Metabolic capabilities are indicated next to representative pictu

Metabolic capabilities are indicated next to representative pictures of symbiont growth in vitro: Plus or minus indicate the ability (+) or inability (−) to grow on the corresponding media. In order to learn more visualize the phylogenetic placement of symbionts and highlight their metabolic capabilities, symbiont strains were connected to their respective hosts with colored lines:

Red lines correspond to strains unable to grow on medium with peptones; green lines correspond to strains unable to grow on ammonium as the only source of nitrogen. Characterization of ‘S. philanthi’ biovars In all antennal samples used for isolation, SCH727965 in vitro the symbiotic Streptomyces showed a characteristic “antennal” phenotype: bacteria looked like individual or relatively short-chained cells, unbranched or with very short side branches, while no well-developed long mycelium was observed (exemplified by biovar ‘triangulum’ in Figure 1A). In culture, the vast majority of biovars developed typical

mycelium. However, two biovars were clearly distinguishable from all other symbiotic Streptomyces due to their “antennal” phenotype also in culture: in liquid medium, the actively growing biovar ‘elongatus’ formed micro-colonies, but in late stage of logarithmic growth or in stationary phase they tended to fall apart into short, often poorly branched fragments. A similar pattern was also observed for the biovar ‘loefflingi’, which could express one or the other phenotype over several transfers and change Pictilisib mouse it by the next passage, although conditions triggering such phenotypic changes remained unknown (Additional file 6: Figure S2). Although all beewolf-associated symbionts were originally assigned to biovars of the same species ‘Ca. S. philanthi’ [21], the morphology of even closely related biovars growing

on the same medium varied strongly (Figure 4). On Grace’s medium, bacteria from the clade ‘S. philanthi’ formed pigmented (yellow or beige) opaque colonies of round or irregular form, flat or gibbous with wave, broken or smooth border, and the surface varied from matte to slightly shiny, from smooth this website to rimmed and rugose. Only biovar ‘multimaculatus’, when grown on the Grace’s insect medium, formed white colonies with well-developed aerial mycelium typical for Streptomyces (Figure 4). Since all isolates were obtained on rich medium (supplemented with the full set of amino acids) imitating insect hemolymph, the next step was to assess the nutrient requirements of the isolated biovars by testing whether they could grow on media containing either an organic (peptones) or inorganic source of nitrogen (ammonium).

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