Brenden S. Holland

Genetics of marine bioinvasions

Anthropogenic biological introductions have captured the attention of marine scientists and resource managers in recent years. Human-mediated marine bioinvasions are presently acknowledged as often ecologically and financially devastating events. Despite recent increases in scientific interest and financial resources devoted to nonindigenous nuisance species globally, fundamental questions pertaining to taxonomic identity, geographic source, introduction vector and invasive population dynamics frequently remain unanswered. Ecological surveys based on photometric and observational methods are unable to identify cryptogenic taxa, source populations, multiple introductions, or genetic diversity. The twofold goal of this paper is to discuss the application of molecular genetic techniques to fundamental ecological questions pertaining to bioinvasions and to demonstrate the utility of DNA technology in providing data useful in the development of predictive models for marine bioinvasion science.

Contrasting phylogeography in three endemic Hawaiian limpets ( Cellana spp.) with similar life histories

The marine environment offers few obvious barriers to dispersal for broadcast‐spawning species, yet population genetic structure can occur on a scale much smaller than the theoretical limits of larval dispersal. Comparative phylogeographical studies of sympatric sister species can illuminate how differences in life history, behaviour, and habitat affinity influence population partitioning. Here we use a mitochondrial DNA marker (612 bp of cytochrome c oxidase subunit I) to investigate population structure of three endemic Hawaiian broadcast‐spawning limpets (Cellana spp.) with planktonic larvae that are competent to settle within 4 days. All three species exhibit significant population structure and isolation by distance, but the spatial scales of partitioning differ among the species. Cellana talcosa (n = 105) exhibits strong population structure between Kauai and the other main Hawaiian Islands (MHI) where the maximum channel width is 117 km, and no shared haplotypes were observed (ΦCT = 0.30, P < 0.001). In contrast, populations of Cellana exarata (n = 149) and Cellana sandwicensis (n = 109) exhibit weaker population structure within the MHI (ΦST = 0.03–0.04, P < 0.05), and between the MHI and the Northwestern Hawaiian Islands (ΦST = 0.03–0.09, P < 0.01), where the maximum channel width is 260 km. Biogeographical range and microhabitat use were correlated with estimates of dispersal, while phylogenetic affiliation and minimum pelagic larval duration were poor predictors of population partitioning. Despite similar life histories, these closely related limpets have contrasting patterns of population structure, illustrating the danger of relying on model species in management initiatives to predict population structure and dispersal in the context of marine protected area delineation.

Invasion Without a Bottleneck: Microsatellite Variation in Natural and Invasive Populations of the Brown Mussel Perna perna (L)

Abstract: Population-level genetic diversity of the brown mussel Perna perna was investigated using nuclear microsatellite markers in 6 natural and 6 invasive populations. A total of 448 individuals from 12 populations spanning the natural and introduced ranges of the brown mussel were scored for 2 polymorphic microsatellite loci. Wrights hierarchical F statistics (FST), Hardy-Weinberg equilibrium, Neis genetic distance, and other descriptive statistics were used to quantify geographic population subdivision, and to estimate the number of migrants per generation. FST values (0.007–0.042) revealed that genetic partitioning among populations was low. Microsatellite data revealed a slight difference in observed heterozygosity and no statistically significant differences in expected heterozygosity or allelic diversity between natural and introduced populations. Effective numbers of migrants (Nem) per generation ranged from 6 to 35 individuals. The potential significance of an invasive species with high genetic variation in terms of the risk of establishment and conservation implications is discussed.

Molecular biogeography and diversification of the endemic terrestrial fauna of the Hawaiian Islands

Oceanic islands have played a central role in biogeography and evolutionary biology. Here, we review molecular studies of the endemic terrestrial fauna of the Hawaiian archipelago. For some groups, monophyly and presumed single origin of the Hawaiian radiations have been confirmed (achatinelline tree snails, drepanidine honeycreepers, drosophilid flies, Havaika spiders, Hylaeus bees, Laupala crickets). Other radiations are derived from multiple colonizations (Tetragnatha and Theridion spiders, succineid snails, possibly Dicranomyia crane flies, Porzana rails). The geographic origins of many invertebrate groups remain obscure, largely because of inadequate sampling of possible source regions. Those of vertebrates are better known, probably because few lineages have radiated, diversity is far lower and morphological taxonomy permits identification of probable source regions. Most birds, and the bat, have New World origins. Within the archipelago, most radiations follow, to some degree, a progression rule pattern, speciating as they colonize newer from older islands sequentially, although speciation often also occurs within islands. Most invertebrates are single-island endemics. However, among multi-island species studied, complex patterns of diversification are exhibited, reflecting heightened dispersal potential (succineids, Dicranomyia). Instances of Hawaiian taxa colonizing other regions are being discovered (Scaptomyza flies, succineids). Taxonomy has also been elucidated by molecular studies (Achatinella snails, drosophilids). While molecular studies on Hawaiian fauna have burgeoned since the mid-1990s, much remains unknown. Yet the Hawaiian fauna is in peril: more than 70 per cent of the birds and possibly 90 per cent of the snails are extinct. Conservation is imperative if this unique fauna is to continue shedding light on profound evolutionary and biogeographic questions.

