Molecular phylogeography and paleodistribution modeling of the boreal tree species <Emphasis Type="Italic">Ulmus lamellosa</Emphasis> (T.Wang et S. L. Chang) (Ulmaceae) in China

The uplift of mountains and climatic oscillations are important for understanding of the demographic history and genetic structure of species. We investigated the biogeographic history of the boreal tree species Ulmus lamellosa (Ulmaceae) in China, by using a combined phylogeographic and paleodistribution modeling approach. In this study, 14 populations of endangered U. lamellosa were analyzed by using chloroplast DNA (cpDNA) sequences. A high level of genetic differentiation (Φ _ST = 86.22%) among populations with a significant phylogeographic pattern (N _ST > G _ST, P < 0.05) was found in U. lamellosa. Ten haplotypes were detected by combining chloroplast DNA data, and haplotype 3 (H3) was found to be common and widespread. The intraspecific divergence of all U. lamellosa cpDNA haplotypes (9.27 Ma; 95% HPD 5.17–13.33 Ma) most probably began in the late Miocene. The pairwise difference among haplotypes and neutrality tests (Tajima’s D and Fu’s Fs statistic) indicated that populations of U. lamellosa, except group I, have not experienced recent sudden expansions. Multiple refuge areas were identified across the entire distribution ranges of U. lamellosa. The low level of gene flow (Nm = 0.14) among populations may have resulted from isolation resulting from distance and complex topography during climatic oscillations; this isolation was probably the major process that shaped the present distribution of haplotypes. These results support the hypothesis that U. lamellosa persisted in situ during glaciations and occupied multiple localized glacial refugia, contrary to the hypotheses of large-scale range contraction and long-distance southward migration.     

Refugia within refugia: the case study of a canopy tree (Eurycorymbus cavaleriei) in subtropical China

Aim Eurycorymbus cavaleriei (Lévl.) Rehd. et Hand.‐Mazz. (Sapindaceae) is a Tertiary relict tree endemic to subtropical China. This area is a centre for speciation and evolution within the East Asia biome and one of its most important refugial locations. In this study we aim to elucidate the phylogeographical patterning in E. cavaleriei, in order to identify the locations of the species’ main refugia and the predominant patterns of migration that have led to the contemporary spatial genetic structure of chloroplast variation. Location Subtropical China. Methods We sampled 18 populations of E. cavaleriei throughout its geographical range. Chloroplast DNA (cpDNA) sequence data from two non‐coding regions ((trnS/trnG and pl20/5′‐rps12) were obtained from 170 individuals for phylogeographical analyses. Relationships among cpDNA haplotypes were determined using median‐joining networks. Genetic structure was examined by spatial analysis of molecular variance (SAMOVA). Population differentiation was estimated by G_ST and N_ST statistics. Results Ten distinct haplotypes were identified. The level of differentiation among populations was relatively high (G_ST = 0.817), and N_ST was significantly higher than G_ST (P < 0.05), indicating that strong phylogeographical structure is exhibited by this species. The SAMOVA revealed five diverging groups of related haplotypes, which coincide with major landscape features in this region. Main conclusions The high differentiation among populations of E. cavaleriei may be a combined effect of historical and contemporary processes, such as the low effective population size for the chloroplast genome of a dioecious species, long‐term range fragmentation and limited seed dispersal for the species. Clear‐cut geographical distributions of ancestral haplotypes of the species suggest multiple potential refugia across subtropical China. The identified refugial regions have long been recognized as centres of plant diversity and endemism for China and have also been suggested as glacial refugia for many other plant species. The combination of these factors means that these locations should be considered as the highest priority for inclusion in conservation policies and sustainable forest management strategies for subtropical China.     

