A sophisticated species conservation strategy for <Emphasis Type="Italic">Nyssa yunnanensis</Emphasis>, a species with extremely small populations in China

Conservation of plant species with extremely small populations (PSESP) is a focus of wild plant conservation in China at present. A relevant strategy for PSESP conservation requires improvement from previous programs for rare and endangered plants and national key protected plants. An integrated strategy for PSESP conservation of the Nyssa yunnanensis was initiated and applied over a 7-year period (2009–2015). Here, we reviewed the processes to implement the strategy: resource inventory, formulation of conservation action plan (CAP), in situ conservation, seedling propagation, near situ conservation, ex situ conservation and scientific research. Major concerns and technical requirements for each action are described and further analyzed within a broad scope to conserve PSESPs. A detailed resource inventory that highlights both the change in population status and the participation of local residents is recommended before the formulation of the CAP. Techniques for determination of the area of a mini-reserve is developed for in situ conservation of N. yunnanensis. Near situ conservation is a novel approach whereby establishing new viable population in contiguous areas with a similar climate, habitat and community, in which techniques for seedling preparation, soil preparation, and early management are introduced. A population-based species conservation strategy for N. yunnanensis may aid additional PSESP conservation, so as to contribute to overall wild plant conservation.     

Conservation genomics of an Australian cycad Cycas calcicola,and the Absence of Key Genotypes in Botanic Gardens

Understanding the genetic diversity of wild populations is fundamental to conserving species in-situ and ex-situ. To aid conservation plans and to inform ex-situ conservation, we examined the genetic diversity of the cycad Cycas calcicola (Cycadaceae). Samples were collected from wild populations in the Litchfield National Park and Katherine regions in the Northern Territory, Australia. Additional samples were obtained from botanic garden plants that were originally collected in the Katherine region, Daly River and Spirit Hills in the Northern Territory, Australia. Using RADseq we recovered 2271 informative genome-wide SNPs, revealing low to moderate levels of gene diversity (uH_e?=?0.037 to 0.135), very low levels of gene flow, and significant levels of inbreeding (mean F_IS?=?0.491). Population structure and multivariate analysis showed that populations fall into two genetic groups (Katherine vs Litchfield?+?Daly River?+?Spirit Hills). Genetic differentiation was twice as high between populations of the Katherine and Litchfield regions (F_ST?~?0.1) compared to within these two regions (F_ST?~?0.05). Increasing population fragmentation together with high levels of inbreeding and very little gene flow are concerning for the future adaptability of this species. The results indicated that the ex-situ collections (1) had significantly lower genetic diversity than the wild populations, and (2) only partly capture the genetic diversity present, particularly because the Litchfield National Park populations are not represented. We recommend that ex-situ collections be expanded to incorporate the genetic diversity found in Litchfield National Park and to increase the number of representatives from Daly River/Spirit Hills, and that in-situ populations from the Katherine and Greater Litchfield regions be conserved as separate management units.

     

Population and genetic status of a critically endangered species in Korea, Euchresta japonica (Leguminosae), and their implications for conservation

The conservation status of Euchresta japonica Hook. f. ex Regel in Korea was investigated, with an emphasis on its genetic diversity. From field surveys, we obtained the only locality record for a wild population in Korea, which contained eight individuals. Genotyping was performed using nine microsatellite markers for all 20 remaining individuals, including those in ex situ collections. Among nine microsatellite loci that amplified within this group, five showed polymorphism with low hererozygosities, and a total of 12 multilocus genotypes were detected. Wild-specific alleles were detected in two individuals, and ex situ-specific alleles were detected in six individuals. Five individuals proved to have individual-specific alleles. The Korean population was also distinguished from the previously reported Japanese population by different alleles and higher diversity. To conserve this species more effectively in Korea, we recommend the following: (1) fencing the remaining wild population; (2) no relocation of wild individuals, as nine ex situ plants are already available; (3) complete ex situ conservation of all genetic diversity via clonal propagation of wild individuals; and (4) continuous protection and monitoring of the wild population.     

