Anastomosis behavior differs between asymbiotic and symbiotic hyphae of Rhizophagus clarus.

The life history of arbuscular mycorrhizal fungi (AMF, Glomeromycota) consists of a short asymbiotic phase when spores germinate and a longer symbiotic phase where hyphae form a network within roots and subsequently in the rhizosphere. Hyphal anastomosis contributes to colony formation, yet this process has been studied mostly in the asymbiotic phase rather than in mycorrhizal plants because of methodological limitations. We sought to compare patterns of anastomosis during each phase of fungal growth by measuring hyphal fusions in genetically identical and different single spore isolates of Rhizophagus clarus from different environments and geographic locations. These isolates were genotyped with two anonymous markers of microsatellite-flanking regions. Anastomosis of hyphae from germinating spores was examined in axenic Petri dishes. A rhizohyphatron consisting of agar-coated glass slides bridging single or paired mycorrhizal sorghum plants allowed evaluation of anastomosis of symbiotic hyphae. Anastomosis of hyphae within a colony, defined here as a mycelium from an individual germinating spore or from mycorrhizal roots of one plant, occurred with similar frequencies (8-38%). However, anastomosis between paired colonies was observed in germinating spores from either genetically identical or different isolates, but it was never detected in symbiotic hyphae. The frequency of anastomosis in asymbiotic hyphae from paired interactions was low, occurring in fewer than 6% of hyphal contacts. These data suggest that anastomosis is relatively unconstrained when interactions occur within a colony but is confined to asymbiotic hyphae when interactions occur between paired colonies. This pattern of behavior suggests that asymbiotic and symbiotic phases of mycelium development by R. clarus may differ in function. Anastomosis in the asymbiotic phase may provide brief opportunities for gene flow between populations of this and possibly other AMF species.

In situ analysis of anastomosis in representative genera of arbuscular mycorrhizal fungi.

Arbuscular mycorrhizal fungi (AMF) form obligate symbiotic associations with plants. As a result, the role of hyphal interactions in the establishment and maintenance of common mycorrhizal networks is poorly understood because of constraints on methods for in situ analysis. We designed a rhizohyphatron that allows the examination of intact mycelia growing from whole mycorrhizal plants. Plants preinoculated with spores were cultivated in a compartment with a connecting tube from which hyphae extend through a fine nylon mesh onto agar-coated slides. Species selected from each of the five AMF genera were used to assess and characterize the anastomosis behavior in the rhizohyphatron. Hyphal networks of Paraglomus occultum, Ambispora leptoticha, Scutellospora heterogama, and Gigaspora gigantea growing on the agar-coated slides showed no evidence of hyphal fusion. In contrast, anastomosis occurred in the hyphal networks of Glomus clarum and Glomus intraradices at an average frequency of less than 15% for both species. The rhizohyphatron developed in this study will provide knowledge of the biology and genetics of self/non-self recognition in AMF and help to better understand Glomeromycotan life history strategies.

Parasitism of arbuscular mycorrhizal fungi: reviewing the evidence.

In order to understand the functioning of mycorrhizal fungi in ecosystems it is necessary to consider the full suite of possible biotic interactions in the soil. While a number of such interactions have recently been shown to be crucially important, parasitism is a highly neglected feature in the ecology of arbuscular mycorrhizal fungi (AMF). A number of studies have classified some interactions between populations of bacteria and fungi with AMF as parasitism, generating discussion about its consequences at both 'parasite' and host population levels. This paper reviews these various publications, and based on a set of criteria that are necessary to demonstrate parasitism, it was concluded that parasitism has not been conclusively shown to exist in AMF, even though some data are highly suggestive of such a relationship. The difficulties in gathering data supportive of parasitism were discussed, and hypotheses for defense were offered. This paper concludes by presenting potential consequences of AMF parasitism at the population/community levels and by discussing applied aspects.