Strong Genetic Differentiation of Primula sikkimensis in the East Himalaya–Hengduan Mountains

The East Himalaya-Hengduan Mountains region is the center of diversity of the genus Primula, and P. sikkimensis is one of the most common members of the genus in the region. In this study, the genetic diversity and structure of P. sikkimensis populations in China were assessed using inter-simple sequence repeat (ISSR) and chloroplast microsatellite markers. The 254 individuals analyzed represented 13 populations. High levels of genetic diversity were revealed by ISSR markers. At the species level, the expected heterozygosity and Shannon’s index were 0.4032 and 0.5576, respectively. AMOVA analysis showed that 50.3% of the total genetic diversity was partitioned among populations. Three pairs of chloroplast microsatellite primers tested yielded a total of 12 size variants and 15 chloroplast haplotypes. Strong cpDNA genetic differentiation (G _ST = 0.697) and evidence for phylogeographic structure were detected (N _ST = 0.788, significantly higher than G _ST). Estimated rates of pollen-mediated gene flow are approximately 27% greater than estimated rates of seed-mediated gene flow in P. sikkimensis. Both seed and pollen dispersal, however, are limited, and gene flow among populations appears to be hindered by the patchiness of the species’ habitats and their geographic isolation. These features may have played important roles in shaping the genetic structure of P. sikkimensis. A minimum-spanning tree of chloroplast DNA haplotypes was constructed, and possible glacial refugia of P. sikkimensis were identified.     

Phylogeography of Podocarpus matudae (Podocarpaceae): pre‐Quaternary relicts in northern Mesoamerican cloud forests

Aim Cloud forests of northern Mesoamerica represent the northern and southern limit of the contact zone between species otherwise characteristic of North or South America. Several phylogeographic studies featuring temperate conifer species have improved our understanding of species responses to environmental changes. In contrast, conifer species that presumably colonized northern Mesoamerica from South America are far less studied. A phylogeographic study of Podocarpus matudae (Podocarpaceae) was conducted to identify any major evolutionary divergences or disjunctions across its range and to determine if its current distribution is associated with pre‐Quaternary climatic and/or long‐distance dispersal events. Location Northern Mesoamerica (Mexico and Guatemala). Methods Sixteen populations (157 individuals) of P. matudae were screened for variation at two plastid DNA markers. The intra‐specific phylogenetic relationships among haplotypes were reconstructed using Bayesian inference. Population genetic analyses were undertaken to gain insight into the evolutionary history of these populations. To test whether genetic divergence among populations occurred at different time‐scales plastid DNA sequence data and fossil‐ and coalescent‐based calibrations were integrated. Results The combination of plastid markers yielded 11 haplotypes. Differentiation among populations based on DNA variation (G_ST) (0.707, SE 0.0807) indicated a clear population structure in P. matudae. Differentiation for ordered alleles (N_ST) (0.811, SE 0.0732) was higher than that for G_ST, indicating phylogeographical structure in P. matudae. Most of the total variation (81.3%, P < 0.0001) was explained by differences among populations. The estimated divergence time between the unique haplotypes from a Guatemalan population and the two most common haplotypes from the Sierra Madre Oriental in Mexico was between 10 and 20 Ma, and further haplotype divergence in the poorly resolved clade of the Sierra Madre Oriental occurred between 3 and 0.5 Ma. Main conclusions Divergence estimations support the hypothesis that extant Podocarpus matudae populations are pre‐Quaternary relicts. This finding is consistent with fossil and pollen data that support a Miocene age for temperate floristic elements in Mesoamerican cloud forests, whereas further haplotype divergence within the Sierra Madre Oriental, Chiapas and Guatemala occurred more recently, coinciding with Pleistocene cloud forest refugia.     

Two genetic divergence centers revealed by chloroplastic DNA variation in populations of <Emphasis Type="Italic">Cinnamomum kanehirae</Emphasis> Hay

