Conforming to the status flow: The influence of altered habitat on fish body-shape characteristics

1. Anthropogenic stream impoundments often result in altered flow regimes and reservoir formations. Functional fish characteristics may change in response to altered flow conditions, and understanding the underlying mechanisms at work (e.g. natural selection and phenotypic plasticity) is of great importance in both evolutionary and conservation contexts. Cyprinella lutrensis is a widely distributed minnow native to stream systems in Missouri, U.S.A. Following impoundment of the Osage River, C. lutrensis currently occurs in the resultant Truman Reservoir and surrounding tributaries. To examine the impacts of this habitat alteration on functional fish characteristics, we hypothesised that C. lutrensis populations in altered systems with no-flow would show an overall less streamlined body shape as compared to those in flowing systems, assuming body streamlining is beneficial for locomotion in flowing environments.
2. We conducted field and laboratory studies to test effects of flow and no-flow on C. lutrensis body shape. Geometric morphometric techniques were used to quantify body shape in field and laboratory individuals, and non-structural lipid extractions were conducted to assess whether fat storage contributes to body shape under flow and no-flow treatments.
3. Morphological comparisons between C. lutrensis from Truman Reservoir and surrounding streams yielded significant differences in body shape, with greater body streamlining in stream populations. A laboratory experiment using artificial stream units revealed similar significant differences in body streamlining between flow and no-flow treatments within a single generation. In addition, individuals under no-flow treatment had significantly higher levels of non-structural lipids compared to flow treatment individuals.
4. Our results indicate that changes in flow regime can alter the evolutionary trajectories (i.e. related to morphology) of fish populations. We suggest that body shape alterations are likely to occur within and across generations in a cumulative manner, therefore, it is plausible that both natural selection and plastic response mechanisms play roles in defining minnow body shape following flow alteration.

     

Isolation and characterization of microsatellite loci from the truffle‐like ectomycorrhizal fungi Rhizopogon occidentalis and Rhizopogon vulgaris

We have isolated and characterized five microsatellite loci from Rhizopogon occidentalis and six loci from Rhizopogon vulgaris (Boletales, Basidiomycota). Microsatellite variation was assessed using 32 R. occidentalis and 48 R. vulgaris individuals from four populations in California. The number of alleles across populations ranged from two to 10 for R. occidentalis and three to eight for R. vulgaris. Expected heterozygosity values within populations ranged from 0.00 to 0.85 for R. occidentalis and 0.00 to 0.75 for R. vulgaris. These are the first microsatellite loci isolated for R. occidentalis and R. vulgaris and will be useful in the examination of their population genetic structure.     

Biogeography of ‘Cyprinella lutrensis’: intensive genetic sampling from the Pecos River ‘melting pot’ reveals a dynamic history and phylogenetic complexity

Thorough sampling is necessary to delineate lineage diversity for polytypic ‘species’ such as Cyprinella lutrensis. We conducted extensive mtDNA sampling (cytochrome b and ND4) from the Pecos River, Rio Grande, and South Canadian River, New Mexico. Our study focussed on the Pecos River due to its complex geological history and potential to harbour multiple lineages. We used geometric–morphometric, morphometric, and meristic analyses to test for phenotypic divergence and combined nucDNA with mtDNA to test for cytonuclear disequilibrium and combined our sequences with published data to conduct a phylogenetic re‐assessment of the entire C. lutrensis clade. We detected five co‐occurring mtDNA lineages in the Pecos River, but no evidence for cytonuclear disequilibrium or phenotypic divergence. Recognized species were interspersed amongst divergent lineages of ‘C. lutrensis’. Allopatric divergence among drainages isolated in the Late Miocene and Pliocene apparently produced several recognized species and major divisions within ‘C. lutrensis’. Pleistocene re‐expansion and subsequent re‐fragmentation of a centralized lineage founded younger, divergent lineages throughout the Rio Grande basin and Edwards Plateau. There is also evidence of recent introductions to the Rio Grande, Pecos and South Canadian Rivers. Nonetheless, deeply divergent lineages have coexisted since the Pleistocene.     

