Genetic signatures of translocations and habitat fragmentation for two evolutionarily significant units of a protected fish species

Genetic population structure was evaluated for the White Sands pupfish (Cyprinodon tularosa), a protected fish species comprised of two Evolutionarily Significant Units (ESUs); the Malpais Spring ESU and the Salt Creek ESU. The Malpais Spring ESU is restricted to Malpais Spring, whereas the Salt Creek ESU includes the native Salt Creek population and two Salt Creek-derived populations at Mound Spring and Lost River; all three of these habitats are physically fragmented. We sampled the upper and lower reaches of the four populations, examining 13 DNA microsatellite loci from 40 individuals per population. As expected, significant genetic structure was observed between the two ESUs; Malpais Spring and Salt Creek. Substantial genetic drift was observed for the introduced Lost River population, with modest genetic drift for the introduced Mound Spring population. Taken together with ecological data, neither of the introduced populations successfully replicates the Salt Creek population. We also report significant reductions in genetic diversity for the upper reaches of both Salt Creek and Lost River, indicating that recent habitat changes have altered the genetic structure of these two populations. We consider these findings along with previously reported ecological data to develop guidelines for managing C. tularosa.     

Tests of phenotypic and genetic concordance and their application to the conservation of Panamanian golden frogs (Anura, Bufonidae)

Evolutionarily significant units (ESUs) differ in the extent to which they capture, or even consider, adaptive variation, and most such designations are based solely on neutral genetic differences that may not capture variation relevant to species' adaptabilities to changing environmental conditions. While concordant patterns of divergence among data sets (i.e. neutral and potentially non-neutral characters) can strengthen ESU designations, determining whether such criteria are met for highly variable taxa is especially challenging. This study tests whether previously defined ESUs for endangered Panamanian golden frogs (Atelopus varius and Atelopus zeteki) exhibit concordant variation among multiple phenotypic traits and mitochondrial DNA sequences, and the extent to which such divergence corresponds to environmental differences. Multivariate analyses identify phenotypic and genetic differentiation consistent with proposed ESUs and support the status of A. varius and A. zeteki as separate species. Moreover, the significant association detected between ESU co-membership and genetic similarity, which remained strong after removing the effect of geographic distance, also indicates that genetic differences are not simply due to isolation by distance. Two phenotypic characters (body size and the extent of dorsal black patterning) that differ among ESUs also co-vary with environmental differences, suggesting that to the extent that these phenotypic differences are heritable, variation may be associated with adaptive divergence. Lastly, discriminant function analyses show that the frogs can be correctly assigned to ESUs based on simultaneous analysis of multiple characters. The study confirms the merit of conserving the previously proposed golden frog ESUs as well as demonstrates the utility and feasibility of combined analyses of ecological, morphological and genetic variation in evaluating ESUs, especially for highly variable taxa.     

Phylogeography,Population Structure,and Conservation of the Javan Gibbon (<Emphasis Type="Italic">Hylobates moloch</Emphasis>)

An understanding of phylogeography and population genetics is needed for a comprehensive long-term conservation management strategy. The Javan gibbon (Hylobates moloch), an Endangered species endemic to the island of Java, has been protected since 1924 but is threatened by ongoing habitat loss, habitat degradation, and the wildlife trade. We studied the phylogeography and population genetic structure of the Javan gibbon, to define the number of Evolutionary Significant Units (ESUs) in the species, and the population genetic structure in each ESU. We sampled 47 individuals, analyzing 35 for variation in mitochondrial DNA control region, 41 for variation in 8 nuclear DNA microsatellites, and 13 for variation in 45 nuclear DNA single nucleotide polymorphisms (SNPs). We found support for two ESUs across the species range: a western ESU, extending from Ujung Kulon to Gunung Gede–Pangrango, and a central ESU, extending from Gunung Masigit–Simpang–Tilu to Gunung Slamet. Analysis of molecular variance and population structure analysis indicate significant structuring in the western ESU between Ujung Kulon and Gunung Halimun–Salak–Gede–Pangrango, and little to moderate structure in the central ESU, underscoring the importance of conserving as many populations as possible to preserve the full array of genetic diversity in this species. Our results will inform future more comprehensive population genetic surveys and the conservation genetic management of the Javan gibbon. This study demonstrates the importance of genetics when designing conservation management strategies for endangered primates.     

