Characterization of microsatellite loci in Sacramento perch (Archoplites interruptus)

We characterized 23 polymorphic tetranucleotide microsatellite loci for Sacramento perch (Archoplites interruptus). This species is extirpated in its native range, the Sacramento–San Joaquin Delta (California, USA), and is therefore targeted for recovery. A concerted effort is currently underway to re‐establish self‐sustaining populations of Sacramento perch in its native range. These microsatellites will be used to analyse the population structure of the species and, in conjunction with life history and physiological data, develop a comprehensive recovery plan.     

Characterization of microsatellite loci for five members of the minnow family Cyprinidae found in the Sacramento–San Joaquin Delta and its tributaries

Two microsatellite‐enriched libraries [(CAGA)_n, (TAGA)_n] were constructed using pooled DNA from three cyprinid species native to the Sacramento–San Joaquin Delta of California: Sacramento splittail (Pogonichthys macrolepidotus); Sacramento pikeminnow (Ptychocheilus grandis); and tui chub (Siphateles bicolor). Primers were designed for 105 loci and tested for levels of polymorphism in five cyprinid species found in the Delta: Sacramento splittail, Sacramento pikeminnow, tui chub, hitch (Lavinia exilicauda), and Sacramento blackfish (Orthodon microlepidotus). Fifty‐one loci were polymorphic for at least one species and 31 loci were polymorphic for multiple species. The number of polymorphic loci per species ranged from 16 to 26.     

Persistence of historical population structure in an endangered species despite near‐complete biome conversion in California's San Joaquin Desert

Genomic responses to habitat conversion can be rapid, providing wildlife managers with time‐limited opportunities to enact recovery efforts that use population connectivity information that reflects predisturbance landscapes. Despite near‐complete biome conversion, such opportunities may still exist for the endemic fauna and flora of California's San Joaquin Desert, but comprehensive genetic data sets are lacking for nearly all species in the region. To fill this knowledge gap, we studied the rangewide population structure of the endangered blunt‐nosed leopard lizard Gambelia sila, a San Joaquin Desert endemic, using restriction site‐associated DNA (RAD), microsatellite and mtDNA data to test whether admixture patterns and estimates of effective migration surfaces (EEMS) can identify land areas with high population connectivity prior to the conversion of native xeric habitats. Clustering and phylogenetic analyses indicate a recent shared history between numerous isolated populations and EEMS reveals latent signals of corridors and barriers to gene flow over areas now replaced by agriculture and urbanization. Conflicting histories between the mtDNA and nuclear genomes are consistent with hybridization with the sister species G. wislizenii, raising important questions about where legal protection should end at the southern range limit of G. sila. Comparative analysis of different data sets also adds to a growing list of advantages in using RAD loci for genetic studies of rare species. We demonstrate how the results of this work can serve as an evolutionary guidance tool for managing endemic, arid‐adapted taxa in one of the world's most compromised landscapes.     

Distribution of algae in the San Joaquin River, California, in relation to nutrient supply, salinity and other environmental factors

