Relict Tree Research Articles

Genetic diversity and differentiation of the riparian relict tree <i>Pterocarya fraxinifolia</i> (Juglandaceae) along altitudinal gradients in the Hyrcanian forest (Iran)

Riparian trees, especially relict trees, are attractive and important for research to understand both past and recent biogeographical and evolutionary processes. Our work is the first study to elucidate the genetic diversity and spatial genetic structure of the canopy-dominating riparian (Juglandaceae) along two altitudinal gradients in different river systems of the Hyrcanian forest, which is one of the most important refugium of relict trees in Western Eurasia. Altitudinal gradients were chosen along two river systems at 100, 400 and 900 m a.s.l. Leaf samples were collected from 116 trees, and the genetic diversity was evaluated with eight SSR markers. Overall, 39 alleles were identified for all of the populations studied. The observed heterozygosity (Ho) varied from 0.79 to 0.87 (with a mean of 0.83). The results of the AMOVA analysis indicated that the variation within populations was 88%, whereas the variation among populations was 12% for all of the gradients. A structure analysis indicated that 93% of the trees were grouped in the same gradient. The genetic distance based on Fst confirmed the structure result and indicated a high rate of gene flow among the investigated populations. Based on high gene flow (low differentiation of the population along the same river) and the clearly distinct genetic structure of the investigated gradients, it can be concluded that rivers are the main seed dispersal vector among populations. The genetic diversity of did not show any trend from upstream to downstream. The high level of gene flow and uniform genetic diversity along each river suggest the “classical” metapopulation structure of the species. Pterocarya fraxinifoliaP. fraxinifoliaP. fraxinifolia

Rangewide determinants of population performance in Prunus lusitanica: Lessons for the contemporary conservation of a Tertiary relict tree

Relict species are an extremely important part of biodiversity and as such studies on the factors that allow their current persistence are required. The aim of this study was to assess the determinants of the distribution and range-wide population performance of the Tertiary relict tree Prunus lusitanica L. This threatened species is confined to Iberia, Northern Morocco and Macaronesia with a fragmented and scattered distribution. Using ecological niche modelling, we calculated the level of range filling across the range and tested its relationship with human impact. We then assessed the relative importance of climatic suitability as obtained through niche modelling, topographic factors and contemporary human impact on range-wide population performance. Results showed that the species occupies only 2.4% of the overall area predicted to be climatically suitable for its presence and the level of range filling varied across regions. A weak negative relationship among range filling and human impact was found. Overall climatic suitability was the strongest predictor of population performance. However, it showed high variability across regions: the effect was positive in Iberia whereas negative but not significant in Macaronesia and Morocco. Human impact showed a significant negative effect and finally topographic factors such as altitude had a minor negative effect. Our results highlight that both climate and human impact play a major role in the current limited range filling and performance of the species. Management plans to minimize anthropogenic disturbances together with reforestation measures are urgently needed in order to conserve this unique species.

Climate extremes and predicted warming threaten Mediterranean Holocene firs forests refugia

Warmer and drier climatic conditions are projected for the 21st century; however, the role played by extreme climatic events on forest vulnerability is still little understood. For example, more severe droughts and heat waves could threaten quaternary relict tree refugia such as Circum-Mediterranean fir forests (CMFF). Using tree-ring data and a process-based model, we characterized the major climate constraints of recent (1950-2010) CMFF growth to project their vulnerability to 21st-century climate. Simulations predict a 30% growth reduction in some fir species with the 2050s business-as-usual emission scenario, whereas growth would increase in moist refugia due to a longer and warmer growing season. Fir populations currently subjected to warm and dry conditions will be the most vulnerable in the late 21st century when climatic conditions will be analogous to the most severe dry/heat spells causing dieback in the late 20th century. Quantification of growth trends based on climate scenarios could allow defining vulnerability thresholds in tree populations. The presented predictions call for conservation strategies to safeguard relict tree populations and anticipate how many refugia could be threatened by 21st-century dry spells.

