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At least 73 records · Page 4Linked to original sources

Monitoring status and trends in genetic diversity for the Convention on Biological Diversity: An ongoing assessment of genetic indicators in nine countries

Recent scientific evidence shows that genetic diversity must be maintained, managed, and monitored to protect biodiversity and nature's contributions to people. Three genetic diversity indicators, two of which do not require DNA-based assessment, have been proposed for reporting to the Convention on Biological Diversity and other conservation and policy initiatives. These indicators allow an approximation of the status and trends of genetic diversity to inform policy, using existing demographic and geographic information. Application of these indicators has been initiated and here we describe ongoing efforts in calculating these indicators with examples. We specifically describe a project underway to apply these indicators in nine countries, provide example calculations, address concerns of policy makers and implementation challenges, and describe a roadmap for further development and deployment, incorporating feedback from the broader community. We also present guidance documents and data collection tools for calculating indicators. We demonstrate that Parties can successfully and cost-effectively report these genetic diversity indicators with existing biodiversity observation data, and, in doing so, better conserve the Earth's biodiversity.

Conservation Letters↗

Development of microsatellite markers for three at risk tiger beetles Cicindela dorsalis dorsalis, C. d. media, and C. puritana

Objective Tiger beetles inhabiting sandy beaches and cliffs along the east coast of the United States are facing increasing habitat loss due to erosion, urbanization, and sea level rise. The northeastern beach tiger beetle Cicindela dorsalis dorsalis and Puritan tiger beetle Cicindela puritana are both listed as threatened under the Endangered Species Act of 1973, while the white beach tiger beetle Cicindela dorsalis media is not listed but has been declining. Extirpation of these beetles, in some cases from entire states, has isolated many populations reducing gene flow and elevating the risk for the loss of genetic variation. To facilitate investigations of population genetic structure, we developed suites of microsatellite loci for conservation genetic studies. Results Shotgun genomic sequencing of all species identified thousands of candidate microsatellite loci, among which 17 loci were optimized and verified to cross-amplify within C. d. media and C. d. dorsalis , and eight separate loci were optimized for C. puritana . Most loci conformed to Hardy–Weinberg equilibrium, showed no evidence of linkage disequilibrium or null alleles, and revealed population genetic characteristics informative for natural resource managers among the populations tested.

