Genetic differences between the two remaining wild populations of the endangered Indian rhinoceros (Rhinoceros unicornis)

The management of rare and endangered species in the wild and in captivity requires an understanding of the characterization of the genetic units within each species and their relationships to each other. The Indian rhinoceros (Rhinoceros unicornis) is an endangered species with a current population size of c. 2800 individuals. We analyzed 26 individuals of known origin kept in captivity and 21 wild ranging individuals of the two remnant large wild populations in Assam (India) and Nepal employing mitochondrial and microsatellite markers to determine whether the two geographically isolated populations show distinct patterns of genetic diversity, and whether the genetic diversity of the populations is influenced by past demographic bottlenecks. We identified 10 different mitochondrial D-loop haplotypes, of which 4 were specific to the Assam population (10 sequences examined) and 6 specific to the Nepal population (19 sequences). Similarly, the microsatellite analysis demonstrated a strong genetic differentiation between the Assam and Nepal populations and allowed to assign each individual to its origin with high confidence. Furthermore, our analyses revealed the occurrence of a bottleneck in the Assam population long before the reported bottleneck in 1908, and it revealed that the Nepal population is a recent (probably post-glacial) colonization. In summary, the extent of genetic divergence between the two remnant R. unicornis populations suggests separate conservation programs (even for captive individuals) as long as the persistence of the entire species is not severely threatened. The microsatellite markers can also be used to determine the origin of confiscated material such as horns.     

Geographic isolation, genetic divergence, and ecological non-exchangeability define ESUs in a threatened sky-island rattlesnake

We utilized nine microsatellite loci and 865 bases from two mtDNA genes to estimate demographic parameters and visualize historic/contemporary connectivity among populations of a sky-island rattlesnake (New Mexico ridge-nosed rattlesnake, Crotalus willardi obscurus). This taxon is listed as threatened under the United States Endangered Species Act (ESA) and is distributed patchily within three borderland mountain ranges [Animas (ANM), Peloncillo (PEL), Sierra San Luis (SSL)] of southeastern Arizona, southwestern New Mexico and north-central México. Molecular data support a hypothesis of northward range expansion from México, with subsequent isolation on sky-islands through vicariant desertification that transformed intervening wooded valleys and low elevation passes into inhospitable habitat. Historic and recent movements have been within rather than among mountains. Although populations are linked via ancestral polymorphism, they do not represent a single mtDNA gene pool. All are genetically bottlenecked, with the Peloncillo reflecting deepest/sharpest declines and fewest captures per unit effort. Most recent population declines occurred ∼4 kybp (thousands of years before present). Thus, population reductions are historic and environmental rather than contemporary and anthropogenic. Our data demonstrate that PEL is ecologically non-exchangable with other sky-island populations, and thus comprises one ‘evolutionary significant unit’ (ESU), while SSL and ANM comprise ‘management units’ (MUs) within a second ESU. All three meet legal criteria for recognition as ‘distinct population segments’ (DPS) under the ESA.     

Genetic diversity of a newly established population of golden eagles on the Channel Islands,California

Gene flow can have profound effects on the genetic diversity of a founding population depending on the number and relationship among colonizers and the duration of the colonization event. Here we used data from nuclear microsatellite and mitochondrial DNA control region loci to assess genetic diversity in golden eagles of the recently colonized Channel Islands, California. Genetic diversity in the Channel Island population was low, similar to signatures observed for other recent colonizing island populations. Differences in levels of genetic diversity and structure observed between mainland California and the islands suggests that few individuals were involved in the initial founding event, and may have comprised a family group. The spatial genetic structure observed between Channel Island and mainland California golden eagle populations across marker types, and genetic signature of population decline observed for the Channel Island population, suggest a single or relatively quick colonization event. Polarity in gene flow estimates based on mtDNA confirm an initial colonization of the Channel Islands by mainland golden eagles, but estimates from microsatellite data suggest that golden eagles on the islands were dispersing more recently to the mainland, possibly after reaching the carrying capacity of the island system. These results illustrate the strength of founding events on the genetic diversity of a population, and confirm that changes to genetic diversity can occur within just a few generations.     

