Threatened gastropods under the evolutionary genetic species concept: redescription and new species of the genus Aylacostoma (Gastropoda: Thiaridae) from High Paraná River (Argentina–Paraguay)

The genus Aylacostoma Spix, 1827, is mainly endemic to South America, and comprises about 30 nominal species, most of which were described based solely on conchological features following the typological approaches of most of the 19^th and the mid‐20^th century authors. Here, we redescribe Aylacostoma chloroticum Hylton Scott, 1954, and describe A ylacostoma brunneum sp. nov . from the High Paraná River (Argentina–Paraguay) by means of morphological and molecular characters. Both are threatened species currently included into an ongoing ex situ conservation programme, as their habitats have disappeared because of damming and the filling up of the Yacyretá Reservoir in the early 1990s. We used DNA sequences from cytochrome b and cytochrome oxidase subunit I (COI) genes to estimate their genetic distances, and the COI sequences were also used to assess their specific status under the evolutionary genetic species concept by means of the K/θ method. Our results clearly demonstrate that both must be recognized as evolutionary genetic species, despite only minor differences in morphological characters other than in the shells. © 2014 The Linnean Society of London     

Genetic diversity in wild and reared populations of the Japanese bitterling <Emphasis Type="Italic">Tanakia tanago</Emphasis> (Cyprinidae)

 The Japanese bitterling Tanakia tanago is an endangered cyprinid species; thus, captive breeding programs are being conducted in various facilities as ex situ conservation. To examine the genetic diversity in one wild and three reared populations, and its changes during the process of captive breeding, sequences of the mitochondrial cytochrome b gene and control region were determined. The wild population, collected in 1993, was monomorphic. Although the reared population that originated from the wild population was almost monomorphic, a rare haplotype, distinct from all others by a relatively large sequence divergence, was also observed. In the other reared populations, some degree of genetic diversity had been maintained. A reared hybrid population, which originated from a mixture of three distinct populations, showed the greatest genetic diversity. These results suggest considerable genetic diversity within and among populations of T. tanago in the past. Although a loss of genetic diversity was observed in some year-classes of reared populations, there was no tendency for genetic diversity to decrease as a result of captive breeding, probably because offspring were obtained from multi-year-class parents in the captive breeding program. Accordingly, this breeding method should be appropriate for conserving the genetic diversity of T. tanago. Received: June 12, 2002 / Revised: December 3, 2002 / Accepted: December 16, 2002     

Chloroplast DNA phylogeography of the endangered Japanese red maple (Acer pycnanthum): the spatial configuration of wetlands shapes genetic diversity

Aim Japanese red maple (Acer pycnanthum K. Koch) is an endangered species which grows in discrete wetland ecosystems within a limited geographical range. It is an important relic of geologic time, an endemic of unique wetland ecosystems and an indicator of hotspots of plant species diversity. However, information on its genetic characteristics across its range is lacking. Our aim was to determine the genetic structure and diversity of the species and make recommendations for conservation. Location Wetlands in central Honshu Island, Japan. Methods We collected leaf samples from 400 individuals of A. pycnanthum in 30 populations, extracted total DNA from each and sequenced three non‐coding regions of chloroplast DNA. Results We identified nine haplotypes. High haplotype diversity (0.81) and the occurrence of rare haplotypes in eight distant populations suggest that wetlands provided multiple, adequate‐size refuges during the Last Glacial Maximum. We found only one to four haplotypes within populations. The high degree of differentiation (G_ST = 0.83) implies that gene flow by seeds among populations is restricted. Eight populations demonstrated a positive contribution to the total genetic diversity owing to occurrence of rare and private haplotypes. Such populations are concentrated in the south‐western part of the species distribution. According to the spatial autocorrelation analysis, there were significant spatial clusters of populations, which were characterized by similar haplotype composition. Using the haplotype distribution, samova and barrier detected nearly identical genetic boundaries. Main conclusion In spite of the species’ limited geographical range, we identified a relatively high number of haplotypes and a clear geographical structure. We propose six management units, which can be used for future conservation activities, such as introduction of new individuals for on‐site conservation projects and seed collection for ex situ conservation.     

