Genetics and extinction

The role of genetic factors in extinction has been a controversial issue, especially since Lande’s paper [Genetics and demography in biological conservation, Science 241 (1988) 1455-1460] paper in Science. Here I review the evidence on the contribution of genetic factors to extinction risk. Inbreeding depression, loss of genetic diversity and mutation accumulation have been hypothesised to increase extinction risk. There is now compelling evidence that inbreeding depression and loss of genetic diversity increase extinction risk in laboratory populations of naturally outbreeding species. There is now clear evidence for inbreeding depression in wild species of naturally outbreeding species and strong grounds from individual case studies and from computer projections for believing that this contributes to extinction risk. Further, most species are not driven to extinction before genetic factors have time to impact. The contributions of mutation accumulation to extinction risk in threatened taxa appear to be small and to require very many generations. Thus, there is now sufficient evidence to regard the controversies regarding the contribution of genetic factors to extinction risk as resolved. If genetic factors are ignored, extinction risk will be underestimated and inappropriate recovery strategies may be used.     

Population genetic structure and the conservation of isolated populations of Acacia raddiana in the Negev Desert

There is much concern over the high mortality of many populations of Acacia raddiana, a keystone tree species in the Negev desert of Israel. We used random amplified polymorphic DNA (RAPD) to assess patterns of genetic variation within and among 12 populations of A. raddiana from the Arava (Syrian-African Rift) valley and western Negev. A high level of genetic polymorphism was recorded within populations. An analysis of molecular variance (AMOVA) showed that about 59.4% of total genetic variance occurred among populations, which is considerably greater population differentiation than that recorded for other outbreeding species. Cluster and principal coordinates analyses and AMOVA indicate that the western Negev and Arava valley populations are highly differentiated. We suggest that there may have been two invasions of A. raddiana into Israel: one across the northern Sinai/Gaza Strip area into the western Negev, with some plants reaching the Dead Sea and a second invasion across the southern part of the Sinai peninsula, or even from Saudi Arabia, up to the Arava valley. From the conservation point of view, each population should be conserved separately because they are genetically highly differentiated and loss of any one population would lead to a dramatic loss of genetic variation. The mixing of genetically distinct populations may give rise to outbreeding depression (particularly because of GXE interactions). An obvious first step to the maintenance of this species' genetic diversity is the separate management of the western Negev and Arava valley populations because of their different evolutionary histories.     

Current genetic isolation and fragmentation contrasts with historical connectivity in an alpine lizard (Cyclodomorphus praealtus) threatened by climate change

Assessing levels of genetic diversity, connectivity and historical demography for threatened species provides important information for conservation management. We used a combination of the mitochondrial ND4 gene and seven microsatellite markers to examine both historical and recent population genetic structure and demography of the threatened alpine she-oak skink, Cyclodomorphus praealtus. This species is restricted to the “sky islands” of the Australian alpine region. Based on mtDNA, the New South Wales and Victorian regions are reciprocally monophyletic and highly divergent, with among population variation of 0.9 and net sequence divergence of 4.28%, which suggests that they should be considered separate Evolutionary Significant Units for management purposes. The mtDNA data also indicate historical connectivity between the three Victorian populations. However, a model-based clustering analysis of microsatellite genotypes identified strong population structure in Victoria, with three distinct populations that have no current inter-population gene flow. This suggests that the Victorian populations are effectively isolated from each other, and is indicative of very low dispersal capacity and a high degree of habitat specialisation. This is reinforced by the substantially lower genetic diversity within the lowest elevation population compared to the other higher elevation populations. We found no genetic signature of major changes in effective population size. These data provide a baseline for assessing future impacts of climate change on the genetic structure of this alpine endemic species.     

Interspecific hybridisation between alien and native plant species in Germany and its consequences for native biodiversity

We explored the extent of interspecific hybridisation between alien and native plant species in Germany with a special focus on the potential threat for native biodiversity. In total we listed 134 hybrids which are interpreted as products of hybridisation between 81 alien and 109 native plant species (including 13 archeophytes) that occur in Germany. Seventy-five of these hybrids have been recorded in Germany, while the remaining 59 hybrids have not been detected in Germany yet, although both parental species currently occur in Germany. Interspecific hybridisation between abundant alien and rare native species can threaten populations of the native species through outbreeding depression and/or through high rates of gene flow swamping native populations. We identified 37 threatened native plant species which hybridise with aliens. Seventeen of these threatened plant species may suffer from outbreeding depression when hybridising with a more abundant alien invader (minority disadvantage). Using hybrid abundance as an indicator of hybrid fitness we argue that introgression of alien genes may affect the gene pool of eight threatened native plant species. Consequently, hybridisation with aliens has to be considered as an additional risk potentially leading to a loss of biodiversity and should be included in the repertoire of causes for rare species extinction in German Red Lists of threatened plant species.     

