Daniel J. Schmidt

Genes in Streams: Using DNA to Understand the Movement of Freshwater Fauna and Their Riverine Habitat

Today, decisions regarding the management and conservation of populations are often informed to some degree by population genetics. A fundamental measure sought by decisionmakers is the degree of connectivity between populations, which, when approached from a genetic perspective, may be influenced by many factors, making it difficult to generalize across taxa, habitats, or life histories. In the case of freshwater-limited fauna, the shared constraint of habitat structure (e.g., a dendritic stream network) imposed on all species in the system simplifies the task. A number of models have been proposed that predict how populations of taxa with different life-history traits and dispersal capabilities interact within structured freshwater habitats of this kind. In this article, we summarize these models and illustrate the general patterns of phylogeographic structure expected to occur under different scenarios of freshwater population connectivity. Additionally, we describe how the genetic structure of stream inhabitants can reflect historical changes in the physical structure of streams and thus open a window on past patterns of connectivity. A greater understanding of these concepts will contribute to an improved multidisciplinary approach to managing freshwater ecosystems.

Mogurnda adspersa microsatellite markers: multiplexing and multi-tailed primer tagging

A set of twelve microsatellite DNA loci were developed for the threatened Australian freshwater fish Mogurnda adspersa (Eleotridae). Primers were tailed with one of four 20-mer oligonucleotides for use in four-colour fluorescent detection and optimised for multiplex PCR. The loci were used to genotype individuals from two populations in the Pioneer River catchment of central Queensland, eastern Australia. Number of alleles per locus ranged from 2 to 33 and per locus heterozygosity ranged from 0.06 to 0.81. Successful cross-species amplification of all loci was achieved in the congener M. mogurnda. These markers will be used to estimate effective population size and to examine the relationship between flow regime and population demographic parameters.

Extreme Genetic Structure in a Small-Bodied Freshwater Fish, the Purple Spotted Gudgeon, Mogurnda adspersa (Eleotridae)

Freshwater fish are a group that is especially susceptible to biodiversity loss as they often exist naturally in small, fragmented populations that are vulnerable to habitat degradation, pollution and introduction of exotic species. Relatively little is known about spatial dynamics of unperturbed populations of small-bodied freshwater fish species. This study examined population genetic structure of the purple spotted gudgeon (Mogurnda adspersa, Eleotridae), a small-bodied freshwater fish that is widely distributed in eastern Australia. The species is threatened in parts of its range but is common in coastal streams of central Queensland where this study took place. Microsatellite (msat) and mitochondrial DNA (mtDNA) variation was assessed for nine sites from four stream sections in two drainage basins. Very high levels of among population structure were observed (msat F ST = 0.18; mtDNA ΦST = 0.85) and evidence for contemporary migration among populations was rare and limited to sites within the same section of stream. Hierarchical structuring of variation was best explained by stream section rather than by drainage basin. Estimates of contemporary effective population size for each site was low (range 28 – 63, Sibship method), but compared favorably with similar estimates for other freshwater fish species, and there was no genetic evidence for inbreeding or recent population bottlenecks. In conclusion, within a stable part of its range, M adspersa exists as a series of small, demographically stable populations that are highly isolated from one another. Complimentary patterns in microsatellites and mtDNA indicate this structuring is the result of long-term processes that have developed over a remarkably small spatial scale. High population structure and limited dispersal mean that recolonisation of locally extinct populations is only likely to occur from closely situated populations within stream sections. Limited potential for recolonisation should be considered as an important factor in conservation and management of this species.

Cytonuclear evidence for hybridogenetic reproduction in natural populations of the Australian carp gudgeon (Hypseleotris: Eleotridae)