Genetic Variation in a Chilean Endangered Endemic: <Emphasis Type="Italic">Gomortega keule</Emphasis> (molina) Baillon

Gomortega keule (Molina) Baillon is an endangered, rare species, the only representative of its genus, and endemic to Central Chile. Populations of this tree are now fragmented and few individuals can be found in any of them. Genetic diversity was studied in 33 individuals from three populations in Cauquenes, a coastal mountain area (35°58'S-72°41'W). Fifteen InterSimple Sequence Repeat primers were used to determine the degree of similarity between and within populations. This revealed that 30% of the variation exhibited was between populations while 70% was within; nevertheless individuals were clearly clustered in a pattern which reflected a narrow base of diversity. Three other species from the Laurales order were used in order to provide an external reference as to the degree of diversity. In addition, an external wild population from the native species, Peumus boldus, was used to verify the utility of the markers. We show that the primers are effective in quickly giving an estimate of the degree of diversity of a population, thus giving important topical information relevant to preserving endangered species. Aspects of the conservation and management policy for the species in order to maintain the remaining populations and to preserve the genetic resources are discussed.     

Genetic diversity and taxonomy: a reassessment of species designation in tuatara (<Emphasis Type="Italic">Sphenodon</Emphasis>: Reptilia)

The identification of species boundaries for allopatric populations is important for setting conservation priorities and can affect conservation management decisions. Tuatara (Sphenodon) are the only living members of the reptile order Sphenodontia and are restricted to islands around New Zealand that are free of introduced mammals. We present new data of microsatellite DNA diversity and substantially increased mtDNA sequence for all 26 sampled tuatara populations. We also re-evaluate existing allozyme data for those populations, and together use them to examine the taxonomic status of those populations. Although one could interpret the data to indicate different taxonomic designations, we conclude that, contrary to current taxonomy, Sphenodon is best described as a single species that contains distinctive and important geographic variants. We also examine amounts of genetic variation within populations and discuss the implications of these findings for the conservation management of this iconic taxon.     

Genetic monitoring of the Amazonian fish matrinchã (Brycon cephalus) using RAPD markers: insights into supportive breeding and conservation programmes

The importance of genetic evaluations in aquaculture programmes has been increased significantly not only to improve effectiveness of hatchery production but also to maintain genetic diversity. In the present study, wild and captive populations of a commercially important neotropical freshwater fish, Brycon cephalus (Amazonian matrinchã), were analyzed in order to evaluate the levels of genetic diversity in a breeding programme at a Brazilian research institute of tropical fish. Random Amplified Polymorphic DNA fingerprinting was used to access the genetic variability of a wild stock from the Amazon River and of three captive stocks that correspond to consecutive generations from the fishery culture. Although farmed stocks showed considerably lower genetic variation than the wild population, a significantly higher level of polymorphism was detected in the third hatchery generation. The results seem to reflect a common breeding practice on several hatchery fish programmes that use a small number of parents as broodstocks, obtaining reproductive success with few non‐identified mating couples. The obtained data were useful for discussing suitable strategies for the genetic management and biodiversity conservation of this species.     

Maintenance of Phenotypic and Genotypic Diversity in Managed Populations of Stenocereus Stellatus (Cactaceae) by Indigenous Peoples in Central Mexico