绵羊mtDNA D-环序列与微卫星DNA遗传多样性和系统发育分析

采用mtDNA D-环序列和微卫星DNA两种标记方法,对9个绵羊群体225只个体进行遗传多样性和系统发育分析。结果表明：两种方法在遗传多样性分析中得出的结论一致,即在研究的9个绵羊群体中青海藏羊的遗传多样性最丰富,而湖羊和岷县黑裘皮羊遗传多样性都较低。但在系统发育分析中,通过mtDNA D-环序列和微卫星DNA座位数据构建系统发育树,结果表现为很大的不同。由于微卫星DNA符合孟德尔遗传规律,能够反映群体间的亲缘关系,构建的系统发育树结构可靠。mtDNA是核外遗传物质,具有母系遗传的特点,在反映群体间的亲缘关系上不具优势。此外,青海细毛羊和甘肃高山细毛羊的育种实践表明它们的遗传来源相似,亲缘关系相近,这与微卫星DNA座位数据构建系统发育树反映的结果一致,因此,在群体的亲缘关系研究上,微卫星DNA数据分析的结果比mtDNA序列分析结果更可信。172个单倍型序列网络关系分析表明研究的9个绵羊群体可能有三个母系起源。     

Low impact of present and historical landscape configuration on the genetics of fragmented Anthyllis vulneraria populations

Decreasing habitat fragment area and increasing isolation may cause loss of plant population genetic diversity and increased genetic differentiation between populations. We studied the relation between the historical and the present landscape configuration (i.e., patch area and patch connectivity), and the present management of calcareous grassland fragments on the one hand, and the within and between population genetic structure of 18 Anthyllis vulneraria populations on the other hand. Despite the long-time fragmentation history and the mainly selfing breeding system of the species, we detected very low genetic differentiation (Φ_st = 0.056) among habitat fragments and no significant isolation-by-distance relation. Average within fragment genetic diversity measured as molecular variance and expected heterozygosity, were relatively high (16.46 and 0.28, respectively), and weakly positively correlated with the current fragment area, most likely because larger fragments contained larger populations. We found no effects of the historical landscape configuration on the genetic diversity of the populations. Our data suggest that the consequences of habitat fragmentation for genetic differentiation and genetic diversity of A. vulneraria are relatively minor which is very likely due to the historical high levels of seed exchange among fragments through grazing and roaming livestock. This study provides indirect evidence that nature management by grazing not only positively affects habitat quality but that it might also mitigate the genetic consequences of habitat fragmentation. From the conservation point of view, this study illustrates the importance of grazing and of the regular transport of livestock between fragments to prevent the long-term effects of fragmentation on the genetic diversity of the populations studied.     

Two genetically distinct units of the Chinese sika deer (cervus nippon): analyses of mitochondrial DNA variation

Chinese sika deer, Cervus nippon, are currently threatened by habitat loss, fragmentation and human hunting, which has led to the extinction of three subspecies in the wild. The remaining subspecies subsist in the narrow regions of Jilin, Heilongjiang, Sichuan, Gansu, Jiangxi, Anhui, and Zhejiang provinces. In order to design effective conservation strategies for the Chinese sika deer, we have investigated genetic diversity, population structure and gene flow in the Chinese sika deer populations by analyzing ≈995 base pairs of the mitochondrial DNA (mtDNA) control region in 59 individuals sampled from the northeast of China, Sichuan, Jiangxi and Zhejiang. Chinese sika deer exhibited low mtDNA diversity and high gene flow among the four populations, and showed no strong geographical structure. The analyses of mtDNA variation among individuals of sika deer identified only two main phylogenetic groups even though three subspecies were sampled. These data singles out the Zhejiang population as being highly genetically distinct and worthy of separate conservation consideration. Therefore, it is recommended that a breeding program for the Zhejiang population be established.     

