Mitochondrial DNA analysis reveals hidden genetic diversity in captive populations of the threatened American crocodile (Crocodylus acutus) in Colombia

Identification of units within species worthy of separate management consideration is an important area within conservation. Mitochondrial DNA (mtDNA) surveys can potentially contribute to this by identifying phylogenetic and population structure below the species level. The American crocodile (Crocodylus acutus) is broadly distributed throughout the Neotropics. Its numbers have been reduced severely with the species threatened throughout much of its distribution. In Colombia, the release of individuals from commercial captive populations has emerged as a possible conservation strategy that could contribute to species recovery. However, no studies have addressed levels of genetic differentiation or diversity within C. acutus in Colombia, thus complicating conservation and management decisions. Here, sequence variation was studied in mtDNA cytochrome b and cytochrome oxidase I gene sequences in three Colombian captive populations of C. acutus. Two distinct lineages were identified: C. acutus‐I, corresponding to haplotypes from Colombia and closely related Central American haplotypes; and C. acutus‐II, corresponding to all remaining haplotypes from Colombia. Comparison with findings from other studies indicates the presence of a single “northern” lineage (corresponding to C. acutus‐I) distributed from North America (southern Florida), through Central America and into northern South America. The absence of C. acutus‐II haplotypes from North and Central America indicates that the C. acutus‐II lineage probably represents a separate South American lineage. There appears to be sufficient divergence between lineages to suggest that they could represent two distinct evolutionary units. We suggest that this differentiation needs to be recognized for conservation purposes because it clearly contributes to the overall genetic diversity of the species. All Colombian captive populations included in this study contained a mixture of representatives of both lineages. As such, we recommend against the use of captive‐bred individuals for conservation strategies until further genetic information is available.     

Subspecies of the Central American Squirrel Monkey (Saimiri oerstedii) as Units for Conservation

The accurate diagnosis of conservation units now typically includes recognition of genetic diversity and unique evolutionary lineages and is necessary to inform the conservation management of endangered species. We evaluated whether the two currently recognized subspecies of the endangered Central American squirrel monkey (Saimiri oerstedii) in Costa Rica are evolutionarily significant units (ESUs) that should be managed separately in conservation efforts. We used previously published sequences of 50 individuals of Saimiri oerstedii for 880 bp of the mtDNA d-loop and genotypes of 244 individuals for 16 microsatellites and conducted novel analyses to characterize genetic differentiation between subspecies of Saimiri oerstedii. We measured sequence differentiation and inferred an intraspecific molecular phylogeny and a haplotype network, and found consistent results supporting statistically significant divergence and reciprocal monophyly between subspecies. A population aggregation analysis also supported Saimiri oerstedii citrinellus and S. o. oerstedii as diagnosably distinct units. These results confirm previous genetic studies with smaller sample sizes and are consistent with other factors including differences in pelage and morphology and divergence at nuclear markers. Conservation managers should manage these subspecies separately to prevent the loss of genetic diversity via artificially induced outbreeding. High levels of genetic diversity may buffer populations against outside extinction pressures, to which Saimiri oerstedii are vulnerable because of their dwindling habitat and small population size.     

Population genetic structure of the endangered Eastern Bristlebird, Dasyornis brachypterus; implications for conservation

For species that are habitat specialists or sedentary, population fragmentation may lead to genetic divergence between populations and reduced genetic diversity within populations, with frequent inbreeding. Hundreds of kilometres separate three geographical regions in which small populations of the endangered Eastern Bristlebird, Dasyornis brachypterus, a small, ground-dwelling passerine that occurs in fire-prone bushland in eastern Australia, are currently found. Here, we use mitochondrial and microsatellite DNA markers to: (i) assess the sub-specific taxonomy designated to northern range-edge, and central and southern range-edge D. brachypterus, respectively, and (ii) assess levels of standing genetic variation and the degree of genetic subdivision of remnant populations. The phylogenetic relationship among mtDNA haplotypes and their spatial distribution did not support the recognised subspecies boundaries. Populations in different regions were highly genetically differentiated, but in addition, the two largest, neighboring populations (located within the central region and separated by ~50 km) were moderately differentiated, and thus are likely closed to migration (microsatellites, F _ST = 0.06; mtDNA, F _ST = 0.12, ?? _ST = 0.08). Birds within these two populations were genotypically diverse and apparently randomly mating. A long-term plan for the conservation of D. brachypterus??s genetic diversity should consider individual populations as separate management units. Moreover, managers should avoid actively mixing birds from different populations or regions, to conserve the genetic integrity of local populations and avoid outbreeding depression, should further translocations be used as a recovery tool for this species.     