Genomic data indicate ubiquitous evolutionary distinctiveness among populations of California metalmark butterflies

Conservation geneticists have argued that evolutionarily significant units (ESUs) must be both genetically distinct and adaptively significant to be recognized for conservation protection. High-throughput DNA approaches can greatly increase the power to identify genetic distinctiveness, even if inferring adaptive significance remains a challenge. Here we present the first genomic evaluation of Lange’s metalmark, Apodemia mormo langei (Lepidoptera: Riodinidae), a U.S. federally endangered subspecies restricted to sand dune habitats in a single National Wildlife Refuge in California. Previous work based on very few genetic markers detected little genetic distinction for Lange’s metalmark. We use several thousand genome-wide single nucleotide polymorphisms to characterize the population structure of the A. mormo complex across California and determine if Lange’s metalmark qualifies as an ESU. We found that Lange’s metalmark is genetically identifiable, but is no more distinct than many other isolated populations across the study area. It remains unclear whether this genetic variation is adaptive, and so conservation efforts would benefit from more ecological characterization to determine conservation priorities.     

The gap between the concept and definitions in the Evolutionarily Significant Unit: the need to integrate neutral genetic variation and adaptive variation

The Evolutionarily Significant Unit (ESU) was conceptualized in 1986 as a conservation unit below the species level, theoretically applicable to a wide range of taxa. The concept has gained support, and various definitions or criteria, some of which are inconsistent with each other, have since been proposed. Recent critiques of the ESU have pointed out the dominance of definitions biased to the identification of long-term isolation or neutral genetic variation, which has largely ignored the adaptive components. We present here the validity of such claims and show how the ESU definitions have actually been applied in research. We surveyed scientific journals for original papers supporting ESU designations and determined who among the proponents of ESU definitions have gained wider support. Our results indicate that indeed there are inconsistencies with the original concept and with the existing definitions. Although the original concept recommended both ecological and genetic data as the basis for identification of ESUs, which reflect true evolutionary variation, recent definitions have become biased to either neutral genetic variation or adaptive variation. The definition which uses genetic data to assess neutral genetic variation (long-term isolation) has gained major support, and therefore validates the earlier claims. To bridge the gap between the original concept and the practical application, we propose the use of partial ESU and full ESU designations. The application of full ESU should be limited solely to when both information about neutral genetic variation and adaptive variation are available. In other cases, in which only a part of the variation is examined, we should use the term partial ESU (e.g., molecular-based ESU) and continue to investigate focal populations from other aspects of variations to designate full ESU.     

Adaptive evolutionary conservation: towards a unified concept for defining conservation units

Recent years have seen a debate over various methods that could objectively prioritize conservation value below the species level. Most prominent among these has been the evolutionarily significant unit (ESU). We reviewed ESU concepts with the aim of proposing a more unified concept that would reconcile opposing views. Like species concepts, conflicting ESU concepts are all essentially aiming to define the same thing: segments of species whose divergence can be measured or evaluated by putting differential emphasis on the role of evolutionary forces at varied temporal scales. Thus, differences between ESU concepts lie more in the criteria used to define the ESUs themselves rather than in their fundamental essence. We provide a context-based framework for delineating ESUs which circumvents much of this situation. Rather than embroil in a befuddled debate over an optimal criterion, the key to a solution is accepting that differing criteria will work more dynamically than others and can be used alone or in combination depending on the situation. These assertions constitute the impetus behind adaptive evolutionary conservation.     

Phylogeographic evidence links the threatened ‘Grampians’ Mountain Dragon (Rankinia diemensis Grampians) with Tasmanian populations: conservation implications in south-eastern Australia