BMC Research Notes↗

Genomics of Arctic cod

The Arctic cod (Boreogadus saida) is an abundant marine fish that plays a vital role in the marine food web. To better understand the population genetic structure and the role of natural selection acting on the maternally-inherited mitochondrial genome (mitogenome), a molecule often associated with adaptations to temperature, we analyzed genetic data collected from 11 biparentally-inherited nuclear microsatellite DNA loci and nucleotide sequence data from from the mitochondrial DNA (mtDNA) cytochrome b (cytb) gene and, for a subset of individuals, the entire mitogenome. In addition, due to potential of species misidentification with morphologically similar Polar cod (Arctogadus glacialis), we used ddRAD-Seq data to determine the level of divergence between species and identify species-specific markers. Based on the findings presented here, Arctic cod across the Pacific Arctic (Bering, Chukchi, and Beaufort Seas) comprise a single panmictic population with high genetic diversity compared to other gadids. High genetic diversity was indicated across all 13 protein-coding genes in the mitogenome. In addition, we found moderate levels of genetic diversity in the nuclear microsatellite loci, with highest diversity found in the Chukchi Sea. Our analyses of markers from both marker classes (nuclear microsatellite fragment data and mtDNA cytb sequence data) failed to uncover a signal of microgeographic genetic structure within Arctic cod across the three regions, within the Alaskan Beaufort Sea, or between near-shore or offshore habitats. Further, data from a subset of mitogenomes revealed no genetic differentiation between Bering, Chukchi, and Beaufort seas populations for Arctic cod, Saffron cod (Eleginus gracilis), or Walleye pollock (Gadus chalcogrammus). However, we uncovered significant differences in the distribution of microsatellite alleles between the southern Chukchi and central and eastern Beaufort Sea samples of Arctic cod. Finally, using ddRAD-Seq data, we identified species-specific markers and in conjunction with mitogenome data, identified an Arctic cod x Polar cod hybrid in western Canadian Beaufort Sea. Overall, the lack of genetic structure among Arctic cod within the Bering, Chukchi and Beaufort seas of Alaska is concordant with the absence of geographic barriers to dispersal and typical among marine fishes. Arctic cod may exhibit a genetic pattern of isolation-by-distance, whereby populations in closer geographic proximity are more genetically similar than more distant populations. As this signal is only found between our two fartherest localities, data from populations elsewhere in the species’ global range are needed to determine if this is a general characteristic. Further, tests for selection suggested a limited role for natural selection acting on the mitochondrial genome of Arctic cod, but do not exclude the possibility of selection on genes involved in nuclear-mitogenome interactions. Unlike previous genetic assessment of Arctic cod sampled from the Chukchi Sea, the high levels of genetic diversity found in Arctic cod assayed in this study, across regions, suggests that the species in the Beaufort and Chukchi seas does not suffer from low levels of genetic variation, at least at neutral genetic markers. The large census size of Arctic cod may allow this species to retain high levels of genetic diversity. In addition, we discovered the presence of hybridization between Arctic and Polar cod (although low in frequency). Hybridization is expected to occur when environmental changes modify species distributions that result in contact between species that were previously separated. In such cases, hybridization may be an evolutionary mechanism that promotes an increase in genetic diversity that may provide species occupying changing environments with locally-adapted genotypes and, therefore, phenotypes. Natural selection can only act on the standing genetic variation present within a population. Therefore, given its higher levels of genetic diversity in combination with a large population size, Arctic cod may be resilient to current and future environmental change, as high genetic diversity is expected to increase opportunities for positive selection to act on genetic variants beneficial in different environments, regardless of the source of that genetic variation.

OCS Study↗

Natural selection after release from a hatchery leads to domestication in steelhead, Oncorhynchus mykiss

Genetic theory and data suggest that sea ranching of anadromous salmonids (Oncorhynchus spp. and Salmo spp.) results in domestication (increased fitness in the hatchery program) accompanied by a loss of fitness for natural production. We tested for genetic differences in growth, survival, and downstream migration of hatchery and wild steelhead (O. mykiss) reared together in a hatchery. We found little or no difference in survival during hatchery rearing but substantial differences in growth and subsequent downstream migration. Intense natural selection after release from the hatchery favored fish that had performed well (e.g. grew fast) in the hatchery. This selection in the natural environment genetically changes (domesticates) the population because at least some of the performance traits are heritable. Domestication should improve the economic efficiency for producing adult hatchery fish but compromise conservation of wild populations when hatchery fish interbreed with wild fish.

Book chapter↗

Genetic structure in Elk persists after translocation

Elk ( Cervus canadensis ) translocation success is thought to be facilitated by high post‐release herd cohesion and limited movements; both should ensure genetic mixing following release. Such mixing is important to reduce potential effects of inbreeding or genetic drift, which can be especially important in small founding populations. We had a natural experiment where we could evaluate genetic mixing of 2 distinct lineages of elk after translocation to the same area. Founding elk ultimately came from north and south of a road barrier at Elk Island National Park (EINPN or EINPS, respectively), Alberta, Canada and the 2 groups were genetically distinct. During 2000 to 2003, elk originating from Elk Island National Park were translocated to Cumberland Mountains, Tennessee (TNCM) and Great Smoky Mountains National Park, North Carolina (GSMNP), USA (some elk spent time at Land Between the Lakes Recreation Area, Kentucky, USA, before their final translocation). At TNCM, translocated elk were hard released, whereas at GSMNP elk were held in pens up to 60 days before release (i.e., soft release). We hypothesized that associations formed in the source population would affect genetic structure in the future population. We predicted that matrilineal groups would stay closer together and have similar movements after translocation. We used 16 microsatellite markers to analyze genetic composition and structure of translocated elk and their offspring in the years after release. Most source elk used for translocation strongly assigned to either EINPN or EINPS (93.2%, n = 204). Evaluating the genetic structure of offspring after translocation, we found the 2 genetic groups mostly persisted ≥11 years following release. We measured the Euclidean distance between all possible pairs of telemetered female elk during each season and year and calculated the maximum distance moved from the release sites for females surviving >1 year. Mean Euclidean distances between pairwise locations of female elk were similar for each genetic cluster for each area. The mean distances for all paired locations (genetic clusters combined) in TNCM were 14.67 km ( n = 4,576 ± 13.23 [SD]) and in GSMNP were 9.30 km ( n = 1,468 ± 9.75). However, when looking at only simultaneous locations <50 m apart, the frequency of occurrence was higher ( P < 0.001) for elk with the same genetic structure (71.1%) compared with those with different structure (28.9%). The maximum distance travelled from the release site was not different for the 2 genetic groups, but EINPN females tended to travel farther. Pairwise female distances were lower in GSMNP where we used a soft release. Release methodology and social structure appear to affect movements and possibly genetic mixing after translocation. Given that restoration success can depend on maintaining genetic diversity and number of founders, our analyses suggest that within‐cluster breeding bias can result in lower genetic variability and a smaller effective population size than previously assumed.

