Population status and distribution modelling of the critically endangered riverine rabbit (Bunolagus monticularis)

The riverine rabbit (Bunolagus monticularis) is one of the most endangered mammals in the world due to fragmentation of its habitat in the semi‐arid Karoo region of South Africa, to which it is endemic. It is an umbrella conservation species for the riparian shrubland associated with the seasonal drainage system of the Karoo, where its presence is an indicator of ecosystem health. In this study, we analysed historical survey data to derive an improved assessment of the current B. monticularis population status and distribution. Geospatial analysis was conducted using geographical information systems, and distribution modelling was performed using Maxent. Extent of occurrence for the species is 54,227 km², and area of occupancy is 2943 km². Estimates of 157–207 mature individuals confirm an alarmingly small species population size, and it appears that no subpopulation has >50 mature individuals. Our findings thus support the continued classification of this species as ‘critically endangered’ under IUCN Red List criteria. However, with most remaining habitat occurring outside of protected areas, and with habitat loss being exacerbated by climate change, a viable conservation plan remains elusive.     

Genetic assessment of a conservation breeding program of the houbara bustard (Chlamydotis undulata undulata) in Morocco,based on pedigree and molecular analyses

Protection and restoration of species in the wild may require conservation breeding programs under genetic management to minimize deleterious effects of genetic changes that occur in captivity, while preserving populations' genetic diversity and evolutionary resilience. Here, through interannual pedigree analyses, we first assessed the efficiency of a 21-year genetic management, including minimization of mean kinship, inbreeding avoidance, and regular addition of founders, of a conservation breeding program targeting on Houbara bustard (Chlamydotis undulata undulata) in Morocco. Secondly, we compared pedigree analyses, the classical way of assessing and managing genetic diversity in captivity, to molecular analyses based on seven microsatellites. Pedigree-based results indicated an efficient maintenance of the genetic diversity (99% of the initial genetic diversity retained) while molecular-based results indicated an increase in allelic richness and an increase in unbiased expected heterozygosity across time. The pedigree-based average inbreeding coefficient F remained low (between 0.0004 and 0.003 in 2017) while the proportion of highly inbred individuals (F > .1) decreased over time and reached 0.2% in 2017. Furthermore, pedigree-based F and molecular-based individual multilocus heterozygosity were weakly negatively correlated, (Pearson's r = −.061 when considering all genotyped individuals), suggesting that they cannot be considered as alternatives, but rather as complementary sources of information. These findings suggest that a strict genetic monitoring and management, based on both pedigree and molecular tools can help mitigate genetic changes and allow to preserve genetic diversity and evolutionary resilience in conservation breeding programs.     

A novel framework for evaluating in situ breeding management strategies in endangered populations

Conservation breeding management aims to reduce inbreeding and maximize the retention of genetic diversity in endangered populations. However, breeding management of wild populations is still rare, and there is a need for approaches that provide data-driven evidence of the likelihood of success of alternative in situ strategies. Here, we provide an analytical framework that uses in silico simulations to evaluate, for real wild populations, (i) the degree of population-level inbreeding avoidance, (ii) the genetic quality of mating pairs, and (iii) the potential genetic benefits of implementing two breeding management strategies. The proposed strategies aim to improve the genetic quality of breeding pairs by splitting detrimental pairs and allowing the members to re-pair in different ways. We apply the framework to the wild population of the Critically Endangered helmeted honeyeater by combining genomic data and field observations to estimate the inbreeding (i.e., pair-kinship) and genetic quality (i.e., Mate Suitability Index) of all mating pairs for seven consecutive breeding seasons. We found no evidence of population-level inbreeding avoidance and that ~91.6% of breeding pairs were detrimental to the genetic health of the population. Furthermore, the framework revealed that neither proposed management strategy would significantly improve the genetic quality or reduce inbreeding of the mating pairs in this population. Our results demonstrate the usefulness of our analytical framework for testing the efficacy of different in situ breeding management strategies and for making evidence-based management decisions.     