Islands within an island: phylogeography and conservation genetics of the endangered Hawaiian tree snail Achatinella mustelina

Mitochondrial DNA (mtDNA) sequences were used to evaluate phylogeographic structure within and among populations of three endangered Hawaiian tree snail species (n = 86). The primary focus of this investigation was on setting conservation priorities for Achatinella mustelina. Limited data sets for two additional endangered Hawaiian tree snails, A. livida and A. sowerbyana, were also developed for comparative purposes. Pairwise genetic distance matrices and phylogenetic trees were generated, and an analysis of molecular variance was performed on 675‐base pair cytochrome oxidase I gene sequences from multiple populations of Hawaiian tree snails. Sequence data were analysed under distance‐based maximum‐likelihood, and maximum‐parsimony optimality criteria. Within the focal species, A. mustelina, numbers of variable and parsimony informative sites were 90 and 69, respectively. Pairwise intraspecific mtDNA sequence divergence ranged from 0 to 5.3% in A. mustelina, from 0 to 1.0% in A. livida and from 0 to 1.9% in A. sowerbyana. For A. mustelina, population genetic structure and mountain topography were strongly correlated. Maximum genetic distances were observed across deep, largely deforested valleys, and steep mountain peaks, independent of geographical distance. However, in certain areas where forest cover is presently fragmented, little mtDNA sequence divergence exists despite large geographical scales (8 km). Genetic data were used to define evolutionarily significant units for conservation purposes including decisions regarding placement of predator exclusion fences, captive propagation, re‐introduction and translocation.

Diversification of sympatric broadcast-spawning limpets (Cellana spp.) within the Hawaiian archipelago

Speciation remains a central enigma in biology, and nowhere is this more apparent than in shallow tropical seas where biodiversity rivals that of tropical rainforests. Obvious barriers to gene flow are few and most marine species have a highly dispersive larval stage, which should greatly decrease opportunities for speciation via geographic isolation. The disparity in the level of geographic isolation for terrestrial and marine species is exemplified in Hawai‘i where opportunities for allopatric speciation abound in the terrestrial realm. In contrast, marine colonizers of Hawai‘i are believed to produce only a single endemic species or population, due to the lack of isolating barriers. To test the assertion that marine species do not diversify within Hawai‘i, we examine the evolutionary origin of three endemic limpets (Cellana exarata, C. sandwicensis and C. talcosa) that are vertically segregated across a steep ecocline on rocky shores. Analyses of three mtDNA loci (12S, 16S, COI; 1565 bp) and two nDNA loci (ATPSβ, H3; 709 bp) in 26 Indo‐Pacific Cellana species (N = 414) indicates that Hawai‘i was colonized once ∼3.4–7.2 Ma from the vicinity of Japan. Trait mapping demonstrates that high‐shore residence is the ancestral character state, such that mid‐ and low‐shore species are the product of subsequent diversification. The Hawaiian Cellana are the first broadcast‐spawners demonstrated to have speciated within any archipelago. The habitat stratification, extensive sympatry, and evolutionary history of these limpets collectively indicate a strong ecological component to speciation and support the growing body of evidence for non‐allopatric speciation in the ocean.

A geographic mosaic of passive dispersal: population structure in the endemic Hawaiian amber snail Succinea caduca (Mighels, 1845).

We used 276 cytochrome c oxidase subunit I (COI, 645 bp) and a subset of 84 16S large ribosomal subunit (16S, 451 bp) sequences to evaluate geographic patterns of genetic variation in 24 populations of the endemic Hawaiian land snail Succinea caduca spanning its range on six islands. Haplotype networks, gene tree topologies, pairwise molecular divergence and FST matrices suggest substantial geographic genetic structuring and complex dispersal patterns. Low nucleotide diversity and low pairwise molecular divergence values within populations coupled with higher between population values suggest multiple founder events. High overall haplotype diversity suggests diversification involving rare interpopulation dispersal, fragmentation by historical lava flows and variation in habitat structure. Within‐island rather than between‐island population comparisons accounted for the majority of molecular variance. Although 98% of 153 COI haplotypes were private by population, a Mantel test showed no evidence for isolation by distance. Mismatch distributions and population partitioning patterns suggest that genetic fragmentation has been driven by punctuated, passive dispersal of groups of closely related haplotypes that subsequently expanded and persisted in isolation for long periods (average > 2 million years ago), and that Pleistocene island connections may have been important in enhancing gene flow. Historical availability of mesic coastal habitat, together with effective dispersal may explain the long‐term persistence and unusual multi‐island distribution of this species, contrasting with the single‐island endemism of much of the Hawaiian biota.