Conserving plant species with extremely small populations (PSESP) in China

Over the past decades, biodiversity conservation in China has achieved a number of successes. However, due to inadequate conservation policies, poor implementation and lack of financial support, wild plant species that are extremely small in population size and therefore seriously threatened have not had the attention they require. But the new concept of plant species with extremely small populations (PSESP), first promulgated in Yunnan Province, is becoming more widely accepted in China. Several national and regional-level conservation strategies and actions for conserving China’s PSESP are being implemented over the next 5 years. With this new policy framework leading the way, plant conservation in China is set to make important new advances.     

Decreasing genetic diversity in wild and captive populations of endangered Itasenpara bittering (Acheilognathus longipinnis) in the Himi region,central Japan,and recommendations for conservation

Biodiversity is increasingly declining as a result of direct human impact and structural alteration of ecosystems resulting from changes in human life styles. Itasenpara bittering (Acheilognathus longipinnis), which has been maintained in floodplain and paddy fields, is a threatened cyprinid fish endemic to central Japan. To aid in the preservation of this species, information on genetic diversity and demographic variables in wild and captive populations was obtained using microsatellite DNA analysis. Temporal changes in genetic diversity and effective population size (N _e) tended to be relatively stable in the wild Moo River population, although lower values were detected in the wild Busshouji River population, suggesting an extremely high risk of extinction in the latter. Captive populations derived from the Busshouji River population demonstrated significant genetic divergence even among intrapopulational cohorts, suggesting the influence of genetic drift caused by geographic isolation and small population size. Active maintenance of genetic diversity in captive population is a necessary part of conservation programs, as are continuous addition of wild individuals and replacement of individuals among captive populations. In addition, increasing or maintaining suitable floodplain areas and artificial habitats such as paddy fields might contribute to the conservation of genetic diversity in the Itasenpara bittering.     

Population genetics and distribution data reveal conservation concerns to the sky island endemic <Emphasis Type="Italic">Pithecopus megacephalus</Emphasis> (Anura,Phyllomedusidae)

Pithecopus megacephalus is a reticulated monkey–frog species endemic to the highlands of the Espinhaço Mountain Range in southeastern Brazil, an important centre of endemism in South America. This species has a discontinuous distribution and is considered “data-deficient” by the IUCN Red List, raising concerns about its conservation. Understanding the historical dynamics and connectivity of P. megacephalus populations can provide guidelines for preservation of this species in the wild. To investigate the population dynamics of P. megacephalus, we obtained multilocus DNA data for 55 individuals from different locations along the species’ known distribution. Spatial population structure, genetic diversity and demographic parameters were evaluated using population genetic and phylogeographical tools. We also evaluated its extent of occurrence and area of occupancy to investigate extinction risk of this species. We found genetic structure along P. megacephalus’ spatial distribution in the South Espinhaço Mountain Range corresponding to three population groups: northern, central and southern. Our results could provide important data on geographic distribution and population dynamics for a Data Deficient species. Therefore, we suggest these population data, together with the species’ limited occurrence in sky island environments could be used for a more accurate classification of P. megacephalus in the IUCN list, and conservation strategies for the species should be planned accordingly.     

Genetic population structure and management units of the endangered Tokyo bitterling, Tanakia tanago (Cyprinidae)

The Tokyo bitterling Tanakia tanago (Cyprinidae) was once found throughout the Kanto Plain, central Japan, but most of their habitats have been lost due to human activities such as urbanization and improvement of paddy fields. Subsequently, conservation efforts, including captive breeding and reintroduction, have been ongoing. However, the genetic relationships among populations of this species including captive and remnant wild populations have been uncertain and thus management units for this species have been unidentified. We examined the population differentiation among 12 populations, including four wild and eight captive populations, and their relative genetic diversities to assist in conservation management decisions. Phylogeographic analyses based on partial mitochondrial cytochrome b gene sequences and microsatellite polymorphisms revealed four geographically associated genetic groups in the populations. Northern Tochigi populations have diverged from other populations (0.77% of d _A ), likely stemming from allopatric fragmentation following a change in the route of the Naka River, which occurred during the middle of the Pleistocene epoch. Microsatellite analysis has revealed that the genetic diversity of each population is generally low, and that most of the populations have experienced genetic bottlenecks. For future in- and ex-situ conservation programs to succeed, the population structure and genetic variability of remnant populations need to be taken into consideration.     