Use of animals with unknown ancestries in scientifically managed breeding programs

Whether to incorporate animals with unknown ancestries as founders into scientifically managed captive breeding programs, can be a difficult decision. If the animals are offspring of known founders, their inclusion in the breeding program will result in an increased incidence of inbreeding in the captive population. If the animals are additional founders, excluding them from the breeding program will result in the loss of valuable genetic variation. In general, the practice in scientifically managed captive breeding programs is to exclude animals with unknown ancestries to avoid possible inbreeding. A method of estimating the cost of making an incorrect decision on whether to use animals of unknown ancestry as founders both in terms of lost genetic variation and increased inbreeding is presented. It was determined that the loss of genetic variation resulting from excluding founders is always greater than the loss of genetic variation caused by unequal founder line representation resulting from including related animals, as if they were founders. In addition, the increased rate of accumulation of inbreeding resulting from excluding founders will eventually overcome the initial inbreeding resulting from including related animals. However, in some cases, it will take a substantial number of generations for this to occur, and the benefits of possible lowered future expected inbreeding may never be realized. The decision concerning whether to use animals with unknown ancestry should, therefore, be based on the estimated relative costs of making an error, in terms of both lost genetic variation and expected future inbreeding, rather than on avoiding the immediate possibility of increased inbreeding alone. Two examples using studbook data are given to show how this method can be practically applied to the management of captive populations. © 1993 Wiley-Liss, Inc.     

DNA fingerprinting in Speke's gazelle: a test for genetic distinctness, and the correlation between relatedness and similarity

In the absence of pedigree information, the determination of genetic distinctness of populations can only be made by genetic methods. Using DNA fingerprinting on the North American captive herd of Speke's gazelle Gazella spekei , we were able to address two hypotheses. First, two new individuals were found to have come from a genetically distinct population ( P = 0.008, permutation test), and represent potential new founders to be added to the population. Secondly, genetic similarity was not significantly correlated with relatedness under extreme inbreeding and very close relationship (coefficient of relationship range 0.304-0.717).     

AlleleRetain: a program to assess management options for conserving allelic diversity in small,isolated populations

Preserving genetic health is an important aspect of species conservation. Allelic diversity is particularly important to conserve, as it provides capacity for adaptation and thus enables long‐term population viability. Allele retention is difficult to predict beyond one generation for real populations with complex demography and life‐history traits, so we developed a computer model to simulate allele retention in small populations. AlleleRetain is an individual‐based model implemented in r and can be applied to assess management options for conserving allelic diversity in small populations of animals with overlapping generations. AlleleRetain remedies the limitations of similar existing software, and its source code is freely available for further modification. AlleleRetain and its supporting materials can be downloaded from https://sites.google.com/site/alleleretain/ or CRAN ( http://cran.r-project.org ).     

Differentiation in DNA fingerprinting among species of the genus Acer L. in Campania (Italy)

Abstract

Natural populations of species in the Acer genus occurring in Campania (southern Italy) were surveyed by screening seven microsatellite loci. Primer pairs for Acer pseudoplatanus L. microsatellite loci were analysed in six different species: Acer lobelii Ten., Acer campestre L., Acer pseudoplatanus L., Acer obtusatum W. et K., Acer neapolitanum Ten. and Acer monspessulanum L. The aim of the present study was to survey the genetic variability and genetic structure of natural populations of the Acer genus in Campania. The high degree of polymorphism observed in six different species of Acer makes these markers useful for investigating genetic variation at various spatial scales, and for the analysis of gene flow and of the mating system.     

Population genetics and breeding system of Tupistra pingbianensis (Liliaceae), a naturally rare plant endemic to SW China

