Haplotype diversity of the mitochondrial DNA D-loop region in Calomys musculinus (Rodentia,Muridae) detected by PCR-RFLP

In order to contribute to knowledge of colonization patterns in the rodent Calomys musculinus, a natural reservoir of the virus producing Argentine hemorrhagic fever (AHF), we studied the haplotype diversity of the mitochondrial DNA D-loop region in five natural populations from central Argentina. Digestion with eight restriction enzymes (RsaI, MseI, Tsp509I, AluI, AciI, HaeIII, NlaIII, and AseI) revealed polymorphism in the 1300 bp fragment amplified by PCR. Twenty different composite haplotypes were detected. Hierarchical analyses indicated that almost all variation (94%) is contained within local populations. Haplotypes 1 and 2, shared by all populations, were the most frequent. Nonsignificant genetic differentiation was found among populations of the endemic and nonendemic areas of AHF: All locations sampled presented exclusive haplotypes in spite of their geographic proximity, which would support previous observations indicating restricted gene flow among C. musculinus populations.     

Restricted gene flow in Calomys musculinus (Rodentia, Muridae), the natural reservoir of Junin virus

Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) markers were used to evaluate the relative contribution of gene flow as a determinant of the population genetic structure of the wild rodent Calomys musculinus (the reservoir of Argentine hemorrhagic fever [AHF]) in central Argentina. One hundred eighty-seven individuals from 13 populations (9 of them from the endemic zone of AHF and 5 from areas outside it) were analyzed using 78 polymorphic RAPD loci. Genetic variation within each population was high; each individual was characterized by a unique RAPD phenotype. C. musculinus populations showed a moderate to high genetic subdivision and a random pattern of differentiation. Populations separated by the same geographic distance showed very different degrees of genetic divergence. The results indicate that populations of C. musculinus have colonized their present ranges relatively recently and differentiation by genetic drift has proceeded faster than homogenization by gene flow at the macrogeographic scale analyzed (10-700 km).     

Inheritance of random amplified polymorphic DNA (RAPD-PCR) markers and their use in population genetic studies of Calomys musculinus (Rodentia,Muridae), the reservoir of Argentine hemorrhagic fever

In order to assess the reliability of RAPD markers in the estimation of the genetic structure of natural populations of the murid rodent Calomys musculinus (reservoir of Junin virus, ethiological agent of Argentine Hemorrhagic Fever), we have analyzed the heritability of RAPD bands in 10 parents and their offspring (33 individuals). Fourteen out of a total of 119 bands obtained were absent in the parental patterns, but consistently amplified in offspring from some families. These bands can be eliminated from analyses. Overall degree of band sharing between individuals, including non-parental bands, correctly grouped members of a family in the same cluster in a UPGMA tree, with a high bootstrap percentage. Results support the usefulness of RAPDs as hereditable markers. One hundred polymorphic RAPD loci were identified in three natural populations of C. musculinus. Mean expected heterozygosity in three natural populations ranged from 0.206 to 0.220. Allele frequency based and phenotype based measures of genetic differentiation among natural populations of C. musculinus gave similar results (Weir and Cockerham's theta = 0.133; Excoffier et al.'s phi = 0.127). These values were considerably higher than those found previously using allozymes as genetic markers, and are compatible with moderate to low levels of gene flow among populations.     

Isolation and characterization of microsatellite markers in Calomys musculinus (Muridae,Sigmodontinae, Phyllotini), the natural reservoir of Junin virus

Calomys musculinus, one of the most abundant rodent species in central Argentina, is the reservoir of Junin virus, the aetiological agent of Argentine hemorrhagic fever. We isolated six polymorphic microsatellite loci for microgeographical studies of population structure in this species. Amplification of these loci in 36 individuals from three natural populations revealed five to 14 alleles per locus and expected heterozygosities from 0.426 to 0.868. Cross‐species amplifications suggest that primers designed for these loci may be useful in other closely related species of the tribe Phyllotini, but not in species of other more distantly related tribes of the subfamily Sigmodontinae.     

