Twelve microsatellite loci were characterized in California mountain lions (Puma concolor) and sufficient polymorphism was found to uniquely genotype 62 animals sampled at necropsy. Microsatellite genotypes obtained using mountain lion faecal DNA matched those from muscle for all of 15 individuals examined. DNA from potential prey species and animals whose faeces could be misidentified as mountain lion faeces were reliably distinguished from mountain lions using this microsatellite panel. In a field application of this technique, 32 faecal samples were collected from hiking trails in the Yosemite Valley region where seven mountain lions previously had been captured, sampled, and released. Twelve samples yielded characteristic mountain lion genotypes, three displayed bobcat-type genotypes, and 17 did not amplify. The genotype of one of the 12 mountain lion faecal samples was identical to one of the mountain lions that previously had been captured. Three of the 12 faecal samples yielded identical genotypes, and eight new genotypes were detected in the remaining samples. This analysis provided a minimum estimate of 16 mountain lions (seven identified by capture and nine identified by faecal DNA) living in or travelling through Yosemite Valley from March 1997 to August 1998. Match probabilities (probabilities that identical DNA genotypes would be drawn at random a second time from the population) indicated that the samples with identical genotypes probably came from the same mountain lion. Our results demonstrate that faecal DNA analysis is an effective method for detecting and identifying individual mountain lions.

Migration is essential for the reproduction and survival of many animals, yet little is understood about its underlying molecular mechanisms. We used the salmonid Oncorhynchus mykiss to gain mechanistic insight into smoltification, which is a morphological, physiological, and behavioral transition undertaken by juveniles in preparation for seaward migration. O. mykiss is experimentally tractable and displays intra- and inter-population variation in migration propensity. Migratory individuals can produce non-migratory progeny and vice versa, indicating a high degree of phenotypic plasticity. One potential way that phenotypic plasticity might be linked to variation in migration-related life history tactics is through epigenetic regulation of gene expression. To explore this, we quantitatively measured genome-scale DNA methylation in fin tissue using reduced representation bisulfite sequencing of F2 siblings produced from a cross between steelhead (migratory) and rainbow trout (non-migratory) lines. We identified 57 differentially methylated regions (DMRs) between smolt and resident O. mykiss juveniles. DMRs were high in magnitude, with up to 62% differential methylation between life history types, and over half of the gene-associated DMRs were in transcriptional regulatory regions. Many of the DMRs encode proteins with activity relevant to migration-related transitions (e.g. circadian rhythm pathway, nervous system development, protein kinase activity). This study provides the first evidence of a relationship between epigenetic variation and life history divergence associated with migration-related traits in any species.

Many populations of lake sturgeon Acipenser fulvescens have decreased in size throughout the Great Lakes basin. To implement management strategies such as stocking, it is important to understand the genetic structure of lake sturgeon spawning populations. Lake sturgeon from 27 spawning locations (25 from the Great Lakes basin and 2 from the Hudson Bay drainage) were analyzed using 12 microsatellite loci. Population structure was detected at different spatial scales. At the largest scale, consistent genetic breaks were observed among three clusters of spawning populations: (1) Hudson Bay–northern Lake Superior, (2) southern Lake Superior, and (3) the rest of the Great Lakes. These clusters were identified using a Bayesian approach that does not define the populations a priori. Within each of the three clusters, sublevels of genetic structure were detected. These sublevel clusters accounted for 8.82% of the genetic variation (P < 0.000), while differences among populations within the clusters accounted for 3.72% of the genetic variation (P < 0.000). At the smallest scale, significant genetic differentiation was detected between most sampled locations through pairwise genetic differentiation index (FST) tests and pairwise contingency tests. Lake sturgeon showed greater genetic differentiation in Lake Superior than elsewhere, which could be due to the lake's bathymetry. The lower genetic resolution observed elsewhere in the Great Lakes could be due to more recent colonization events. The results can be used to delineate management units and to select appropriate donor populations for supplementation or reintroductions.

No abstract

Eleven tri- and tetra-meric motif microsatellite loci were identified in a lake sturgeon genomic library and were tested against the six North American species of Acipenser (lake sturgeon, A. fulvescens; shortnose sturgeon, A. brevirostrum; white sturgeon, A. transmontanus; green sturgeon, A. medirostris; Atlantic sturgeon, A. oxyrhynchus oxyrhynchus; gulf sturgeon, A. o. desotoi) and the two species of Scaphirhynchus (pallid sturgeon, S. albus; shovelnose sturgeon, S. platorhynchus) using four to six individuals of each species. Eight loci were amplified equally well in all eight species, and the remaining three loci were amplified in two, four, and seven species, respectively. Of the eight loci that worked in all species, one was monomorphic in all species, while the other seven were polymorphic in three to eight species. Single repeat differences in these tri- and tetra-meric repeat motifs can be readily scored on 4% Metaphor agarose gels stained with ethidium bromide. In addition, dosage for those loci exhibiting four gene doses can also be readily scored with this technique. These loci provide a much needed group of genetic markers, detectable with non-invasive sampling (blood, barbel, or fin), that should work well in threatened and endangered species of sturgeon worldwide.

Low levels of genetic variation at traditional molecular markers have hampered genetic research within the family Acipenseridae. In an effort to develop a large set of polymorphic genetic markers, 172 clones were sequenced from three subgenomic libraries of shovelnose sturgeon Scaphirhynchus platorynchus; the libraries were enriched for two dinucleotide and one tetranucleotide microsatellite motifs (CA, GA, and TAGA). Primers were designed for 113 of the sequences and tested against shovelnose sturgeon, pallid sturgeon S. albus, white sturgeon Acipenser transmontanus, lake sturgeon A. fulvescens, and green sturgeon A. medirostris. Of the 113 primer sets tested, 96% amplified in one or more species (58 dimeric and 50 tetrameric). In Scaphirhynchus species, 93% of all loci amplified, and 76% were polymorphic. Within the individual Acipenser species, 65–80% of loci amplified, with 42–58% being polymorphic. Polymorphic systems for Scaphirhynchus species predominately displayed simple, disomic banding patterns, while those for Acipenser species typically displayed banding patterns characteristic of tetraploid or higher polyploid levels. These new microsatellite loci provide a group of genetic markers that are detectable with noninvasive sampling and that should prove useful in the preservation of threatened and endangered sturgeon species worldwide.

No abstract

Nine tetramer motif (GATA)n microsatellite systems were developed for use in the white sturgeon, Acipenser transmontanus. We report inheritance patterns for these nine systems, which range from one possible disomic system to tetrasomy and octosomy, with some systems containing null alleles. Because of the complex modes of inheritance underlying these systems and the highly duplicated nature of the genome, we propose each allele be scored as its own dominant marker, similar to AFLPs or RAPDs. The utility of this method is validated by the observation that individual alleles within a microsatellite system generally fit the expectation for independent transmission and fit the expected transmission frequency for single copy nuclear markers.