National inventories of crop wild relatives and wild harvested plants: case-study for Portugal

Crop wild relatives (CWR) and wild harvested plant species (WHP) constitute an important element of a nation’s plant genetic resources (PGR) available for utilisation. Although their natural populations are threatened like other wild species by habitat lost and fragmentation, little attention has generally been paid to their systematic conservation. The development of checklists and inventories is considered by the convention on biological diversity (CBD) and the global strategy for plant conservation (GSPC) as the first step in any national strategy for conservation and sustainable use of plant diversity. Methodological approaches to the development of a national inventory of wild PGR are discussed in the light of a case-study for the CWR and WHP growing in mainland Portugal. The resultant inventory comprises 2319 taxa, of which 97.5% are CWR, 21.4% are WHP and 19.0% are both CWR and WHP. Approximately 6.1% are endemic to mainland Portugal; 24.1% occur in 1 to 4 Portuguese administrative regions; 15.6% are threatened, but only 5.9% are covered by legislative protection. Taxonomic misalignments and the dispersed nature of biological literature were the major impediments to the production of the national inventory, but once the inventory was established it has proven to be a powerful tool in conservation management.     

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.     

Neutral DNA markers fail to detect genetic divergence in an ecologically important trait

The development of strategies for in situ, ex situ conservation and reforestation of the monkey puzzle tree (Araucaria araucana), a vulnerable tree endemic to southern South America, has led to an interest in the level and distribution of the genetic diversity of the species. Neutral DNA markers (RAPDs) and quantitative genetic techniques were used to characterise genetic heterogeneity within and among populations from throughout the natural range of the species. Both the level and pattern of genetic variation estimated using the different techniques were essentially uncorrelated. An important discrepancy was found with the neutral markers failing to detect an important quantitative genetic divergence across the Andean Range relating to drought tolerance. This study clearly demonstrates the potential problems associated with making recommendations for conserving the genetic resource of threatened species based solely on neutral marker studies. Alternative approaches are discussed, including a stronger focus on ecologically important traits and the potential use of surrogate measures of genetic variability.     

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.     

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.     

The extinction of genetic resources of Asian wild rice, Oryza rufipogon Griff.: A case study in Thailand

Natural populations of wild rice, Oryza rufipogon Griff., are now threatened with the disturbance of their natural habitats by various human activities. To obtain basic information on genetic erosion or loss of genetic diversity in wild rice, we investigated how environmental changes of habitat affected the genetic structure of its natural population at a study site in the central plain of Thailand. During 10 years from 1985 to 1994, the wild-rice population at this site was seriously destroyed and fragmented. Using two sets of seed sample collected in 1985 and 1994 from the same population, allozyme variability at 17 loci of 11 enzymes were examined. Isozyme genotypes of mother plants of seed samples were estimated by the segregation in each progeny, and we calculated genetic parameters for the population. Gene diversity severely decreased in the 1994 sample compared with the 1985 sample. It is supposed that declining and fragmentation of the wild rice population, which happened during the 10 years, caused loss of genetic variability and forced the habitually outbreeding plants to inbreed, accelerating a reduction in gene variability. Pgi1-1 allele which was common in Indica rice cultivars of this region was found in the wild rice plants growing at the side of rice fields. Probably, introgression has occurred between wild and cultivated rice plants, and consequently the intrinsic nature of wild rice was gradually blurred by cultivar genes. We must realize that the genetic erosion of wild rice is rapidly proceeding and that an action for their conservation in natural environment, so called in situ conservation, is urgently needed.     