The levels and partitioning of genetic diversity and inbreeding depression were investigated in Tupistra pingbianensis, a narrow endemic of South-east Yunnan, China, characterized by a naturally fragmented distribution due to extreme specialization on a rare habitat type. Here genetic diversity and patterns of genetic variation within and among 11 populations were analyzed using AFLP markers with 97 individuals across its whole geographical range. High levels of genetic variation were revealed both at the species level (P₉₉ = 96.012%; H_t = 0.302) and at the population level (P₉₉ = 51.41%; H_s = 0.224). Strong genetic differentiation among populations was also detected (F_ST = 0.2961; ⍬^嘦= 0.281), which corresponded to results reported for typical animal-pollinated, mixed selfing and outcrossing plant species. This result was consistent with mating patterns detected by our pollination experiments. The indirect estimate of gene flow based on ⍬^嘦 was low (N_m = 0.64). Special habitat and its life history traits may play an important role in shaping the genetic diversity and the genetic structure of this species. A pollination experiment also fail to detect significant inbreeding depression upon F₁ fruit set, seed weight and germinate rate fitness-traits. Since naturally rare species T. pingbianensis is not seriously genetically impoverished and likely to have adapted to tolerating a high level of inbreeding early in its history, we propose this species need only periodic monitoring to ensure their continued persistence but not intervention to remain viable.     

DNA fingerprinting in horses using a simple (TG)n probe and its application to population comparisons

A synthetic polynucleotide (TG)_n was hybridized to equine DNA digested with HinfI and hypervariable hybridization patterns were obtained. Mendelian inheritance of these DNA fingerprinting patterns was confirmed by pedigree analysis. Estimates of the probabilities of identical band patterns in unrelated individuals of different breeds (Swedish Trotters, North Swedish Trotters, Thoroughbreds and Arabians) were in the range 1 times 10-⁴-7 times 10-⁶ The variability derived with the (TG)_n, probe in horses was higher than what we obtained with several other commmonly used probes for DNA fingerprinting. Individuals within breeds tended to be more similar to each other with regard to DNA fingerprint pattern than to individuals of other breeds. Moreover, a parsimony analysis made on the basis of the hybridization patterns gave clustering of individuals within breeds. The possibility of using hypervariable probes for the identification of breed-specific characters is discussed.     

Temporal dynamics of genetic variability in a mountain goat (Oreamnos americanus) population

The association between population dynamics and genetic variability is of fundamental importance for both evolutionary and conservation biology. We combined long-term population monitoring and molecular genetic data from 123 offspring and their parents at 28 microsatellite loci to investigate changes in genetic diversity over 14 cohorts in a small and relatively isolated population of mountain goats (Oreamnos americanus) during a period of demographic increase. Offspring heterozygosity decreased while parental genetic similarity and inbreeding coefficients (F(IS) ) increased over the study period (1995-2008). Immigrants introduced three novel alleles into the population and matings between residents and immigrants produced more heterozygous offspring than local crosses, suggesting that immigration can increase population genetic variability. The population experienced genetic drift over the study period, reflected by a reduced allelic richness over time and an 'isolation-by-time' pattern of genetic structure. The temporal decline of individual genetic diversity despite increasing population size probably resulted from a combination of genetic drift due to small effective population size, inbreeding and insufficient counterbalancing by immigration. This study highlights the importance of long-term genetic monitoring to understand how demographic processes influence temporal changes of genetic diversity in long-lived organisms.     

Population genetics and breeding system of Tupistra pingbianensis (Liliaceae),a naturally rare plant endemic to SW China

The levels and partitioning of genetic diversity and inbreeding depression were investigated in Tupistra pingbianensis, a narrow endemic of southeast Yunnan, China, characterized by a naturally fragmented distribution due to extreme specialization on a rare habitat type. Here genetic diversity and patterns of genetic variation within and among 11 populations were analyzed using amplified fragment length polymorphism markers with 97 individuals across its whole geographical range. High levels of genetic variation were revealed both at the species level (P_(99)=96.012%; H_t=0.302) and at the population level (P_(99)=51.41%; H_8= 0.224). Strong genetic differentiation among populations was also detected (F_(ST)= 0.2961; θ~Ⅱ= 0.281), which corresponded to results reported for typical animal-pollinated, mixed selfing, and outcrossing plant species. This result was consistent with mating patterns detected by our pollination experiments. The indirect estimate of gene flow based on θ~Ⅱwas low (N_m=0.64). Special habitat and its life history traits might play an important role in shaping the genetic diversity and the genetic structure of this species. A pollination experiment also failed to detect significant inbreeding depression upon F_1 fruit set, seed weight, and germinate rate fitness-traits. As a naturally rare species, T. pingbianensis is not seriously genetically impoverished and likely to have adapted to tolerating a high level of inbreeding early in its history, we propose this species need only periodic monitoring to ensure their continued persistence, but not intervention to remain viable.     

