Fig 1.
a) Map showing the Adriatic Sea, reprinted from GeoMapApp (www.geomapapp.org) under a CC BY license, with permission from William B. F. Ryan, original copyright 2009 [53], b) Detailed drawing of the coastline around Piran with special emphasis on the three sampling sites.
Fig 2.
Colonies of Zoothamnium mariella sp. nov. attached to wood.
Table 1.
Site, substrate, and date of samples used for various methods.
Table 2.
Probes used for fluorescence in situ hybrization.
Fig 3.
Zoothamnium mariella sp. nov. Detailed drawing of an entire colony and a microzooid, both in expanded condition. mi: microzooid, ma: macrozooid, t: terminal zooid, br: branch, st: stalk, sp: spasmoneme, ad: adhesive disc, am1: adoral membranelle 1 (polykinety), pm: paroral membrane (haplokinety), in: infundibulum, cp: cytopharynx, fv: food vacuole, man: macronucleus, min: micronucleus, tl: telotrochal band, b: bacteria.
Fig 4.
Micrographs of living and silver-stained Zoothamnium mariella sp. nov. specimens.
a) expanded colony, b) contracted colony, c) most basal part of the stalk, where the spasmoneme splits into bands, which bundle towards the basal end of the colony, d) ellipsoid terminal zooid on the verge of cell division, e) macrozooid, f) microzooid, g–m) oral ciliature of the microzooids, n) microzooid with emphasize on its telotrochal band. a–d) living specimens. e–n) silver-stained specimens. mi: microzooid, ma: macrozooid, t: terminal zooid, st: stalk, sp: spasmoneme, ad: adhesive disc, man: macronucles, oc: oral ciliature, pm: paroral membrane (haplokinety), am1–3: adoral membranelles 1–3 (polykineties), s: stomatogenic kinety, tl: telotrochal band.
Fig 5.
Schematic illustration of the oral ciliature in Zoothamnium mariella sp. nov., viewed from the oral pole.
pm: paroral membrane (haplokinety), am1–3: adoral membranelles 1–3 (polykineties), s: stomatogenic kinety.
Fig 6.
Maximum Likelihood tree inferred from the nucleotide sequences of the small subunit rRNA genes of the monophyletic clade II of Zoothamnidae.
The numbers in parentheses are the NCBI accession numbers for each species and the numbers at the nodes represent Maximum Likelihood and Maximum Parsimony bootstrap values greater than or equal to 70%. The scale bar corresponds to 1 substitution per 200 nt positions.
Fig 7.
Maximum Likelihood tree inferred from the nucleotide sequences of gamma proteobacterial small subunit rRNA genes displaying the phylogeny of Ca. Fusimicrobium zoothamnicola gen. nov., sp. nov.
The numbers in parentheses are the NCBI accession numbers for each species and the numbers at the nodes represent Maximum Likelihood and Maximum Parsimony bootstrap values greater than or equal to 70%. The scale bar corresponds to 5 substitutions per 100 nt positions.
Fig 8.
Epifluorescent micrographs showing the Z. mariella sp. nov. symbiosis.
a, b, c) LIVE/DEAD staining, d,h) DAPI (blue), e, i) EUB mix in Cy5 (green), f, j) probe ZMS152 and ZMS1239 in Cy3 (red), g, k) overlay of the three colours, mi: microzooid, ma: macrozooid, t: terminal zooid, st: stalk, br: branch.
Fig 9.
Scanning electron micrographs of Zoothamnium mariella sp. nov.
a) entire colony in contracted state, b) stalk with bacterial coat, c) branch with microzooids and a terminal zooid, d) microzooid, e) macrozooid, f) bacteria colonizing macrozooid, g) barely colonized macrozooid, h) barely colonized microzooid. mi: microzooid, t: terminal zooid, st: stalk, br: branch, sc: somatic ciliature.
Table 3.
Comparison between Zoothamnium mariella sp. nov, Z. ignavum, and Z. alternans.