Effects of Climate Change on the Potentially Suitable Climatic Geographical Range of Liriodendron chinense

Identifying the potentially suitable climatic geographical range for Liriodendron chinense (L. chinense) and predicting its responses to climate change is urgently necessary, as L. chinense is an important tertiary relict tree species. In this study, we simulated the potentially suitable climatic habitat of L. chinense in China using maximum entropy (MaxEnt) modeling. We found that the MaxEnt model was highly accurate with an average training Area Under the Curve (AUC) value of 0.912. Annual precipitation and mean temperature of the driest quarter are the main factors controlling the geographical distribution of L. chinense. Currently, the suitable climatic habitat of L. chinense is mainly located in Southeastern China. Forecasted patterns of predicted suitable climatic habitat show a significant change by the 2050s and 2070s, suggesting that the suitable climatic habitat of L. chinense would shift north with future climate change, based on four Representative Concentrations Pathways for carbon dioxide (CO2) emissions. The southern extent of the current distribution would become unsuitable for L. chinense, pointing to a threat of extinction and highlighting the urgent need for conservation within the next half century. The potentially suitable climatic habitat of L. chinense was predicted to move further north, but those habitat gains may be inaccessible because of dispersal limitations. Our unique findings offer a climatic suitability map for L. chinense in China, which can help to identify locations where L. chinense may already exist, but has not yet been detected; to recognize locations where L. chinense is likely to spread in the future given forecasted climate change; and to select priority areas for its introduction, cultivation, and conservation.

Variation in the Concentrations of Major Secondary Metabolites in Ginkgo Leaves from Different Geographical Populations

Ginkgo biloba L. is a well-known relict tree species and an important medicinal plant. Ginkgo is rich in secondary metabolites (SMs), mainly including flavonoids, lactones, and ginkgolic acid. The aim of this study was to determine variations in the concentrations of these SMs in Ginkgo leaves from different geographical populations. The SMs in the leaves of 298 clones from 10 geographical populations grafted under the same conditions were extracted and measured by high performance liquid chromatography (HPLC). The results showed that there were significant differences in concentrations of SMs in leaves from different populations (p < 0.01). The concentrations of both flavonoids and lactones were significantly negatively correlated with that of ginkgolic acid. Altitude and annual rainfall were important factors influencing the concentrations of lactones, and the frost-free period influenced the concentration of isorhamnetin. Population Yingdianjie (YDJ) was ideal for the plantations from which medicinal flavonoids and lactones are extracted, followed by populations Xiaopu (XP), Anlu (AL) and Wuchuan (WC). As variations within each population were found, attention should be paid to selection within populations.

Phylogeography and population genetics of the riparian relict tree Pterocarya fraxinifolia (Juglandaceae) in the South Caucasus

We aimed to (i) assess the extant genetic diversity of the riparian relict tree Pterocarya fraxinifolia across its current distribution range in the South Caucasus, including the past refugial areas Colchis and Hyrcan, and (ii) test if a separation of these areas is reflected in its phylogeographic history. Genetic diversity of natural populations was examined using nuclear microsatellite and plastid DNA markers. Spatial genetic structure was evaluated using Bayesian clustering methods and the reconstruction of plastid DNA networks. Divergence times of Colchic and Hyrcanian populations were estimated via divergence dating using a relaxed molecular clock. Allelic richness, private allelic richness, and expected heterozygosity were significantly higher in Hyrcan than in Colchis and the Greater Caucasus, and significant genetic differentiation was revealed between the two groups. Whereas only two plastid haplotypes were detected for the Colchic and Caucasian populations, the Hyrcanian populations displayed 11 different haplotypes. Significant isolation by distance was detected in Hyrcan. The most recent common ancestor of all P. fraxinifolia haplotypes was dated to a time well before a suggested glaciation period in the Caucasus during the late Pliocene (5.98 Ma [11.3–2.48 Ma HPD]). The widespread Colchic haplotype that also occurs along the southern slope of the Greater Caucasus and reaches south-eastern Azerbaijan has appeared more recently (0.24 Ma [1.41–0 Ma HPD]). This diversification pattern of Colchic haplotypes from ancient Hyrcanian haplotypes suggests a colonization of the region from south-east to north-west that predates the last glacial maximum (LGM). Natural populations of P. fraxinifolia show low-to-intermediate levels of genetic diversity and a significant decrease of diversity from Hyrcan to Colchis. However, the genetic differentiation between Colchic-Caucasian and Hyrcanian populations for nuclear markers suggests that independent gene pools existed in both areas at least since the LGM. Particular attention to conservation seems justified for the more diverse Hyrcanian populations.