Taxonomic investigation using DNA fingerprinting in Geitlerinema species (Oscillatoriales, Cyanobacteria)

The taxonomic study of 14 strains of Geitlerinema amphibium (Ag. ex Gom.) Anagnostidis and Geitlerinema unigranulatum (R.N. Singh) Komárek and Azevedo, coming from several localities was undertaken. Use was made of morphological data and molecular data were obtained by means of the DNA fingerprinting technique using highly iterated palindrome (HIP1) sequences. The employed morphological characteristics were those used for species taxonomic identification belonging to the Geitlerinema genus, namely, cell dimensions, shape of the apical cell, motility, number and localization of cyanophycin granules in the cell. The two species revealed as polymorphic were discriminated only by means of the average cellular diameters. In spite of this, minima and maxima values of the cellular diameters overlapped. It was found from molecular analysis that a high genetic diversity and the formation of two clusters consisted of G. amphibium and G. unigranulatum, plus a sole strain keeping itself isolated from the remaining. Also, these clusters were not related to the geographic location; they encompassed strains from water bodies distant from each other by as much as 3500 km, or Brazilian and Spanish strains. Molecular and morphological data support the possibility that G. unigranulatum could be considered a synonym for G. amphibium. HIP1 fingerprinting is a powerful tool for the study of genetic of cyanobacteria closely related taxa. This study points to the necessity of using other than morphological data in the taxonomic revision of cyanobacteria, as well as in the proposition of new taxons.     

Polymorphic microsatellite loci in the European rabbit (Oryctolagus cuniculus) are also amplified in other lagomorph species

Six polymorphic microsatellite markers developed for the European wild rabbit ( Oryctolagus cuniculus ) were amplified with 20 other species of lagomorphs, representing both commercially important species and species important from a conservation perspective. Successful amplification of a number of these loci has provided an important set of new molecular markers for this mammalian order.     

COANCESTRY: a program for simulating, estimating and analysing relatedness and inbreeding coefficients

The software package COANCESTRY implements seven relatedness estimators and three inbreeding estimators to estimate relatedness and inbreeding coefficients from multilocus genotype data. Two likelihood estimators that allow for inbred individuals and account for genotyping errors are for the first time included in this user-friendly program for PCs running Windows operating system. A simulation module is built in the program to simulate multilocus genotype data of individuals with a predefined relationship, and to compare the estimators and the simulated relatedness values to facilitate the selection of the best estimator in a particular situation. Bootstrapping and permutations are used to obtain the 95% confidence intervals of each relatedness or inbreeding estimate, and to test the difference in averages between groups.     

Use of DNA fingerprinting for human population genetic studies

DNA fingerprinting techniques have been used in population genetic studies on many different kinds of organisms. Here, we present new applications for multilocus DNA fingerprint probes in population studies and demonstrate the applicability of DNA fingerprinting to human population genetics, using M13 phage DNA as a probe. The new approach, which is based on a factor method of numerical coding of non-quantitative data (factor correspondence analysis-FCA), shows good agreement between population position, as indicated by the three principal factors, and ethnogenetic proximity.     

中国鸭茅主栽品种DNA指纹图谱构建

利用SSR标记和SCoT标记构建了我国主栽的21个鸭茅品种的DNA指纹图谱。从180对SSR引物和80个SCoT引物中,筛选出多态性高、谱带清晰的SSR引物和SCoT引物各24个。24对SSR引物在供试材料中共检测到186个条带,其中多态性条带为175个,品种特异条带6个,平均多态性比率94.03%,多态性信息量均值0.845,Shannon指数变幅0.4479~0.6549,基因多样性指数变幅0.2946~0.4633,可鉴别的品种数2~21个;利用24个SCoT引物在供试材料中共检测到321个条带,其中多态性条带为249个,品种特异条带6个,平均多态性比率76.33%,多态性信息量均值0.907,Shannon指数变幅0.2588~0.6329,基因多样性指数变幅0.1695~0.4451,可鉴别的品种数1~21个;5对SSR引物和5个SCoT引物在10个品种上具有唯一特征谱带,最终综合各项指标筛选出5个引物(A01E14、A01K14、B03E14、D02K13和SCoT23)上的37个条带用于鸭茅品种DNA指纹图谱构建,数据库中每个品种均具有唯一DNA指纹编码,构建的DNA指纹数据可用于鸭茅品种真伪鉴定,为品种权保护提供了科学依据。     

