Marine Ecology Progress Series 362:139
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Vol. 362: 139–147, 2008 MARINE ECOLOGY PROGRESS SERIES Published June 30 doi: 10.3354/meps07464 Mar Ecol Prog Ser Phototrophic nutrition and symbiont diversity of two Caribbean sponge–cyanobacteria symbioses Patrick M. Erwin, Robert W. Thacker* University of Alabama at Birmingham, 109 Campbell Hall, 1300 University Boulevard, Birmingham, Alabama 35294-1170, USA ABSTRACT: Coral reef sponges frequently harbor symbiotic cyanobacteria that can potentially ben- efit their hosts by providing supplemental nutrition. Although nutritional mutualisms have been doc- umented for Indo-Pacific sponges, previous investigations suggest that Caribbean sponges obtain no nutritional benefits from their symbionts. We conducted field-based shading experiments to deter- mine the effects of reduced irradiance on symbiont abundance and host growth in the Caribbean coral reef sponges Aplysina fulva and Neopetrosia subtriangularis, both of which host the unicellular cyanobacterium Synechococcus spongiarum. For both host species, 6 wk of shading significantly reduced chlorophyll a concentrations to less than half of those in control sponges. Shaded A. fulva individuals exhibited less than half the growth of control sponges. For N. subtriangularis, no signifi- cant differences in growth rates were observed between shaded and control sponges. Measurements of gross photosynthesis to respiration (P:R) ratios predicted that both host sponges were heterotrophic at low irradiances (<250 µmol m–2 s–1) and phototrophic at high irradiances (>500 µmol m–2 s–1), with measurements obtained at high irradiances likely to be more indicative of natural conditions. Phylo- genetic analyses of cyanobacterial internal transcribed spacer (ITS) ribosomal RNA gene sequences revealed 3 genetically distinct clades of S. spongiarum (designated A, B and C). Clade A symbionts were isolated exclusively from A. fulva, Clade B symbionts from both host sponge species and Clade C symbionts exclusively from N. subtriangularis. These findings demonstrate that Caribbean sponges associate with diverse clades of cyanobacterial symbionts that may confer variable nutritional bene- fits; in some cases, these symbionts appear to substantially enhance host growth rates. KEY WORDS: Symbiosis · Sponge · Cyanobacteria · Primary productivity · Respiration Resale or republication not permitted without written consent of the publisher INTRODUCTION fied by mutualistic coral–dinoflagellate associations, whose evolution yielded the formation of coral reef Coral reefs are among Earth’s most productive habitats (Stanley 2006), and span a wide range of ecosystems and support a remarkable diversity and invertebrate host taxa, including tridacnid clams, abundance of marine life; however, these habitats are anemones, ascidians and sponges (Rützler 1990). generally characterized by low nutrient availability, Coral reef sponges most commonly harbor cyano- intense competition for space and light, and heavy bacteria as photosynthetic symbionts (Rützler 1990), grazing pressures (Birkeland 1997). To cope with these but also form associations with filamentous algae (Car- environmental demands, many sessile invertebrates ballo & Ávila 2004) and dinoflagellates (Schönberg & form symbioses with photosynthetic organisms, in Loh 2005). Although cyanobacterial symbionts can which autotrophic symbionts contribute to the meta- comprise 25 to 50% of a sponge’s cellular volume bolic and energetic needs of the host (Muscatine (Rützler 1990), relatively little is known about the & Porter 1977), thereby enhancing host growth rates metabolic exchanges and ecological interactions and competitive ability. These symbioses are exempli- between sponges and their microbial symbionts (Tay- *Email: [email protected] © Inter-Research 2008 · www.int-res.com 140 Mar Ecol Prog Ser 362: 139–147, 2008 lor et al. 2007a). Symbiotic cyanobacteria may benefit We assessed the nutritional contribution of symbiotic host sponges through supplemental nutrition (Arillo et cyanobacteria to Caribbean coral reef sponges by al. 1993), nitrogen fixation (Wilkinson & Fay 1979), experimentally shading 2 common sponge species, production of secondary metabolites (Flatt et al. 2005) Aplysina fulva and Neopetrosia subtriangularis, and and UV protection (Regoli et al. 2000), suggesting key monitoring chlorophyll a (chl a) concentrations and roles for these symbionts in the evolutionary history holobiont growth over a 6 wk period. These empirical and ecological success of coral reef sponges. data were compared to predictions of symbiont- Although sponge-associated cyanobacteria span at derived nutritional benefits based on post-experiment least 6 genera (Steindler et al. 2005, Taylor et al. 2007a), measurements