Land Snail Models in Island Biogeography: A Tale of Two Snails*

Abstract: Oceanic islands have long been important in evolutionary biology. Land snails are a major component of oceanic island biotas and have much to offer as systems for addressing major questions in evolution and biogeography. We review patterns of within-archipelago biogeography and diversification in two large Hawaiian land snail groups, the Succineidae and the Achatinellinae. Molecular studies suggest that long-distance oceanic dispersal and colonization of the Hawaiian Islands has been rare but between-island dispersal has been far more common. Long-distance oceanic dispersal is the most important driver for deep phylogenetic divergence. Dispersal is also important within the archipelago, while among-island vicariant processes result in only a portion of tip clade diversity. The Achatinellinae are monophyletic but there is evidence of a deep phylogenetic split between the two Hawaiian succineid clades, a result of two independent colonizations reflecting two oceanic dispersal events. Hawaiian succineids have also dispersed to Samoa and Tahiti. Dispersal is an important biogeographical phenomenon, and its role in shaping distributions of island lineages should not be underestimated. Because of their relatively sedentary nature, yet a proclivity for long-distance passive dispersal, island snails can facilitate insights into mechanisms of evolutionary diversification. Important phylogenetic lessons are emerging from studies of island snails and such studies will eventually allow estimation of ages of species groups, speciation rates, timing of the processes involved in community assembly, and other dynamics, all of which are important contributions to the overall understanding of evolution.

Long-term fluctuations in circalunar Beach aggregations of the box jellyfish Alatina moseri in Hawaii, with links to environmental variability.

The box jellyfish Alatina moseri forms monthly aggregations at Waikiki Beach 8–12 days after each full moon, posing a recurrent hazard to swimmers due to painful stings. We present an analysis of long-term (14 years: Jan 1998– Dec 2011) changes in box jellyfish abundance at Waikiki Beach. We tested the relationship of beach counts to climate and biogeochemical variables over time in the North Pacific Sub-tropical Gyre (NPSG). Generalized Additive Models (GAM), Change-Point Analysis (CPA), and General Regression Models (GRM) were used to characterize patterns in box jellyfish arrival at Waikiki Beach 8–12 days following 173 consecutive full moons. Variation in box jellyfish abundance lacked seasonality, but exhibited dramatic differences among months and among years, and followed an oscillating pattern with significant periods of increase (1998–2001; 2006–2011) and decrease (2001–2006). Of three climatic and 12 biogeochemical variables examined, box jellyfish showed a strong, positive relationship with primary production, >2 mm zooplankton biomass, and the North Pacific Gyre Oscillation (NPGO) index. It is clear that that the moon cycle plays a key role in synchronizing timing of the arrival of Alatina moseri medusae to shore. We propose that bottom-up processes, likely initiated by inter-annual regional climatic fluctuations influence primary production, secondary production, and ultimately regulate food availability, and are therefore important in controlling the inter-annual changes in box jellyfish abundance observed at Waikiki Beach.

Molecular Systematics of the Endangered O'ahu Tree Snail Achatinella mustelina : Synonymization of Subspecies and Estimation of Gene Flow between Chiral Morphs

ABSTRACT The single-island endemic O‘ahu tree snail Achatinella mustelina Mighels, 1845 is an endangered species with dimorphic shell chirality, persisting in small populations restricted to upper-elevation native forest in the Wai‘anae Mountains. We used an intraspecific molecular phylogeny (n = 21 populations) to evaluate the validity of subspecies, most of them introduced by Welch in 1938 on the basis of shell characters, by determining whether the nominal subspecies examined correspond to detectable molecular partitions and to examine the possibility that opposing shell chirality acts as a reproductive isolating mechanism. We mapped the nominal subspecies and shell chiralities onto a mitochondrial DNA (mtDNA) phylogram based on 86 cytochrome c oxidase I gene fragments and the extant range of the species. Although clear genetic breaks and haplotype clusters with well-defined boundaries exist and correspond to topographic features, each of the five monophyletic clades in the gene tree contains multiple supposed subspecies, haplotypes are shared between different subspecies, and none of the 13 nominal subspecies exhibits monophyly. Furthermore the mtDNA clades in the gene tree do not correspond to observed patterns in shell chirality, and both chiralities occur in all clades. Thus, the subspecies are not taxonomically valid and have no relevance for conserving genetic diversity, and chirality differences do not appear to impart a reproductive barrier in this species. Therefore, all subspecies of A. mustelina are herein synonymized. (The) extraordinary Hawaiian biota would have continued its remarkable adaptive radiation at a rapid rate had man not caused its recent decimation. —E. C. Zimmerman (1979:38)