Cinnamomum kanehirae Hayata (Lauraceae), the most valuable subtropical and temperate broadleaf timber tree in Taiwan, is rapidly disappearing from the wild. Taking advantage of a scion garden established by the Taiwan Forestry Research Institute, we examined patterns of chloroplast DNA (cpDNA) variations in 19 populations including 94 individuals. By sequencing two cpDNA fragments using universal primers (the trnL-trnF and petG-trnP intergenic spacers), we found eight polymorphic sites, six haplotypes, and extremely low nucleotide diversity (π = 0.00016) from 792 bp aligned sequences. The ancestral haplotype is widely distributed. Among the populations studied, three separated populations, at Yungfeng, Fuli, and Tahu have high nucleotide diversity. No phylogeographical structures of haplotypes were revealed because the tests of N _ST−G _ST for populations did not differ from zero in any situations; a ‘star-like’ genealogy is characteristic when all haplotypes rapidly coalesce and is a general outcome of population expansion. The neutrality test also suggested demographic expansion. The genetic divergence and diversity analyses suggested that two potential refugia existed during the last glaciation with a major one located in southeastern Taiwan and a minor one located in Tahu in north-central Taiwan in the Hsuehshan Range, west of the Central Mountain Range.     

Genetic structure and phylogeography of Pyrus pashia L. (Rosaceae) in Yunnan Province, China, revealed by chloroplast DNA analyses

In this study, we investigated two chloroplast intergenic spacers (accD-psaI and trnL-trnF) for a total of 266 individuals over 22 populations of Pyrus pashia to detect the genetic diversity, genetic, and phylogeographic structure and provide needed information for the development of conservation strategies. Thirteen haplotypes (H1–H13) were recognized. A high level of total diversity was detected (H _T?=?0.746). Analysis of molecular variance indicated that the genetic variation mainly existed within populations, representing 59.61 % of the total variation. Genetic differentiation among populations was high (F _st?=?0.404). A Mantel test did not show a correlation between the genetic and geographic distances (r?=?0.139, P?=?0.09, 1,000 permutations), implying that geographic distance was not critical to gene flow among populations. A significantly higher N _st than G _st (N _st?=?0.420, G _st?=?0.402, P?<?0.05) reflected the phylogeographic structure in P. pashia. While nested clade analyses of clade 2-2 showed that restricted gene flow existed among populations, clades 1-2 and 2-1, and the total cladogram exhibited contiguous range expansion events in P. pashia. Both sum of square deviations and the raggedness index failed to reject the sudden demographic expansion model. The overall population expansion of P. pashia was estimated to occur between 621,000 and 209,000 years ago.     

Phylogeography of the temperate tree species Quercus acutissima in China: Inferences from chloroplast DNA variations

Quercus acutissima is one of the most widespread temperate deciduous tree species in China. To study its phylogeographical pattern and demographic history, three chloroplast DNA fragments (atpB-rbcL, psbA-trnH and trnS-trnG) from 401 individuals representing 30 populations were sequenced. A total of 19 haplotypes were identified, and these showed a weak phylogeographical structure (N_ST = 0.689 > G_ST = 0.630, P > 0.05) at the species level. The Q. acutissima population harboured a high level of genetic diversity (H_T = 0.791), and the genetic variation mainly resided among populations (59.54%). The unimodal mismatch distribution and significantly negative Fu's F_S value indicate that the Q. acutissima population experienced rapid range expansion, which probably occurred between 0.37 and 0.12 Ma. Molecular phylogeography and ecological niche modelling (ENM) data suggest the existence of multiple localized glacial refugia in central China (e.g., the Qinling, Dabashan and Dabieshan mountain ranges) and southwestern China (Yunnan-Guizhou Plateau and its adjacent regions) during the Quaternary glaciations. Our study showed that geographical heterogeneity and climate changes may have shaped the genetic structure and phylogeographical pattern of this tree species in China.     

Polymorphism of Chloroplast DNA and Phylogeography of Green Alder (<Emphasis Type="Italic">Alnus alnobetula</Emphasis> (Ehrh.) K. Koch s. l.) in Asiatic Russia

This paper studies the phylogeography of Alnus alnobetula s. l. in the Asian part of the Russian range with the use of chloroplast DNA (cpDNA) sequences (intergenic spacers trnH-psbA, trnS-trnG, and psaA-trnS). Nine haplotypes were identified as a result of a combination of polymorphism of all sites in 241 plants in 20 populations of A. alnobetula. High interpopulation differentiation (G_ST = 0.914, N_ST = 0.928) and a significant phylogeographic structure (N_ST > G_ST, p < 0.05) were found. Reconstruction of genealogical relationships of green alder haplotypes revealed five highly divergent genetic lineages: one geographically widespread throughout Siberia and the Urals and several allopatric lines in the Far East of Russia, namely, the Chukotka Peninsula and the mouth of the Yana River, Magadan oblast, Primorye and the central part of Sakhalin Island, and the southern part of Sakhalin Island. Our data confirm that in the past the range of this species in the Far East was fragmented in several refugia that were isolated for a long time in various ice-free regions. This could have contributed to a high level of intraspecific polymorphism of green alder as a result of the divergence of endemic haplotypes, with almost non-overlapping distribution. However, the level of their molecular divergence does not correspond to the level of morphological differences.     