A measure of genetic diversity of goldenseal (Hydrastis canadensis L.) by RAPD analysis

Goldenseal (Hydrastis canadensis L.) is a medicinal plant valued for the treatment of sore eyes and mouths. Although cultivation of the plant has helped meet growing demand, goldenseal is still considered a threatened or endangered species throughout much of its range in North America. In an effort to assess possible conservation strategies for goldenseal genetic resources, levels of genetic diversity within and among cultivated and wild populations were quantified. RAPD analysis was used to examine six cultivated and 11 wild populations sampled from North Carolina, Ohio, Pennsylvania, and West Virginia. The average percentage of polymorphic bands in cultivated and wild populations was low (16.8 and 15.5 %, respectively), and geographic range did not predict the level of genetic diversity. Most of the genetic variation (81.2 %) was within populations; only 3.6 % was partitioned between cultivated and wild populations. Our results differed from a previous study which concluded that genetic differences were greater among than within populations. The results of the current study indicate that, although goldenseal grows clonally and in dense patches, a mixed mating system in which both selfing and outcrossing occur is also operating. We therefore suggest that the ex situ conservation of individual plants within populations, chosen carefully to account for clonal propagation in situ, is an appropriate strategy for sustaining the genetic diversity of goldenseal.     

Genetic Diversity in Tunisian Ceratonia siliqua L. (Caesalpinioideae) Natural Populations

The last two centuries witnessed the human-caused fragmentation of Tunisian Ceratonia siliqua L. (Caesalpinoideae) populations which were often represented by scattered individuals. Seventeen populations growing in four bioclimatic zones: sub-humid, upper semi-arid, mean semi-arid and lower semi-arid zones, were sampled for allozyme diversity to assess their genetic diversity and structuration using eight isozymes revealed by starch gel electrophoresis. The species showed high diversity within populations. The average number of alleles per polymorphic locus was 1.98, the percentage of polymorphic loci was 83.4% and the mean observed heterozygosity (Ho) and expected heterozygosity (He) under Hardy–Weinberg equilibrium were respectively 0.247 and 0.316. A substantial level of inbreeding within populations induced by Wahlund effect, was observed (F_IS = 0.231). High diversity resulted from the great number of genotypes in the ancestral population before fragmentation, favoured by the outbreeding of the species. High differentiation and low gene flow were detected among populations (F_ST = 0.200) and among pairs of ecological zones (0.113< F_ST <0.198). However, the differentiation coefficient of the four zones was low (F_ST = 0.085) and similar to the average F_ST for forest trees. Population structuration depends on geographic distance between sites rather than bioclimate, indicating that structuration has occurred slowly and that climatic conditions have had little effect. Nei's genetic distances (D) between populations were low and ranged from 0.004 to 0.201. Mean D value for all population was 0.087. The UPGMA clustering established for all populations through Nei's genetic distances did not clearly show that, for the majority of populations, grouping had resulted from ecological factors or geographic location. The substantial differentiation and the high genetic similarities between populations indicate that populations have been recently isolated as a result of anthropic pressure. In-situ conservation strategies must first focus on populations with a high level of genetic diversity and rare alleles. Appropriate conservation action should take account of bioclimatic zones. Ex-situ preservation should be based on a maximum number of individuals collected within populations in each ecological group and their propagation in different bioclimates by means of cuttings.     

Conservation of the red-winged grasshopper, Oedipoda germanica (Latr.): the influence of reproductive behaviour

Reproductive isolation can function as a mechanism to maintain locally adapted gene complexes while decreasing the heterozygosity in distinct populations. As a result, reproductive behaviour should be considered a fundamental factor influencing reproductive isolation. This is of interest to conservation biology when one desires to regulate gene flow between two populations either by creating opportunities for increased dispersal, by relocation of individuals, or by re-introduction of a species to its natural habitat. Reproductive behaviour can also influence the effective population size and the actual population size through the Allee effect. We investigated the reproductive behaviour of individuals from two isolated populations of the red winged grasshopper, Oedipoda germanica, an endangered species in Central Europe. We detail several methods to show how several aspects of the reproductive behaviour of this species interact with the conservation of this species. Foreign males were not disfavoured in mate choice and male body size was also unimportant in mating success. Heterogamic matings were as productive as homogamic matings in terms of total number of eggs per female, egg hatching rate, or nymph survival. Therefore, we suggest that cross-matings of individuals from different populations do not positively influence population size by heterosis effects nor act they negatively in the form of outbreeding depression. We found that female O. germanica were able to store viable sperm for extended periods but egg pods showed a decrease in hatching rate when these females were deprived of additional mating opportunities. Multiple mated females laid more eggs than once-mated females. Hence, females are capable of founding new populations even after only one mating but their reproductive output may be reduced. Present-day gene flow between populations of O. germanica probably does not occur. Consequently, there is an urgent need to pursue habitat management and release programmes that maintain current population sizes of this species. With regard to the reproductive behaviour of the red-winged grasshopper, we suggest that relocation programmes release males and female together and at an early adult stage. Also, because female reproductive output increases with mating activity, the initial release should involve excess females. Because males have a shorter lifespan than females a subsequent release of males at a later time may enhance the overall success of local conservation efforts by resulting in more offspring per female.     