毛茛叶报春高分化的谱系地理结构

毛茛叶报春（Primula ranunculoides Chen）是我国特有的珍稀濒危花卉,其野生种质资源已十分稀少。为了揭示其谱系地理结构,本文对11个居群114个个体的叶绿体trnL-F和trnS-G基因片段进行了测序分析。结果共检测到10个单倍型,除YJPA和YJPB两居群共享一个单倍型（H9）外,其余各居群均仅有一个特有单倍型,居群内不存在单倍型多样性。毛茛叶报春具高分化的谱系地理结构,居群间的总平均遗传距离达0.015,单倍型间的平均突变次数高达31.2步,两者均显著高于同属的近缘种。其种子散布能力弱、生境片断化和地理隔离明显及两年生的生活史可能是导致其高分化谱系地理结构产生的主要原因。Samova、Structure以及TCS分析均支持将毛茛叶报春11个居群的遗传结构分为6组,它们与地形（山脉）关系密切对应,彼此间分化明显,因此建议不同山体的居群应视为不同的保护单元进行遗传管理。     

Inferring the population structure and demographic history of the tick, Amblyomma americanum Linnaeus.

A hierarchial population genetic study was conducted on 703 individual Amblyomma americanum from nine populations in Georgia, U.S.A. Populations were sampled from the Coastal Plain, midland Piedmont region, and the upper Piedmont region. Twenty-nine distinct haplotypes were found. A minimum spanning tree was constructed that indicated these haplotypes comprised two lineages, the root of which was distinctly star-like. The majority of the variation found was among ticks within each population, indicating high amounts of gene flow and little genetic differentiation between the three regions. An overall F(ST) value of 0.006 supported the lack of genetic structuring between collection sites in Georgia. Mantel regression analysis revealed no isolation by distance. Signatures of population expansion were detected in the shapes of the mismatch distribution and tests of neutrality. The absence of genetic differentiation combined with the rejection of the null model of isolation by distance may indicate recent range expansion in Georgia or insufficient time to reach an equilibrium where genetic drift may have affected allele frequencies. Alternatively, the high degree of panmixia found within A. americanum in Georgia may be due to bird-mediated dispersal of ticks increasing the genetic similarity between geographically separated populations.     

Connecting the dots: Bridging genetic and spatial differentiation of the genus Eumerus (Diptera: Syrphidae) in the Mediterranean Basin and Balkans

In this study, we explored intraspecific genetic differentiation of hoverfly species of the genus Eumerus with regard to landscape discontinuities (due to paleogeological events), isolation‐by‐distance, evolutionary processes, and Quaternary climatic oscillations. We unveil genetically diverging regions and discuss the potential driving forces that gave rise to these spatial genetic patterns. We generated mitochondrial DNA (mtDNA) barcodes for 274 individuals of nine Eumerus species, sampled from 58 localities in the Mediterranean and Balkans. Spatially explicit Bayesian clustering, correlation tests between geographic and genetic distances (presence of isolation‐by‐distance), median neighbor‐joining haplotype networks, and landscape shape interpolation analyses were employed to investigate spatial genetic patterns. Bayesian clustering generated one to three genetic clusters with high posterior probability values. We also observed high mtDNA haplotype diversity consisting of unique and shared haplotypes, as well as starlike mtDNA haplotype patterns. The mtDNA haplotype network was consistent with species distributions and Bayesian clustering for four tested species. The Mantel tests confirmed the absence of isolation‐by‐distance in seven species. We identified genetically diverging areas through our landscape shape interpolation analyses. Five species displayed neither spatial genetic patterns nor evidence of isolation‐by‐distance, indicative of relict taxa. Our study is the first broad‐ and large‐scale study of Eumerus species in the Mediterranean and Balkans; it reveals spatial genetic clusters in four species and identifies the potential factors driving those patterns.     

Combining geometric morphometrics and pattern recognition to identify interspecific patterns of skull variation: case study in sympatric Argentinian species of the genus Calomys (Rodentia: Cricetidae: Sigmodontinae)