Phylogeography of two hybridizing southern beeches (Nothofagus spp.) with different adaptive abilities

In phylogenetically related plant species, hybridization can influence their current genetic structure. Long-lasting hybridization may be related to persistence in shared glacial refugia, where the differential abilities of each species to survive could have provided adaptations to changing environmental conditions. In temperate South American forests at the Patagonia region, the pattern of Quaternary glaciations offered several opportunities for refuge. At mid-latitudes (42° to 44° S), particular topographic characteristics determined different glaciation patterns, defining the existence of a transitional zone. We studied two widespread Nothofagus species (Nothofagus pumilio, Nothofagus antarctica) characterized by contrasting plasticity. We screened 40 coupled populations with three cpDNA markers and found 14 different haplotypes. Both species presented significant phylogeographic structure (N _ST????G _ST, p?>?0.001), with two geographically segregated lineages (north?Csouth). A latitudinal cline in the distribution of genetic diversity was determined, with most variable populations in the north (35°?C41°?S). Population diversity diminished to southern latitudes, but a particular situation occurs between 42°S and 44°S. The transition zone, a putative refuge area, presented unique haplotypes. The more plastic species, N. antarctica, probably persisted in more refuge areas, which could be reflected in its higher levels of diversity. In these species, sympatric distribution explains introgression (IG?>?IG _e), but the differential levels of haplotype sharing between N. pumilio and N. antarctica at population level are relevant to the understanding of phylogeographic patterns. Hybridization may have facilitated recruitment in the onset of postglacial colonization by middle to long-distance pollen dispersal. In the current scenario of climate change, the presence of hybrids with different plastic responses is of remarkable importance.     

High Genetic Diversity and Population Differentiation in the Critically Endangered Plant Species Trailliaedoxa gracilis (Rubiaceae)

Trailliaedoxa gracilis W. W. Smith et Forrest (Rubiaceae), a Chinese endemic monotypic genus belonging to the Alberteae (Rubiaceae), exhibits a narrow distribution in the dry valleys of the Jinsha River and Red River drainage area in southwestern China. The few sites at which T. gracilis occurs are fragmented and isolated, and several are highly vulnerable to human disturbance. As T. gracilis is a protected plant with a second-degree national priority, the genetic diversity and structure of the populations of this species should be investigated to determine the most suitable conservation strategy. In this study, two chloroplast regions and one nuclear region were used to investigate the genetic diversity, genetic structure, and demographic history of T. gracilis. We observed a high total genetic diversity (H _T?=?0.952 and 0.966) and low average within-population diversity (H _S?=?0.07 and 0.489) based on cpDNA and nDNA analyses. Thus, a strong genetic structure (F _ST?=?0.98049 and 0.59731) was detected. A phylogeographic structure was detected by nuclear DNA analysis (N _ST?>?G _ST, P?<?0.05); however, the chloroplast data did not show a significant phylogeographic structure (N _ST?<?G _ST, P?>?0.05). The Bayesian skyline plot and isolation with migration analysis were used to estimate the demographic history of T. gracilis. The results indicated that a marked bottleneck effect occurred during the glacial-interglacial of the Pleistocene. Among the extant populations of T. gracilis, the population found in ChunJiang, LuQuan, and YuXi showed the highest haplotype diversity based on cpDNA sequences and should be given priority for protection. According to the nDNA analysis, every population presented a high level of diversity and a high content of private haplotypes. Therefore, every population should be protected.