Population management of threatened taxa in captivity within their natural ranges: Lessons from Andean bears (Tremarctos ornatus) in Venezuela

Populations of threatened taxa in captivity within their natural ranges can make important contributions to conservation, but these may be compromised by the inappropriate application of population management goals developed in other contexts. We conducted demographic, genetic, and population viability analyses on the captive population of Andean bears (Tremarctos ornatus) in Venezuela to investigate the management of within-range captive populations in general, and to better integrate this population into the conservation of Andean bears in particular. We found that although the present population is very small and not internally self-sustaining, incorporation of confiscated wild individuals has resulted in a low average number of generations in captivity and low inbreeding, with moderate gene diversity and a high probability of future population persistence. However, past imports from extra-range populations have been from over-represented lineages of unknown origin, which have mixed with under-represented Venezuelan ones, reducing the future value of the Venezuelan population as a source for founder stock. Our analyses indicate that the rate of incorporation of wild recruits is a major factor influencing proxy measures of conservation value, and distinguishing within- from extra-range populations. This implies that, contrary to conventional wisdom, internal self-sustainability can be a misguided goal in within-range populations, which furthermore may not be suitable destinations for surplus animals from captive populations elsewhere.     

Genetic diversity in wild cereals: regional and local studies and their bearing on conservation ex situ and in situ

The current alarming global crisis and extinction of biodiversity affect negatively the planet's biosphere. Conservation of biodiversity is one attempt to alleviate the pending extinction of the biosphere by humans. Genetic diversity, the basis of evolution by natural selection, is gravely threatened in the progenitors of cultivated plants and its exploration, evaluation, conservation in situ and ex situ is imperative to guarantee sustainable development. This is illustrated by the population genetics and ecology of two important progenitors of cereals wild, wheat and barley. The wild cereals are rich in adaptive genetic diversity in the Fertile Crescent, primarily in Israel, which is their center of origin and diversity. The 40–55% intrapopulation diversity level in the wild cereals contrasts sharply with the average of 80% in outbreeders. Genetic diversity in wild wheat and barley is structured, particularly in wild emmer wheat, as an 'archipelago' ecological and genetic structure. These include central, semi-isolated and ecologically peripheral and marginal isolated populations, where specific alleles and allele combinations predominate as coadapted blocks of genes, adaptive to diverse ecological stresses. These involve both physical (climatic and edaphic) and biotic (pathogens and parasites) stresses at macro- and microgeographical scales. Complementary in situ and ex situ conservation is imperative across the geographic range of these species, to safeguard their immensely important genetic resources for crop improvement.     

Isozyme analysis of genetic diversity in wild Sicilian populations of Brassica sect. Brassica in view of genetic resources management

In Sicily and in the small surrounding islands the section Brassica of the genus Brassica comprises five species, B. insularis Moris, B. incana Ten., B. macrocarpa Guss., B. rupestris Raf. and B. villosa Biv. These taxa represent a genetic resource as relatives of kale crops but some populations are endangered or threatened, thus isozyme analyses were performed to assess the genetic diversity degree at population and species levels in order to assist the design of conservation management programs.Eleven loci from five enzyme systems (aconitase, leucine aminopeptidase, 6-phosphogluconate dehydrogenase, phosphoglucoisomerase phosphoglucomutase) were analyzed in sixteen natural population (fifteen from Sicily, one from Calabria). Mean within-population genetic diversity was moderate (P = 41%, A = 1.54, H = 0.16). In some cases a great number of heterozygous individuals were detected, in other cases fixation index (F) deviated significantly from Hardy-Weinberg genotypic expectations.A total of 37 alleles was recognized, six of which resulted exclusive to single populations. The among-population component of the total genetic diversity (Gst mean values) for each species was 0.30–0.37, indicating genetic differentiation among populations.Among B. villosa and B. rupestris populations genetic distance values resulted rather low and they resulted high with B. incana and B. macrocarpa populations.The results are discussed with regard to the distribution of the genetic diversity level and the genetic resources management.     