of P:R ratios. Symbiont identities were a sponge-specific lineage of Synechococcus represents confirmed and fine-scale variability between host the most prevalent symbiont group (Erwin & Thacker sponge populations was assessed using cyanobacterial 2007). The initial description of this species, ‘Candidatus 16S rRNA and 16S–23S rRNA internal transcribed Synechococcus spongiarum’, was based on the distinct spacer (ITS) gene sequences. morphology and unique 16S ribosomal RNA (rRNA) gene sequences of cyanobacteria associated with the host sponge Chondrilla nucula (Usher et al. 2004). Sim- MATERIALS AND METHODS ilar ribotypes have been characterized from at least 20 sponge species, which represent 12 families and 8 Sponge collection and experimental design. The sponge orders, and inhabit tropical and temperate reefs sponges Aplysina fulva and Neopetrosia subtriangu- around the world (Steindler et al. 2005, Thacker 2005). laris were collected from STRI Point, Bocas del Toro, Phylogenetic analyses of S. spongiarum 16S rRNA se- Panama (9° 21’ 7’’ N, 82° 15’ 32’’ W) at depths of 2 to quences from these host species revealed a genetically 3 m. A. fulva and N. subtriangularis are abundant distinct symbiont clade clearly differentiated from free- members of the reef sponge community in these shal- living cyanobacteria, but with little consistent genetic low waters, representing the 2nd and 5th most abun- differentiation within this clade based on host taxonomy dant sponge species, respectively (Erwin & Thacker or geographic location (Steindler et al. 2005, Thacker 2007). Both species exhibit a rope-like morphology 2005). Similarly, comparative morphological analyses of with occasional bifurcating branches. A. fulva indi- S. spongiarum using electron microscopy reported indis- viduals inhabiting the reefs of Bocas del Toro range tinguishable symbiont populations inhabiting disparate in color from yellow to reddish brown, with a com- hosts from distant geographic regions (Usher et al. 2006). pressible consistency. N. subtriangularis individuals Taken together, these studies suggest that S. spongia- range in color from yellowish-brown to dark brown, rum may represent a generalist symbiont capable of with a brittle, hard consistency. A total of 10 individu- horizontal transmission among hosts and widespread als per species were collected; each individual was dispersal across the oceans. divided into 2 small pieces (approximately 3 cm long) The contribution of cyanobacterial symbionts to host and 4 large pieces (approximately 10 cm long). One sponge nutrition has been estimated by measuring pho- small piece was preserved in RNAlater (Ambion) for tosynthetic rates and gross primary productivity to respi- genetic characterization of symbionts, while the other ration (P:R) ratios of intact holobionts (Wilkinson 1987). was used for initial chl a quantification (see ‘Chl a Symbioses with a P:R ratio exceeding 1.5 were defined quantification’). From each source individual, 2 large as phototrophic sponges, receiving supplemental nutri- pieces were randomly assigned to the control treat- tion from the products of symbiont photosynthesis ment and 2 to the shaded treatment; each pair was (Wilkinson 1987). Although phototrophic species were housed in a separate canopy, for a total of 10 control common on Pacific coral reefs, previous investigations and 10 treatment canopies. suggested that they were absent in the Caribbean Canopies consisted of a 15 cm × 15 cm square plastic (Wilkinson 1987). Since photosynthetic rates and meta- grating base attached by plastic cable ties to a 30 cm × bolic exchanges are not necessarily correlated, addi- 30 cm × 3 mm acrylic (Lucite-ES, INEOS Acrylics) roof tional measurements are often required to infer the with 4 PVC pipes of length 10 cm located in the cor- nutritional contribution of symbionts to their hosts (Mus- ners. Acrylic canopies were either transparent to vis- catine et al. 1981). Field-based shading experiments pro- ible light for control treatments or opaque for shaded vide direct evidence for the ultimate role of photosym- treatments, which reduced the light availability to bionts by testing the effects of reduced sunlight exposure approximately 7% of ambient irradiance. Each sponge on symbiont load and host sponge growth and survival. was measured for initial wet mass (after a standardized Such experiments have been conducted for Indo-Pacific blotting with paper towels) and displacement volume, (e.g. Thacker 2005) and Mediterranean sponges (e.g. then attached to respective control or shaded canopies Arillo et al. 1993). and deployed in the field for 6 wk. Erwin & Thacker: Caribbean sponge–cyanobacteria symbioses 141 One sponge per canopy was sampled each week to After incubation, sponges were removed from the monitor chl a concentrations;