     

The effects of Quaternary glaciations in Patagonia as evidenced by chloroplast DNA phylogeography of Southern beech Nothofagus obliqua

Climatic oscillations during the Quaternary strongly affected the distribution of warm-temperate tree species, which experienced local restrictions into ice-free areas and posterior expansions. To evaluate the impact of these range movements on the genetic structure of populations, we performed a phylogeographical analysis of the species Nothofagus obliqua with chloroplast DNA markers. A total of 27 populations covering the whole natural distribution range were analyzed using polymerase chain reaction-restriction fragment length polymorphism. Diversity (h _T, h _S), allelic richness (r _g), and differentiation among populations for unordered (G _ST) and ordered alleles (N _ST) were calculated. The relationships among haplotypes were evaluated by the construction of a minimum spanning network. The spatial distribution of the genetic variation was analyzed through a Mantel test and with a nested analysis of molecular variance to differentiate between geographic regions. The screening of 11 non-coding regions allowed the identification of 14 haplotypes. A high genetic differentiation was detected (N _ST = 0.875 and G _ST = 0.824) with the existence of phylogeographic structure (p < 0.05). The distribution of the genetic variation was partially explained by the topography of the region when the populations were divided longitudinally into Coastal Mountains, Central Valley, and Andes Mountains (ϕ _RT = 0.093, p = 0.001). In agreement with pollen records, our results support the hypothesis of Coastal refuges since the region harbors high diversity together with older and private haplotypes. Long-lasting persistence of some Coastal populations without contribution to re-colonization is proposed. Additional refuges are also postulated along the Andes and Longitudinal Valley. Survival in multiple glacial refuges is discussed together with the possible migratory routes.     

Molecular evidence for glacial expansion and interglacial retreat during Quaternary climatic changes in a montane temperate pine (Pinus kwangtungensis Chun ex Tsiang) in southern China

It has been suggested that the eastern Asian temperate flora have responded to Quaternary climatic changes in a different way compared with temperate plants in Europe and Northern America. However, knowledge about their phylogeographic structure and evolutionary history is still limited. In this study we investigated mitochondrial DNA variation in 17 populations of Pinus kwangtungensis, a five-needled pine inhabiting in isolated mountains of southern China and northern Vietnam. A total of ten mitochondrial haplotypes (mitotypes) were characterized by the polymorphisms of two fragments (cox-1-2 and nad7 intron1), and total genetic diversity was high (h _T = 0.847). The construction of phylogenetic relationships of the ten mitotypes detected three major, well-distinct clades, largely corresponding to four population groups identified by SAMOVA analysis. SAMOVA also indicated that most genetic variance should be attributed to among-group differentiation (F _CT = 0.868), consistent with the substantial genetic structure found within P. kwangtungensis (G _ST = 0.751). The genetic distances of P. kwangtungensis weakly but significantly correlated with geographical distances (R = 0.228, P = 0.03), revealing a pattern of isolation-by-distance. Demographic analysis did not detect any bottleneck events in the recent history of P. kwangtungensis. These results clearly suggested that there were three major refugia for this montane temperate conifer during warm stages (along with other minor refugia), and that the species would have tracked Quaternary climatic changes by expanding to nearby lowlands in the glacial periods, and by retreating to mountaintops (the refugia) during interglacial warmer stages, experiencing fragmentation and isolation among refugia.     

Phylogeographic analysis of the endemic species Sibiraea angustata reveals a marginal refugium in the Qinghai–Tibet Plateau

The phylogeography of Sibiraea angustata, an endemic shrub species, was studied in the Qinghai–Tibet plateau (QTP). We investigated 466 individuals of S. angustata from 39 populations basically covering its total distribution area. Eight haplotypes (A–H) were detected by sequencing the intergenic chloroplast spacer trnS–trnG (600 bp), and one ancestral haplotype (A) was found to be widely distributed. The level of differentiation among populations was very high (G_ST=0.768; N_ST=0.850) and a significant phylogeographical structure was revealed (N_ST>G_ST). Analysis of molecular variance (AMOVA) similarily revealed a high level of differentiation among populations (84%, F_ST=0.842), indicating that little gene flow has occurred among populations mutually isolated by high mountains and rivers in the QTP. On the QTP platform there was only one widespread haplotype (A) in most populations, while populations along the eastern and southeastern edges had high diversity and unique haplotypes. Our results suggest that a glacial refugium may have been located on the eastern or southeastern edges of QTP during the last glaciation, and that interglacial and postglacial range expansion occurred from that refugium. Nested clade analysis (NCA) also suggests this scenario, which indicates that the current spatial distribution of cpDNA haplotypes and populations mainly resulted from long distance colonization, possibly coupled with subsequent or past fragmentation followed by range expansion and allopatric fragmentation.     