1. The taxonomic composition and biomass of the phytoplankton and the taxonomic composition of the phytobenthos of the San Joaquin River and its major tributaries were examined in relation to water chemistry, habitat and flow regime. Agricultural drainage and subsurface flow contribute to a complex gradient of salinity and nutrients in this eutrophic, ‘lowland type’ river. 2. Because of light‐limiting conditions for growth, maintenance demands of the algae exceed production during summer and autumn in the San Joaquin River where there is no inflow from tributaries. In contrast to substantial gains in concentration of inorganic nitrogen and soluble reactive phosphorus during the summer of normal‐flow years, net losses of algal biomass (2–4 μg L^?1 day^?1 chlorophyll a) occurred in a mid‐river segment with no significant tributary inflow. However, downstream of a large tributary draining the Sierra Nevada, a substantial net gain in algal biomass (6–11 μg L^?1 day^?1) occurred in the summer, but not in the spring (loss of 1–6 μg L^?1 day^?1) or autumn (loss of 2–5 μg L^?1 day^?1). 3. The phytoplankton was dominated in summer by ‘r‐selected’ centric diatoms (Thalassiosirales), species both tolerant of variable salinity and widely distributed in the San Joaquin River. Pennate diatoms were proportionally more abundant (in biomass) in the winter, spring and autumn. Abundant taxa included the diatoms Cyclotella meneghiniana, Skeletonema cf. potamos, Cyclostephanos invisitatus, Thalassiosira weissflogii, Nitzschia acicularis, N. palea and N. reversa, and the chlorophytes Chlamydomonas sp. and Scenesdesmus quadricauda. Patterns in the abundance of species indicated that assembly of the phytoplankton is limited more by light and flow regime than by nutrient supply. 4. The phytobenthos was dominated by larger, more slowly reproducing pennate diatoms. Few of the abundant species are euryhaline. The diatoms Navicula recens and Nitzschia inconspicua and cyanophytes, Oscillatoria spp., were the principal late‐summer benthic species upstream in the mainstem and in drainages of the San Joaquin Valley. Many of the other abundant diatoms (Amphora veneta, Bacillaria paxillifer, Navicula symmetrica, Nitzschia amphibia, N. fonticola, N. palea, Pleurosigma salinarum) of late‐summer assemblages in these segments also are motile species. While many of these species also were abundant in segments downstream of confluences with rivers draining the Sierra Nevada, the relative abundance of prostrate (Cocconeis placentula var. euglypta, Navicula minima) and erect or stalked (Achnanthidium deflexum, Achnanthes lanceolata, Gomphonema kobayasii, G. parvulum var. lagenula) diatoms and Stigeoclonium sp. was greater in these lower San Joaquin River segments. 5. A weighted‐averaging regression model, based on salinity and benthic‐algal abundance in the San Joaquin River and segments of its major tributaries within the San Joaquin Valley, yielded a highly significant coefficient‐of‐determination (r²=0.84) and low prediction error between salinity inferred from the species and that observed, indicating that salinity tolerance is a primary constraint on growth and assembly of the phytobenthos. The same measures of predictability indicated poor performance of a model based on inorganic nitrogen. However, with a greater representation of tributaries (including segments within the Sierra Nevada foothills) in the sample set, an inorganic nitrogen model also yielded a highly significant coefficient‐of‐determination (r²=0.87) and low prediction error between the species‐inferred and the observed concentration. As with the salinity model (r²=0.94) for the enlarged data set, a systematic difference (increased deviation of residuals) existed at high inorganic nitrogen concentrations. These results indicate substantial interaction between salinity and inorganic nitrogen as constraints on the structure of benthic‐algal communities of the San Joaquin River basin.     

Complex phylogeography and historical hybridization between sister taxa of freshwater sculpin (Cottus)

Species ranges that span different geographic landscapes frequently contain cryptic species‐ or population‐level structure. Identifying these possible diversification factors can often be accomplished under a comparative phylogeographic framework. However, comparisons suffer if previous studies are limited to a particular group or habitat type. In California, a complex landscape has led to several phylogeographic breaks, primarily in terrestrial species. However, two sister taxa of freshwater fish, riffle sculpin (Cottus gulosus) and Pit sculpin (Cottus pitensis), display ranges based on morphological identifications that do not coincide with these breaks. Using a comprehensive sampling and nuclear, mitochondrial and microsatellite markers, we hypothesized that proposed species ranges are erroneous based on potential hybridization/gene flow between species. Results identified a phylogeographic signature consistent with this hypothesis, with breaks at the Coast Range Mountains and Sacramento/San Joaquin River confluence. Coastal locations of C. gulosus represent a unique lineage, and ‘true’ C. gulosus were limited to the San Joaquin basin, both regions under strong anthropogenic influence and potential conservation targets. C. pitensis limits extended historically throughout the Sacramento/Pit River basin but currently are restricted to the Pit River. Interestingly, locations in the Sacramento River contained low levels of ancestral hybridization and gene flow from C. gulosus but now appear to be a distinct population. The remaining population structure was strongly correlated with Sierra Nevada presence (high) or absence (low). This study stresses the importance of testing phylogeographic breaks across multiple taxa/habitats before conservation decisions are made, but also the potential impact of different geographic landscapes on evolutionary diversification.     