Kentucky, Tennessee↗

Identification of Neosho Smallmouth Bass (Micropterus dolomieu velox) stocks for possible introduction into Grand Lake, Oklahoma

Stocking black basses (Micropterus spp.) is a common practice used to increase angling opportunities in impoundments; however, when non-native black basses are introduced they often invade riverine habitats where they threaten the persistence of other fishes, including native black basses. Neosho Smallmouth Bass (M. dolomieu velox) is endemic to portions of the Ozark Highlands and Boston Mountains ecoregions and is threatened by introductions of non-native Smallmouth Bass (“SMB”) forms. Because of recent interest in stocking SMB into Grand Lake o’ the Cherokees, we assessed the suitability of local Neosho SMB populations as potential broodstock sources by assessing introgression with non-native SMB forms, as well as characterizing population structure and genetic diversity. The majority of Neosho SMB populations contained low, but non-negligible, genomic proportions of two genetically distinct non-native SMB forms. Introgression was highest in the Illinois River upstream of Lake Tenkiller, where Tennessee ‘lake strain’ SMB were stocked in the early 1990’s. We recovered three genetically distinct clusters of Neosho SMB at the uppermost hierarchical level of population structure: a distinct Illinois River cluster and two Grand River clusters that appear to naturally mix at some sites. Genetic diversity measures generally increased with stream size, and smaller populations with low diversity measures may benefit from immigration of novel genetic material. Overall, introgression with non-native SMB forms appears to pose a prominent threat to Neosho SMB; however, relatively intact populations of Neosho SMB exist in some Grand Lake o’ the Cherokees tributaries. Results could be used in developing a stocking program that promotes and sustains existing genetic diversity within and among Neosho SMB populations.

Cooperator Science Series↗

Connectivity of mule deer (Odocoileus hemionus) populations in southern California: A genetic survey of a mobile ungulate in a highly fragmented urban landscape

Urbanization is a substantial force shaping the genetic and demographic structure of natural populations. Urban development and major highways can limit animal movements, and thus gene flow, even in highly mobile species. Characterizing varying species responses to human activity and fragmentation is important for maintaining genetic continuity in wild animals and for preserving biodiversity. As one of the only common and wide-ranging large wild herbivores in much of urban North America, deer play an important ecological role in urban ecosystems, yet the genetic impacts of development on deer are not well known.