Restricted gene flow and genetic drift in recently fragmented populations of an endangered steppe bird

Identifying the genetic processes derived from habitat fragmentation is critical for the conservation of endangered species. We conducted an integrated analysis of genetic patterns in the endangered Dupont’s lark (Chersophilus duponti), a circum-Mediterranean songbird threatened by the loss and fragmentation of natural steppes in recent decades. After sampling all the remaining Spanish populations and the two closest North African ones, we found that the Mediterranean Sea acts as a major barrier against gene flow and that recent habitat fragmentation is isolating Spanish populations at different spatial scales. While we found a historical signal of gene flow among Spanish regions, a coalescent model supported that the ancestral panmictic population is evolving into several different units in the absence of current gene flow, genetic drift being more intense in the smaller and more isolated populations. Moreover, small-scale spatial autocorrelation analyses showed that genetic differentiation is also acting within populations. The spatial genetic structure, significant levels of inbreeding and high relatedness within patches raise concerns on the viability of most of the extant populations. We highlight the urgency for steppe patches to be protected, expanded and reconnected, considering the genetic clusters identified here rather than the previously considered eco-geographic regions occupied by the species. Meanwhile, translocations could be considered as a complementary, faster management action to attenuate the crowding and genetic effects of population fragmentation and the extinction risk of small populations without compromising the current local adaptations, culture diversity and genetic clusters already known for the species.     

Widespread selective sweeps affecting microsatellites in Drosophila populations adapting to captivity: Implications for captive breeding programs

Many threatened species are being maintained in captivity to save them from extinction, often with the eventual aim of reintroduction. The objective of genetic management in captivity is to ‘freeze’ evolution i.e. to avoid genetic adaptation to captivity and to retain genetic diversity. Most current genetic management of threatened species addresses the latter, but does not explicitly address the former. The theory underlying current genetic management and its practical implementation assumes neutrality of loci. However, genetic adaptation in captive populations may cause non-neutral behavior at neutral loci due to selective sweeps (hitchhiking) caused by rapid allele frequency changes at linked fitness loci. We compared changes in microsatellite genetic diversity at eight non-coding loci with neutral predictions in 23 pedigreed captive populations of Drosophila melanogaster maintained with effective sizes of 25 (eight replicates), 50 (6), 100 (4), 250 (3) and 500 (2) for 48 generations. Loss of microsatellite heterozygosity was significantly faster (by 12%) than predicted by neutral theory, as assessed by regressing proportion of heterozygosity retained on pedigree inbreeding coefficients. Further, greater than neutral changes were observed for both variances in allele frequencies across replicates (by 25%), and for temporal changes in allele frequencies (by 33%). All eight microsatellite loci showed signals of selectively-driven changes. Rather than having their evolution ‘frozen’, captive populations are undergoing major genome-wide selective sweeps that affect not only fitness loci but linked neutral loci. Captive genetic management for threatened species destined for reintroduction requires modification to explicitly minimize genetic adaptation to captivity.     

Possible cryptic stock structure for minke whales in the North Atlantic: Implications for conservation and management

The minke whale is the last of the great whale species to be hunted in significant numbers. Effective management must include an understanding of how genetic diversity is divided and distributed among putative local populations, and as for many migratory species, this is complicated for the minke whale by large-scale seasonal movement among geographic regions. The problem is that the geographic identity of breeding populations is not known, and instead these whales are predictably found and hunted where different breeding stocks may mix on seasonal feeding grounds. Here we use microsatellite DNA and mtDNA markers to investigate minke whale population structure across the species’ range in the North Atlantic. We found no evidence of geographic structure comparing putative populations in recognized management areas, though some limited structure had been indicated in earlier studies. However, using individual genotypes and likelihood assignment methods, we identified two putative cryptic stocks distributed across the North Atlantic in similar proportions in different regions. Some differences in the proportional representation of these populations may explain some of the apparent differentiation between regions detected previously. The implication would be that minke whales range extensively across the North Atlantic seasonally, but segregate to some extent on at least two breeding grounds. This means that established stock boundaries in the North Atlantic, currently used for management, should be re-considered to ensure the effective conservation of genetic diversity.     

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

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

Enzyme polymorphism analyses in three endangered species of Himalayan poppy, Meconopsis (Papaveraceae)

Populations of Meconopsis paniculata and M. sinuata inhabit open alpine slopes, while those of M. simplicifolia are restricted to alpine meadows of Sikkim Himalaya. These species are threatened and represented by very small populations. Three enzymes namely acid phosphatase, esterase, and glutamate dehydrogenase were tested for six populations each of M. paniculata and M. simplicifolia and single population of M. sinuata to analyse the genetic diversity of these species. In addition, alkaline phosphatases, glucose 6-phosphate dehydrogenase and malate dehydrogenase were tested for M. paniculata populations. The results on the electrophoretic variations revealed that genetic polymorphism is low to absent in all the populations of the three species of Meconopsis investigated, and the species are characterized by genetic homogeneity with fixed alleles.     