Upscaling hypotheses on herbivore damage in plants facing environmental stress: Variation among scales and plant enemies in a relict tree

The few existing analyses of broad-scale variation in plant–herbivore interactions are largely exploratory and not based on hypotheses on the effect of varying environmental conditions on the patterns of herbivore damage. However, there are two established hypotheses that relate herbivore damage and environmental stress mostly tested at the within-population level: the Plant Stress Hypothesis (PSH) and the Plant Vigor Hypothesis (PVH). To examine if support for the PSH holds true at broader scales, we tested it at the micro- (among individuals), meso- (between habitats) and macroscale (between geographical regions, Iberia and Macaronesia). We measured plant stress and leaf damage by different natural enemies (chewers, miners and pathogens) in 12 populations across the range of the relict tree Prunus lusitanica. Results regarding the proportion of leaves attacked by any enemy showed no support for the PSH at either micro or mesoscale, observing instead higher damage in less stressed individuals within populations and in high-quality habitats. However, the latter was true exclusively within the Macaronesian region, highlighting the important effect of the type of abiotic stress on plant susceptibility to herbivory. At the macroscale, alternatively, results showed support for the PSH: we observed a higher proportion of leaves attacked in Macaronesia than in Iberia, where plants unexpectedly exhibited greater stress symptoms. Among-scale inconsistencies may respond to the effect of distinct climatic factors governing both plant and herbivore dynamics at large scales, particularly in species with distributional ranges across contrasting climatic regions such as P. lusitanica. Finally, different plant enemies showed different patterns of damage, corroborating the need to study them separately to achieve consistent generalizations of these hypotheses.

Geographical variation and the role of climate in leaf traits of a relict tree species across its distribution in China.

Intraspecific trait variation and trait-climate relationships are crucial for understanding a species' response to climate change. However, these phenomena have rarely been studied for tree species. Euptelea pleiospermum is a relict tree species with a wide distribution in China that offers a novel opportunity to examine such relationships. Here, we measured 13 leaf traits of E.pleiospermum in 20 sites across its natural distribution in China. We investigated the extent of trait variation at local and regional scales, and developed geographic and climate models to explain trait variation at the regional scale. We documented intraspecific trait variation among leaf traits of E.pleiospermum at local and regional scales. Five traits exhibited relatively high trait variation: leaf area, leaf density and three leaf economic traits (leaf dry matter content, specific leaf area [SLA] and leaf phosphorus concentration). Significant trait-geography correlations were mediated by local climate. Most leaf trait variation could be explained (from 24% to 64%) by geographic or climate variables, except leaf width, leaf thickness, leaf dry matter content and leaf length-width ratio. Latitude and temperature were the strongest predictors of trait variation throughout the distribution of E.pleiospermum in China, and temperature explained more leaf trait variation than precipitation. In particular, we showed that leaves had longer petiole lengths, higher SLA and lower densities in northern E.pleiospermum populations. We suggest that northern E.pleiospermum populations are adapting to higher latitudinal environments via high growth rate (higher SLA) and low construction investment strategies (lower leaf densities), benefitting northern migration. Overall, we demonstrate that intraspecific trait variation reflects E.pleiospermum response to the local environment. We call for consideration of intraspecific trait variation to examine specific climate response questions. In addition, provenance experiments using widely distributed species are needed to separate trait variation resulting from genetic differentiation and plastic responses to environmental change.

Potential effects of climate change on geographic distribution of the Tertiary relict tree species Davidia involucrata in China

This study, using species distribution modeling (involving a new approach that allows for uncertainty), predicts the distribution of climatically suitable areas prevailing during the mid-Holocene, the Last Glacial Maximum (LGM), and at present, and estimates the potential formation of new habitats in 2070 of the endangered and rare Tertiary relict tree Davidia involucrata Baill. The results regarding the mid-Holocene and the LGM demonstrate that south-central and southwestern China have been long-term stable refugia, and that the current distribution is limited to the prehistoric refugia. Given future distribution under six possible climate scenarios, only some parts of the current range of D. involucrata in the mid-high mountains of south-central and southwestern China would be maintained, while some shift west into higher mountains would occur. Our results show that the predicted suitable area offering high probability (0.5‒1) accounts for an average of only 29.2% among the models predicted for the future (2070), making D. involucrata highly vulnerable. We assess and propose priority protected areas in light of climate change. The information provided will also be relevant in planning conservation of other paleoendemic species having ecological traits and distribution ranges comparable to those of D. involucrata.