Although most vertebrates reproduce sexually, a small number of fishes, amphibians and reptiles are known in which reproduction is asexual, i.e. without meiotic recombination. In fishes, these so‐called unisexual lineages usually comprise only females and utilize co‐occurring males of a related sexual species to reproduce via gynogenesis or hybridogenesis. Here, we examine patterns of microsatellite and mitochondrial DNA (mtDNA) variation in a widespread group of freshwater fishes (carp gudgeons; Hypseleotris spp.) to investigate a long‐standing proposal that this group includes unisexual forms. We show that the mtDNA genome of most carp gudgeons in tributaries of the Goulburn River belongs to one of two deeply divided clades (∼10% cyt b divergence) and that nuclear variation divides the same individuals into four distinct groups. Group 1 exhibits the genotypic proportions of a random mating population and has a 1:1 sex ratio. Two other groups are extremely sex‐biased (98% male, 96% female), exhibit excess heterozygosity at most loci and share at least one allele per locus with group 1. We propose that these two groups represent ‘unisexual’ hybridogenetic lineages and that both utilize co‐occurring group 1 as sexual host. Interestingly, the fourth distinct group appears to represent hybrid offspring of the two putative hybridogenetic lineages. The propagation of clonal haploid genomes by both males and females and the ability of these clones to unite and form sexually mature diploid hybrid offspring may represent a novel mechanism that contributes to the dynamics of coexistence between hybridogenetic lineages and their sexual hosts.

Genetic analysis of threatened Australian grayling Prototroctes maraena suggests recruitment to coastal rivers from an unstructured marine larval source population

Population genetic variation of Australian grayling Prototroctes maraena was examined to determine whether the dispersal strategy of this amphidromous species favours retention of larvae and juveniles in close proximity to their natal river, or mixing of populations via marine dispersal. Variation in microsatellite and mitochondrial DNA markers was unstructured and differentiation was indistinguishable from zero across four coastal rivers spanning approximately one-quarter of the continental range of the species. This result indicates that the marine larval and juvenile phase probably facilitates extensive gene flow among coastal rivers and agrees with a previous analysis of otolith chemistry that suggested larvae probably move into the sea rather than remain in estuaries. It appears likely that the dispersal strategy of P. maraena would enable recolonization of rivers that experience localized extinction provided that connectivity between freshwater habitats and the sea is sufficient to permit migration and that enough source populations remain intact to support viability of the wider population.

Population genetic structure in stream insects: what have we learned?

This chapter synthesizes data from 41 studies on 32 species of stream insects in an attempt to answer three questions: First, what is the major dispersal mechanism in aquatic insects, i.e. stream drift or adult flight? If it is stream drift, then genetic variation would be expected to fit the Stream Hierarchy Model of Meffe and Vrijenhoek (1988). If it is adult flight, populations would be expected to be panmictic at small scales, i.e. among populations in neighbouring catchments. Most stream insects with an adult flight stage do not fit the Stream Hierarchy Model, suggesting that adult flight is the major mechanism of dispersal. Second, at what scale are populations of stream insects structured? Across all studies, there was a signifi cant positive relationship between FST and geographic distance for studies using mitochondrial DNA. The isolation-by-distance relationship for allozyme studies was significant only when studies with high numbers of Hardy Weinberg Equilibrium (HWE) deviations were excluded. Third, what can recent analysis of DNA sequence data contribute to our understanding of historical processes affecting stream insects? Several recent phylogeographic studies using mitochondrial DNA sequence data provide evidence of population and range expansions and contractions, along with past fragmentation, all estimated to have occurred during the Pleistocene.

Low interbasin connectivity in a facultatively diadromous fish: evidence from genetics and otolith chemistry.

Southern smelts (Retropinna spp.) in coastal rivers of Australia are facultatively diadromous, with populations potentially containing individuals with diadromous or wholly freshwater life histories. The presence of diadromous individuals is expected to reduce genetic structuring between river basins due to larval dispersal via the sea. We use otolith chemistry to distinguish between diadromous and nondiadromous life histories and population genetics to examine interbasin connectivity resulting from diadromy. Otolith strontium isotope (87Sr:86Sr) transects identified three main life history patterns: amphidromy, freshwater residency and estuarine/marine residency. Despite the potential for interbasin connectivity via larval mixing in the marine environment, we found unprecedented levels of genetic structure for an amphidromous species. Strong hierarchical structure along putative taxonomic boundaries was detected, along with highly structured populations within groups using microsatellites (FST = 0.046–0.181), and mtDNA (ΦST = 0.498–0.816). The presence of strong genetic subdivision, despite the fact that many individuals reside in saline water during their early life history, appears incongruous. However, analysis of multielemental signatures in the otolith cores of diadromous fish revealed strong discrimination between river basins, suggesting that diadromous fish spend their early lives within chemically distinct estuaries rather than the more homogenous marine environment, thus avoiding dispersal and maintaining genetic structure.