三种笛鲷的野生群体和养殖群体遗传多样性的微卫星分析

摘要：利用微卫星标记技术,采用10对微卫星引物对南海海域红鳍笛鲷、星点笛鲷、紫红笛鲷3种笛鲷鱼的野生种群（HYE、XYE、ZYE）和养殖种群（HYA、XYA、ZYA）进行了遗传多样性分析。10对微卫星引物在6个群体中共检测到78个等位基因,每个位点的等位基因数目在1~8个之间。6个群体的观测杂合度(Ho)为0.2500~1.0000,平均观测杂合度为：ZYE(0.9550)> ZYA(0.8900),HYE(0.8950)> HYA(0.8400),XYE(0.8450)>XYA(0.8100) ;平均多态信息含量(PIC) 为0.3648~0.7964,各野生群体均大于养殖群体;三种笛鲷鱼的野生群体和养殖群体遗传距离HYE与HYA,XYE与XYA,ZYE与ZYA分别为0.1029,0.0371,0.0135,基因分化系数分别为0.0371,0.0211,0.0352。群体遗传结构分析结果表明：红鳍笛鲷、星点笛鲷和紫红笛鲷的遗传多样性较丰富,养殖群体的遗传多样性较野生群体均有一定程度的降低,野生群体和养殖群体分化较弱。     

Using SSR markers to determine the population genetic structure of wild apricot (<Emphasis Type="Italic">Prunus armeniaca</Emphasis> L.) in the Ily Valley of West China

Genetic structure of three wild populations (Xinyuan, Gongliu and Daxigou) of apricot in the Ily Valley, Xinjiang Uygur Autonomous Region of China, was investigated with microsatellite (simple sequence repeat, SSR) markers. The higher polymorphism and greater transportability of these markers between Prunus species proved SSR markers were much efficient for conducting genetic diversity studies in wild apricot. Nei's gene diversity (He) and Shannon's index of diversity (I) were 0.287 and 0.458, respectively. This indicated that the wild apricot in the Ily Valley still maintained a relatively high level of diversity. The Gst of 0.137 and Fst of 0.164 revealed that genetic variation mainly resided among individuals within populations (83.6–86.3%). Population differentiation could also be found according to the distribution of SSR alleles between the populations. Mantel test showed the genetic distance between populations was significantly correlated to the geographical distance. The modest amount of gene flow (2.684) would reduce the disjunction between wild apricots. The long-distance dispersal of pollen by insects was probably the main way of gene flow between populations. Based on the study of population genetic structure, an effective conservation strategy of the species was discussed.     

Unexpected high levels of genetic variability and the population structure of an island endemic rodent (Oryzomys couesi cozumelae)

Oryzomys couesi cozumelae is an endemic, threatened rodent from Cozumel Island, Mexico. We estimated its genetic diversity and structure by analyzing microsatellite loci in 228 samples from 12 sampling sites widely distributed throughout the island. Unexpected high levels of genetic and allelic diversity were found: a total of 54 alleles, an average of 10.8 alleles per locus, and high heterozygosity values (mean H_O = 0.624, H_E = 0.690 and H_Nei = 0.689). These values are higher than those reported for small sized insular mammals, higher than that found in 37 individuals of the mainland O. couesi from southern Mexico (H_O = 0.578) that we analyzed for comparative purposes, and similar to those of other mainland small mammal populations. Despite factors that affect Cozumel’s biota, such as exotic predators and competitors, hurricanes, seasonal population fluctuations and anthropogenic activities, no evidence of genetic bottlenecks was found. A significant population structure was observed and a model of isolation-by-distance was supported. Our findings render O. c. cozumelae a high conservation value, not only for its high genetic diversity and structure, but because available data suggests that its population has declined significantly in recent years. Further habitat fragmentation and population isolation could result in a higher genetic structure and loss of genetic diversity. The protection of habitat, the maintenance of habitat connectivity and the removal of introduced competitors and predators are a conservation priority. Acknowledging that the genetic structure of populations has crucial conservation implications, the present genetic information should be taken into account in management plans for the conservation of O. c. cozumelae.     