The importance of protecting genetic diversity within a species is increasingly being recognised by conservation management authorities. However, discrepancies in conservation policy between authorities, such as state versus national bodies, can have significant implications for species management when they cross state boundaries. We conducted a phylogeographic study of the south-eastern Australian lizard Rankinia diemensis to identify evolutionary significant units (ESUs), including the endangered population from the Grampians National Park in western Victoria. Phylogenetic analyses of two gene regions (mtDNA: ND2; nuclear: RAG1) revealed high levels of genetic divergence between populations, indicating isolation over long evolutionary time frames. Based on criteria of genetic divergence and isolation, R. diemensis contains at least two ESUs that require specific management. We found that R. diemensis from the Grampians are closely related to Tasmanian populations, but that the divergence between these regions is great enough (3.7 % mtDNA) that they should be considered separate ESUs. However, we believe the close evolutionary ties between these two regions needs to be taken into account; yet under current practises, conservation management of subspecific ESUs relies on state-level efforts. We argue that another population that occurs on the Victorian coast also qualifies as an ESU and requires targeted conservation action. Rankinia diemensis provides a case-in-point of the discrepancy between the state-level approach of maintaining genetic variation within a species and the more conservative Commonwealth focus on conserving biodiversity at the species level.     

Microsatellite variation reveals weak genetic structure and retention of genetic variability in threatened Chinook salmon (<Emphasis Type="Italic">Oncorhynchus tshawytscha</Emphasis>) within a Snake River watershed

Pacific salmon (Oncorhynchus spp.) have been central to the development of management concepts associated with evolutionarily significant units (ESUs), yet there are still relatively few studies of genetic diversity within threatened and endangered ESUs for salmon or other species. We analyzed genetic variation at 10 microsatellite loci to evaluate spatial population structure and genetic variability in indigenous Chinook salmon (Oncorhynchus tshawytscha) across a large wilderness basin within a Snake River ESU. Despite dramatic 20th century declines in abundance, these populations retained robust levels of genetic variability. No significant genetic bottlenecks were found, although the bottleneck metric (M ratio) was significantly correlated with average population size and variability. Weak but significant genetic structure existed among tributaries despite evidence of high levels of gene flow, with the strongest genetic differentiation mirroring the physical segregation of fish from two sub-basins. Despite the more recent colonization of one sub-basin and differences between sub-basins in the natural level of fragmentation, gene diversity and genetic differentiation were similar between sub-basins. Various factors, such as the (unknown) genetic contribution of precocial males, genetic compensation, lack of hatchery influence, and high levels of current gene flow may have contributed to the persistence of genetic variability in this system in spite of historical declines. This unique study of indigenous Chinook salmon underscores the importance of maintaining natural populations in interconnected and complex habitats to minimize losses of genetic diversity within ESUs.     

Differences in Responses of Chinook Salmon to Climate Shifts: Implications for Conservation

Understanding how organisms respond to climate is critical for focusing the debate about ways to recover imperiled or manage exploited species. However, efforts to understand climate effects on biota are complicated by differences among species in life history and physiology. Even within a species it is not clear if different populations will react similarly to large-scale climate trends. Climate regimes exhibit basin-wide effects similar to the El Ni no Southern Oscillation but persist for decades. In the North Pacific Ocean, two regime shifts (abrupt changes from one regime to another) occurred in 1976–1977 and 1989–1990 and had wide ranging effects on many species. We examined the response of chinook salmon from 9 evolutionary significant units (ESUs) to the regime shifts. While there was an average decline in spawner numbers associated with the regime shifts, ESUs did not respond in a uniform manner: some ESUs declined, some did not respond and one may have increased. Four ESUs currently listed under the Endangered Species Act may have declined more across regime boundaries than did the five non-listed ones. Interpretation of this result depends on two ESUs: the Snake River spring/summer run and the Central Valley fall run. The Snake River ESU had the largest decline and most sampling effort. If this ESU was excluded from the analysis, there was no evidence that listed and non-listed stocks responded differently to the regimes. The Central Valley ESU is currently a candidate for listing. If this ESU is considered to be a threatened or endangered, then listed ESUs declined more on average than did non-listed ESUs across the regime boundaries regardless of the Snake River ESU. As a whole, these results suggest that long-term climate trends are important to the dynamics of chinook populations and that sub-units of a species (here ESUs) can respond differently to these regimes.     