Improved genetic markers for monitoring recruitment dynamics in the endangered Mary River cod (Maccullochella mariensis)

The Mary River cod (Maccullochella mariensis) is a large predatory freshwater fish identified as a potential flagship for freshwater ecosystem conservation in Australia. The species is endemic to the Mary River catchment in eastern Australia, and is listed as Endangered. Previous conservation genetic assessment of Mary River cod was based on a small set of microsatellite loci developed for congeneric Maccullochella species. Here we develop a novel set of 15 microsatellite loci specific to M. mariensis, and demonstrate that these markers exhibit higher variability than those used previously. Mean number of alleles per locus was 4 and mean expected heterozygosity was 0.57. We genotyped 35 Mary River cod larvae belonging to a single cohort using the 15 novel loci and eight previously used loci, and found 10 full‐sib family groups along with clear genetic differentiation between individuals collected from the two sub‐catchments – Tinana Creek and Mary River. Microsatellites presented here will be useful for cost‐effective monitoring of genetic diversity and recruitment dynamics in this endangered fish species.     

Evolution of a perfect simple sequence repeat locus in the context of its flanking sequence

Microsatellites, which have rapidly become the preferred markers in population genetics, reliably assign individual chinook salmon to the winter, fall, late-fall, or spring chinook runs in the Sacramento River in California's Central Valley (Banks et al. 2000. Can. J. Fish. Aquat. Sci. 57:915-927). A substantial proportion of this discriminatory power comes from Ots-2, a simple CA repeat, which is expected to evolve rapidly under the stepwise mutation model. We have sequenced a 300-bp region around this locus and typed 668 microsatellite-flanking sequence haplotypes to explore further the basis of this microsatellite divergence. Three sites of nucleotide polymorphism in the Ots-2 flanking sequence define five haplotypes that are shared by the Californian and Canadian populations. The Ots-2 microsatellite alleles are nonrandomly distributed among these five haplotypes in a pattern of gametic disequilibrium that is also shared among populations. Divergence between the winter run and other Central Valley stocks appears to be caused by a combination of surprisingly static evolution at Ots-2 within a context of more rapidly changing haplotype frequencies.     

Microsatellite development for the endangered Yangtze sturgeon (Acipenser dabryanus Duméril, 1869) using 454 sequencing

The Yangtze sturgeon (Acipenser dabryanus) is an endemic species in China. Using 454 sequencing, eight polymorphic tri‐, tetra‐, penta‐, and hexanucleotide microsatellite loci were isolated in this study. The raw sequence data from a one‐eighth run of 454 sequencings were 38.0 Mbp containing 94 222 reads/sequences. Of 80 microsatellite loci, only eight loci were polymorphic in a population of 30 individuals. The number of alleles per locus ranged from 4 to 14 (mean 7.62), and the observed heterozygosities varied between 0.46 and 0.88 (mean 0.74). Cross amplification was tested in congeneric species Acipenser sturio and Acipenser sinensis. These new microsatellite markers will be useful for further studies on genetic variation, parentage analysis, and conservation management for this critically endangered species.     

Isolation and characterization of microsatellite loci in the ant Myrmica scabrinodis

Five polymorphic microsatellite loci were developed for the ant Myrmica scabrinodis using a magnetic bead hybridization selection protocol. The number of alleles per locus varied between three and six. Cross‐species amplification of four of the loci yielded positive amplification products in four Myrmica species, suggesting their general suitability for microsatellite analysis within this taxonomic group.     