DNA fingerprinting of captive breeding pairs of lesser white-fronted geese (Anser erythropus) with unknown pedigrees

For a number of decades, the lesser white-fronted goose (Anser erythropus) has been almost-absent from the Fennoscandian fauna and has a current population size of only about 60 breeding pairs, with fewer than 10 pairs in Sweden. During the period 1981–1991 more than 200 young have been reintroduced in northern Sweden. However, the origin and possible relatedness of lesser white-fronted individuals were unknown when the breeding program started. We have used DNA fingerprinting to assess the similarity of 18 individuals, i.e., the entire captive population used for breeding in 1991 and about 60% of the captive population used in 1981–1991. Minisatellite probe 33.15 provided an index for an average similarity of 0.39 between the mates of the 12 breeding pairs used for producing offspring for reintroduction. This is a higher similarity than in natural populations of birds in general but lower than in populations that have passed through serious population bottlenecks. Individuals originating from different breeders are more dissimilar than those from the same breeder. However, the close relationships (similarity, 0.5–0.6) found in a group of five individuals from different breeders show that selecting individuals from different breeding groups is not sufficient to prevent mating between closely related individuals.     

Assessment of genomic variability through DNA fingerprinting within and between chicken lines divergently selected for residual food consumption

DNA fingerprints obtained by multi-locus probes have been shown to be convenient tools to assess genetic variability and genetic distances in animal populations. This method has been applied in the present study to two Rhode Island Red lines of chickens divergently selected for residual food consumption (RFC) for 16 generations (R-: low RFC; R+: high RFC). Within each line, individuals were sampled from each tail of the phenotypic distribution of RFC for males and for females. The three probes used were two minisatellite probes, R18-1 and 33-6, and the endogenous avian retroviral element probe. Altogether, 101 bands were scored. Band-sharing levels obtained from analysis of pooled DNA were extremely high within-Line (> 0.9) and still high between the lines (0.82 to 0.86). When band frequencies were compared, correlation coefficients were lower between lines than between extreme groups within lines regardless of the performance level. Although the selected lines differed widely in the selected trait RFC and a few correlated traits, it was concluded that they are probably different by only a low proportion of their genome.     

Mating strategies with genomic information reduce rates of inbreeding in animal breeding schemes without compromising genetic gain

We tested the hypothesis that mating strategies with genomic information realise lower rates of inbreeding (∆F) than with pedigree information without compromising rates of genetic gain (∆G). We used stochastic simulation to compare ∆F and ∆G realised by two mating strategies with pedigree and genomic information in five breeding schemes. The two mating strategies were minimum-coancestry mating (MC) and minimising the covariance between ancestral genetic contributions (MCAC). We also simulated random mating (RAND) as a reference point. Generations were discrete. Animals were truncation-selected for a single trait that was controlled by 2000 quantitative trait loci, and the trait was observed for all selection candidates before selection. The criterion for selection was genomic-breeding values predicted by a ridge-regression model. Our results showed that MC and MCAC with genomic information realised 6% to 22% less ∆F than MC and MCAC with pedigree information without compromising ∆G across breeding schemes. MC and MCAC realised similar ∆F and ∆G. In turn, MC and MCAC with genomic information realised 28% to 44% less ∆F and up to 14% higher ∆G than RAND. These results indicated that MC and MCAC with genomic information are more effective than with pedigree information in controlling rates of inbreeding. This implies that genomic information should be applied to more than just prediction of breeding values in breeding schemes with truncation selection.