Chloroplast DNA Microsatellites Reveal Contrasting Phylogeographic Structure in Mahogany (Swietenia macrophylla King, Meliaceae) from Amazonia and Central America

Big-leaf mahogany (Swietenia macrophylla King) is one of the most valuable and overharvested timber trees of tropical America. In order to better characterize geographic patterns of genetic variation, we performed a phylogeographic analysis of S. macrophylla based on six polymorphic chloroplast genome simple sequence repeat loci (cpSSRs) analyzed in 16 populations (N?=?245 individuals) distributed across Central America and the Brazilian Amazon. Of the 31 total cpDNA haplotypes identified, 16 occurred in Central America and 15 in Amazonia with no single haplotype shared between the two regions. Populations from Central America showed moderate differentiation (F _ST ?=?0.36) while within population genetic diversity was generally high (mean Nei’s H _E ?=?0.639). In contrast, the Amazonian populations were strongly differentiated (F _ST ?=?0.91) and contained relatively low genetic diversity (mean H _E ?=?0.176), except for one highly diverse population (H _E ?=?0.925) from eastern Amazonia. Spatial analysis of molecular variance (SAMOVA) identified a single Central American phylogroup and four Amazonian phylogroups, indicating stronger phylogeographic structure within Amazonia. The results demonstrate distinctive regional patterns of S. macrophylla differentiation, and the first evidence of a strong phylogeographic break between Central American and South American mahogany populations. We suggest that the frequent occurrence of hurricanes in Central America, the differences in the glacial histories and in the duration and intensity of anthropogenic disturbance during the late Holocene may have played important roles in the geographic structuring of cpDNA lineages in the two regions. The high private haplotype diversity in Brazilian populations suggests that cpSSRs can be used as DNA barcodes for regional timber certification.     

Phylogeographic analyses suggest that a deciduous species (Ostryopsis davidiana Decne., Betulaceae) survived in northern China during the Last Glacial Maximum

Aim Palaeontologial data suggest that all temperate forest species in northern China migrated southwards during the Last Glacial Maximum (LGM) and recolonized post‐glacially within the Holocene. We tested this assumption using phylogeographical studies of a temperate deciduous shrub species (Ostryopsis davidiana Decne., Betulaceae), which has a wide distribution in northern China. Location Northern China. Methods We sequenced two chloroplast DNA (cpDNA) fragments (trnL–trnF and psbA–trnH, together about 1300 bp in length) of 294 plants from 21 populations across the total distribution range of this species. We used maximum parsimony and haplotype network methods to construct phylogenetic relationships among haplotypes. Results The analysis of cpDNA variation identified eight haplotypes. A single haplotype was fixed in all populations except for one population that was polymorphic, having two haplotypes. The population subdivisions were extremely high (G_ST = 0.972 and N_ST = 0.974), suggesting very low gene flow between populations. Haplotypes clustered into two tentative clades, both of which occur in the southern region of the species’ range but only one of which occurs in the northern region. Across the sampled populations, the haplotype distributions were differentiated geographically. Main conclusions Our analyses suggest that multiple refugia were maintained across the range of O. davidiana in both northern (north of the Qing Mountains) and southern (south of the Qing Mountains) regions during the LGM rather than that the species survived only in the south and subsequently colonized northwards. The extremely low within‐population diversity of this species suggests strong bottleneck or founder effects within each fragmented region during the Quaternary climatic oscillations. These findings provide important clues for understanding range shifts and changes in within‐ and/or between‐population genetic diversity of temperate forests in response to past climatic oscillations in northern China.     