The intersection of conservation and horticulture: bird-pollinated Lotus species from the Canary Islands (Leguminosae)

This paper critically reviews the status of the ??lotus vines?? (Lotus section Rhyncholotus) in the wild and in cultivation. The literature concerning the conservation status of this group is scattered and in many places is not updated. Besides, the most recent efforts in conservation on this group have not been published in widely available sources and are mainly published in Spanish. Our aims in this paper are to compile the most recent information regarding the conservation status, studies on genetic diversity and the latest strategies for the conservation of this group. The four species are all critically endangered in the wild. Two species (Lotus berthelotii and L. maculatus) are widely cultivated in Europe and North America but the range of genotypes in cultivation is likely to be small, as the plants are propagated by cuttings. Hybrids between these two species are also in cultivation, and some have been patented. Commercial horticulture has considerable implications for overall conservation strategies, and these implications are discussed. We suggest that a circa situm conservation could improve the conservation strategies of these four species in the Canary Islands.     

Genetic diversity and structure of Pinus dabeshanensis revealed by expressed sequence tag-simple sequence repeat (EST-SSR) markers

Assessing patterns of genetic variation in rare endangered species is critical for developing both in situ and ex situ conservation strategies. Pinus dabeshanensis Cheng et Law is an endangered species endemic to the Dabieshan Mountains of eastern China. To obtain fundamental information of genetic diversity, population history, effective population size, and gene flow in this species, we explored patterns of genetic variation of natural populations, in addition to an ex situ conserved population, using expressed sequence tag-simple sequence repeats (EST-SSR) markers. Our results revealed moderate levels of genetic diversity (e.g., H_E = 0.458 vs. H_E = 0.423) and a low level of genetic differentiation (F_ST = 0.028) among natural and conserved populations relative to other conifers. Both contemporary and historical migration rates among populations were high. Bayesian coalescent-based analyses suggested that 3 populations underwent reductions in population size ca. 10,000 yr ago, and that two populations may have experienced recent genetic bottlenecks under the TPM. Bayesian clustering revealed that individuals from the ex situ population were largely assigned to the ‘red’ cluster. Additionally, our results identified private alleles in the natural populations but not in the ex situ population, suggesting that the ex situ conserved population insufficiently represents the genetic diversity present in the species. Past decline in population size is likely to be due to Holocene climate change. Based on the genetic information obtained for P. dabeshanensis, we propose some suggestions for the conservation and efficient management of this endangered species.     

Genetic structuring in wild populations of two important medicinal plant species as inferred from AFLP markers

Systematic analysis of germplasm diversity and genetic relationship among cultivars is critical for development of appropriate conservation and breeding strategies. This approach has been applied to gain an insight about the molecular variance that exists in wild population of two important medicinal plant species of India that have a long history of therapeutic usage in herbal medicine. Adhatoda vasica and Andrographis paniculata, members of the family Acanthaceae, have wide geographical and climatic distribution across India suggesting a large amount of genetic diversity available for resource management and breeding programs. In this study we have assessed the genetic diversity of both these species distributed in five varied geo-environmental regions, using Amplified Fragment Length Polymorphism (AFLP) fingerprinting with selected primer combinations and statistical analysis. Cluster analysis and analysis of molecular variance also suggested a very high genetic variation. Detailed analyses of the results predict that the genetic variation found in A. vasica was more discrete that reflected strongly in the populations studied, whereas the genetic variation in A. paniculata was relatively uniform. Considering significantly large sample size and distinctive characteristics of the selected populations, this work contributes valuable insights that can be used to engineer conservation and utilization strategies for these species.     