Sympatric species of vesper mice Calomys laucha and Calomys musculinus are difficult to discriminate, especially in natural history collections where they are identified by external body measurements and cranial characteristics. Accurate identification of these two species can be important because only one of them, C. musculinus , is a Junin virus reservoir, the aetiological agent of the Argentine Hemorragic Fever. Research has focused into the development of molecular techniques to unambiguously identify these species. We apply statistical procedures from the field of pattern recognition to three-dimensional geometric morphometric data based on skull landmarks to identify sympatric species C. laucha , C. musculinus and Calomys venustus . Pattern recognition techniques correctly identified the three species without any prior information on specimen identity. By contrast to expectations, C. venustus differed from the other two species mainly on the basis of shape and not by its centroid size. The main sources of difference between C. laucha and C. musculinus were of shape, specifically localized at the landmarks defined by: (1) the sutures between the premaxillaries, the nasals and the frontals; (2) the sutures between the parietals, the frontals and the squamosals; and (3) the suture between the parietals and the interparietal. Nevertheless, allometries dominate the patterns of interspecific variation between these latter species and may partly explain past identification difficulties. Morphological evolution is discussed. The need for objective methods to define phenotypic clusters is highlighted with respect to the need for fast and precise biodiversity assessments.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 365–378.     

Gene Flow Among Calomys Musculinus (Rodentia, Muridae) Populations in Argentina

Calomys musculinus is a Sigmodontinae rodent inhabiting periodically disturbed habitats in the central eastern plains of Argentina. It is the natural reservoir host of Junin virus, the etiological agent of Argentine Hemorrhagic Fever (AHF). In order to analyze the levels of gene flow among populations of this species, allozymic variability at 26 loci was studied in 291 individuals from the endemic zone of AHF and localities outside it. All populations showed high levels of polymorphism (H _e between 0.107 and 0.144; P_95% between 38 and 54%). Individual loci f values were in most cases negative, although not significantly different from zero. Mean genetic differentiation among populations was low, but statistically significant (=0.020; P<0.01). There was no correlation between genetic and geographic distances between pairs of populations, and scatter of the pairwise points suggests that, at the regional scale, genetic drift is more influential than gene flow. This result can be interpreted as indicative of a relatively recent expansion of C. musculinus habitat and restricted on-going gene flow, which would be compatible with a relatively slow expansion rate of AHF.     

Spatial variation in abundance of the junin virus hosts in endemic and nonendemic Argentine haemorrhagic fever zones

Abstract Argentine haemorrhagic fever (AHF) is caused by Junin (JUN) virus, which is hosted by the drylands vesper mouse (Calomys musculinus). In this work we monitored population abundance of C. musculinus and rodent assemblages for 3 years in and outside the AHF endemic zones (northern Buenos Aires, southern Córdoba and Santa Fe Provinces, Argentina). The study area was divided into endemic and nonendemic zones. In the endemic zone epidemic sites were recognized, characterized by recent emergence and maintenance of AHF cases, and also historical sites, characterized by decreased incidence or disappearance of AHF human cases. In the nonendemic zone AHF has never been recognized. Although differences were statistically significant only during some periods, population abundance of C. musculinus was usually lower in the nonendemic sites. The pattern and magnitude of seasonal fluctuations in C. musculinus populations were also distinct in the nonendemic sites as compared to endemic sites. The relative abundance of C. musculinus in rodent assemblage was lower in nonendemic sites than in the endemic sites. The lower population densities and dampened seasonal dynamics may be at least partly responsible for the absence of AHF cases in the nonendemic zone. It is suggested that the balance between intra and interspecific interactions might be the cause of the pattern of incidence and prevalence of pathogens in the host species.     

Genetic structure of the star sea squirt, Botryllus schlosseri, introduced in southern European harbours

The introduction of new genetic variants or species is often caused by maritime transport between harbours. Botryllus schlosseri is a cosmopolitan ascidian species that is found in both harbours and open shore habitats. In order to determine the influence of ship traffic on the genetic structure and phylogeography of B. schlosseri in southern Europe, we analyzed the variability of a fragment of the mitochondrial gene cytochrome c oxidase subunit I (COI). We sampled seven Atlanto-Mediterranean harbour populations and three open-shore populations. In addition, we sequenced some colonies from the US-Atlantic coast and from other Mediterranean localities to perform phylogenetic analyses. Although the number of polymorphic sites recorded (25.8%) was within the range observed in other population studies based on ascidian COI sequences, the haplotypic diversity (16 haplotypes out of 181 sequences) was much lower. Moreover, a lack of intermediate haplotypes was observed. This pattern of high nucleotide diversity and low haplotype diversity was consistent with introduction events of a few divergent haplotypes. We found a strong genetic structure in the study populations. Gene flow was only appreciable between some harbour populations. Harbour- and open-shore populations were well differentiated, although there was no evidence for isolation by distance. A nested clade analysis pointed to long-distance colonization, possibly coupled with subsequent fragmentation, as the underlying process. Our results suggest that B. schlosseri entered the study area via harbour-hopping, possibly through recurrent introduction events. The haplotypes from North America and most of the European ones were grouped in the same phylogenetic clade. This suggests occasional gene flow between both continents, probably through ship transport.     