Conservation of the critically endangered Rumex rothschildianus as implied from AFLP diversity

Rumex rothschildianus (Polygonaceae) is a small dioecious annual endemic to the central coastal plain of Israel. This very rare sorrel is on the verge of global extinction, as its total gene pool consists of two surviving populations only: one small and the other relatively large and flourishing. The other ten populations of this species recorded during the 20th century (all in Israel) are now extinct. Amplified fragment length polymorphism (AFLP) markers were used to evaluate the population genetic structure of the two remaining populations. Genetic diversity in R. rothschildianus was H=0.17 with 48.9% polymorphic loci. Gene flow values suggest that the differentiation between the two populations is moderate (mean F_st=0.09). Considering these genetic data, two conservation management strategies should be applied: (1) Large scale reseeding in a neighbouring nature reserve to maintain the genetic diversity detected in this study. Seeds from diverse sources should be used for repeated reseedings in order to prevent the reintroduced populations from going through a genetic bottleneck or suffering genetic drift. (2) To maintain the genetic diversity in the two remaining wild populations, and to avoid risks such as possible outbreeding depression, only intrapopulation reinforcement should be considered. These populations should be carefully monitored and protected.     

Inbreeding and outbreeding reduces cocoon production in the earthworm Eisenia andrei

Earthworms are animals with reciprocal insemination. Eisenia andrei Bouché, 1972 is a simultaneous hermaphroditic earthworm that lives in manure heaps at high densities, with low opportunities of dispersal, thus very close inbreeding is expected. As the negative effects of inbreeding and outbreeding may be severe, we studied whether E. andrei adjusts its breeding effort according to the degree of mate relatedness. To test this, we performed laboratory experiments in which earthworms were mated with their sibs and with non-sibs from the same population and no-sibs from a geographically isolated population. Inbreeding and outbreeding matings caused a strong reduction of cocoon production, especially in genetic lines with high reproductive rates. As far as we know, this is the first study that indicates reproductive adjustment in earthworms according to the genetic divergence of their partners. Optimal outbreeding should be considered a crucial point in the management of breeding populations for applied purposes.     

Isozyme polymorphism and geographic differentiation in a collection of French perennial ryegrass populations

Summary A sample of 60 natural populations of perennial ryegrass from France has been studied for allelic variation at 7 polymorphic enzyme loci. Population genetic statistics are of the same magnitude than those previously reported for other outbreeding, short-lived perennial species (P = 64%, A = 2.75, H = 0.270).Genotype frequencies at most collection sites do not deviate significantly from Hardy-Weinberg expectations, with however a slight deficit of heterozygotes which may be accounted for by a Wahlund effect. Gene diversity is mainly explained by the within population component. The between population differentiation F_st averaged on 4 loci is only 0.054, which accounts for only 6% of the whole diversity.When mapped, most allele frequencies do not show any special structure. Only five alleles present a clinal trend from North to South. These 5 alleles are probably related to some climatic factors such as average temperature or potential evapotranspiration. The causal hypotheses about the low level of between-population differentiation and spatial structure are discussed with reference to the literature.The consequences of the found population structure for sampling and conservation strategies of natural populations for genetic resources are presented and discussed.     

Genetic variation in the endangered Rutaceae species <Emphasis Type="Italic">Citrus hongheensis</Emphasis> based on ISSR fingerprinting

Citrus hongheensis is a critically endangered species endemic to the Honghe river region in southeastern Yunnan, China. Its genetic diversity and differentiation were investigated using Inter-Simple Sequence Repeat (ISSR) markers. One hundred primers were screened, and a total of 245 loci were amplified from seven natural populations by 13 informative and reliable primers. Of these 245 ISSR loci, 233 were polymorphic and the detected variations revealed a relatively high level of intraspecific genetic diversity. At the population level, the mean percentage of polymorphic loci (PPB) was 36.50%, while the average expected heterozygosity (He) and Shannon diversity index (Ho) were 0.1327 and 0.1972, respectively. At the species level (across all populations), PPB was 95.10%, while He and Ho were 0.3520 and 0.5195, respectively. A high Gst value (0.6247) indicated that there is significant differentiation among populations, which was confirmed by AMOVA analysis (Φst = 0.6420). Pairwise genetic identity (I) values among populations ranged from 0.6341 to 0.7675, with a mean of 0.7008. We propose that the high level of genetic differentiation may be the result of habitat fragmentation and limited gene flow (Nm = 0.1502). For effective in situ conservation and population restoration of C. hongheensis it will be important to maintain historical processes, including high outbreeding rates, sufficient gene flow, and large effective population sizes.