Comparative Genetic Diversity of Wild and Captive Populations of the Bare-Faced Curassow (Crax fasciolata) Based on Cross-Species Microsatellite Markers: Implications for Conservation and Management

The bare-faced curassow (Crax fasciolata) is a large Neotropical bird that suffers anthropogenic pressure across much of its range. A captive population is maintained for conservation management, although there has been no genetic screening of stocks. Based on the six microsatellite markers developed for Crax globulosa, the genetic variability of C. fasciolata and possible differences between a wild and a captive population were investigated. Only three loci were polymorphic, with a total of 27 alleles. More than half of these alleles were private to the wild (n = 8) or captive (n = 7) populations. Significant deviations from Hardy–Weinberg equilibrium were restricted to the captive population. Despite the number of private alleles, genetic drift has probably promoted differentiation between populations. Our results indicate that wild C. fasciolata populations are genetically impoverished and structured, but species-specific microsatellite markers will be necessary for a more reliable assessment of the species’ genetic diversity.     

Conservation implications of genetic structure in the narrowest endemic quillwort from the Eastern Amazon

The quillwort Isoëtes cangae is a critically endangered species occurring in a single lake in Serra dos Carajás, Eastern Amazon. Low genetic diversity and small effective population sizes (N _e) are expected for narrow endemic species (NES). Conservation biology studies centered in a single species show some limitations, but they are still useful considering the limited time and resources available for protection of species at risk of extinction. Here, we evaluated the genetic diversity, population structure, N _e, and minimum viable population (MVP) of I. cangae to provide information for effective conservation programs. Our analyses were based on 55 individuals collected from the Amendoim Lake and 35,638 neutral SNPs. Our results indicated a single panmictic population, moderate levels of genetic diversity, and N _e in the order of thousands, contrasting the expected for NES. Negative F_IS values were also found, suggesting that I. cangae is not under risk of inbreeding depression. Our findings imply that I. cangae contains enough genetic diversity to ensure evolutionary potential and that all individuals should be treated as one demographic unit. These results provide essential information to optimize ex situ conservation efforts and genetic diversity monitoring, which are currently applied to guide I. cangae conservation plans.     

Wild European Apple (Malus sylvestris (L.) Mill.) Population Dynamics: Insight from Genetics and Ecology in the Rhine Valley. Priorities for a Future Conservation Programme

The increasing fragmentation of forest habitats and the omnipresence of cultivars potentially threaten the genetic integrity of the European wild apple (Malus sylvestris (L.) Mill). However, the conservation status of this species remains unclear in Europe, other than in Belgium and the Czech Republic, where it has been declared an endangered species. The population density of M. sylvestris is higher in the forests of the upper Rhine Valley (France) than in most European forests, with an unbalanced age-structure, an overrepresentation of adults and a tendency to clump. We characterize here the ecology, age-structure and genetic diversity of wild apple populations in the Rhine Valley. We use these data to highlight links to the history of this species and to propose guidelines for future conservation strategies. In total, 255 individual wild apple trees from six forest stands (five floodplain forests and one forest growing in drier conditions) were analysed in the field, collected and genotyped on the basis of data for 15 microsatellite markers. Genetic analyses showed no escaped cultivars and few hybrids with the cultivated apple. Excluding the hybrids, the genetically “pure” populations displayed high levels of genetic diversity and a weak population structure. Age-structure and ecology studies of wild apple populations identified four categories that were not randomly distributed across the forests, reflecting the history of the Rhine forest over the last century. The Rhine wild apple populations, with their ecological strategies, high genetic diversity, and weak traces of crop-to-wild gene flow associated with the history of these floodplain forests, constitute candidate populations for inclusion in future conservation programmes for European wild apple.     