Phylogeography and conservation genetics of the Iowa pleistocene snail

The Iowa Pleistocene snail, Discus macclintocki, is an endangered species that survives only in relictual populations on algific (cold-air) talus slopes in northeast Iowa and northwest Illinois in the central region of the USA. These populations are believed to have been isolated since the temperatures began to warm at the end of the last glacial period around 16 500 years ago. DNA sequencing of the 16s rRNA gene of the mitochondria was used to determine the genetic relationship among 10 populations and the genetic diversity within these populations. Genetic diversity is extremely high within this species with 40 haplotypes spread across the 10 populations sampled within a 4000 km2 region. Phylogenetic analyses showed that haplotypes formed monophyletic groups by the watershed on which they were found, suggesting that watersheds were important historical avenues of gene flow. Genetic distances were strongly related to the geographical distance among all populations, but this relationship was dependent on the scale being considered.     

Phylogeography of the common ivy (Hedera sp.) in Europe: genetic differentiation through space and time

We studied the phylogeography of ivy (Hedera sp.), a liana widespread in Europe, throughout its natural range. The populations sampled belong to four closely related species differing by ploidy levels and morphological characters. Chloroplast (cp) markers were used and 13 haplotypes were detected, usually shared across species, contrary to ribosomal internal transcribed spacer (ITS) variants. We demonstrated the existence of a strong overall cpDNA phylogeographical structure. Several methods of data analysis were conducted to describe how this structure and the genetic diversity change through space and time. Southern populations, especially those from Spain, are the most divergent. Pairwise estimates of differentiation point to isolation by distance, and the existence of a latitudinal gradient of divergence was demonstrated using a regression procedure. Similarly, latitudinal differences in haplotype richness and diversity exist, as shown by population permutations ('differentiation through space'). Finally, we measured differentiation by taking into account successive levels of divergence between haplotypes ('differentiation through time'). Genetic differentiation turns out to be much greater when differences between closely related haplotypes are not considered. Further, these results indicate that the phylogeographical structure is essentially due to the relative distribution of the most similar haplotypes. Diversity decreases from south to north, whereas haplotype frequencies change longitudinally. It appears that Hedera survived in Spanish and Balkan refugia during the last ice age. A third refugium must have been present in the Alps or in Italy. During the northward expansion, the decrease in overall diversity was attenuated by some mixing of lineages at intermediate latitudes, resulting in comparatively higher levels of differentiation in the south.     

The interplay between habitat availability and population differentiation: a case study on genetic and morphological structure in an inland wader (Charadriiformes)

Long‐distance migrant waders breeding in the Arctic often have globally structured populations, largely because they were isolated in glacial or interstadial refugia or were restricted to fragmented coastal wetlands in winter. Conversely, inland species using continentally distributed wetlands appear to be less structured (more often panmictic), presumably because they are less likely to have been isolated by multiple refugia or by current events. We analyzed genetic variation in a widely distributed inland species, the ruff (Philomachus pugnax), sampled from seven Eurasian breeding localities, and from migration routes and wintering areas in Europe and Africa. One mitochondrial marker (N = 118) and eight nuclear microsatellites (N = 170) showed: (1) high genetic variation; (2) large genetic distances among mitochondrial (private) haplotypes within breeding populations; (3) the absence of a signature of isolation‐by‐distance; and (4) a distribution of private microsatellite alleles indicating dispersal between Scandinavia and Siberia but not between western and eastern Siberia. These results were consistent with a large refugial population during the Last Glacial Maximum, and postglacial long range expansions spreading ancestral polymorphisms, and not with a stepping‐stone model of gene flow. The divergence between breeding populations in Europe and Siberia was dated to approximately 12 000 years ago. Although genetic population structure is presently statistically non‐existent, support for evolving population structure came from analyses of geographical variation in two relevant phenotypic traits: wing length and the timing of migration. Analysis of 6077 individuals sampled on migration in 2002–08 revealed that, in each year, shorter‐winged birds migrated through significantly later than longer‐winged birds. The late‐passing birds were associated with more westerly breeding localities. In conclusion, the lack of genetic structuring in ruffs (and other inland species we examined) contrasts with strong structuring in many coastal species. This suggests that the ability to use more widely available inland habitat influences the evolution of genetic structure and the maintenance of genetic variation in waders. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106 , 641–656.     