Do lowland habitats represent barriers to dispersal for a rainforest mayfly, Bungona narilla, in south-east Queensland?

Long-distance dispersal might be an important mechanism for the maintenance of aquatic insect populations in heterogeneous landscapes. However, these events can be difficult to measure by direct observation because the techniques can be time-consuming, expensive and technically difficult. When dispersal results in gene flow within and between populations, patterns of variation can be detected by genetic methods. The levels of population genetic structuring and the relationship between gene flow and geographical distance were assessed in the mayfly species Bungona narilla (Harker, 1957) in rainforest streams in south-east Queensland that are separated by lowland habitats. An analysis of molecular variance based on mitochondrial DNA data, using a fragment of the cytochrome oxidase I gene, revealed significant differentiation between regions, suggesting that maternal gene flow was restricted. A nested clade analysis revealed patterns of historical (contiguous) range expansions and recent restricted gene flow along with some long-distance dispersal events. Our analyses have shown that populations of B. narilla are significantly structured throughout the species range in south-east Queensland and that the low elevation habitats separating the northern and southern populations are restricting gene flow to some extent.

Migration history and stock structure of two putatively diadromous teleost fishes, as determined by genetic and otolith chemistry analyses

Abstract: Migratory life cycles and population structure of 2 putatively diadromous Australian fishes were examined using otolith chemistry (87Sr/86Sr) and genetics (microsatellites and mitochondrial deoxyribonucleic acid [mtDNA]). Australian whitebait (Lovettia sealii) is widely considered to be one of only a few anadromous fish species in the southern hemisphere. The congolli or tupong (Pseudaphritis urvillii) is reported to undertake an unusual form of sexually segregated catadromous migration, where females switch habitats between marine and freshwater, while males remain in marine or estuarine environments. Sr-isotope profiles of L. sealii showed this species does not move into fully freshwater habitats during its life cycle, suggesting it should be considered semianadromous or estuarine-dependent, rather than truly anadromous. This life-history strategy is unique among the Galaxiidae. Lovettia sealii is regionally divided into at least 3 well differentiated genetic stocks: northern and southern Tasmanian coasts and mainland Australia. Sr-isotope profiles of P. urvillii showed that females are catadromous, with the early life history spent in the marine environment and a single migratory transition from marine to freshwater occurring at an early point in the life history. Lack of bidirectional adult migration between freshwater and the sea suggests that female P. urvillii are semelparous, returning to the marine habitat to mate with resident males after an extended period of freshwater residence. Pseudaphritis exhibit weak genetic structure across their mainland range. An isolation-by-distance relationship describes the genetic structure of this species, a pattern it shares with several other nearshore-restricted catadromous fishes.

Speciation in chestnut-shouldered fairy-wrens (Malurus spp.) and rapid phenotypic divergence in variegated fairy-wrens (Malurus lamberti): A multilocus approach

The chestnut-shouldered fairy-wrens comprise a subgroup of four species in the genus Malurus (Passeriformes: Maluridae). Collectively, they are widespread across the Australian continent but phenotypic variation is strongly structured geographically in just one species, M. lamberti. Earlier phylogenetic analyses of this group have been limited to one or two individuals for each species and have not represented all currently recognised subspecies of M. lamberti. Historically, the taxonomy and nomenclature of the M. lamberti complex has been debated, in part because of morphological similarities among its subspecies and another member of the group, M. amabilis. We reconstructed the phylogeny of all four species of chestnut-shouldered fairy-wrens including all four subspecies of M. lamberti using a mitochondrial gene (ND2), five anonymous nuclear loci and three nuclear introns. Phylogenetic analysis of the mitochondrial ND2 gene nests M. amabilis within M. lamberti rendering the latter paraphyletic. Individual nuclear gene trees failed to reliably resolve each of the species boundaries or the phylogenetic relationships found in the mtDNA tree. When combined, however, a strongly supported overall topology was resolved supporting the monophyly of M. lamberti and its sister species relationship to M. amabilis. Current subspecific taxonomy of M. lamberti was not concordant with all evolutionary lineages of M. lamberti, nominotypical M. l. lamberti being the only subspecies recovered as a monophyletic group from mtDNA. Some genetic structuring is evident and potential barriers to gene flow are discussed.