Effect of developmental stage and peloton morphology on success in isolation of mycorrhizal fungi in Caladenia formosa (Orchidaceae)

Abstract

Six developmental stages (leafing, budding, flowering, fruiting, senescence and dormancy) were chosen in the threatened terrestrial orchid Caladenia formosa G.W.Carr to optimise isolation of effective fungi. Loose (undigested) pelotons were observed by scanning electron microscopy in the old tuber and collar, suggesting a role in infection of new tissue. In collars collected at early life stages (leafing, budding, flowering), pelotons had loosely coiled hyphae that were uniformly fine (1–2 μm diameter), with or without monilioid cells. In collars collected from older life stages (fruiting, senescence), pelotons had increasing proportions (up to 94%) of clumped fine hyphae. Coarser hyphae (4–6 μm diameter) were also present in the fruiting stage in one year. Only fungi isolated from single pelotons in collars of early life stages (leafing, budding, flowering) had fine hyphae with monilioid cells and induced seed germination (to green leaf production). Sectioned protocorms had pelotons of fine, loosely coiled hyphae with monilioid cells, as in field-collected material from early life stages. This suggests that the most effective fungi for conservation of this orchid are likely to be isolated from pelotons of loose fine hyphae with monilioid cells from leafing to flowering stages.