Figure 1.
Co-occurrence of Type N and Type S B. neritina.
Colonies collected together in (A) Beaufort, NC, and (B) Oyster, VA. Blue arrows indicate Type N colonies; red indicate Type S.
Table 1.
Bugula neritina symbiont frequency along North American Atlantic coast.
Table 2.
Primers and probes used in this study.
Figure 2.
B. neritina sibling species collected by haphazard sampling.
Proportions at each site indicated by blue (Type N) and red (Type S) in charts.
Figure 3.
Symbiotic status of sampled colonies.
Proportions at each site indicated by black (symbiotic) and gray (aposymbiotic) in charts. Results from both haphazardly collected and targeted samples are included. Number of colonies sampled shown beside each graph.
Figure 4.
Bugula neritina sibling species-specific PCR.
Reactions were performed with gDNA (10 ng) extracted from pooled larvae collected from >200 adult colonies as template. Control DNA was extracted from adult B. neritina colonies of both phylotypes to serve as 100% Type N or S cytochrome c oxidase templates.
Figure 5.
Confirmation of B. neritina symbiotic status via fluorescence in situ hybridization (FISH).
White arrows indicate the circular pallial sinus on the aboral surface of the larva. (A–D) FISH micrographs of larvae using the Eub338 probe. (E–H) FISH micrographs of larvae using the symbiont-specific Bn1253r probe. Diameter of pallial sinus is approximately 150 µm.
Figure 6.
Molecular characterization of B. neritina and HPLC analysis of crude larval extract.
Agarose gel showing DdeI and HhaI restriction digestion of B. neritina adult (top two samples) or pooled larval (bottom sample) cytochrome c oxidase I; amplification of bryS to demonstrate presence of symbiont; and absorbance at 229 nm (which is diagnostic of bryostatins) of extract collected from larvae represented by this gDNA.
Table 3.
Percent identity of marker sequences among B. neritina sibling species.