Ten new microsatellite loci for analysis of genetic diversity in isolated populations of the Alpine land snail <Emphasis Type="Italic">Cylindrus obtusus</Emphasis>

In this study we characterized 10 polymorphic microsatellite markers for the land snail Cylindrus obtusus, an endemism of the Austrian Alps with a distribution in isolated populations above approximately 1,600 m. The microsatellite loci were analyzed in 44 individuals from two populations. Number of alleles per locus ranged between two and eight. Observed heterozygosity ranged between 0.00 and 1.00, and expected heterozygosity between 0.09 and 0.72. No significant linkage disequilibrium was found between pairs of loci. One of the sampled populations (Dachstein) showed no deviation from Hardy–Weinberg equilibrium and no presence of null alleles, whereas the other one (Schneeberg) did. These diverging results probably reflect differences in population structure rather than characteristics of the microsatellite loci and underline the usefulness of these markers for studying genetic diversity, population structure and differentiation in C. obtusus.     

Development and optimization of microsatellite DNA primers for boreal owls (Aegolius funereus)

We developed 22 microsatellite loci for boreal owls (Aegolius funereus). We genotyped 275 unrelated boreal owls (Aegolius f. richardsoni) and 36 unrelated Tengmalm's owls (Aegolius f. funereus) using seven loci that were polymorphic and did not have detectable null alleles. Among North American and Scandinavian boreal owls, respectively, allelic diversity ranged from three to 11 alleles and from one to 11 alleles, observed heterozygosity ranged from 0.31 to 0.80 and from 0.00 to 0.81, and expected heterozygosity ranged from 0.28 to 0.81 and from 0.00 to 0.81. These markers appeared to amplify DNA in six other Strigidae species.     

Identification and characterization of eight microsatellite loci in Machilus pseudokobu (Lauraceae), an endemic species of the Bonin Islands

Eight microsatellite loci were identified and characterized for the endangered Machilus pseudokobu (Lauraceae), an endemic tree species of the Bonin Islands. The observed number of alleles at each locus ranged from 1 to 20 with an average of 6.2, and the expected heterozygosity ranged from 0.00 to 0.83 with an average of 0.47. All eight loci were screened in cross-amplification tests for two other endemic Machilus species that also inhabit the Bonin Islands. All loci were successfully amplified in these species.     

Current and historical hybridization with differential introgression among three species of cyprinid fishes (genus <Emphasis Type="Italic">Cyprinella</Emphasis>)

Hybridization is common among freshwater fishes, particular among the Cyprinidae. We used two mitochondrial genes and one nuclear gene to characterize hybridization among two species pairs of Cyprinella in southwestern North America. Genalogical patterns revealed that C. lutrensis and C. venusta are currently hybridizing in several localities producing apparent F₁, F₂ and backcross generations, yet there was no evidence for introgression outside of local hybrid zones. Alternatively, mitochondrial haplotypes from C. lutrensis appear to have introgressed into a C. lepida population in the Nueces River completely replacing the native C. lepida haplotype. There was no evidence of introgression of nuclear DNA and there does not appear to be ongoing hybridization. The population of C. lepida from the nearby Frio River exhibits no evidence of hybridization with C. lutrensis. Thus, contact between C. lutrensis and C. venusta results in the formation of localized hybrid swarms, while contact between C. lutrensis and C. lepida has resulted in complete mitochondrial introgression in the Nueces River or no apparent hybridization in the Frio River. The three different outcomes of contact between these species illustrate the variable nature of interspecific reproductive interactions and provide an excellent system in which to better understand the factors influencing hybridization among freshwater fishes.     

Microsatellite loci for the stingless bee Melipona rufiventris (Hymenoptera: Apidae)

Eight microsatellite primers were developed from ISSR (intersimple sequence repeats) markers for the stingless bee Melipona rufiventris. These primers were tested in 20 M. rufiventris workers, representing a single population from Minas Gerais state. The number of alleles per locus ranged from 2 to 5 (mean = 2.63) and the observed and expected heterozygosity values ranged from 0.00 to 0.44 (mean = 0.20) and from 0.05 to 0.68 (mean = 0.31), respectively. Several loci were also polymorphic in M. quadrifasciata, M. bicolor, M. mandacaia and Partamona helleri and should prove useful in population studies of other stingless bees.     