Phylogeography of the bushmaster (Lachesis tnuta: Viperidae): implications for neotropical biogeography, systematics, and conservation

We used mitochondrial gene sequences to reconstruct phylogenetic relationships among subspecies of the bushmaster, Lachesis muta. These large vipers are widely distributed in lowland tropical forests in Central and South America, where three of four allopatric subspecies are separated by montane barriers. Our phylogeny indicates that the four subspecies belong to two clades, the Central American and South American lineages. We use published molecular studies of other taxa to estimate a 'reptilian mtDNA rate' and thus temporal boundaries for major lineage divergences in Lachesis. We estimate that the Central and South American forms diverged 18-6 Mya, perhaps due to the uplifting of the Andes, whereas the two Central American subspecies may have diverged 11-4 Mya with the uprising of the Cordillera de Talamanca that separates them today. South American bushmasters from the Amazon Basin and the Atlantic Forest are not strongly differentiated, perhaps due to episodic gene flow during the Pleistocene, when suitable habitat for this species was at times more continuous. Our results agree with previous evidence that genetic divergence among some neotropical vertebrates pre-dated Pleistocene forest fragmentation cycles and the appearance of the Panamanian Isthmus. Based on morphological, behavioral, and molecular evidence, we recognize three species of Lachesis. In addition to L. muta, the widespread South American form, the Central American forms are treated as distinct species (L. meknocephak and L. stenophrys), each deserving of special conservation status due to restricted distribution and habitat destruction.     

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.     

The Complex Biogeography of the Plant Pathogen Xylella fastidiosa: Genetic Evidence of Introductions and Subspecific Introgression in Central America

The bacterium Xylella fastidiosa is a plant pathogen with a history of economically damaging introductions of subspecies to regions where its other subspecies are native. Genetic evidence is presented demonstrating the introduction of two new taxa into Central America and their introgression into the native subspecies, X. fastidiosa subsp. fastidiosa. The data are from 10 genetic outliers detected by multilocus sequence typing (MLST) of isolates from Costa Rica. Six (five from oleander, one from coffee) defined a new sequence type (ST53) that carried alleles at six of the eight loci sequenced (five of the seven MLST loci) diagnostic of the South American subspecies Xylella fastidiosa subsp. pauca which causes two economically damaging plant diseases, citrus variegated chlorosis and coffee leaf scorch. The two remaining loci of ST53 carried alleles from what appears to be a new South American form of X. fastidiosa. Four isolates, classified as X. fastidiosa subsp. fastidiosa, showed a low level of introgression of non-native DNA. One grapevine isolate showed introgression of an allele from X. fastidiosa subsp. pauca while the other three (from citrus and coffee) showed introgression of an allele with similar ancestry to the alleles of unknown origin in ST53. The presence of X. fastidiosa subsp. pauca in Central America is troubling given its disease potential, and establishes another route for the introduction of this economically damaging subspecies into the US or elsewhere, a threat potentially compounded by the presence of a previously unknown form of X. fastidiosa.     

Genetic variation of complete mitochondrial genome sequences of the Sumatran rhinoceros (<Emphasis Type="Italic">Dicerorhinus sumatrensis</Emphasis>)

The Sumatran rhinoceros (Dicerorhinus sumatrensis) is the smallest and one of the most endangered rhinoceros species, with less than 100 individuals estimated to live in the wild. It was originally divided into three subspecies but only two have survived, D. sumatrensis sumatrensis (Sumatran subspecies), and D. s. harrissoni (Bornean). Questions regarding whether populations of the Sumatran rhinoceros should be treated as different management units to preserve genetic diversity have been raised, particularly in light of its severe decline in the wild and low breeding success in captivity. This work aims to characterize genetic differentiation between Sumatran rhinoceros subspecies using complete mitochondrial genomes, in order to unravel their maternal evolutionary history and evaluate their status as separate management units. We identified three major phylogenetic groups with moderate genetic differentiation: two distinct haplogroups comprising individuals from both the Malay Peninsula and Sumatra, and a third group from Borneo. Estimates of divergence time indicate that the most recent common ancestor of the Sumatran rhinoceros occurred approximately 360,000 years ago. The three mitochondrial haplogroups showed a common divergence time about 80,000 years ago corresponding with a major biogeographic event in the Sundaland region. Patterns of mitochondrial genetic differentiation may suggest considering Sumatran rhinoceros subspecies as different conservation units. However, the management of subspecies as part of a metapopulation may appear as the last resource to save this species from extinction, imposing a conservation dilemma.     