Fruit consumption and seed dispersal by birds in native vs. ex situ individuals of the endangered Chinese yew,Taxus chinensis

The conservation success of endangered trees may depend on re-establishing or replacing the mutualisms that were important in their native habitats. In this study, we quantified avian frugivore diversity on individuals of the endangered Chinese yew (Taxus chinensis) in a botanical garden and at a natural site. We found that frugivore species diversity was lower in the botanical garden than in the natural site. In spite of the relatively low frugivore diversity, however, the ex-situ population of Chinese yew attracted similar disperser species to those in the natural population, and trees in the ex-situ population were visited more frequently than those in the natural population. Furthermore, the different perching behavior of dispersers resulted in different dispersal efficiencies. Although the Chinese bulbul (Pycnonotus sinensis) was the most common forager in the botanical garden, its dispersal efficiency was the lowest, and thus it could not like the role played by the mountain bulbul (Hypsipetes maclellandii) in the natural site. Only the red-billed blue magpie (Urocissa erythrorhyncha) provided a high-quality dispersal service in both sites. Our results highlight the ability of the Chinese yew to recruit seed dispersal agents in new habitats. However, if the newly recruited species is a low-quality disperser, the plants will depend more heavily on other avian vectors for regeneration.     

Genetic variability,management, and conservation implications of the critically endangered Brazilian pitviper Bothrops insularis

Information on demographic, genetic, and environmental parameters of wild and captive animal populations has proven to be crucial to conservation programs and strategies. Genetic approaches in conservation programs of Brazilian snakes remain scarce despite their importance for critically endangered species, such as Bothrops insularis, the golden lancehead, which is endemic to Ilha da Queimada Grande, coast of São Paulo State, Brazil. This study aims to (a) characterize the genetic diversity of ex situ and in situ populations of B. insularis using heterologous microsatellites; (b) investigate genetic structure among and within these populations; and (c) provide data for the conservation program of the species. Twelve informative microsatellites obtained from three species of the B. neuwiedi group were used to access genetic diversity indexes of ex situ and in situ populations. Low‐to‐medium genetic diversity parameters were found. Both populations showed low—albeit significant—values of system of mating inbreeding coefficient, whereas only the in situ population showed a significant value of pedigree inbreeding coefficient. Significant values of genetic differentiation indexes suggest a small differentiation between the two populations. Discriminant analysis of principal components (DAPC) recovered five clusters. No geographic relationship was found in the island, suggesting the occurrence of gene flow. Also, our data allowed the establishment of six preferential breeding couples, aiming to minimize inbreeding and elucidate uncertain parental relationships in the captive population. In a conservation perspective, continuous monitoring of both populations is demanded: it involves the incorporation of new individuals from the island into the captive population to avoid inbreeding and to achieve the recommended allelic similarity between the two populations. At last, we recommend that the genetic data support researches as a base to maintain a viable and healthy captive population, highly genetically similar to the in situ one, which is crucial for considering a reintroduction process into the island.     

Genetic diversity and population structure of Schizopygopsis younghusbandi Regan in the Yarlung Tsangpo River inferred from mitochondrial DNA sequence analysis

The genetic diversity and population structure of Schizopygopsis younghusbandi from six sites in the middle reach of the Yarlung Tsangpo River were examined based on mitochondrial DNA cytochrome b (Cyt b) gene and control region (D-loop) sequences. 1141 bp of complete Cyt b sequences and 737 bp of partial D-loop sequences for 153 individuals of S. younghusbandi were obtained by using PCR amplification and sequencing. The results showed that S. younghusbandi populations had high haplotype diversity and low nucleotide diversity, and the population genetic diversity of this species was at a low level. Analysis of molecular variance revealed that most genetic variation occurred within populations, and that genetic differentiation was at low or moderate levels. The network of haplotypes based on Cyt b gene showed that there were two groups within the examined populations. Neutrality tests and mismatch distribution analysis suggested that this species might have undergone a population expansion and the expansion time was estimated to be 0.25–0.46 Ma BP. All these results would be crucial to establish scientific strategies for effective conservation and sustainable exploitation of wild resources of S. younghusbandi.     