The columnar cactus Stenocereus stellatus is used in Central Mexico for its edible fruits which are harvested in wild, managed in situ and cultivated populations. Management in situ of wild populations is conducted by selectively sparing and enhancing the abundance of plants with desirable phenotypes when fields are cleared for agricultural use. Cultivation of desirable phenotypes is carried out by vegetative propagation in homegardens. Effects of human management on morphological and genetic variation of S. stellatus were analyzed by comparing morphological diversity indices (MD, based on Simpson’s index) and expected (H_e) heterozygosity indices from allozyme analysis, in wild, managed in situ, and cultivated populations from La Mixteca and the Tehuacán Valley regions. Morphological diversity was similar among regions, but populations from the wetter La Mixteca region averaged higher genetic variation (H_e = 0.279) than populations from Tehuacán (H_e = 0.265). On average, populations manipulated by people had higher levels of variation (MD = 0.479 ± 0.012, H_e = 0.289 in cultivated populations; MD = 0.461 ± 0.014, H_e = 0.270 in managed in situ populations) than wild populations (MD = 0.408 ± 0.017, H_e = 0.253), which is apparently due to a continual introduction and replacement of plant materials in the manipulated populations. The results illustrate that human management may not only maintain but also increase both morphological and genetic diversity of manipulated plant populations in relation to that existing in the wild. Managed in situ and cultivated populations of S. stellatus are important reservoirs of variation, and are crucial for the general maintenance of diversity in wild populations. These populations may play a principal role in designing strategies for the conservation of variation of this cactus.     

Genetic variation of wild litchi (<Emphasis Type="Italic">Litchi chinensis</Emphasis> Sonn. subsp. <Emphasis Type="Italic">chinensis</Emphasis>) revealed by microsatellites

Understanding the amount and distribution of genetic diversity in natural populations can inform the conservation strategy for the species in question. In this study, genetic variation at eight nuclear microsatellite loci was used to investigate genetic diversity and population structure of wild litchi (Litchi chinensis Sonn. subsp. chinensis). Totally 215 individuals were sampled, representing nine populations of wild litchi. All eight loci were polymorphic, with a total of 51 alleles. The expected heterozygosity in the nine populations ranged from 0.367 to 0.638 with an average value of 0.526. Inbreeding within wild litchi populations was indicated by a strong heterozygote defect. Significant bottleneck events were detected in the populations from Yunnan and Vietnam, which could be responsible for lower levels of genetic diversity in these populations. Measures of genetic differentiation (F _ST = 0.269) indicated strong differentiation among wild litchi populations. Significant correlation was found between genetic differentiation and geographical distance (r = 0.655, P = 0.002), indicating a strong isolation by distance in these populations. Bayesian clustering suggested genetic separation among three regional groups, namely, the western group, the central group and the eastern group. Some conservation strategies for wild litchi populations were also proposed based on our results.     

Strategies for the retention of high genetic variability in European flat oyster (<Emphasis Type="Italic">Ostrea edulis</Emphasis>) restoration programmes

The native European flat oyster Ostrea edulis is listed in the OSPAR Convention for the Protection of the Marine Environment of the North-East Atlantic (species and habitat protection) and in the UK Biodiversity Action Plan. Once extremely abundant in the nineteenth century, European stocks of O. edulis have declined during the twentieth century to rare, small, localised populations due to overexploitation, habitat degradation and, most recently, the parasitic disease bonamiosis. Selective breeding programmes for resistance to bonamiosis have been initiated in France and Ireland. High genetic diversity and bonamiosis-resistance would be important features of any sustainable restoration programmes for O. edulis. Oysters were sampled across Europe from four hatchery sources, four pond-cultured sources and four wild, but managed fisheries and were genotyped at five microsatellite loci. Hatchery-produced populations from small numbers of broodstock showed a significant loss of genetic diversity relative to wild populations and pedigree reconstruction revealed that they were each composed of a single large full-sib family and several small full-sib families. This extremely low effective population size highlights the variance in reproductive success among the potential breeders. Pond-cultured oysters were intermediate in genetic diversity and effective population size between hatchery and wild populations. Controlled hatchery production allows the development of bonamiosis-resistant strains, but at the expense of genetic diversity. Large scale pond culture on the other hand can provide a good level of genetic diversity. A mixture of these two approaches is required to ensure a healthy and sustainable restoration programme for O. edulis in Europe.     