Regional differentiation among populations of the Diamondback terrapin (Malaclemys terrapin)

The Diamondback terrapin (Malaclemys terrapin) is a brackish-water turtle species whose populations have been fragmented due to anthropogenic activity such as development of coastal habitat and entrapment in commercial blue crab (Callinectes sapidus) fishing gear. Genetic analyses can improve conservation efforts for the long-term protection of the species. We used microsatellite DNA analysis to investigate levels of gene flow among and genetic variability within 21 geographically separate collections of the species distributed from Massachusetts to Texas. Quantified levels of genetic variability (allelic diversity, genotypic frequencies, and heterozygosity) revealed three zones of genetic discontinuity, resulting in four discrete populations: Northeast Atlantic, Coastal Mid-Atlantic, Florida and Texas/Louisiana. The average number of alleles and expected heterozygosity for the four genetic clusters were N_A = 6.54 and H_E = 0.050, respectively. However, the geographic boundaries of the populations did not correspond to accepted terrapin subspecies limits. Our results illuminate not only the need to sample terrapins in additional sites, specifically in the southeast, but also the necessity for allowing uninterrupted gene flow among population groupings to preserve current levels of genetic diversity.     

Molecular phylogeny of Central and South American slider turtles: implications for biogeography and systematics (Testudines: Emydidae: Trachemys)

We analyse phylogeny, systematics and biogeography of slider turtles (Trachemys spp.) using sequence data of four mitochondrial genes (3242 bp) and five nuclear loci (3396 bp) of most South American and southern Central American taxa and representatives of northern Central American, West Indian and North American slider species (16 species and subspecies) and allied North American species (genera Chrysemys, Deirochelys, Graptemys, Malaclemys, Pseudemys). By applying maximum likelihood, relaxed molecular clock and ancestral range analyses, we provide evidence for two successive colonizations of South America by slider turtles. In addition, we show that the current species delineation of Central and South American slider turtles is incorrect. Our data suggest that Trachemys grayi is a distinct polytypic species that embraces, besides the nominotypical subspecies, T. g. emolli and T. g. panamensis. Trachemys ornata is also polytypic with the subspecies T. o. ornata, T. o. callirostris, T. o. cataspila, T. o. chichiriviche and T. o. venusta. Moreover, T. adiutrix should be regarded as a subspecies of T. dorbigni. All studied Trachemys species are inferred to have originated in the Late Miocene to Early Pliocene. The ancestor of the two subspecies of T. dorbigni colonized South America most probably prior to the establishment of the land bridge connecting Central and South America, whereas the two South American subspecies of T. ornata represent a younger independent immigration wave from Central America.     

Rhinanthus minor population genetic structure and subspecies: Potential seed sources of a keystone species in grassland restoration projects

In the last few decades unimproved semi-natural grasslands have been affected by intensification of land use and habitat fragmentation. Because of their biodiversity these species-rich grasslands are of high conservation importance and efforts are under way to restore such habitats. Detailed knowledge of within species diversity will aid deciding on the optimal seed source for such restoration projects, e.g. local genotypes or ecotypes. Rhinanthus minor is a species that is typically found in semi-natural grasslands and is commonly used in grassland restoration projects. This is because R. minor is a hemiparasitic plant that takes minerals and nutrients from its host, which in turn decreases the host's biomass and leads to opportunities for less competitive species in the vegetation. Here, we investigate genetic diversity within and between R. minor populations. This allowed us to test whether the six different subspecies of R. minor that have been described in the UK, based on their morphology, flowering time, and habitat, can be differentiated using molecular markers. We identified moderate levels of genetic differentiation between R. minor populations within the UK. In addition, R. minor individuals from the UK appear to be distinct from R. minor and Rhinanthus angustifolius individuals from other European countries based on microsatellite genotyping and DNA sequencing of cpDNA and rDNA ITS. The molecular markers used in the current study did not separate populations of R. minor based on either their subspecies or habitat. The implication for the use of R. minor in grassland restoration projects seems to be that it is not necessary to use local seeds or seeds from the same subspecies.     

