Figure 1.
Map of the collection sites for this study.
The numbers refer to the species locations detailed in Table 1.
Table 1.
List of species/specimens used in this work according to the classification of Systema Porifera [89] and the recent updates added to the World Porifera Database [9].
Table 2.
Three sets of non-molecular characters-states for each species/specimen: ecology/geography, external morphology and associated microbes.
Table 3.
Histology, cytology and embryology morphological characters-states for each species/specimen.
Table 4.
Identity values between members of clade D for mitochondrial markers.
Table 5.
Nucleotide diversity (π) of Oscarella lobularis, O. tuberculata and D3 members compared to Demospongiae species for the atp6 marker.
Figure 2.
Phylogenetic analysis of nuclear markers.
(A) 18S rDNA and (B) 28S rDNA. The topologies presented are posterior consensus trees obtained by the BI analysis using MrBayes. Similar topologies were obtained in ML analysis with PhyML. The numbers are posterior probabilities for BI and bootstrap values (>50) for ML.
Figure 3.
Phylogenetic analysis of mitochondrial markers.
(A) atp6 and (B) tatC. The topologies presented are posterior consensus trees obtained by the BI analysis using MrBayes. Similar topologies were obtained in ML analysis with PhyML. The numbers are posterior probabilities for BI and bootstrap values (>50) for ML.
Figure 4.
Oscarellidae relationships based on the analyses of concatenated sequences.
(A) 18S rDNA +28S rDNA nuclear markers, (B) atp6+ tatC mitochondrial markers and (C) nuclear (18S rDNA +28S rDNA)+mitochondrial (atp6+ tatC) markers. The topologies presented are posterior consensus trees obtained by the BI analysis using MrBayes. Similar topologies were obtained in ML analysis with PhyML. The numbers are posterior probabilities for BI and bootstrap values (>50) for ML.
Figure 5.
Schemas of the secondary structure predictions of the 18S rDNA V4 region mapped on the 18S rDNA tree topology.
Elements composing the structures and included in the characters matrix are circled in a specific color. Characters that can be considered as synapomorphies are mentioned next to the corresponding node.
Figure 6.
Oscarella bergenensis sp. nov.
(A) External morphology in vivo. (B) General anatomy, observed with light microscopy. (C) TEM of exopinacocyte. (D) TEM of endopinacocyte. (E) TEM of apopylar cell. (F) TEM of choanocyte. (G) TEM of archaeocyte. (H) TEM of vacuolar cells. (I) TEM of granular cells. (J) TEM of symbiotic bacteria. (b) Symbiotic bacteria; (bm) Basement membrane; (cc) Choanocyte chamber; (ec) Ectosome; (exc) Exhalant canal; (exp) Exopinacodem; (f) Flagellum; (gc) Glycocalyx; (gr) Granules; (mv) Microvilli; (n) Nucleus; (o) Osculum; (v) Vacuole. Scale bar: J = 0.5 µm.
Figure 7.
(A) External morphology in vivo. (B) General anatomy, observed with light microscopy. (C) Light microscopy, details of hermaphrodite sponge during reproduction. (D) TEM of exopinacocyte. (E) TEM of endopinacocyte. (F) TEM of apopylar cell. (G) TEM of choanocyte. (H) TEM of archaeocyte. (I) TEM of granular cell. (J) TEM of spherulous cells with paracrystalline inclusions. (K) TEM of symbiotic bacteria. (b) Symbiotic bacteria; (bm) Basement membrane; (cc) Choanocyte chamber; (cpi) Spherulous cells with paracrystalline inclusions; (ec) Ectosome; (em) Embryos; (exc) Exhalant canal; (exp) Exopinacoderm; (f) Flagellum; (fo) Follicle; (gc) Glycocalyx; (gr) Granules; (n) Nucleus; (o) Osculum; (oo) Oocyte; (pcr) Spherules with paracrystalline inclusions; (sp) Spermatic cyst. Scale bar: J = 0.5 µm.
Figure 8.
Simplified consensus tree based on tatC+atp6 molecular phylogenies.
All robust nodes (BP>50+ PP>0.5) were conserved. Polytomy was prioritized for weakly-supported nodes (BP<50 or PP<0.5). Molecular and non-molecular characters that are synapomorphies/diagnostic characters of the clades are indicated on the corresponding nodes. The absence of vacuolar cells (1) is diagnostic of A. The presence of cortex (2), leuconoid aquiferous system (3) and diplodal chambers (4) are specific characters of Oscarella jarrei. The presence of vacuolar cells in O. balibaloi is a reversal character (5) The two new species names are indicated in bold in the tree. In light grey are specimen/species for which uncertainties remain (new species or not). Schemas of morphological characters are modified from [60] or are new.