Fine-scale genetic structure, estuarine colonization and incipient speciation in the marine silverside fish Odontesthes argentinensis

The identification of incipient ecological species represents an opportunity to investigate current evolutionary process where adaptive divergence and reproductive isolation are associated. In this study we analysed the genetic structure of marine and estuarine populations of the silverside fish Odontesthes argentinensis using nine microsatellite loci and 396 bp of the mitochondrial DNA (mtDNA) control region. Our main objective was to investigate the relationship among estuarine colonization, divergent selection and speciation in silversides. Significant genetic structure was detected among all marine and estuarine populations. Despite the low phylogeographic structure in mtDNA haplotypes, there was clear signal of local radiations of haplotypes in more ancient populations. Divergence among marine populations was interpreted as a combined result of homing behaviour, isolation by distance and drift. On the other hand, ecological shifts due to the colonization of estuarine habitats seem to have promoted rapid adaptive divergence and reproductive isolation in estuarine populations, which were considered as incipient ecological species. This conclusion is supported by the existence of a set of environmental factors required for successful reproduction of estuarine ecotypes. The pattern of genetic structure indicates that phenotypic and reproductive divergence evolved in the face of potential gene flow between populations. We suggest that the 'divergence-with-gene-flow' model of speciation may account for the diversification of estuarine populations. The approach used can potentially identify 'incipient estuarine species', being relevant to the investigation of the evolutionary relationships of silversides in several coastal regions of the world.     

Spatial analysis of nuclear and mitochondrial RFLP genotypes in populations of the chestnut blight fungus, Cryphonectria parasitica

Spatial structure of both nuclear and mitochondrial RFLPs were studied in several populations of the chestnut blight fungus, Cryphonectria parasitica, using a variety of spatial autocorrelation tests designed to detect nonrandom patterns. Fungal individuals were sampled from cankers on infected chestnut trees, and the location of each tree was mapped. Single-locus nuclear RFLPs, nuclear fingerprints, and mitochondrial DNA haplotypes were determined for each individual. Individuals with the same DNA fingerprint genotypes occurred closer together than would be expected at random in four of the five plots, while mitochondrial DNA haplotypes were aggregated in all five plots. Genetic distances between individuals, expressed as one minus the proportion of shared restriction fragment size classes for fingerprints and mitochondrial haplotypes, were significantly correlated with Euclidean distances between individuals in four of the five populations, but these correlations were very weak (r < 0.18). The same DNA fingerprint and single-copy nuclear RFLP alleles occurred on the same trees or immediately neighbouring trees more often than would be expected at random. Most of the aggregation for all three genetic markers occurred among individuals within the same cluster of chestnut stems or on neighbouring trees. Lack of spatial autocorrelation in one population was probably due to sampling on a larger scale that was too coarse to detect any patterns. Significant aggregation of genotypes in C. parasitica is most likely caused by some degree of restricted dispersal within populations. The implications of restricted dispersal are discussed in relation to the breeding system and isolation by distance in populations of. C. parasitica.     

Distribution of chloroplast DNA diversity within and among populations in gynodioecious Beta vulgaris ssp. maritima (Chenopodiaceae)

With the recent technical advances in molecular biology, chloroplast DNA (cpDNA) has become a marker used for the study of cytoplasmic differentiation of natural populations of plants. As chloroplasts are maternally inherited in most plant species, the seed component of gene flow is thus made accessible. We present here a study of cpDNA polymorphism within the maritima subspecies of the gynodioecious Beta vulgaris in which we try to assess the impact of such a reproductive system on seed flow. One hundred and eighty-eight wild beets were sampled from 20 hermaphroditic and 20 gynodioecious (i.e. containing both hermaphroditic and female plants) populations from the Atlantic coast of Europe. cpDNA variability in these populations was characterized with a rapid restriction fragment length polymorphism (RFLP) method. Eight cpDNA haplotypes were found. Strong differentiation among populations was observed ( F _ST = 0.43) and was consistent with isolation by distance, although most of the cpDNA haplotypes were ubiquitous. Gynodioecy seems to affect the distribution of cpDNA diversity: gynodioecious populations of Beta vulgaris ssp. maritima contained a greater number of cpDNA types but were less differentiated among themselves than hermaphroditic ones.     