Patterns of geographic distribution have a considerable influence on population genetic structure in one common and two rare species of Rhododendron (Ericaceae)

Genetic diversity is essential for species to sustain their populations and evolutionary potential. In order to develop effective conservation strategies for rare species, it is necessary to understand differences in patterns of genetic diversity between common and rare species. Data about population genetic structure is important to design effective conservation strategies for rare species. In this study, we compared the genetic diversity and population genetic structure of a common species, Rhododendron weyrichii, to those of two rare species, Rhododendron sanctum and Rhododendron amagianum, with different geographic distributions. We analyzed five microsatellite loci in 16 populations of R. weyrichii, 9 populations of R. sanctum, and 6 populations of R. amagianum. As expected, the level of genetic diversity indicated by allelic richness and gene diversity was lower for the rare species R. sanctum than for the common species R. weyrichii. However, there was no statistically significant difference in genetic diversity between R. weyrichii and the other rare species, R. amagianum. Analyses of the isolation-by-distance pattern, neighbor-joining trees, and Bayesian clustering indicated that R. sanctum had a strong population genetic structure whereas R. amagianum exhibited very weak genetic structure among populations and that there was moderate population genetic structure for R. weyrichii. Therefore, the degree and pattern of population genetic structure in each species was unrelated to its rarity and instead merely reflected its geographic distribution.     

Restricted gene flow in fragmented populations of a wind-pollinated tree

Fragmentation of natural populations can have negative effects at the genetic level, thus threatening their evolutionary potential. Many of the negative genetic impacts of population fragmentation can be ameliorated by gene flow and it has been suggested that in wind-pollinated tree species, high or even increased levels of gene flow are a feature of fragmented populations, although several studies have disputed this. We have used a combination of nuclear microsatellites and allele-specific PCR (AS-PCR) analysis of chloroplast single nucleotide polymorphisms (SNPs) to examine the levels and patterns of genetic diversity and population differentiation in fragmented populations of juniper (Juniperus communis) in Ireland and inform conservation programs for the species. Significant population differentiation was found for both chloroplast and nuclear markers, indicating restricted gene flow, particularly over larger geographic scales. For conservation purposes, the existence of genetically distinct clusters and geographically localised chloroplast haplotypes suggests that the concept of provenance should be taken into account when formulating augmentation or reintroduction strategies. Furthermore, the potential lack of seed dispersal and seedling establishment means that ex-situ approaches to seed and seedling management may have to be considered.     

Genetic diversity and differentiation of the Korean starry flounder (Platichthys stellatus) between and within cultured stocks and wild populations inferred from microsatellite DNA analysis

The Korean starry flounder, Platichthys stellatus, is economically valuable coastal resident fish species. However, the annual catch of this fish has fluctuated and suffered major declines in Korea. We examined the genetic diversity and population structure for four wild populations and three hatchery stocks of Korean starry flounder to protect its genetic integrity using nine microsatellites. A group of 339 genotypes belonging to seven populations were screened. High degrees of polymorphism at the microsatellite loci were observed within both the wild and hatchery populations. Compared to the wild populations, genetic changes, including reduced genetic diversity and highly significant differentiation, have occurred in cultured stocks. Significant population differentiation was also observed in wild starry flounder populations. Similar degrees of inbreeding and significant Hardy–Weinberg equilibrium deviations were detected in both the wild and the hatchery populations. The genetic connectivity pattern identified four distinct metapopulations of starry flounder in Korea by clustering in the phylogenetic tree, Bayesian analyses, molecular variance analysis, PCA and multidimensional scaling analysis. A pattern of isolation-by-distance was not significant. This genetic differentiation may be the result of the co-effects of various factors, such as historic dispersal, local environment or anthropogenic activities. These results provide useful information for the genetic monitoring of P. stellatus hatchery stocks, for the genetic improvement of this species by selective breeding and for designing suitable management guidelines for the conservation of this species.