The repeated failures reported in cultivating some microbial lineages are a major challenge in microbial ecology and probably linked, in the case of Frankia microsymbionts to atypical patterns of auxotrophy. Comparative genomics of the so far uncultured cluster-2 Candidatus Frankia datiscae Dg1, with cultivated Frankiae has revealed genome reduction, but no obvious physiological impairments. A direct physiological assay on nodule tissues from Coriaria myrtifolia infected with a closely-related strain permitted the identification of a requirement for alkaline conditions. A high pH growth medium permitted the recovery of a slow-growing actinobacterium. The strain obtained, called BMG5.1, has short hyphae, produced diazovesicles in nitrogen-free media and fulfilled Koch’s postulates by inducing effective nodules on axenically grown Coriaria spp. and Datisca glomerata. Analysis of the draft genome confirmed its close proximity to the Candidatus Frankia datiscae Dg1 genome with the absence of 38 genes (trehalose synthase, fumarylacetoacetase, etc) in BMG5.1 and the presence of 77 other genes (CRISPR, lanthionine synthase, glutathione synthetase, catalase, Na+/H+ antiporter, etc) not found in Dg1. A multi-gene phylogeny placed the two cluster-2 strains together at the root of the Frankia radiation.
(2014). The host ranges of conifer-associated Tricholoma matsutake, Fagaceae-associated T. bakamatsutake and T. fulvocastaneum are wider in vitro than in nature. Mycologia: Vol. 106, No. 3, pp. 397-406.
Miyake-Jima, a round island of about 8 km in diameter, is located about 200 km south of Tokyo, Japan (34°08′ N, 139°53′ E). In July 2000, the central cone of this island collapsed and a new eruption started with the falling of volcanic ash and SO2 emission. In October 2001, we measured the distribution of bacteria, and Frankia, a symbiotic nitrogen-fixing actinomycete that forms root nodules, and ectomycorrhizal fungi associated with alder in newly deposited ash and its underlying soil by cultivation of Alnus sieboldiana seedlings in these soils. There were fewer bacteria growing on the nutrient broth agar and fewer bacteria growing on 100-fold diluted nutrient broth agar in newly deposited volcanic ash than there were growing in the buried old soil. In four out of five sites, little or no nodulation was observed in newly deposited ash; abundant root nodules were formed in the underlying soil. Ectomycorrhizae formed in the seedlings cultivated in the underlying soil. In May 2003, the distribution of Frankia and ectomycorrhizal fungi in soil at different depths (up to 160 cm) was also investigated. Frankia and ectomycorrhizal fungi were relatively abundant in surface soil layers.
The role of tetrapartite associations among Frankia, Gigaspora margarita (an arbuscular mycorrhizal fungus), Pseudomonas putida (rhizobacterium), and Alnus sieboldiana in growth, nitrogen fixation, and mineral acquisition of A. sieboldiana was investigated. Seedlings of A. sieboldiana were inoculated with Frankia isolated from root nodules of alder, followed by inoculation of G. margarita and P. putida, and were grown for 5 months in a greenhouse. The seedlings inoculated with Frankia and G. margarita together produced the highest biomass of shoots and root nodules. Nitrogen-fixation activity, measured by acetylene reduction assay, was observed when Frankia was inoculated. The activity, on a per-nodule gram basis, decreased after G. margarita inoculation, but on a per-plant basis there was no significant difference in the activity among inoculation treatments. The mineral content in the seedlings changed after inoculation with Frankia, but not after inoculation with P. putida and/or G. margarita. The results showed a synergistic interaction among Frankia, the mycorrhizal fungus, and the rhizobacterium on the growth of A. sieboldiana.