Fish conservation in the land of steppe and sky: Evolutionarily significant units of threatened salmonid species in Mongolia mirror major river basins

Mongolia's salmonids are suffering extensive population declines; thus, more comprehensive fisheries management and conservation strategies are required. To assist with their development, a better understanding of the genetic structure and diversity of these threatened species would allow a more targeted approach for preserving genetic variation and ultimately improve long‐term species recoveries. It is hypothesized that the unfragmented river basins that have persisted across Mongolia provide unobstructed connectivity for resident salmonid species. Thus, genetic structure is expected to be primarily segregated between major river basins. We tested this hypothesis by investigating the population structure for three salmonid genera (Hucho, Brachymystax and Thymallus) using different genetic markers to identify evolutionarily significant units (ESUs) and priority rivers to focus conservation efforts. Fish were assigned to separate ESUs when the combined evidence of mitochondrial and nuclear data indicated genetic isolation. Hucho taimen exhibited a dichotomous population structure forming two ESUs, with five priority rivers. Within the Brachymystax genus, there were three B. lenokESUs and one B. tumensisESU, along with six priority rivers. While B. tumensiswas confirmed to display divergent mtDNA haplotypes, haplotype sharing between these two congeneric species was also identified. For T. baicalensis,only a single ESU was assigned, with five priority rivers identified plus Lake Hovsgol. Additionally, we confirmed that T. nigrescens from Lake Hovsgol is a synonym of T. baicalensis. Across all species, the most prominent pattern was strong differentiation among major river basins with low differentiation and weak patterns of isolation by distance within river basins, which corroborated our hypothesis of high within‐basin connectivity across Mongolia. This new genetic information provides authorities the opportunity to distribute resources for management between ESUs while assigning additional protection for the more genetically valuable salmonid rivers so that the greatest adaptive potential within each species can be preserved.     

The endangered Sonoran topminnow: Examination of species and ESUs using three mtDNA genes

There has been controversy over the species status of Sonoran topminnows and debate about the presence of ESUs in the Gila topminnow. From examination of sequence variation at 2626 base pairs over three mtDNA genes, we found a 29 (1.1%) nucleotide genetic difference between Gila and Yaqui topminnows. This provides strong support that these two taxa are separate species, Poeciliopsis occidentalis (Gila topminnow) and P. sonoriensis (Yaqui topminnow) and have been separated for approximately one million years. All the Gila topminnows within Arizona have the same sequence for the three mtDNA genes, that is, there is not reciprocal monophyly for mtDNA sequence data for the two previously designated ESUs. However, evidence of the unique habitat for Monkey Spring, its long-term isolation from other Gila topminnow habitats, and the presence of unique fish and invertebrate taxa in Monkey Spring support the designation of the Monkey Spring topminnows as an ESU. Finally, theoretical considerations using molecular data and estimates of heterozygosity and genetic distance for nuclear genes between populations of the Gila topminnow show that the lack of mtDNA variation is not inconsistent with the level and pattern of nuclear genetic variation observed.     

Variability in fish size/otolith radius relationships among populations of Chinook salmon

Back-calculation of growth trajectories from otolith microstructure is a valuable tool for understanding mechanisms underlying variability in growth among fish populations. We analyzed fish length/otolith radius relationships for Snake River spring/summer Chinook and Snake River fall Chinook salmon, listed as separate “Evolutionarily Significant Units” (ESUs) under the US Endangered Species Act, to determine whether these ESUs shared relationships. In addition, we analyzed otoliths from seven separate populations within the Snake River spring/summer Chinook ESU to assess the variability in relationships among populations, which are much more closely related than ESUs. We also examined several potential functional forms for the equations. We found that the separate ESUs had significantly different fish length/otolith radius relationships, but that variability in otolith growth rate could not explain the difference. Relationships among populations within the spring/summer Chinook ESU did not vary nearly as much as those between ESUs. The quadratic model and the power model fit the data equally well, and constraining these models to pass through a biological intercept (estimated fish length and otolith radius at hatching) resulted in only a slight decrease in model fit. To test the ability of the models to back-calculate fish lengths, we predicted the length at tagging for 17 PIT-tagged fall Chinook that were measured at release and at recapture. The back-calculation demonstrated little bias (<1 mm FL, on average) and relatively small standard deviation (～3.5 mm) for the best model. When we repeated the back-calculation with data from both ESUs combined, bias increased substantially (to 15 mm FL), demonstrating the importance of determining the proper taxonomic level at which to combine data within a species.     