Long forsaken species diversity in the Middle American lizard Holcosus undulatus (Teiidae)

Numerous reptile species have been divided into subspecies. Although this classification may capture the morphological variation within species, it often conceals significant species diversity because many subspecies actually represent species under lineage‐based species concepts. The lizard Holcosus undulatus is a common, widely distributed, monotypic species in Middle America. However, 12 subspecies of this taxon were recognized until the early 1970s. We used two lineage‐based methods for species delimitation to re‐evaluate species limits within H. undulatus with DNA sequence and morphological data. We included all the previously recognized subspecies of H. undulatus except H. u. miadis. Holcosus undulatus was exclusive. In addition, H. u. amphigrammus, H. u. gaigeae, H. u. hartwegi, H. u. parvus, H. u. pulcher, H. u. sinister, H. u. stuarti, H. u. thomasi and H. u. undulatus were supported as distinct evolutionary lineages based on the molecular and morphological evidence. We therefore elevate all of these subspecies to species rank. In addition, two separate mitochondrial lineages may represent cryptic, undescribed species within H. undulatus. The morphological distinctness and allopatry of H. u. miadis and H. u. pulcher, as well as the high genetic divergence of the latter species, suggest that they also represent distinct evolutionary species. Our results also suggest that additional species diversity may still be hidden within the H. u. amphigrammus, H. u. parvus, H. u. sinister and H. u. undulatus lineages. This work supports resurrection of overlooked diversity within Holcosus, which has important implications for the conservation of this genus in Middle America. © 2015 The Linnean Society of London     

Conservation genetics of the endangered San Francisco Bay endemic salt marsh harvest mouse (<Emphasis Type="Italic">Reithrodontomys raviventris</Emphasis>)

The salt marsh harvest mouse (SMHM, Reithrodontomys raviventris) is an endangered species endemic to the San Francisco Bay region of California, USA, where habitat loss and fragmentation over the past century have reduced the mouse’s distribution to <25 % of its historical range. To aid in conservation prioritization, we first investigated the possibility of hybridization with the morphologically similar western harvest mouse (WHM, R. megalotis) in areas of sympatry and developed genetic tools to differentiate the two species. We then investigated the phylogeography and genetic structure of the SMHM, including support for currently recognized SMHM subspecies designations. Lastly, we evaluated the morphological criteria currently used for the identification of species in the field. Analyses using mtDNA cytochrome b sequences and 11 microsatellites from 142 mice indicated complete and substantial separation of the SMHM and WHM, with no evidence of hybridization. These genetic markers as well as the mtDNA control region also identified a deep genetic division within the SMHM concordant with the current subspecies designations, R. r. raviventris and R. r. halicoetes. We identified the lowest genetic diversity within the southern subspecies, which inhabits a much reduced and highly fragmented portion of the species range. Morphological field identification of harvest mouse species was more successful at identifying SMHM (92 %) than WHM (44 %), with a large portion of WHM being incorrectly identified as SMHM. Field identification of harvest mouse species in the range of the southern SMHM subspecies was just above 50 %, indicating that current methods for morphological differentiation of species in that area are insufficient. Our confirmation of genetically distinct SMHM subspecies highlights the importance of determining the status and genetic composition of relict populations in the remaining patches of marshland in the central San Francisco Bay where the two subspecies may occur, as well as developing better tools for the discrimination of species, particularly in the range of the southern subspecies     

A tale of two clades: Comparative study of Glyptodon Owen and Glyptotherium Osborn (Xenarthra,Cingulata, Glyptodontidae)

Glyptodon and Glyptotherium represent the most conspicuous taxa of late Neogene and Pleistocene glyptodonts in South America and North America, respectively. The earliest records of Glyptodon in South America are 1.07 Ma (late early Pleistocene, Calabrian), although the possibility that “Paraglyptodon uquiensis” represents a Pliocene specimen of Glyptodon cannot be rejected. Glyptotherium originated from South American ancestry in northern South America or Central America about 3.9 Ma (early late Pliocene, Zanclean) or earlier. The diversity of South American Glyptodon is currently under study, but preliminary evidence would indicate that no more than three species (G. munizi, G. elongatus and G. reticulatus) are valid, plus a possible new Andean species. In turn, according to the updated taxonomy proposed herein, Glyptotherium includes two chronospecies. The earliest species, Gl. texanum, differs only slightly from the latest species, Gl. cylindricum. The relationship of Glyptodon and Glyptotherium has been problematical since the discovery of the North American lineage, at first identified as various species of Glyptodon and later considered a separate genus. Glyptodon is recognized as a natural group and recent taxonomic and phylogenetic revisions place all North American glyptodontines into Glyptotherium. In this paper, we propose a detailed morphological comparison between the southern South American species of Glyptodon and Glyptotherium in order to identify diagnostic differences and potential synapomorphies. Both genera can be distinguished mainly by differences in the skull, mandible, dentition, dorsal carapace, and caudal armor, Glyptodon being somewhat larger than Glyptotherium. Both clades show a highly conservative evolution, which could be interpreted as an anagenesis. The scarce records of glyptodonts in Central America show more morphological affinity with Glyptotherium than with Glyptodon.     