Conservation of genetic diversity hotspots of the high‐valued relic yellowhorn (Xanthoceras sorbifolium) considering climate change predictions

Genetic structure and major climate factors may contribute to the distribution of genetic diversity of a highly valued oil tree species Xanthoceras sorbifolium (yellowhorn). Long‐term over utilization along with climate change is affecting the viability of yellowhorn wild populations. To preserve the species known and unknown valuable gene pools, the identification of genetic diversity “hotspots” is a prerequisite for their consideration as in situ conservation high priority. Chloroplast DNA (cpDNA) diversity was high among 38 natural populations (H_d = 0.717, K = 4.616, Tajmas’ D = ?0.22) and characterized by high genetic divergence (F_ST = 0.765) and relatively low gene flow (N_m = 0.03), indicating populations isolation reflecting the species’ habitat fragmentation and inbreeding depression. Six out of the studied 38 populations are defined as genetic diversity “hotspots.” The number and geographic direction of cpDNA mutation steps supported the species southwest to northeast migration history. Climatic factors such as extreme minimum temperature over 30 years indicated that the identified genetic “hotspots” are expected to experience 5°C temperature increase in next following 50 years. The results identified vulnerable genetic diversity “hotspots” and provided fundamental information for the species’ future conservation and breeding activities under the anticipated climate change. More specifically, the role of breeding as a component of a gene resource management strategy aimed at fulfilling both utilization and conservation goals.     

Population structure and genetic diversity of wild <Emphasis Type="Italic">Helianthus</Emphasis> species from Mozambique

The production of sunflower suffered a major decline in Mozambique after its independence in 1975. Civil war, human activities and environmental damage subjected the species to an ecological stress contributing to reduce the number and size of wild populations. As this reduction is often related to a loss of genetic variation we estimated the genetic diversity within and among populations of wild Helianthus from five districts of Mozambique using RAPD markers. The 44 accessions studied grouped into four major clusters exhibiting structured variability with regard to geographic origin. A high level of genetic diversity (He = 0.350 and I = 0.527) was retained at the population level. The genetic variation among populations was high (59.7%), which is consistent with low gene flow (Nm = 0.338). The proportion of total genetic diversity residing among these populations should be kept in mind to devise different conservation strategies in order to preserve these populations. Currently wild Helianthus genetic resources present in Maputo and Sofala are on the edge of extinction mainly due to excessive urbanization. Therefore, conservation of what remains of this plant genetic diversity is essential for sustainable utilization and can be useful for breeding programs.     

Population genetics of Galápagos land iguana (genus Conolophus) remnant populations

The Galápagos land iguanas (genus Conolophus) have faced significant anthropogenic disturbances since the 17th century, leading to severe reduction of some populations and the extinction of others. Conservation activities, including the repatriation of captive‐bred animals to depleted areas, have been ongoing since the late 1970s, but genetic information has not been extensively incorporated. Here we use nine species‐specific microsatellite loci of 703 land iguanas from the six islands where the species occur today to characterize the genetic diversity within, and the levels of genetic differentiation among, current populations as well as test previous hypotheses about accidental translocations associated with early conservation efforts. Our analyses indicate that (i) five populations of iguanas represent distinct conservation units (one of them being the recently discovered rosada form) and could warrant species status, (ii) some individuals from North Seymour previously assumed to be from the natural Baltra population appear related to both Isabela and Santa Cruz populations, and (iii) the five different management units exhibit considerably different levels of intrapopulation genetic diversity, with the Plaza Sur and Santa Fe populations particularly low. Although the initial captive breeding programmes, coupled with intensive efforts to eradicate introduced species, saved several land iguana populations from extinction, our molecular results provide objective data for improving continuing in situ species survival plans and population management for this spectacular and emblematic reptile.     

Partnering conservation actions. <Emphasis Type="Italic">Inter situ</Emphasis> solutions to recover threatened species in South West Western Australia

South West Western Australia has a rich endemic flora of global significance. The threats facing this floral diversity are increasing in type, severity and scale, demonstrated by the rising numbers of species threatened with extinction. In particular, the root-rot pathogen Phytophthora cinnamomi is causing widespread destruction, threatening the survival of many of the region’s unique plants. In situ conservation of wild plants is considered the most essential component of a flora conservation program, but the ability to conserve some species adequately is often unachievable in the short term and urgent management intervention is required to prevent extinction. We present data on the status and management of wild populations of four threatened species from the region, including an ex situ program, and describe our efforts to bridge the gap between these two components. Such inter situ conservation recovery work enables monitoring of biological attributes, research into reproductive biology and collection of genetic material for further ex situ conservation, and provides the source of material for future restoration of wild populations.     