Mitochondrial DNA sequence, morphology and ecology yield contrasting conservation implications for two threatened buckmoths (Hemileuca: Saturniidae)

Taxa of conservation interest are frequently identified using morphological or ecological characters. These characters are assumed to represent evolutionary importance, population structure and/or phylogenetic relationships in such organisms. We tested this assumption using two species complexes of the moth genus Hemileuca (Saturniidae). Both have populations threatened by habitat loss and need conservation protection. Legislation protects one taxon with apparent ecological differences. We sequenced 624 base pairs of mtDNA from the COI gene for geographically distant populations of the Hemileuca maia species complex and the H. electra species complex. Resultant phylogenies contradict prior assumptions about relationships in both species complexes. The legislatively protected Bog Buckmoth is paraphyletic with widespread H. maia, and its use of a novel hostplant seems to be a local adaptation. Divergent morphology and hostplant use among H. electra subspecies are associated with modest genetic divergence (0.48%). However, a group of unrecognized populations that are morphologically similar and geographically close to H. electra electra have mtDNA that is divergent by an average of 4.1%. There is disagreement regarding prioritization of ecological divergence over neutral genetic distance in conservation. We place ecological variation in a phylogenetic context and recommend that exploration of genetic relationships be undertaken when populations are threatened. Adaptive ecological variation should be evaluated in a phylogenetic context to understand its conservation importance. This study illustrates the importance both of phylogenetic context and the use of independent characters in assessing biodiversity for conservation prioritization.     

From genetic diversity and structure to conservation: Genetic signature of recent population declines in three mouse lemur species (Microcebus spp.)

The exceptional biodiversity of Madagascar is threatened by anthropogenic landscape changes that took place during the 2000 years of human colonization. This study focuses on the influence of geographic distance and forest fragmentation on genetic diversity and population differentiation of three rare, nocturnal, arboreal lemur species in northwestern Madagascar. Historic declines in population sizes as a consequence of forest fragmentation are quantified and dated. Eighteen sites were visited, and a total of 205 Microcebus ravelobensis, 45 M. bongolavensis and 78 M. danfossi were genotyped with eight microsatellite loci. Genetic differentiation among the sites, as measured by F_ST, ranged from 0.01 to 0.19. These values were significant in almost all cases and indicated genetic structure in the samples. Isolation-by-distance was detected in one species and a STRUCTURE analysis indicated that fragmentation further promoted genetic differentiation. Bayesian methods revealed that populations from all three species underwent a major demographic collapse of around two orders of magnitude. This decrease probably began after the arrival of humans, most likely within the last 500 years. This result suggests that anthropogenic changes may have been limited during the first 1500 years of human colonization in all three ranges. Two of the study species (M. danfossi, M. bongolavensis) lack effectively protected areas in their ranges. Consequently, quick conservation actions are now needed in order to secure the remaining genetic diversity of these species.     

Distinguishing historical fragmentation from a recent population decline - shrinking or pre-shrunk skink from New Zealand?

Species that are rare when first described present a practical management problem because it may be unclear whether the taxon is in the final stages of an anthropogenic decline, or is naturally uncommon, and each scenario dictates a distinct approach to management. We analysed mitochondrial and microsatellite DNA data with population genetic and phylogeographic tools to distinguish between these possibilities in a rare lizard from southern New Zealand. Grand skinks, Oligosoma grande, are large rock-dwelling lizards that have a fragmented distribution consisting of a western and eastern cluster of populations separated by ca. 120 km. This distribution could result from human disturbance, pre-human climatic and vegetation changes, or both. All populations were highly genetically structured (overall F_ST 0.171, R_ST 0.235), indicating that populations were demographically independent and skinks are unlikely to expand their range without human intervention. In addition, the current fragmented distribution is likely to have both historical and recent anthropogenic elements. Two eastern populations showed evidence of being historically large (high θ mtDNA genetic diversity), although they are now small, supporting anecdotal data that grand skinks have declined in historical times. However, eastern and western populations were reciprocally monophyletic for mtDNA lineages, suggesting long independent evolutionary histories that predate the arrival of humans in New Zealand. Eastern and western populations fulfil many criteria to be considered as evolutionarily significant units, but such a classification must be balanced against addressing more immediate threats to the species’ survival, such as introduced predators.     