More genetic data are needed before populations are mixed: response to ‘Sourcing Eurasian beaver Castor fiber stock for reintroductions in Great Britain and Western Europe’

Halley described three options for sourcing Eurasian beavers Castor fiber for reintroduction in Britain and Western Europe: (i) use animals from populations within a single western evolutionarily significant unit (ESU); (ii) mix animals from two or three western ESUs; and (iii) mix animals from eastern and western ESUs. Option three contravenes International Union for Conservation of Nature guidelines, so debate should focus on options one and two. We believe there is a need for further genetic analysis, before a final decision can be made. This decision should not be heavily influenced by cost or be determined by the genotypes of existing captive or escaped populations.     

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.     

Combined definition of seed transfer guidelines for ecological restoration in the French Pyrenees

Question: Can genetic tools combined with phytogeography help to define local plants and how geographically close the source population should be to the restoration site? Location: Subalpine and alpine French Pyrenees. Methods: The main phytogeographic boundaries in the French Pyrenees described by different authors were studied and this geographic pattern was compared with the results of genetic analysis for the four Pyrenean plants studied (Trifolium alpinum, Festuca eskia, Festuca gautieri and Rumex scutatus), based on random amplified polymorphic DNA (RAPD) marker analysis, unweighted pair‐group method with arithmetic averages (UPGMA) analysis and Mantel correlograms comparing geographic and genetic distances. Results: The genetic analysis allowed definition of two main evolutionarily significant units (ESUs) for the plants under study. Although the limit between the two zones was slightly variable according to the species considered, an eastern and a western ESU was consistently observed. This delineation was concordant with the main phytogeographic boundaries of the French Pyrenees. Conclusion: RAPD markers and associated Mantel correlograms can be useful to draw ESUs for individual species when the sampling intensity is relatively dense, and similarities were revealed between species sharing the same distribution range. This delineation allowed integration of infraspecific plant variation in the management of natural resources for revegetation in the Pyrenees. Nevertheless, caution is needed for the establishment of seed pools in order to maximize genetic diversity in each of the pools during collection and production.     

Tigers of Sundarbans in India: Is the Population a Separate Conservation Unit?

The Sundarbans tiger inhabits a unique mangrove habitat and are morphologically distinct from the recognized tiger subspecies in terms of skull morphometrics and body size. Thus, there is an urgent need to assess their ecological and genetic distinctiveness and determine if Sundarbans tigers should be defined and managed as separate conservation unit. We utilized nine microsatellites and 3 kb from four mitochondrial DNA (mtDNA) genes to estimate genetic variability, population structure, demographic parameters and visualize historic and contemporary connectivity among tiger populations from Sundarbans and mainland India. We also evaluated the traits that determine exchangeability or adaptive differences among tiger populations. Data from both markers suggest that Sundarbans tiger is not a separate tiger subspecies and should be regarded as Bengal tiger (P. t. tigris) subspecies. Maximum likelihood phylogenetic analyses of the mtDNA data revealed reciprocal monophyly. Genetic differentiation was found stronger for mtDNA than nuclear DNA. Microsatellite markers indicated low genetic variation in Sundarbans tigers (He= 0.58) as compared to other mainland populations, such as northern and Peninsular (Hebetween 0.67- 0.70). Molecular data supports migration between mainland and Sundarbans populations until very recent times. We attribute this reduction in gene flow to accelerated fragmentation and habitat alteration in the landscape over the past few centuries. Demographic analyses suggest that Sundarbans tigers have diverged recently from peninsular tiger population within last 2000 years. Sundarbans tigers are the most divergent group of Bengal tigers, and ecologically non-exchangeable with other tiger populations, and thus should be managed as a separate “evolutionarily significant unit” (ESU) following the adaptive evolutionary conservation (AEC) concept.     