Landscape Ecology↗

Influence of barriers to movement on within-watershed genetic variation of coastal cutthroat trout

Because human land use activities often result in increased fragmentation of aquatic and terrestrial habitats, a better understanding of the effects of fragmentation on the genetic heterogeneity of animal populations may be useful for effective management. We used eight microsatellites to examine the genetic structure of coastal cutthroat trout (Oncorhynchus clarki clarki) in Camp Creek, an isolated headwater stream in western Oregon. Our objectives were to determine if coastal cutthroat trout were genetically structured within streams and to assess the effects of natural and anthropogenic barriers on coastal cutthroat trout genetic variation. Fish sampling occurred at 10 locations, and allele frequencies differed significantly among all sampling sections. Dispersal barriers strongly influenced coastal cutthroat trout genetic structure and were associated with reduced genetic diversity and increased genetic differentiation. Results indicate that Camp Creek coastal cutthroat trout exist as many small, partially independent populations that are strongly affected by genetic drift. In headwater streams, barriers to movement can result in genetic and demographic isolation leading to reduced coastal cutthroat trout genetic diversity, and potentially compromising long-term population persistence. When habitat fragmentation eliminates gene flow among small populations, similar results may occur in other species.

Oregon↗

Newly documented population extends geographic range and genetic diversity for the Leaf-toed Gecko (Phyllodactylus nocticolus) into the Transverse Ranges of southern California

Between 19 – 30 May 2018, one of us [AW] discovered a disjunct population of Peninsula leaf-toed geckos, Phyllodactylus nocticolus (Phyllodactylidae) on the northern edge of the Coachella Valley in the Little San Bernardino Mountains of the Transverse Ranges (Fig. 1a). The previously northernmost location for the species is Tahquitz Canyon, Riverside Co. (MVZ 212205) in the Peninsular Ranges 20 km to the south. Southern California has many herp enthusiasts and it is possible that this population is of anthropogenic origin through accidental or misguided purposeful introduction. The apparent barrier to dispersal suggests that, if of natural origin, the dispersal may have occurred at a time when the aeolian sand barrier was less severe. Thus, if this newly discovered population is of natural origin, we expect the genetic data of the disjunct Transverse Ranges population to differ from that of any population sampled in the Peninsular Ranges. Herein we describe this newly discovered population of P. nocticolus, we analyze genetic diversity from the new population, and we compare it with genetic data gathered from populations of P. nocticolus throughout southern California to help determine if this isolated gecko population was of natural dispersal or the result of human intervention. This is particularly noteworthy given the apparent strength of the Coachella Valley’s sand fields as a barrier to dispersal of highly saxicolous lizard faunas.

California↗

Genetic similarity between mates and extra-pair parentage in three species of shorebirds

Matings between close relatives often reduce the fitness of offspring, probably because homozygosity leads to the expression of recessive deleterious alleles 1 , 2 , 3 , 4 , 5 . Studies of several animals have shown that reproductive success is lower when genetic similarity between parents is high 4 , 5 , 6 , 7 , and that survival and other measures of fitness increase with individual levels of genetic diversity 8 , 9 , 10 , 11 . These studies indicate that natural selection may favour the avoidance of matings with genetically similar individuals. But constraints on social mate choice, such as a lack of alternatives, can lead to pairing with genetically similar mates. In such cases, it has been suggested that females may seek extra-pair copulations with less related males 4 , but the evidence is weak or lacking 4 , 5 . Here we report a strong positive relationship between the genetic similarity of social pair members and the occurrence of extra-pair paternity and maternity (‘quasi-parasitism’) in three species of shorebirds. We propose that extra-pair parentage may represent adaptive behavioural strategies to avoid the negative effects of pairing with a genetically similar mate.

Nature↗

Integrated geochemical approach to determine the source of methane in gas hydrate from Green Canyon Block 955 in the Gulf of Mexico