Restriction fragment length polymorphisms of different DNA regions as genetic markers in the hoverfly Episyrphus balteatus (Diptera: Syrphidae)

A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis using mitochondrial (A+T-rich region; mtDNA) and genomic (zen-region; nDNA) DNA was performed on 182 female individuals of Episyrphus balteatus (DeGeer), a widespread aphidophagous hoverfly with supposed migratory behaviour. Specimens originated from 13 sampling sites in six European countries. The analyses revealed 12 and 18 haplotypes, respectively, for the two DNA types, several of them with a wide distribution, although seven and eight haplotypes, respectively, occurred only in one location. In contrast to other studies on mobile insects, the genetic diversity was relatively high. However, lack of population subdivision, low genetic distances between populations, the very high gene flow rates, and the complete lack of isolation by distance suggest that E. balteatus populations are largely connected and that there is an absence of large-scale geographic structuring. These results support the hypothesis that E. balteatus is a migratory hoverfly species, capable of moving over large distances. These findings related to the seasonal migrations of this species are discussed.     

Phylogeography and monophyly of the swordtail fish species Xiphophorus birchmanni (Cyprinodontiformes, Poeciliidae)

We used sequences of the mitochondria control region to assess the distribution of genetic variation within and among populations of the poeciliid fish species Xiphophorus birchmanni . We collected 122 X. birchmanni samples from 11 sites in three drainage systems comprising the distribution of the species. We found low levels of polymorphism among aligned sequences and low levels of genetic variation within populations but high levels of genetic differentiation among populations. Haplotypes are exclusive to three river drainages (Los Hules, Calabozo and San Pedro). Mantel tests revealed correlations between geographical (both straight-line and river distances) and genetic distance, consistent with an isolation by distance scenario, while nested clade analysis suggested allopatric fragmentation between haplotypes from two of the major drainages, and isolation by distance with restricted gene flow within those drainages. Finally, monophyly of X. birchmanni is strongly supported while the previous hypothesis of the evolutionary origin of this species from X. malinche is not.     

Phylogeography of Camellia taliensis (Theaceae) inferred from chloroplast and nuclear DNA: insights into evolutionary history and conservation

ABSTRACT: BACKGROUND: As one of the most important but seriously endangered wild relatives of the cultivated tea, Camellia taliensis harbors valuable gene resources for tea tree improvement in the future. The knowledge of genetic variation and population structure may provide insights into evolutionary history and germplasm conservation of the species. RESULTS: Here, we sampled 21 natural populations from the species' range in China and performed the phylogeography of C. taliensis by using the nuclear PAL gene fragment and chloroplast rpl32-trnL intergenic spacer. Levels of haplotype diversity and nucleotide diversity detected at rpl32-trnL (h = 0.841; pi = 0.00314) were almost as high as at PAL (h = 0.836; pi = 0.00417). Significant chloroplast DNA population subdivision was detected (GST = 0.988; NST = 0.989), suggesting fairly high genetic differentiation and low levels of recurrent gene flow through seeds among populations. Nested clade phylogeographic analysis of chlorotypes suggests that population genetic structure in C. taliensis has been affected by habitat fragmentation in the past. However, the detection of a moderate nrDNA population subdivision (GST = 0.222; NST = 0.301) provided the evidence of efficient pollen-mediated gene flow among populations and significant phylogeographical structure (NST > GST; P < 0.01). The analysis of PAL haplotypes indicates that phylogeographical pattern of nrDNA haplotypes might be caused by restricted gene flow with isolation by distance, which was also supported by Mantel's test of nrDNA haplotypes (r = 0.234, P < 0.001). We found that chlorotype C1 was fixed in seven populations of Lancang River Region, implying that the Lancang River might have provided a corridor for the long-distance dispersal of the species. CONCLUSIONS: We found that C. taliensis showed fairly high genetic differentiation resulting from restricted gene flow and habitat fragmentation. This phylogeographical study gives us deep insights into population structure of the species and conservation strategies for germplasm sampling and developing in situ conservation of natural populations.