High genetic diversity in an endangered medicinal plant, <Emphasis Type="Italic">Saussurea involucrata</Emphasis> (<Emphasis Type="Italic">Saussurea</Emphasis>, Asteraceae), in western Tianshan Mountains,China

Saussurea involucrata (Asteraceae) is a medicinal and second-degree national priority endangered plant that is mainly distributed in the high latitude region of the western Tianshan Mountains. The population is fragmented and isolated, and extensive human impact merits a suitable and specific conservation strategy, which can be compiled based on the genetic diversity, population structure, and demographic history. Phylogeographic studies were conducted on a total of five natural populations and 150 individuals were sampled. Data from three cpDNA intergenic spacer regions (trnL-F, matK, and ndhF-rpl32) and nrDNA ITS sequences showed that twelve haplotypes in cpDNA and five haplotypes in nrDNA indicated high genetic diversity among populations sampled (H _T?=?0.820 and 0.756) and within populations sampled (H _S?=?0.792 and 0.721). Additionally, the high genetic diversity did not mirror genetic structure in either cpDNA (F _ST?=?0.03153, G _ST?>?N _ST, p?<?0.05) or nrDNA (F _ST?=?0.03666, meaningless G _ST and N _ST). Two groups (north and south) were determined for a SAMOVA analysis. Based on this analysis, the demographic history was conducted with a Bayesian Skyline Plot and Isolation with Migration analysis, which showed sustainable and stable extension without a marked bottleneck. Divergence time was indicated at c. 6.25 Mya (90%HPD: 15.30–0.22 Mya) in the Miocene, which is consistent with the formation of the Kelasu section of Tianshan. The southern populations in the Bayanbulak and Gonglu regions require additional attention and transplanting would be an effective way to restore rare cpDNA haplotypes, increase effective population size, and migration rate. Our results suggested that in situ conservation of S. involucrata in western Tianshan should be the main strategy for protection and recovery of the species.     

Phylogeographic structure of Hippophae tibetana (Elaeagnaceae) highlights the highest microrefugia and the rapid uplift of the Qinghai‐Tibetan Plateau

The uplift of the Qinghai‐Tibetan Plateau (QTP) dramatically changed the topography and climate of Asia and affected the biodiversity of the plateau and its adjacent areas. However, the effects of the uplift on the dispersal, differentiation and adaptation of plants remain a puzzle when the date and processes of the uplift cannot be determined with certainty and the impacts of the Quaternary glaciations on plants on the QTP are unknown. To clarify the relationships among plants on the QTP with the plateau uplift and the Quaternary glaciations, the cpDNA trnT‐trnF regions of 891 individuals from 37 populations of Hippophae tibetana, endemic to the QTP, were sequenced in the present study. A total of 50 haplotypes were found and a strong phylogeographic structure was revealed (N_ST = 0.854, G_ST = 0.611, N_ST > G_ST, P < 0.01). The results show that three main lineages of the present populations of H. tibetana occupy the western, the middle, and the eastern geographical range, respectively, and their divergence time dates back to 3.15 Ma before present. Of 50 haplotypes, 33 (66%) are private haplotypes, which are restricted to single populations. These private haplotypes are scattered throughout the present geographical range of H. tibetana and originated from multiple differentiations in many lineages during more than 1.0 Ma period, strongly suggesting that multiple microrefugia of H. tibetana existed throughout the present geographical range during the last glacial maximum (LGM) and even earlier glaciations. Additionally, the average elevation of present populations is over 4500 m in the west and the equilibrium‐line of glaciers in the LGM was 500–300 m lower than present in the major interior part of the plateau suggesting that at most sites in the west, LGM microrefugia of H. tibetana may have been above 4000 m above sea level, the highest of all known refugia. Moreover, the divergence times among and within the three lineages and their distinct distributions as well as dispersal barriers support the theory of the recent and rapid uplift of the QTP. The rapid uplift of the plateau within the last 3.4 Ma and the associated environmental changes may have affected the dispersal and differentiation of H .tibetana and shaped its phylogeographic structure.     