Isolation and characterization of six microsatellites in the Mexican beaded lizard Heloderma horridum

Six polymorphic microsatellite loci were isolated and characterized in the threatened Mexican beaded lizard, Heloderma horridum. The number of alleles per locus ranged from three to 12, with observed heterozygosity estimates ranging from 0.00 to 0.77, and expected heterozygosity estimates ranging from 0.00 to 0.73. These microsatellites will provide a valuable tool for the investigation of the genetic variation and structure of both wild and captive H. horridum populations.     

Development of microsatellite markers for the endangered semi‐shrub Chimaphila umbellata (Ericaceae)

Based on the DNA sequencing reads obtained using 454 pyrosequencing, primers amplifying 16 microsatellite loci were developed for the endangered semi‐shrub Chimaphila umbellata, which occurs sporadically in the Japanese Archipelago. These 16 loci were polymorphic in the populations sampled from the Hokkaido and Tohoku Districts; the mean number of alleles was 3.31 and 3.44, and the mean expected heterozygosity was 0.42 and 0.44, respectively. These loci were not linked to each other and contained no null alleles. Amplification using these primers was also tested in the congeneric species C. japonica, but only three of them successfully amplified DNA of the species. These markers will be used to examine genetic diversity and genetic differentiation in populations of C. umbellata.     

Isolation and characterization of polymorphic microsatellite markers in <Emphasis Type="Italic">Cupressus </Emphasis><Emphasis Type="Italic">chenggiana</Emphasis> S. Y. Hu (Cupressaceae)

Cupressus chenggiana S. Y. Hu (Cupressaceae) is an endemic and endangered conifer species in southwest China. In order to study the population genetics and design the effective conservation methods, we aimed to develop microsatellite primers for this species in the present study. We developed eight new microsatellite loci for this species through biotin capture method. Polymorphism of each locus was further assessed in 18 individuals from three geographically distant populations. The number of alleles per locus ranged from 6 to 11 with an average of 8.13. The observed and expected heterozygosities ranged from 0.219 to 0.296 and from 0.374 to 0.470, with averages of 0.254 and 0.417, respectively. We further found that three of nine microsatellite loci developed previously for another congeneric species showed polymorphic banding patters. We performed primer-crossing tests of these loci in the other two congeneric species which are closely related to C. chenggiana (C. gigantea and C. duclouxiana). These microsatellite markers would be effective for analyzing genetic diversity and population genetic structure of this species and its morphological differentiation with the close relatives.     

Genetic diversity and structure of the endemic Caesalpinia hintonii complex (Leguminosae: Caesalpinioideae) in Mexico

The Caesalpinia hintonii complex is formed by five endemic species (C. hintonii, C. laxa, C. macvaughii, C. melanadenia and C. epifanioi) occurring in central Mexico. This species complex is under incipient genetic divergence as by-product of local adaptations in reproductive and morphological traits to different habitats. We estimate the genetic variation and structure of populations of this species complex to assess the extent of genetic differentiation among populations and related species along its geographic distribution. Estimations of genetic diversity and structure were done based on ten enzymes and 18 loci. Mean number of alleles per locus ranged from 1.5 to 1.9. Polymorphic loci ranged from 42.1 to 68.4. Observed (H_o: range 0.191–0.275) and expected (H_e: range 0.205–0.317) heterozygosities in this complex were higher compared with other endemic and legume species. Neis genetic diversity estimates showed that most genetic variation was found within (H_S = 0.325) rather than among populations (D_ST = 0.085). Populations of the species C. hintonii showed a considerable genetic differentiation (F_ST = 0.207). The results of genetic diversity and structure within and among populations are in accord with the great morphological differentiation described for this species complex.     

Eight microsatellite markers for the San Esteban chuckwalla,Sauromalus varius

We characterize eight polymorphic microsatellite loci for the San Esteban chuckwalla, Sauromalus varius, which are being used as markers in a study of the genetic results from a decade of captive breeding in this endangered species. Observed heterozygosity estimates for S. varius ranged from 0.00 to 0.30, whereas expected heterozygosity estimates ranged from 0.06 to 0.58.     