Characterization of the complete chloroplast genome of Emmenopterys henryi (Gentianales: Rubiaceae), an endangered relict tree species endemic to China

The chloroplast genome sequence of an endangered relict tree species endemic to China, Emmenopterys henryi, was determined based on the Illumina data. The chloroplast genome of E. henryis was sequenced (155,379 in length, GenBank accession number KY273445). It contains 62.36% A+T base pairs and consists of a pair of inverted repeats (IRs, 25,790 bp) separated by the large single-copy (LSC, 85,554 bp) and small single-copy (SSC, 18,245 bp) regions. The chloroplast genome contains 132 unique genes, including 87 protein-coding genes, 37 transfer RNA genes and 8 ribosomal RNA genes, respectively. The phylogenetic analysis based on chloroplast genomes indicates that E. Henryi is most closely related to coffea arabica and coffea canephora.

Intraspecific variation in heritable secondary metabolites and defensive strategies in a relict tree

Intraspecific variation in heritable secondary metabolites and defensive strategies in a relict tree

Effects of Isoflavonoids from Maackia Amurensis Roots on the Metabolic Reactions of the Liver in Experimental Toxic Hepatitis

A complex of isoflavonoids containing more than 78% isoflavone and pterocarpan glycosides was obtained from an alcoholic extract of the roots of the Amur maackia, a far eastern relict tree (Maackia amurensis Ruper et Maxim.). This complex contained the 7-O-gentiobiosides of daidzein, genistein, afromosin, pseudobaptigenin, formononetin, and 5-O-methylgenistein, the 3-O-gentiobiosides of maackiain and medicarpin, the compounds daidzin, and genistein, 7-O-primverosylformononetin, and the novel 7-O-primverosylpseudobaptigenin. Studies using a model of toxic hepatitis induced by poisoning of rats with carbon tetrachloride addressed the effects of the complex of isoflavonoids on the state of carbohydrate metabolism in the rat liver. Administration of this complex to animals with CCl4 hepatitis decreased the activity of marker enzymes for cytolysis and the specific weight of the liver, promoted maintenance of the blood glucose and oxidized nicotinamide coenzyme (NAD+) levels, and normalized pyruvate and lactate levels in the animals’ livers. The isoflavonoid complex extracted from Maackia amurensis was more effective in restoring the reactions of liver carbohydrate metabolism (gluconeogenesis, Krebs cycle) than the reference hepatoprotector Legalon®.

Moderate Genetic Diversity and Genetic Differentiation in the Relict Tree Liquidambar formosana Hance Revealed by Genic Simple Sequence Repeat Markers.

Chinese sweetgum (Liquidambar formosana) is a relatively fast-growing ecological pioneer species. It is widely used for multiple purposes. To assess the genetic diversity and genetic differentiation of the species, genic SSR markers were mined from transcriptome data for subsequent analysis of the genetic diversity and population structure of natural populations. A total of 10645 potential genic SSR loci were identified in 80482 unigenes. The average frequency was one SSR per 5.12 kb, and the dinucleotide unit was the most abundant motif. A total of 67 alleles were found, with a mean of 6.091 alleles per locus and a mean polymorphism information content of 0.390. Moreover, the species exhibited a relatively moderate level of genetic diversity (He = 0.399), with the highest was found in population XY (He = 0.469). At the regional level, the southwestern region displayed the highest genetic diversity (He = 0.435) and the largest number of private alleles (n = 5), which indicated that the Southwestern region may be the diversity hot spot of L. formosana. The AMOVA results showed that variation within populations (94.02%) was significantly higher than among populations (5.98%), which was in agreement with the coefficient of genetic differentiation (Fst = 0.076). According to the UPGMA analysis and principal coordinate analysis and confirmed by the assignment test, 25 populations could be divided into three groups, and there were different degrees of introgression among populations. No correlation was found between genetic distance and geographic distance (P > 0.05). These results provided further evidence that geographic isolation was not the primary factor leading to the moderate genetic differentiation of L. formosana. As most of the genetic diversity of L. formosana exists among individuals within a population, individual plant selection would be an effective way to use natural variation in genetic improvement programs. This would be helpful to not only protect the genetic resources but also attain effective management and exploit genetic resources.