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 Communities of the Sacramento River Basin: Implications for Conservation of Native Fishes in the Central Valley, California

The associations of resident fish communities with environmental variables and stream condition were evaluated at representative sites within the Sacramento River Basin, California between 1996 and 1998 using multivariate ordination techniques and by calculating six fish community metrics. In addition, the results of the current study were compared with recent studies in the San Joaquin River drainage to provide a wider perspective of the condition of resident fish communities in the Central Valley of California as a whole. Within the Sacramento drainage, species distributions were correlated with elevational and substrate size gradients; however, the elevation of a sampling site was correlated with a suite of water-quality and habitat variables that are indicative of land use effects on physio-chemical stream parameters. Four fish community metrics – percentage of native fish, percentage of intolerant fish, number of tolerant species, and percentage of fish with external anomalies – were responsive to environmental quality. Comparisons between the current study and recent studies in the San Joaquin River drainage suggested that differences in water-management practices may have significant effects on native species fish community structure. Additionally, the results of the current study suggest that index of biotic integrity-type indices can be developed for the Sacramento River Basin and possibly the entire Central Valley, California. The protection of native fish communities in the Central Valley and other arid environments continues to be a conflict between human needs for water resources and the requirements of aquatic ecosystems; preservation of these ecosystems will require innovative management strategies.     

New polymorphic microsatellite loci,cross‐species amplification and PCR multiplexing in the black aphid,Aphis fabae Scopoli

Aphis fabae includes four morphological cryptic subspecies, which are mostly identified by their partially distinct secondary host range. To determine the extent of gene flow and isolation between these four taxa, we isolated and characterized 12 microsatellite loci from Aphis fabae fabae and tested cross‐species amplification of eight loci from the closely related species Aphis gossypii. Using eight previously described microsatellite loci, we have developed the polymerase chain reaction (PCR) multiplexing of 24 loci, which were separated in tree sets and five PCRs. These sets of microsatellite loci provide high throughput capacity for large‐scale population genetic studies at a minimum cost.     

Isolation and characterization of 15 microsatellite loci from mango (Mangifera indica L.) and cross‐species amplification in closely related taxa

We report here on the development and characterization of 15 microsatellite loci isolated from Mangifera indica L. These markers were evaluated using 59 Florida cultivars and four related species from the USDA germplasm collection for mango. Two loci were monomorphic and 13 polymorphic, with two to seven alleles per locus. Four loci departed significantly from Hardy–Weinberg equilibrium and have significant heterozygote deficiency. Nine loci exhibited significant linkage disequilibrium. Cross‐species amplification was successful in four related species. These loci are being used to investigate patterns of genetic variation within M. indica and between closely related species.     

Characterization of microsatellite loci in the creek chub (Semotilus atromaculatus)

Thirty‐two polymorphic microsatellite loci (29 novel, 3 cross‐species) are characterized in the creek chub, Semotilus atromaculatus, a widespread and abundant North American minnow. Novel repeat‐containing sequences were isolated using a standard microsatellite enrichment procedure. A set of di‐, tri‐, and tetranucleotide markers with allele numbers ranging from two 16 are now available for estimating relatedness within and among populations in this ecologically important species.     

Isolation and characterization of polymorphic microsatellites in the tropical plant‐ant Cataulacus mckeyi (Formicidae: Myrmicinae)

Eleven microsatellite loci were isolated from the plant‐ant Cataulacus mckeyi (Hymenoptera: Myrmicinae) and their polymorphism was characterized. High levels of within‐population variation were observed at most loci, with number of alleles ranging from one to 16, and heterozygosity from 0 to 0.929 per population sample. Cross‐species amplification of these loci was also tested in four other species of the ant genus Cataulacus.     

