Species discovery in marine planktonic invertebrates through global molecular screening

Species discovery through large‐scale sampling of mitochondrial diversity, as advocated under DNA barcoding, has been widely criticized. Two of the primary weaknesses of this approach, the use of a single gene marker for species delineation and the possible co‐amplification of nuclear pseudogenes, can be circumvented through incorporation of multiple data sources. Here I show that for taxonomic groups with poorly characterized systematics, large‐scale genetic screening using a mitochondrial DNA marker can be a very effective approach to species discovery. Global sampling (120 locations) of 1295 individuals of 22 described species of eucalanid copepods identified 15 novel evolutionarily significant units (ESUs) within this marine holoplanktonic family. Species limits were tested under reciprocal monophyly at the mitochondrial (mt) gene 16S rRNA, and 13 of 15 lineages were reciprocally monophyletic under three phylogenetic inference methods. Five of these mitochondrial ESUs also received moderate support for reciprocal monophyly at the independently‐inherited nuclear gene, internal transcribed spacer 2 (ITS2). Additional support for the utility of mt DNA as a proxy for species boundaries in this taxon is discussed, including results from related morphological and biogeographic studies. Minimal overlap of intra‐ESU and inter‐ESU 16S rRNA genetic distances was observed, suggesting that this mt marker performs well for species discovery via molecular screening. Sampling coverage required for the discovery of new ESUs was found to be in the range of >50 individuals/species, well above the sampling intensity of most current DNA Barcoding studies. Large‐scale genetic screening can provide critical first data on the presence of cryptic species, and should be used as an approach to generate systematic hypotheses in groups with incomplete taxonomies.     

Dispersal-limited zooplankton distribution and community composition in new ponds

Twelve new experimental ponds were constructed identically, filled simultaneously, had similar physical and chemical properties, and were maintained with minimal manipulation. Colonizing zooplankton communities were sampled bi-weekly for one year. Rotifers dominated zooplankton communities in densities, biomass, and species number (47 of 61 observed species were rotifers). Only 14 species were observed in all 12 ponds; 9 were rotifers. Twenty-nine species (26 rotifers) were recorded in <-6 ponds. Species with high vagility exhibited greater viability. Ponds differed in zooplankton community composition throughout the year, due to differences in both vagility and viability among colonizing species.     

Selective feeding of Arctodiaptomus salinus (Copepoda, Calanoida) on co-occurring sibling rotifer species

1. Using two‐ and three‐dimensional video recordings, we examined the steps involved in predation that lead to the differential vulnerability of three sympatric rotifer sibling species (Brachionus plicatilis, B. ibericus and B. rotundiformis) to a co‐occurring, predatory, calanoid copepod (Arctodiaptomus salinus). 2. Brachionus rotundiformis, the smallest prey tested, was the most vulnerable with the highest encounter rate, probability of attack, capture and ingestion, and the lowest handling time. 3. Comparison of our results with those of a previous study shows that A. salinus is a more efficient predator than a co‐occurring cyclopoid copepod (Diacyclops bicuspidatus odessanus) feeding on these same rotifer species. However, despite its higher capture rates, A. salinus seems to be less selective than D. b. odessanus based on attack distances and prey handling times. 4. The differential vulnerability to both calanoid and cyclopoid copepod predation can help explain the coexistence and seasonal succession of these co‐occurring rotifer species.     

Coexistence of cryptic rotifer species: ecological and genetic characterisation of Brachionus plicatilis

1. The coexistence of five cryptic species of the rotifer species complex Brachionus plicatilis was investigated in four coastal Mediterranean ponds. Monthly sampling was undertaken for 15 months and species were characterised using allozyme electrophoresis. 2. We describe species‐diagnostic allozyme loci that can be used for rapid identification of these species. 3. The five species overlapped to some extent in their temporal use of the ponds, although some seasonal segregation was observed. 4. The match between temporal and spatial distribution and limnological conditions suggested ecological specialisation in some cases, although we found striking examples of extensive seasonal overlap. 5. Our results indicate that sympatry of cryptic rotifer species is largely because of seasonal ecological specialisation, which allows seasonal succession and partitioning of resources. The processes that might be involved in the long periods of overlapping seasonal distributions of species which are potentially competitors are discussed. This example illustrates that the ‘paradox of the plankton’ is more the rule than the exception.     