The Role of Geographical and Ecological Factors on Population Divergence of the Neotropical otter <Emphasis Type="Italic">Lontra longicaudis</Emphasis> (Carnivora,Mustelidae)

The geographic distribution of the populations of a species are influenced by the spatial structure of the ecosystems, the environmental factors and the presence of geographic barriers. The Neotropical otter, Lontra longicaudis, is widely distributed throughout the Americas, where a wide range of environmental conditions and geographical features could promote genetic and morphological variation on the three currently recognized subspecies. In this study, we combined phylogeographic, morphometric and environmental niche modelling analyses to examine whether: (1) genetic variation is associated with the presence of barriers to gene flow and/or hydrography; (2) genetic and morphologic variation are associated with environmental variation; and (3) the observed variation in L. longicaudis populations corresponds to the previously defined subspecies. We found strong phylogeographic structure between the northern (L. l. annectens) and the two-southern subspecies (L. l. longicaudis and L. l. enudris), and although shallower, we also detected genetic differentiation between the two South American subspecies. Such genetic differentiation corresponds to the hydrography and to the geographical barriers characteristic of the distributional area of the species. We found a correlation between the shape of the skull and mandible with the environmental variation through the distribution of the species, and we rejected the hypothesis of niche equivalency and similarity between the three identified genetic lineages, suggesting adaptations to different environmental conditions. Our results support that the variation in environmental conditions, in concert with geographical barriers to gene flow and hydrography, have led to population divergence of L. longicaudis along the Neotropics. These results have important taxonomic implications for the species and its conservation.     

Low genetic diversity and evidence of population structure among subspecies of Nerodia harteri, a threatened water snake endemic to Texas

Nerodia harteri is a threatened small-bodied water snake that occupies one of the most restricted ranges of any snake within the continental United States. It is found closely associated with rivers and tributaries in the Colorado and Brazos river basins, which flow through north-central Texas. Nerodia harteri has been at the center of debate owing to conflicts between conservation efforts and the construction of dams that change or destroy its preferred habitat. Additionally, its taxonomic status has also been under contention with some authors recognizing two subspecies, the Brazos water snake (N. h. harteri) and the Concho water snake (N. h. paucimaculata), whereas other authors consider each separate species. Despite its relatively recent discovery during the 1940s, N. harteri has been the subject of several ecological studies, yet no population genetic assessment of either subspecies has been performed to date. We first evaluated the phylogenetic placement of both subspecies among other North American Nerodia using partial sequence data from the mitochondrial gene cytochrome-b. We then tested for population subdivision among four rivers encompassing the range of N. harteri and tested for the presence of admixture between river basins using mitochondrial sequence data (920?bp of cyt-b) and five cross-species amplified microsatellite loci. We found low mitochondrial haplotype diversity represented by two unique haplotypes in each river basin, which were separated by no more than four nucleotide changes. Nuclear loci showed low genetic diversity and population structuring within and among river basins. We did not find conclusive evidence of admixture between basins, and we support the presence of two separate evolutionarily significant units and two separate management units corresponding to each major river basin. Given increasing natural and anthropogenic threats, we recommend continued ecological and genetic monitoring of both subspecies.     