PCR primers for polymorphic microsatellite loci in the plant‐ant Azteca ulei cordiae (Formicidae: Dolichoderinae)

Twelve microsatellite loci were isolated from the Peruvian tropical plant‐ant, Azteca ulei cordiae (Hymenoptera: Dolichoderinae). High levels of within‐population variation were observed at most loci, with number of alleles ranging from four to 18, and heterozygosity from 0.118 to 1 per population sample. Cross‐species amplification of these loci was also successfully tested in several other species of the same ant genus Azteca.     

Isolation of polymorphic microsatellite loci in the genus Clusia (Clusiaceae)

Microsatellite flanking region sequences may provide phylogenetically useful information. We isolated 13 polymorphic microsatellite loci from two species, Clusia minor (five loci) and Clusia nemorosa (eight loci), to aid in the determination of phylogenetic relationships within the genus Clusia. Eleven loci amplified across all 17 Clusia species tested, while two loci amplified in 10 out of 17 species. The extensive cross‐species amplification suggests that these loci may be useful for an examination of phylogenetic relationships in this genus.     

Novel tetranucleotide microsatellite markers from the Del Norte salamander (Plethodon elongatus) with application to its sister species the Siskiyou Mountain salamander (P. stormi)

Eleven tetranucleotide microsatellite loci were developed for the Del Norte salamander (Plethodon elongatus). The loci were variably polymorphic, ranging from two to 20 alleles per locus, with expected heterozygosities ranging from 0.07 to 0.86. The loci also amplified in a congener, the Siskiyou Mountain salamander (P. stormi). The microsatellite loci will be used to assess the utility of highly polymorphic markers to assay within‐ and between‐species differentiation between these two closely related species.     

Identifying sources of dissolved organic carbon in agriculturally dominated rivers using radiocarbon age dating: Sacramento–San Joaquin River Basin, California

We used radiocarbon measurements of dissolved organic carbon (DOC) to resolve sources of riverine carbon within agriculturally dominated landscapes in California. During 2003 and 2004, average Δ¹⁴C for DOC was ?254‰ in agricultural drains in the Sacramento–San Joaquin Delta, ?218‰ in the San Joaquin River, ?175‰ in the California State Water Project and ?152‰ in the Sacramento River. The age of bulk DOC transiting the rivers of California’s Central Valley is the oldest reported for large rivers and suggests wide-spread loss of soil organic matter caused by agriculture and urbanization. Using DAX 8 adsorbent, we isolated and measured ¹⁴C concentrations in hydrophobic acid fractions (HPOA); river samples showed evidence of bomb-pulse carbon with average Δ¹⁴C of 91 and 76‰ for the San Joaquin and Sacramento Rivers, respectively, with older HPOA, ?204‰, observed in agricultural drains. An operationally defined non-HPOA fraction of DOC was observed in the San Joaquin River with seasonally computed Δ¹⁴C values of between ?275 and ?687‰; the source of this aged material was hypothesized to be physically protected organic-matter in high clay-content soils and agrochemicals (i.e., radiocarbon-dead material) applied to farmlands. Mixing models suggest that the Sacramento River contributes about 50% of the DOC load in the California State Water Project, and agricultural drains contribute approximately one-third of the load. In contrast to studies showing stabilization of soil carbon pools within one or two decades following land conversion, sustained loss of soil organic matter, occurring many decades after the initial agricultural-land conversion, was observed in California’s Central Valley.     

Major histocompatibility complex differentiation in Sacramento River chinook salmon

The chinook salmon of the Sacramento River, California, have been reduced to a fraction of their former abundance because of human impact and use of the river system. Here we examine the genetic variation at a major histocompatibility complex class II exon in the four Sacramento chinook salmon runs. Examination of the alleles found in these and other chinook salmon revealed nucleotide patterns consistent with selection for amino acid replacement at the putative antigen-binding sites. We found a significant amount of variation in each of the runs, including the federally endangered winter run. All of the samples were in Hardy-Weinberg proportions. A significant amount of genetic differentiation between runs was revealed by several measures of differentiation. Winter run was the most genetically divergent, while the spring, late-fall, and fall runs were less differentiated.