A range wide geographic pattern of genetic diversity and population structure of Castanea mollissima populations inferred from nuclear and chloroplast microsatellites

Understanding geographical pattern of genetic diversity and population structure is of great importance for formulating conservation and utilization strategies. In this study, we investigated the genetic diversity and population structure of 28 natural populations of Castanea mollissima in China using eight nuclear and six chloroplast microsatellite makers (nSSRs and cpSSRs). Populations from central China harbored the highest genetic diversity at both nSSR and cpSSR markers (nSSR: H _E?=?0.705; cpSSR: H?=?0.461). The standardized measure of genetic differentiation estimated as G′ _ST was 0.447 for nSSR and 0.803 for cpSSR, respectively. The G′_ST-based pollen to seed flow ratio is 3.043, indicating that pollen flow is not extensive among C. mollissima populations. No obvious population genetic structure by geographical locations was found by STRUCTURE analysis based on nSSR data, and similarly, no signal of phylogeographic structure was detected for cpSSR analysis. Five boundaries defining zones of maximum genetic differences within the network of the C. mollissima populations were found, and the locations of those barriers were consistent with those of four mountains, i.e., Daloushan Mountain, Dabashan Mountain, Wushan Mountain, and Qingliangfeng Mountain, indicating that those mountains might act as genetic barriers obstructing the genetic exchange among natural C. mollissima populations. These results provide valuable baseline data for conservation and utilization of this species.     

Effects of life-history traits and species distribution on genetic structure at maternally inherited markers in European trees and shrubs

Aim To examine relationships between life‐history traits, ecological and chorological characteristics of woody plant species and patterns of genetic differentiation among populations as assessed by chloroplast DNA (cpDNA) markers, and to compare them with patterns previously described from nuclear markers. Location Europe. Methods Data on cpDNA variation were compiled for 29 temperate European broad‐leaved tree and shrub species. Six qualitative and three quantitative characters of the species were tested for their relationship with two parameters of genetic population differentiation (G_ST and N_ST). Both direct species comparisons and phylogenetically independent contrast analyses were performed. Results When the phylogeny was not taken into account, five characters were significantly related to levels of population differentiation. The relationship disappeared in all but two cases (distribution type and seed mass) when analyses controlled for phylogenetic relationships among species. Main conclusions The correlation between distribution type (boreal‐temperate or temperate) and cpDNA differentiation of temperate European woody plant species suggests that their Quaternary history, in particular the location and isolation of their glacial refugia, is an important determinant of their present‐day level of genetic structure. By contrast, the relationship between life‐history traits and genetic differentiation at maternally inherited markers is weaker, especially when phylogenetic effects are controlled for.     

Distribution and polymorphism of Mariner-like elements in the Bambusoideae subfamily

Picea omorika (Pan?.) Purk. is a relict from the Arcto-Tertiary flora with its entire current natural range confined to an area of only 10,000 km² within the Balkans, a region well known as a Quaternary refugium. We have amplified the second intron of the mitochondrial NADH dehydrogenase subunit1 gene in 200 trees originating from ten natural populations to assess the phylogeographic structure and history of this conifer. Five haplotypes harbouring different numbers of 34-bp minisatellites were detected, revealing haplotypic richness of 3.007 and gene diversities H _S = 0.075 and H _T = 0.225. More interestingly, despite the very small distribution range of P. omorika and its dispersal by wind, non-random distribution of haplotypes was observed, resulting in an unexpectedly high estimate of population differentiation (G _ST = 0.668), and 56.8% of molecular variation assigned to variation among populations. Those findings suggest substantial isolation of populations and their partitioning into two gene pools characterized by different history and levels of genetic diversity, and very limited seed flow in this species (Nm = 0.25). They support the hypothesised early arrival of P. omorika in the Balkan region, and residence within this refugium during several ice ages at least. We demonstrate that the assessment of genetic diversity and structuring are not straightforward in species confined to refugial regions, and that past microvicariance might bias formal phylogeographic (G _ST = N _ST = 0.668) and isolation-by-distance analysis (r = 0.028, P > 0.05).