Ecological and genetic measurements of dispersal in a threatened dragonfly

Leucorrhinia caudalis is a rare dragonfly, threatened throughout its European distribution. The species was formerly widespread in the Swiss lowlands, but only a single population remained in the 1980s. However, a spread has recently been observed, with additional ponds being colonised, sometimes at considerable distance. Despite this evidence of recent long-distance dispersal, it is unknown whether L. caudalis regularly moves among ponds or whether this is a rather rare event. A combination of an ecological mark-resight and a population genetic study was applied to investigate contemporary dispersal and the genetic footprint of the recent population history of L. caudalis in Switzerland. DNA for genetic microsatellite analysis was extracted from exuviae. The mark-resight study and the genetic analysis gave congruent results. They showed that L. caudalis is mostly a sedentary species, with only a few contemporary dispersal events over distances up to 5 km being observed. The genetic analysis was in agreement with the recent population history of the Swiss populations. The oldest and largest population showed large genetic diversity and acted as source population for the recent spread of L. caudalis in Switzerland. Recurrent gene flow among this source population and close populations caused substantial local genetic variation in the latter, as well as low population differentiation. The two recently founded distant populations (?30 km distance) were genetically less diverse and highly differentiated. These distant populations and another recently colonised population also expressed signatures of genetic bottlenecks.     

Spatial genetic variation of a riparian wolf spider Pardosa agricola (Thorell, 1856) on lowland river banks: The importance of functional connectivity in linear spatial systems

Habitat patches along river systems are often highly isolated and characterised by a high degree of heterogeneity at different spatio-temporal scales. The connectivity between river bank arthropod populations directly adjoining the river channel is often greatly disturbed due to river regulation. While flight-active arthropods easily disperse upstream, less mobile species are expected to show predominant downstream dispersal unless specific upward movements are prevalent. In linear river ecosystems, downstream drift of organisms may therefore prevail with subsequently strong asymmetrical gene flow. We analysed patterns of genetic variation within and among nine spatially structured populations of the highly stenotopic riparian wolf spider (Lycosidae) Pardosa agricola (Thorell, 1856) using Amplified Fragment Length Polymorphism (AFLP) markers. No evidence was found for downstream accumulation of genetic diversity. The high genetic diversity is hypothesised to be the result of historical drift processes. Nearby populations on the same river shore were less genetically differentiated compared with populations further away and/or on the opposite shore. This indicates that short-distance dispersal still occurs on river banks during low water flow levels, but at the same time that the river channel constitutes a physical barrier for species exchange between opposite shores. The rehabilitation of the riparian corridor will increase the amount of suitable habitat, the functional connectivity during periods of low flow-discharges and eventually riparian spider population persistence.     

Genetic conservation of South African wattled cranes

The wattled crane (Grus carunculatus), a species highly dependent on wetlands, is the largest and rarest of the six African crane species. The once vast range of the wattled crane now consists of only three disjunct populations. The South African population has shown dramatic declines and supplementation of this population using eggs from south-central Africa has been proposed. The objectives of this study were to compare levels of genetic variation in South African and south-central African populations to determine if such supplementation is needed, and if so, whether the south-central African populations represent a genetically similar source for supplementation. We surveyed genetic variation in samples from South Africa, Zimbabwe, and Botswana using 12 microsatellite DNA loci and a 400-bp fragment of the mitochondrial D-loop. Samples from Zimbabwe and Botswana were deemed genetically similar and pooled to increase sample size. Subsequent analyses indicate that the pooled south-central and South African populations show differentiation in microsatellite DNA genotypes, as well as mitochondrial DNA. As the results from both genetic markers indicate genetic isolation, these populations should be managed as separate entities. As no indication was seen from either microsatellite or mtDNA data that significant loss of genetic diversity has occurred within South African wattled cranes, supplementation from outside populations may not be necessary at this time.