The memory remains: application of historical DNA for scaling biodiversity loss

Few species worldwide have attracted as much attention in relation to conservation and sustainable management as Pacific salmon. Most populations have suffered significant reductions, many have disappeared, and even entire evolutionary significant units (ESUs) are believed to have been lost. Until now, no ‘smoking gun’ in terms of direct genetic evidence of the loss of a salmon ESU has been produced. In this issue of Molecular Ecology, Iwamoto et al. (2012) use microsatellite analysis of historical scale samples of Columbia River sockeye salmon (Oncorhynchus nerka) from 1924 ( Fig. 1 ) to ask the pertinent question: Do the historical samples contain salmon from extirpated populations or ESUs? They identified four genetic groups in the historical samples of which two were almost genetically identical to contemporary ESUs in the river, one showed genetic relationship with a third ESU, but one group was not related to any of the contemporary populations. In association with ecological data, the genetic results suggest that an early migrating Columbia River headwater sockeye salmon ESU has been extirpated. The study has significant importance for conservation and reestablishment of sockeye populations in the Columbia River, but also underpins the general significance of shifting baselines in conservation biology, and how to assess loss of genetic biodiversity. The results clearly illustrate the huge and versatile potential of using historical DNA in population and conservation genetics. Because of the extraordinarily plentiful historical samples and rapid advances in fish genomics, fishes are likely to spearhead future studies of temporal ecological and population genomics in non‐model organisms.

Figure 1 Open in figure viewer PowerPoint (a) Kokanee sampling site between Columbia and Windermere lakes on the upper Columbia River at Fairmont Hot Springs, British Columbia, Canada. (b) Bureau of Fisheries scale books that contained sockeye salmon (locally called ‘blueback’ salmon) scales collected from commercial fisheries during the 1920s in the lower Columbia River. (c) Kokanee on spawning beds in Kuskanax Creek, a tributary to Upper Arrow Lake, British Columbia. Photo credit Rick Gustafson and Jim Myers.     

Using microsatellite and MHC variation to identify species, ESUs, and MUs in the endangered Sonoran topminnow

Highly variable loci can provide insight into the recognition of species, evolutionarily significant units (ESUs) and management units (MUs). In general, the ESU and MU categories are thought to be reflective of adaptive differences between them. Here we examine this premise by presenting a comprehensive examination of genetic variation for both microsatellite loci and a major histocompatibility complex (MHC) locus, thought to be of adaptive significance, in the endangered Sonoran topminnow. The extent of variation for the microsatellite loci and the MHC gene within the 13 populations of the Gila topminnow is highly correlated, suggesting that nonselective factors have played an important role in influencing variation within and between populations for the MHC locus. Therefore, using all of these loci, we found that the eight natural populations of the Gila topminnow fell into two different ESUs, one of which had four different MUs. The source of the Boyce Thompson sample, a population that was used extensively for restocking, appeared to be Monkey Spring. The source of the Watson Wash population also appeared to be Monkey Spring (or Boyce Thompson). The newly colonized Santa Cruz River population, which had the most genetic variation of any Gila topminnow population, appeared to descend primarily from Sonoita Creek populations. The Yaqui topminnow, presently considered another subspecies of the Sonoran topminnow, was very distinct for both microsatellite (only two of 25 alleles found in the Yaqui were in any of the Gila topminnow samples) and MHC alleles (nonoverlapping sets of alleles for the two groups). As a result, it appeared that the taxonomic status of the two subspecies should be re-evaluated and that full species status for Gila and Yaqui topminnows was appropriate. There was evidence for the importance of long-term selection at the MHC locus in the higher rate of nonsynonymous than synonymous substitution. In addition, there appeared to have been a duplication of the MHC locus that was present in most of the fish in six of the natural populations of the Gila topminnow.     

Cryptic genetic diversity of <Emphasis Type="Italic">Neverita didyma</Emphasis> in the coast of China revealed by phylogeographic analysis: implications for management and conservation

Revealing cryptic biodiversity and understanding the processes that promote lineage diversification will provide valuable insights into management and protection of exploitable species. Neverita didyma is one of the most common marine species along the coast of China and possesses highly economic and nutritional value. Despite being heavily harvested each year, the genetic diversity of this species has never been assessed in the coastal areas of China. Here, we analyzed the diversity of this species based on the barcode region of the mitochondrial gene Cytochrome Oxidase subunit I (COI) and utilized different species delineation approaches to infer evolutionarily significant units (ESUs). Three distinct ESUs, with high genetic distance among each, were identified. Divergence time estimates suggested that the high genetic distances were probably associated with historical isolation of the marginal seas during Pleistocene low sea level periods. The three ESUs did not map to distinct geographical distribution, possibly attributing to the repeated isolation in different refugia and random postglacial recolonization. Moreover, N. didyma in Haizhou Bay deserves priority protection due to its unique ESU. To improve management regulations in the marine realm, our research also stresses the need for more empirical studies on genetic diversity of commercially exploited species in coastal environments of China.