Microsatellite loci from Russula brevipes,a common ectomycorrhizal associate of several tree species in North America

Six microsatellite loci were isolated and characterized from genomic libraries enriched for (CA)_n (GA)_n (ATG)_n, and (CAG)_n, microsatellite motifs from Russula brevipes, a common ectomycorrhizal fungus that forms mutualisms with several species of trees in North America. The polymerase chain reaction primers were tested on 27 sporocarps of R. brevipes sampled in Douglas fir (Pseudotsuga menziesii), grey pine (Pinus sabiniana), and Sitka spruce (Picea sitchensis) stands. The number of alleles per locus ranged from two to 14 with expected heterozygosity values from 0.00 to 0.92 within populations. These are the first six microsatellite loci characterized from Russula brevipes that can be used for estimating genotypic diversity and population structure.     

Assessment of allozyme variation among New Zealand populations of Gracilaria chilensis (Gracialiares,Rhodophyta) using starch-gel electrophoresis

New Zealand populations of Gracilaria chilensis are uniform in anatomical reproductive characteristics but vary morphologically and have been found to separate into two distinct groups with respect to agar methylation level, namely low (24–30%) and high (43–47%). To investigate the genetic variation within New Zealand populations of this species, 14 isozyme loci detected by starch-gel electrophoresis were examined in 17 wild populations from a wide range of localities, and in cultures derived from these populations. Five of these loci were polymorphic, but the genetic variation within populations was low: of the 17 populations examined, 15 were fixed at all loci (heterozygosity 0.000) and in the remaining two populations the observed heterozygosity was still low (0.004 and 0.011). The genetic distances between the populations ranged from 0.00 to 0.43. UPGMA cluster analysis separated the populations into two groups, a northern group and a group found throughout the country. Although these two groups do not correlate with the two groups based on agar methylation level at every locality, the correlation is sufficiently striking to merit further investigation.     

Polymorphic microsatellite loci in the sponge Crambe crambe (Porifera: Poecilosclerida) and their variation in two distant populations

Seven polymorphic microsatellite loci were characterized in the marine encrusting sponge Crambe crambe from a partial genomic DNA‐enriched library. Preliminary data on allelic variation of these loci in two distant populations of C. crambe are presented to assess their potential utility as high‐resolution genetic markers for this species. The number of alleles per locus ranged between three and 16 and the distributions of allele frequencies differed considerably between the two populations, indicating a marked genetic differentiation between them. These are the first microsatellite loci reported from any species in the phylum.     

Identification of 20 polymorphic microsatellite loci in European crow (Corvus corone) from existing passerine loci

The European crow (Corvus corone) occurs in two subspecies (or species) with distinct plumage coloration: the black carrion crow (C. c. corone) and the grey and black hooded crow (C. c. cornix). We tested 42 passerine microsatellite loci for amplification in the European crow and identified 20 loci that were both polymorphic and easy to score. In 50 individuals sampled in the Danish part of the species’ pan‐European hybrid zone, the number of alleles ranged between two and 21. One locus deviated from Hardy–Weinberg equilibrium and had a high estimated null allele frequency. These 20 loci were highly successful in amplifying polymorphic products also in other crow populations and in another Corvidae species, the rook (Corvus frugilegus).     

Isolation of polymorphic microsatellite loci from the endangered mudflat crab Chasmagnathus convexus (Crustacea: Decapoda: Varunidae)

Chasmagnathus convexus is a mudflat crab that is distributed throughout East Asia. However, most populations of this species in Korea are endangered due to continuing seashore developments and habitat fragmentations. In this study, we isolated and characterized nine microsatellite loci from C. convexus. The number of alleles per locus ranged from 3 to 23. The expected heterozygosities ranged from 0.145 to 0.974 and the observed heterozygosities ranged from 0.150 to 0.947. Cross-species amplifications for 10 other Varunid crab species were performed. These microsatellite markers could be useful for planning conservation strategies for C. convexus and related species.     

Development and characterization of the first microsatellite markers in the mangrove species Pelliciera rhizophorae Triana & Planchon

Pelliciera rhizophorae is a unique Neotropical mangrove species belonging to Pelliciera genus. We isolated eight microsatellite loci from this species. All loci were polymorphic and showed three to nine alleles per locus in Colombian Pacific and Caribbean populations. Polymorphic information content ranged from 0.46 to 0.69. Two loci (PeRh‐14 and PeRh‐19) showed null alleles on the Caribbean coast, which suggest genetic differentiation between Pacific and Caribbean populations of P. rhizophorae. Development of these microsatellite loci constitutes a new molecular tool to carry out studies in the genome of the species and to evaluate its population dynamics.