Massive volumes of gas are sequestered within gas hydrate in subsurface marine sediments in the Gulf of Mexico. Methane associated with gas hydrate is a potentially important economic resource and a significant reservoir of carbon within the global carbon cycle. Nevertheless, uncertainties remain about the genetic source (e.g., microbial, thermogenic) and possible migration history of natural gas incorporated into hydrate. Previous studies have primarily used the hydrocarbon molecular (CH 4 /C 2 H 6 +) and isotopic (δ 13 C-CH 4 , δ 2 H-CH 4 ) compositions of natural gas to address these uncertainties. However, hydrocarbon tracers are altered by mixing, oxidation, secondary methanogenesis, or fluid migration, which presents challenges when deciphering the mechanisms responsible for methane formation and accumulation. To evaluate the genetic source of natural gases from Green Canyon Block 955 (GC 955), east of the Sigsbee escarpment, we collected and analyzed samples from the first pressurized hydrate-bearing sediment cores collected from a coarse-grained hydrate reservoir in the Gulf of Mexico. Gas samples were analyzed for hydrocarbon gas (C 1 –C 5 ), major gas (e.g., N 2 , CO 2 ), and noble gas (He-Xe) abundance and isotopic (e.g., δ 13 C-CH 4 , δ 2 H-CH 4 , δ 13 C-CO 2 , δ 15 N-N 2 , 3 He/ 4 He, 4 He/ 20 Ne) compositions. We determined that natural gas in hydrates from this location are predominantly of primary microbial origin (conservatively at least 76%) and are formed by the hydrogenotrophic (CO 2 reduction) methanogenesis pathway. We also note increased thermogenic proportions (∼6%) in a hydrate-bearing layer below the main hydrate-bearing interval (separated by a 5-m water-bearing layer). Our results suggest that microbial methane may be abundant below the base of gas hydrate stability at GC 955.

Louisiana↗

Microsatellite variation reveals weak genetic structure and retention of genetic variability in threatened Chinook salmon (Oncorhynchus tshawytscha) 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.

Idaho↗

Parentage-based tagging using mothers balances accuracy and cost for discriminating between natural and stocked recruitment for inland fisheries

Hatchery programmes are frequently used to supplement inland fisheries, yet achieving successful management outcomes often requires information on stocked versus naturally reproduced fish abundance. Parentage-based tagging – genetically assigning offspring to their parents – has potential to be an effective approach for distinguishing stocked and naturally reproduced fish. However, several challenges may limit its application to inland fisheries, including genetic relatedness among populations that can affect identification accuracy and high costs of genotyping large broodstocks. Here, we demonstrate the efficacy of parentage-based tagging based on broodmothers in the Lake Ontario Chinook Salmon fishery, which uses thousands of broodparents and has potential for substantial relatedness between stocked and naturally reproduced fish. Restricting parent sampling to broodmothers reduced costs by two-thirds, was logistically pragmatic, and achieved >95% accuracy in distinguishing stocked from naturally reproduced salmon. Combined, these results highlight the potential wide applicability of parentage-based tagging for assessing stocking programmes in inland waters.

New York, Ontario↗

Genetic changes from artificial propagation of Pacific salmon affect the productivity and viability of supplemented populations

Although several studies have shown genetic differences between hatchery and wild anadromous Pacific salmon (Oncorhynchus spp.), none has provided compelling evidence that artificial propagation poses a genetic threat to conservation of naturally spawning populations. When the published studies and three studies in progress are considered collectively, however, they provide strong evidence that the fitness for natural spawning and rearing can be rapidly and substantially reduced by artificial propagation. This issue takes on great importance in the Pacific Northwest where supplementation of wild salmon populations with hatchery fish has been identified as an important tool for restoring these populations. Recognition of negative aspects may lead to restricted use of supplementation, and better conservation, better evaluation, and greater benefits when supplementation is used.

Conference Paper↗

Genetic and environmental influences on leaf phenology and cold hardiness of native and introduced riparian trees

To explore the roles of plasticity and genetic variation in the response to spatial and temporal climate variation, we established a common garden consisting of paired collections of native and introduced riparian trees sampled along a latitudinal gradient. The garden in Fort Collins, Colorado (latitude 40.6°N), included 681 native plains cottonwood ( Populus deltoides subsp. monilifera ) and introduced saltcedar ( Tamarix ramosissima , T. chinensis and hybrids) collected from 15 sites at 29.2–47.6°N in the central United States. In the common garden both species showed latitudinal variation in fall, but not spring, leaf phenology, suggesting that the latitudinal gradient in fall phenology observed in the field results at least in part from inherited variation in the critical photoperiod, while the latitudinal gradient in spring phenology observed in the field is largely a plastic response to the temperature gradient. Populations from higher latitudes exhibited earlier bud set and leaf senescence. Cold hardiness varied latitudinally in both fall and spring for both species. For cottonwood, cold hardiness began earlier and ended later in northern than in southern populations. For saltcedar northern populations were hardier throughout the cold season than southern populations. Although cottonwood was hardier than saltcedar in midwinter, the reverse was true in late fall and early spring. The latitudinal variation in fall phenology and cold hardiness of saltcedar appears to have developed as a result of multiple introductions of genetically distinct populations, hybridization and natural selection in the 150 years since introduction.

