Population Connectivity of the Corallivorous Gastropod Coralliophila Abbreviata: Larval Dispersal Potential and Genetic Structure
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Population connectivity of the corallivorous gastropod Coralliophila abbreviata: larval dispersal potential and genetic structure Lyza Johnston University of Miami Rosenstiel School of Marine and Atmospheric Science 4600 Rickenbacker Causeway Miami, FL 33149 NOAA Award Number: NA07NMF4630096 Reporting Period: 7/1/07 - 12/31/09 Final Report Abstract. Coralliophila abbreviata (Lamarck) is a coral-eating gastropod found throughout the Caribbean and Western Atlantic. These snails cause substantial and chronic mortality of the two ESA listed ‘Threatened’ scleractinian coral species, Acropora palmata and A. cervicornis. The objectives of this project were to 1) determine the species status of Coralliophila abbreviata 2) estimate the dispersal potential of C. abbreviata veligers by determining the adult reproductive season and frequency, planktonic larval duration, larval settlement behavior and 3) estimate actual gene flow among C. abbreviata populations on a Caribbean-wide scale, using novel, highly polymorphic molecular markers (microsatellites). Eight polymorphic microsatellite loci of C. abbreviata were isolated and then characterized using 60 C. abbreviata from two geographically disparate populations (Key Largo, FL USA and the Grenadines). All loci were highly polymorphic with an average number of alleles per locus of 24 (range 13-34). Observed and expected heterozygosity values ranged from 0.375 - 0.969 and 0.877 - 0.981, respectively. Three loci deviated significantly from Hardy-Weinberg equilibrium in both populations, presumably due to null alleles. No linkage disequilibrium between pairs of loci was detected A total of 211 C. abbreviata from three coral host taxa and five geographical locations, spanning the species range, have been genotyped using the five loci that conformed to HW expectations. The mitochondrial Cytochrome b gene was sequenced for 82 individuals from two coral host taxa and three geographical locations. No significant genetic structure among hosts or geographically disparate populations was found, indicating that C. abbreviata is one plastic species with high population connectivity. Settlement assays were unsuccessful as culturing and raising C. abbreviata veliger larvae to competence and metamorphosis proved difficult. However, we did maintain a batch of larvae swimming in the water column for approximately 32 days. Little shell growth and no metamorphosis were observed. Such a protracted pelagic larval duration may be suitable to explain the lack of genetic structure observed for this species. Monthly in situ sampling of female C. abbreviata to ascertain reproductive patterns was abandoned as we wished to collect only one or two females from each coral colony throughout the study period to avoid complications with socially induced sex change of these sequential hermaphrodites and we could not locate enough infested Acropora palmata colonies in the study area support this sampling scheme. Therefore, we were only able to collect females every one to two months for a six month period. Females reproduced throughout the observed six month period and, although the degree of difference was variable, the proportion of females brooding egg capsules was consistently lower for snail populations found on Montastraea spp. than on A. palmata. These data support the hypothesis that snail populations feeding on acroporid corals have higher fitness than those feeding on other corals. This project has generated valuable information on the population genetic structure, reproductive patterns and early life-history of C. abbreviata, which to our knowledge, has not been previously reported. Furthermore, the first microsatellite markers were developed for C. abbreviata and thus constitute a valuable tool set that can be used to address a multitude of ecological and evolutionary questions for this species. NARRATIVE Reproductive Season and Frequency Population dynamics, connectivity, and genetic structure of a species are all highly dependent on adult reproductive biology (mode, season, and frequency of reproduction, and fecundity). For C. abbreviata, brood size increases as a power function with female shell length (Johnston and Miller 2007). Females feeding on acroporid corals reach larger sizes than those feeding on other prey species (Hayes 1990; Bruckner et al. 1997; Baums et al. 2003; Johnston and Miller 2007) and thus, on average, produce more offspring (Johnston and Miller 2007). However, very little is known about the reproductive patterns of C. abbreviata such as seasonality or frequency of reproduction or the variation in these patterns among snail populations on different coral hosts. The objective of this component of the project was to assess seasonal and host specific patterns in reproduction of C. abbreviata. The goal was to collect female snails monthly for 12 months from two coral host taxa (A. palmata and