A survey of equid mitochondrial DNA: Implications for the evolution, genetic diversity and conservation of Equus

The evolution, taxonomy and conservation of the genus Equuswere investigated by examining the mitochondrial DNA sequences of thecontrol region and 12S rRNA gene. The phylogenetic analysis of thesesequences provides further evidence that the deepest node in thephylogeny of the extant species is a divergence between twolineages; one leading to the ancestor of modern horses (E.ferus, domestic and przewalskii) and the other to thezebra and ass ancestor, with the later speciation events of the zebrasand asses occurring either as one or more rapid radiations, or withextensive secondary contact after speciation. Examination of the geneticdiversity within species suggested that two of the E. hemionussubspecies (E. h. onager and E. h. kulan) onlyrecently diverged, and perhaps, are insufficiently different to beclassified as separate subspecies. The genetic divergence betweendomestic and wild forms of E. ferus (horse) and E.africanus (African ass) was no greater than expected within anequid species. In E. burchelli (plains zebra) there was anindication of mtDNA divergence between populations increasing withdistance. The implications of these results for equid conservation arediscussed and recommendations are made for conservation action.     

Is there more than one Crocodile Lizard? An Integrative Taxonomic Approach Reveals Vietnamese and Chinese Shinisaurus crocodilurus Represent Separate Conservation and Taxonomic Units

The Crocodile lizard Shinisaurus crocodilurus, the only living representative of the family Shinisauridae, is a habitat specialist adapted to remote freshwater habitats within evergreen broadleaf forests. Its current distribution is restricted to few small and isolated remnant occurrences in South China and North Vietnam. Multiple anthropogenic threats such as massive habitat destruction and unsustainable over-collection for the international pet trade brought the species to the brink of extinction. We herein employed an integrative taxonomic approach including comprehensive molecular comparisons based on fragments of mitochondrial genes (cytochrome b, partial ND6, and partial tRNA-Glu) in concert with in-depth morphological and ecological analyses in order to determine the status of the extant populations. Based on molecular, morphological, and ecological differences, we herein describe a new subspecies, Shinisaurus crocodilurus vietnamensis ssp. n., from Vietnam. Our findings emphasize the importance of improved in situ conservation measures in both countries, as both China and Vietnam harbor unique Crocodile lizard forms. We also recommend additional ex situ conservation measures, i.e., separate conservation breeding management of the subspecies in order to maintain genetic integrity and adjust husbandry conditions according to detected differences in ecological niche occupation.     

The study of genetic diversity patterns of Coffea commersoniana, an endangered coffee species from Madagascar: a model for conservation of other littoral forest species

Madagascar has 59 described species of Coffea, of which 42 are listed as critically endangered, endangered, or vulnerable by the criteria of the Red List Category system of the World Conservation Union (IUCN). The littoral forest of Madagascar is a distinctive type of humid evergreen forest restricted to unconsolidated sand located within a few kilometers of the Indian Ocean, now persisting only as small fragments with ca. 10 % of its original range remaining. In an attempt to understand the genetic diversity of Madagascan coffee species, we studied ex situ and in situ populations of Coffea commersoniana, an endemic species of the littoral forests of southeastern Madagascar and soon to be impacted by mining activities in that region. The in situ populations studied showed higher genetic diversity than the ex situ population. The genetic partitioning among the two in situ populations of C. commersoniana was high enough to necessitate keeping the two populations separate for restoration purposes. Based on these findings, recommendations for conservation management (in situ and ex situ) are made.     

Population genetics of the Federally Threatened Miccosukee gooseberry (Ribes echinellum), an endemic North American species

Ribes echinellum (Coville) Rehder (Miccosukee gooseberry; Grossulariaceae) is a Federally Threatened species known from only two localities: Jefferson County (Florida, FL) and McCormick County (South Carolina, SC). This perennial shrub, ca. 1 m tall, is deciduous, and reproduces both vegetatively (clonal growth) and sexually (seed production). Recent surveys of the FL population revealed a dramatic decline in plant numbers. To assist in conservation and management of this species in FL and SC populations, microsatellite genetic markers were used to identify genotypes and assess the genetic structure of R. echinellum. We genotyped seven microsatellite loci in 102 individuals: 74 collected in FL and 28 in SC. Unbiased heterozygosity was between 0.28 and 0.53. All seven loci were polymorphic, showing a range of 1.52–2.13 effective number of alleles per locus (mean = 1.75). The two populations of R. echinellum show low genetic diversity, especially in SC. Clonality was not widespread, but was higher in the SC population. Both populations show signatures of bottlenecks but isolation by distance was not evident. We found significant deviation from HW equilibrium, with higher number of heterozygotes than expected. However when HW test was done for the combined populations as two separate groups, only FL showed a significant HW test and for SC the test was non-significant. Bayesian analysis and F_ST values suggest high genetic divergence between the populations. These results are important for developing a recovery plan and an ex situ and reintroduction conservation programs.