International Journal of Biometeorology↗

Conservation genetics of evolutionary lineages of the endangered mountain yellow-legged frog, Rana muscosa (Amphibia: Ranidae), in southern California

Severe population declines led to the listing of southern California Rana muscosa (Ranidae) as endangered in 2002. Nine small populations inhabit watersheds in three isolated mountain ranges, the San Gabriel, San Bernardino and San Jacinto. One population from the Dark Canyon tributary in the San Jacinto Mountains has been used to establish a captive breeding population at the San Diego Zoo Institute for Conservation Research. Because these populations may still be declining, it is critical to gather information on how genetic variation is structured in these populations and what historical inter-population connectivity existed between populations. Additionally, it is not clear whether these populations are rapidly losing genetic diversity due to population bottlenecks. Using mitochondrial and microsatellite data, we examine patterns of genetic variation in southern California and one of the last remaining populations of R. muscosa in the southern Sierra Nevada. We find low levels of genetic variation within each population and evidence of genetic bottlenecks. Additionally, substantial population structure is evident, suggesting a high degree of historical isolation within and between mountain ranges. Based on estimates from a multi-population isolation with migration analysis, these populations diversified during glacial episodes of the Pleistocene, with little gene flow during population divergence. Our data demonstrate that unique evolutionary lineages of R. muscosa occupy each mountain range in southern California and should be managed separately. The captive breeding program at Dark Canyon is promising, although mitigating the loss of neutral genetic diversity relative to the natural population might require additional breeding frogs.

California↗

Genetic analysis of Missouri’s Topeka Shiners with implications for the propagation of understudied small-bodied freshwater fishes

Objective Best practices for conservation hatcheries to conserve genetic diversity and minimize adaptation to captivity have been established for decades, but how to apply them is not clear in every circumstance. As a growing number of aquatic species are propagated in captive settings, addressing the fit of these practices to each system will help managers operate optimally while conserving hatchery resources. Small-bodied freshwater fish present a unique set of traits compared with species that are typically considered for propagation (i.e., salmonids), including a patchy distribution within a watercourse. We examine the propagation and reintroduction program that supports the Topeka Shiner Miniellus topeka , an endangered minnow in the Midwestern USA. Methods We genotyped shiners from groups with different histories (two reintroduced, three captive, and two remnant populations) at 11 microsatellite loci and compared genetic diversity, genetic structure, effective population size, and evidence of population bottlenecks. We also looked at the breeding structure by genetically assigning hatchery-reared young ( n = 148) to candidate parents. Results We documented high levels of genetic structure among the two natural populations in our study. We also noted lower diversity and evidence of bottlenecks in hatchery-reared groups. However, hatcheries may support sufficient (>50) effective population sizes with minimal space. Conclusions Hatcheries may avoid bottlenecks in other small-bodied freshwater fish by collecting wild fish from a broad area and frequently incorporating them into the captive population. Within the hatchery, we emphasize the need to reduce generational overlap by stocking all production fish and/or subdividing the captive populations.

Missouri↗

Molecular genetics at the Fort Collins Science Center

The Fort Collins Science Center operates a molecular genetic and systematics research facility (FORT Molecular Ecology Laboratory) that uses molecular genetic tools to provide genetic information needed to inform natural resource management decisions. For many wildlife species, the data generated have become increasingly important in the development of their long-term management strategies, leading to a better understanding of species diversity, population dynamics and ecology, and future conservation and management needs. The Molecular Ecology Lab serves Federal research and resource management agencies by developing scientifically rigorous research programs using nuclear, mitochondrial and chloroplast DNA to help address many of today's conservation biology and natural resource management issues.

Fact Sheet↗