Montastraea spp.) from multiple sites in the Florida Keys National Marine Sanctuary (FKNMS) to determine their reproductive status. However, to avoid complications with socially induced sex change of these sequential hermaphrodites, we did not want to resample groups on individual coral colonies during the study period. We thus had to depart from the planned monthly sampling scheme as we could not locate enough Acropora palmata colonies in the study area that harbored snails that were not part of a long term monitoring project that requires that snail populations remain in tact. Female snails were thus only collected in June, August, September, and November of 2007, from 10 sites in the FKNMS (Table 1). At each site, SCUBA divers searched Acropora palmata and Montastraea spp. colonies for snails. C. abbreviata are protandrous hermaphrodites that exhibit host and sex specific variation in size frequency distributions: snails on A. palmata are larger than snails on Montastraea spp., and females are larger than males on both host taxa. Thus, to increase the likelihood of collecting females, only snails larger than 30 and 20 mm shell length were collected from A. palmata and Montastraea spp., respectively (size ranges are based on data from Johnston and Miller 2007). Only 1-2 snails were collected from any one coral colony. Because female C. abbreviata often expel egg capsules during collection, snails were taken off the coral and placed immediately into individual containers for transport back to the laboratory. At the laboratory, shell length was measured with vernier calipers to the nearest 0.01 mm. Shells were crushed and the soft tissue was examined to confirm sex (absence of penis above the right eye stock or presence of egg capsules in the mantle cavity). The presence and number of egg capsules was recorded along with the developmental stage of the embryos. The following criteria were used to classify embryonic development into four sequential stages: 1.) yellow egg mass concentrated at one pole of the transparent egg capsule (early); 2.) larger yellow embryos diffuse throughout the egg capsule (mid-early); 3.) orange embryos (mid-late); 4.) dark orange/brown swimming embryos filling the egg capsule (late). Stages 1 and 2 and 3 and 4 were then grouped into early and late stage embryos, respectively (Fig. 1). The proportion of females brooding egg capsules was averaged over sites for each coral host taxa and sampling time and compared between hosts and among months using two-way analysis of variance (ANOVA; Fig. 1). Female snails from both coral hosts brooded egg capsules at every sampling time. There was no significant effect of sampling date (p = 0.562), but there was a significant effect of coral host (p < 0.01) and a significant host*date interaction (p = 0.037) on the proportion of females brooding egg capsules (Table 2; Fig. 2). Although higher frequency sampling combined with non-destructive monitoring of individual snails is needed to fully elucidate the reproductive season, number of broods per year, and the length of intracapsular development for C. abbreviata, these data provide some first insight on host-specific reproductive patterns of C. abbreviata. Females reproduced throughout the observed six month period and, although the degree of difference was variable, the proportion of females brooding egg capsules was consistently lower for snail populations found on Montastraea spp. than on A. palmata. These data support the hypothesis that snail populations feeding on acroporid corals have higher fitness than those feeding on other corals. A.palmata Montastraea spp. Table 1 Number of female Brooding Brooding C. abbreviata (Nf) collected Date Site Nf (%) Nf (%) from two coral host taxa (A. 6/20/2007 EL 7 100 4 75 palmata and Montastraea DR 4 100 5 60 spp.) at each site and LG 5 100 6 66.67 sampling time and percent of ML 3 100 5 60 those females brooding egg 8/23/2007 EL 6 100 5 60 capsules. Reef sites in the GR 6 66.67 4 100 FKNMS include Elbow CF 4 50 4 75 (EL), Key Largo Dry Rocks TR 6 83.33 4 50 (DR), Little Grecian (LG), 9/22/2007 GR 3 100 5 60 Molasses (ML), Grecian SI 6 100 4 50 Rocks (GR), Carysfort (CF), TR 0 N/A 4 50 Turtle Rocks (TR), Sand WA 4 100 4 25 Island (SI), and French (FR). 11/30/2007 DR 1 100 4 75 EL 4 100 4 25 FR 3 66.67 5 40 LG 0 N/A 4 75 Fig. 1 Percent of embryos in early (blue) and late (red) stages of development. Embryos from female C. abbreviata found on Montastraea spp. (A) and A. palmata (B) corals. SS DF MS F p Table 2 Two-way ANOVA. The effects of Date 558.2 3 186.1 0.6979 0.562488 coral host and date on the 8515.1 1 8515.1 31.9381 0.000008 Host proportion of female C. Date*Host 2650.4 3 883.5 3.3137 0.037016 abbreviata brooding egg Error 6398.7 24 266.6 capsules. Fig. 2 Percent of female C. abbreviata from each coral host taxa, brooding egg capsules (averaged across sites ± 1 StdErr). See Table 2 for ANOVA results. Larval Growth and Pelagic Duration Connectivity of populations of benthic marine organisms is often dependent on the dispersal of planktonic larvae.