nuclei in perfect fusions showed active protoplasm flow both within and between lineages. In pairings between different lineages, post-fusion incompatible reactions occurred in 6–48% of hyphal contacts and pre-fusion incompatibility in 2–17%. Molecular fingerprinting profiles showed genetic divergence among lineages, with overall Jaccard similarity indices ranging from 0.85 to 0.95. Here, phenotypic divergence among the five F. mosseae lineages was demonstrated by the reduction of their ability to form anastomosis and the detection of high levels of vegetative incompatibility. Our data suggest that potential genetic divergence may occur in AMF over only 20 years and represent the basis for detailed studies on the relationship between genes regulating anastomosis formation and hyphal compatibility in AMF

Compatibility and incompatibility in hyphal anastomosis of arbuscular mycorrhizal fungi

Arbuscular mycorrhizal fungi (AMF), which live in symbiosis with 80 % of plants, are not able to grow when separated from their hosts. Spore germination is not host-regulated and germling growth is shortly arrested in the absence of host roots. Germling survival chances may be increased by hyphal fusions (anastomoses), which allow access to nutrients flowing in the extraradical mycelium (ERM). Perfect anastomoses, occurring with high frequency among germlings and the ERM of the same isolate, show protoplasm continuity and disappearance of hyphal walls. A low frequency of perfect fusions has been detected among co-specific genetically different isolates, although fungal nuclei have been consistently detected in all perfect fusions, suggesting active nuclear migration. When plants of different taxa establish symbioses with the same AMF species, anastomoses between ERM spreading from single root systems establish a common mycelium, which is an essential element to plant nutrition and communication. The interaction among mycelia produced by different isolates may also lead to pre-fusion incompatibility which hinders anastomosis formation, or to incompatibility after fusion, which separates the hyphal compartments. Results reported here, obtained by analyses of hyphal compatibility/incompatibility in AMF, suggest that anastomosis formation and establishment of protoplasm flow, fundamental to the maintenance of mycelial physiological and genetic continuity, may affect the fitness of these ecologically important biotrophic fungi

Compatibility and incompatibility in hyphal anastomosis of arbuscular mycorrhizal fungi

ABSTRACT: Arbuscular mycorrhizal fungi (AMF), which live in symbiosis with 80 % of plants, are not able to grow when separated from their hosts. Spore germination is not host-regulated and germling growth is shortly arrested in the absence of host roots. Germling survival chances may be increased by hyphal fusions (anastomoses), which allow access to nutrients flowing in the extraradical mycelium (ERM). Perfect anastomoses, occurring with high frequency among germlings and the ERM of the same isolate, show protoplasm continuity and disappearance of hyphal walls. A low frequency of perfect fusions has been detected among co-specific genetically different isolates, although fungal nuclei have been consistently detected in all perfect fusions, suggesting active nuclear migration. When plants of different taxa establish symbioses with the same AMF species, anastomoses between ERM spreading from single root systems establish a common mycelium, which is an essential element to plant nutrition and communication. The interaction among mycelia produced by different isolates may also lead to pre-fusion incompatibility which hinders anastomosis formation, or to incompatibility after fusion, which separates the hyphal compartments. Results reported here, obtained by analyses of hyphal compatibility/incompatibility in AMF, suggest that anastomosis formation and establishment of protoplasm flow, fundamental to the maintenance of mycelial physiological and genetic continuity, may affect the fitness of these ecologically important biotrophic fungi