Fish as vectors in the dispersal of Bythotrephes cederstroemi: diapausing eggs survive passage through the gut

• 1 Bythotrephes cederstroemi (Crustacea: Onychopoda: Cercopagidae) is an introduced invertebrate predator currently spreading through the Laurentian Great Lakes region of North America. We examined a previously unsuspected way in which B. cederstroemi may be dispersed by fish by their consumption of diapausing eggs.
• 2 Ninety‐four percentage of the mature B. cederstroemi diapausing eggs consumed by fish were egested apparently intact. This proportion is considerably above previous estimates for the ephippial eggs of Daphnia. The hatching success of diapausing eggs was compared among four categories: (a) eggs released naturally by B. cederstroemi (control, 73% hatched) (b) eggs released during ‘stressful confinement’ (46% hatched) (c) eggs dissected from dead females (13% hatched) and (d) eggs recovered from faecal pellets following consumption by fish (viable gut passage experiment, 41% hatched).
• 3 Samples of small fish and B. cederstroemi were collected simultaneously. Examination of gut contents revealed that fish contained B. cederstroemi diapausing eggs and that B. cederstroemi bearing resting eggs were consumed selectively over those without eggs. Moreover, fish selected B. cederstroemi bearing mature rather than immature diapausing eggs.
• 4 The fact that diapausing eggs survive gut passage is important for the dispersal of B. cederstroemi. Fish often move between the pelagic and littoral zones of lakes and may thus disperse diapausing eggs widely. Fish may also move between lakes connected by river systems and can be caught and passively dispersed by anglers or piscivorous birds. Our results demonstrate the potential for fish to act as vectors in the spread of B. cederstroemi.

     

Deeply divergent lineages of the widespread New Zealand amphipod Paracalliope fluviatilis revealed using allozyme and mitochondrial DNA analyses

1. We evaluated the population genetic structure of the common New Zealand amphipod Paracalliope fluviatilis using eight allozyme loci, and the mitochondrial cytochrome oxidase c subunit I (COI) gene locus. Morphological analyses were also conducted to evaluate any phenotypic differences. Individuals belonging to P. fluviatilis were collected from a total of 14 freshwater fluvial habitats on the North and South Islands, New Zealand. 2. We found evidence for strong genetic differentiation among locations (Wright's F_ST > 0.25), and fixed differences (non‐shared alleles) at two of the eight allozyme loci indicating the possibility of previously unknown species. Analysis of a 545‐bp fragment of the COI locus was mostly congruent with the allozyme data and revealed the same deeply divergent lineages (sequence divergences up to 26%). 3. Clear genetic breaks were identified between North Island and South Island populations. North Island populations separated by <100 km also showed genetic differences between east and west draining watersheds (sequence divergence >12%). Accordingly, present‐day dispersal among hydrologically isolated habitats appears minimal for this taxon. 4. Although population differences were clearly shown by allozyme and mtDNA analyses, individuals were morphologically indistinguishable. This suggests that, as in North American and European taxa (e.g. Hyalella and Gammarus), morphological conservatism may be prevalent among New Zealand's freshwater amphipods. We conclude that molecular techniques, particularly the COI gene locus, may be powerful tools for resolving species that show no distinctive morphological differences.     

Birds introduced in new areas show rest disorders

All colonizing individuals have to settle in a novel, conspecific-free environment. The introduction process should be poorly compatible with a good rest. We compared the resting behaviour of radio-tagged house sparrows (Passer domesticus) experimentally translocated into new, conspecific-free areas (introduced individuals, n = 10), with that of translocated sparrows that settled in naturally established populations (controls, n = 5). Resting habits of introduced sparrows markedly differed from those of control birds: they did not vocalize before going to roost, they changed their roosting habitat and they roosted 24 ± 7 min later and departed 13 ± 4 min earlier from the roost, resulting in a 5% rest debt. Because colonizing a new environment is expected to require heightened cognitive and physical activities, which in turn are constrained by the quality and duration of rest, we hypothesize that rest disorders and resulting cognitive impairments of newly released individuals could functionally contribute to the low post-release survival observed in (re)introduction attempts.     