Ex Situ Conservation Priorities for the Wild Relatives of Potato (Solanum L. Section Petota)

Crop wild relatives have a long history of use in potato breeding, particularly for pest and disease resistance, and are expected to be increasingly used in the search for tolerance to biotic and abiotic stresses. Their current and future use in crop improvement depends on their availability in ex situ germplasm collections. As these plants are impacted in the wild by habitat destruction and climate change, actions to ensure their conservation ex situ become ever more urgent. We analyzed the state of ex situ conservation of 73 of the closest wild relatives of potato (Solanum section Petota) with the aim of establishing priorities for further collecting to fill important gaps in germplasm collections. A total of 32 species (43.8%), were assigned high priority for further collecting due to severe gaps in their ex situ collections. Such gaps are most pronounced in the geographic center of diversity of the wild relatives in Peru. A total of 20 and 18 species were assessed as medium and low priority for further collecting, respectively, with only three species determined to be sufficiently represented currently. Priorities for further collecting include: (i) species completely lacking representation in germplasm collections; (ii) other high priority taxa, with geographic emphasis on the center of species diversity; (iii) medium priority species. Such collecting efforts combined with further emphasis on improving ex situ conservation technologies and methods, performing genotypic and phenotypic characterization of wild relative diversity, monitoring wild populations in situ, and making conserved wild relatives and their associated data accessible to the global research community, represent key steps in ensuring the long-term availability of the wild genetic resources of this important crop.     

Genetic diversity in the US ex situ populations of the endangered Puerto Rican Boa,Chilabothrus inornatus

The Puerto Rican Boa (Chilabothrus inornatus) was placed on the US Endangered Species List in 1970. Progress has been made since to clarify the recovery status of this species, though the design of a new recovery plan must include information regarding genetic variation within and among populations of this species. While measures of genetic diversity in wild populations of this species are finally becoming available, relative genetic diversity represented in ex situ populations is unknown, which hampers efforts to develop an ex situ species management plan. Here, we provide an analysis of genetic diversity in US public and private collections (zoos and breeders) using mitochondrial sequence data and five highly polymorphic nuclear microsatellite loci. We analyzed 50 boas from the US ex situ population and determined overall genetic diversity and relatedness among these individuals. We then compared these data to mitochondrial and microsatellite data obtained from 176 individuals from wild populations across the native range of the species. We found little inbreeding and a large amount of retained genetic diversity in the US ex situ population of C. inornatus relative to wild populations. Genetic diversity in the ex situ population is similar to that found in wild populations. Ours is only the second explicit attempt to characterize genetic diversity at the molecular level in ex situ populations of boid snakes. We anticipate that these results will inform current breeding strategies as well as offer additional information that will facilitate the continuation of ex situ conservation breeding or management in boas.     

Genetic diversity of North American captive‐born gorillas (Gorilla gorilla gorilla)

Western lowland gorillas (Gorilla gorilla gorilla) are designated as critically endangered and wild populations are dramatically declining as a result of habitat destruction, fragmentation, diseases (e.g., Ebola) and the illegal bushmeat trade. As wild populations continue to decline, the genetic management of the North American captive western lowland gorilla population will be an important component of the long‐term conservation of the species. We genotyped 26 individuals from the North American captive gorilla collection at 11 autosomal microsatellite loci in order to compare levels of genetic diversity to wild populations, investigate genetic signatures of a population bottleneck and identify the genetic structure of the captive‐born population. Captive gorillas had significantly higher levels of allelic diversity (t₇ = 4.49, P = 0.002) and heterozygosity (t₇ = 4.15, P = 0.004) than comparative wild populations, yet the population has lost significant allelic diversity while in captivity when compared to founders (t₇ = 2.44, P = 0.04). Analyses suggested no genetic evidence for a population bottleneck of the captive population. Genetic structure results supported the management of North American captive gorillas as a single population. Our results highlight the utility of genetic management approaches for endangered nonhuman primate species.     