Shifting dispersal modes at an expanding species’ range margin

While it is generally recognized that noncontiguous (long‐distance) dispersal of small numbers of individuals is important for range expansion over large geographic areas, it is often assumed that colonization on more local scales proceeds by population expansion and diffusion dispersal (larger numbers of individuals colonizing adjacent sites). There are few empirical studies of dispersal modes at the front of expanding ranges, and very little information is available on dispersal dynamics at smaller geographic scales where we expect contiguous (diffusion) dispersal to be prevalent. We used highly polymorphic genetic markers to characterize dispersal modes at a local geographic scale for populations at the edge of the range of a newly invasive grass species (Brachypodium sylvaticum) that is undergoing rapid range expansion in the Pacific Northwest of North America. Comparisons of Bayesian clustering of populations, patterns of genetic diversity, and gametic disequilibrium indicate that new populations are colonized ahead of the invasion front by noncontiguous dispersal from source populations, with admixture occurring as populations age. This pattern of noncontiguous colonization was maintained even at a local scale. Absence of evidence for dispersal among adjacent pioneer sites at the edge of the expanding range of this species suggests that pioneer populations undergo an establishment phase during which they do not contribute emigrants for colonization of neighbouring sites. Our data indicate that dispersal modes change as the invasion matures: initial colonization processes appear to be dominated by noncontiguous dispersal from only a few sources, while contiguous dispersal may play a greater role once populations become established.     

Evolution in caves: Darwin’s ‘wrecks of ancient life’ in the molecular era

Cave animals have historically attracted the attention of evolutionary biologists because of their bizarre ‘regressive’ characters and convergent evolution. However, understanding of their biogeographic and evolutionary history, including mechanisms of speciation, has remained elusive. In the last decade, molecular data have been obtained for subterranean taxa and their surface relatives, which have allowed some of the classical debates on the evolution of cave fauna to be revisited. Here, we review some of the major studies, focusing on the contribution of phylogeography in the following areas: biogeographic history and the relative roles of dispersal and vicariance, colonization history, cryptic species diversity and modes of speciation of cave animals. We further consider the limitations of current research and prospects for the future. Phylogeographic studies have confirmed that cave species are often cryptic, with highly restricted distributions, but have also shown that their divergence and potential speciation may occur despite the presence of gene flow from surface populations. Significantly, phylogeographic studies have provided evidence for speciation and adaptive evolution within the confines of cave environments, questioning the assumption that cave species evolved directly from surface ancestors. Recent technical developments involving ‘next generation’ DNA sequencing and theoretical developments in coalescent and population modelling are likely to revolutionize the field further, particularly in the study of speciation and the genetic basis of adaptation and convergent evolution within subterranean habitats. In summary, phylogeographic studies have provided an unprecedented insight into the evolution of these unique fauna, and the future of the field should be inspiring and data rich.     

潜生体定植的原位观察技术研究

对小鼠结肠组织原位标本进行潜生体定植的实验研究。染色采用美蓝初染后,高锰酸钾复染的方法。方法简便、有效。     

Dispersal strategies, secondary range expansion and invasion genetics of the nonindigenous round goby, Neogobius melanostomus, in Great Lakes tributaries

Dispersal strategies are important mechanisms underlying the spatial distribution and colonizing ability of all mobile species. In the current study, we use highly polymorphic microsatellite markers to evaluate local dispersal and colonization dynamics of the round goby (Neogobius melanostomus), an aquatic invader expanding its range from lake to river environments in its introduced North American range. Genetic structure, genotype assignment and genetic diversity were compared among 1262 round gobies from 20 river and four lake sites in three Great Lakes tributaries. Our results indicate that a combination of short-distance diffusion and long-distance dispersal, collectively referred to as 'stratified dispersal', is facilitating river colonization. Colonization proceeded upstream yearly (approximately 500 m/year; 2005-2009) in one of two temporal replicates while genetic structure was temporally stable. Contiguous dispersal from the lake was observed in all three rivers with a substantial portion of river fish (7.3%) identified as migrants. Genotype assignment indicated a separate introduction occurred upstream of the invasion front in one river. Genetic diversity was similar and relatively high among lake and recently colonized river populations, indicating that founder effects are mitigated through a dual-dispersal strategy. The remarkable success of round goby as an aquatic invader stresses the need for better diffusion models of secondary range expansion for presumably sessile invasive species.