A genetically distinct lion (Panthera leo) population from Ethiopia

Lion (Panthera leo) numbers are in serious decline and two of only a handful of evolutionary significant units have already become extinct in the wild. However, there is continued debate about the genetic distinctiveness of different lion populations, a discussion delaying the initiation of conservation actions for endangered populations. Some lions from Ethiopia are phenotypically distinct from other extant lions in that the males possess an extensive dark mane. In this study, we investigated the microsatellite variation over ten loci in 15 lions from Addis Ababa Zoo in Ethiopia. A comparison with six wild lion populations identifies the Addis Ababa lions as being not only phenotypically but also genetically distinct from other lions. In addition, a comparison of the mitochondrial cytochrome b (CytB) gene sequence of these lions to sequences of wild lions of different origins supports the notion of their genetic uniqueness. Our examination of the genetic diversity of this captive lion population shows little effect of inbreeding. Immediate conservation actions, including a captive breeding programme designed to conserve genetic diversity and maintain the lineage, are urgently needed to preserve this unique lion population.     

A comparative study of Odonata (Insecta) assemblages along an altitudinal gradient in the sierra de Coalcomán Mountains, Michoacán, Mexico

Odonate diversity in the Coalcomán Mountain Range (CMR), Michoacán State, Mexico, was surveyed, and samplings were made during 2 years in eight streams along an altitudinal gradient. Presence–absence data were analyzed using non-parametric and parametric methods. Beta and gamma diversities were estimated using Whittaker’s and Lande’s formulae, respectively. A total of 2,526 adults and 489 larvae were captured, yielding 116 species (γ diversity), 44 genera and 9 families. Five new species were discovered. The genus Argia was the most important contributor to Zygoptera diversity and total richness (γ diversity), yielding 40.4 and 14.7%, respectively. The non-parametric estimator Chao2 provided the closest theoretical estimate of species richness, and Clench’s model fit the data well (R ² ranged from 99.44 to 99.99) to explain a high proportion of the variance (98.8). We conclude that beta diversity is important at the landscape scale, supporting the hypothesis that Mexico is a beta diverse country. Our results triple the number of known species of Odonata for Michoacán. Given the considerable richness of odonates at local and landscape scales, our results support the proposal of the Coalcomán Mountain Range as a priority area for conservation and related research.     

Introgression between native and prehistorically naturalized (archaeophytic) wild pear (<Emphasis Type="Italic">Pyrus</Emphasis> spp.) populations in Northern Tohoku,Northeast Japan

Hybridization between native and cultivated species is a concern in conservation biology. However, detecting such hybridization and distinguishing true natives from prehistorically naturalized species based on phenotypic characteristics is difficult. Here, we report on introgression between native and prehistorically introduced pear (Pyrus) species in Northern Tohoku (northern end of Honshu Island), Japan. We analyzed 20 microsatellites in 226 wild, seemingly wild, or cultivated materials. Phenetic analysis showed that wild Japanese populations of P. ussuriensis var. ussuriensis in Northern Tohoku, previously considered true natives based on morphology and phytogeography, differed from those in continental Asia, confirming their nativeness. However, Bayesian inference of population structures showed that Japanese P. ussuriensis was genetically admixed with two genetic clusters: true native P. ussuriensis var. ussuriensis and prehistorically introduced P. pyrifolia. Even in the Kitakami Mountains, where true native populations of var. ussuriensis are believed to persist, most wild trees were at least somewhat admixed. Prehistorically introduced then naturalized plants are treated as natives in Japan’s conservation management, and some are considered endangered. However, introgression of prehistorically naturalized P. pyrifolia into threatened native P. ussuriensis var. ussuriensis has occurred. This paper examines the implications for conservation management.