Phylogenetic relationships of Pterocarya (Juglandaceae) with an emphasis on the taxonomic status of Iranian populations using ITS and trnH-psbA sequence data

Pterocarya fraxinifolia (Lam.) Spach., a relict tree species of the Juglandaceae family, is native to the Great Caucasus, Anatolia, and to the Hyrcanian forests of the southern Azerbaijan and Northern Iran. In this study, the phylogenetic relationship of the species, sampled in selected Iranian populations, and the global biogeography of the genus Pterocarya were addressed. Leaves were collected from 8 to 10 trees from three geographically isolated habitats. The samples were analyzed with nuclear (internal transcribed spacer [ITS] regions) and chloroplast (trnH-psbA) DNA markers. The obtained results were compared and analyzed with the data registered in NCBI GenBank. It is reported that the ITS regions varied from 644 to 652 for Pterocarya genus, but we did not observe polymorphisms for Iranian Pterocarya. The phylogenetic tree divided the Pterocarya genus in three clades: clade 1 grouping exclusively the samples P. fraxinifolia, clearly separated from the East Asiatic taxa; clade 2 that includes the species P. hupehensis and P. macroptera; clade 3 clustering P. stenoptera and P. tonkinensis. Although the Iranian Pterocarya samples and P. fraxinifolia from the Caucasus were in the same clade, they presented two different secondary structures. The Iranian populations showed the maximum genetic distance with P. stenoptera and P. tonkinensis. Our analysis demonstrates that the traditional division of all the six species sampled throughout their distribution area as well as the phylogeny of the genus Pterocarya needs to be reviewed.

Towards sustainable rural landscapes? a multivariate analysis of the structure of traditional tree cropping systems along a human pressure gradient in a mediterranean region

The worldwide increase of human pressure in rural areas has resulted in a progressive fragmentation of agro-forest landscapes. Relatively few studies have identified processes of landscape modification and patch fragmentation reflecting long-term human activity from those caused by recent urbanization stimulated by economic development, population growth and improved accessibility of rural areas. In rural districts dominated by high-quality crop mosaics and exposed to increasing levels of human pressure, urbanization-driven changes in the use of land have determined complex and non-linear processes of landscape fragmentation which deserve further investigation. Multidimensional procedures and data mining are appropriate to explore such transformations, basing on a comprehensive assessment of landscape metrics. The present study investigates changes in land-use and landscape structure (2000–2008) in three districts of Latium (central Italy) featuring different cropping systems, physiographic attributes and level of human pressure. A principal component analysis was run to identify a core set of metrics aimed at (i) evaluating the role of traditional cropping systems in the preservation of traditional agro-forest systems and (ii) inferring the relationship between spatial heterogeneity in rural landscapes and socio-ecological processes of change. Our results indicate that crop intensification has contributed to landscape homogeneity and simplification in all areas studied, in contrast with the effects of urban expansion resulting in a more fragmented and diversified landscape with relict tree crop patches. The findings of this study clarify the contribution of traditional tree crop systems to sustainable structures and functions of rural landscapes by preserving the place-specific eco-mosaic complexity.

The complete chloroplast genome of a Tertiary relict species Liriodendron chinense (Magnoliaceae)

Liriodendron chinense is a rare Tertiary relict tree species. In this study, we report the complete chloroplast genome of a Chinese tulip tree, L. chinense. The total genome size was 159,429 bp in length, containing a pair of inverted repeats (IRs) of 26,333 bp, separated by large single copy (LSC) and small single copy (SSC) of 87,766 and 18,997 bp, respectively. The chloroplast genome contained the conservative quadripartite structure present in most Magnoliaceae chloroplasts composing of 79 protein-coding genes, 30 tRNAs and 4 rRNAs. Among these genes, 15 harbored a single intron, and 2 contained a couple of introns. The overall G+C content of the cpDNA is 39.2 %, while the corresponding values of the LSC, SSC and IR regions are 37.8, 34.3 and 43.2 % respectively. A maximum likelihood phylogenomic analysis showed that L. chinense is closely related to L. tulipifera that belongs to the tribe Magnoliaceae.

Genetic evidence for central‐marginal hypothesis in a Cenozoic relict tree species across its distribution in China

AbstractAimThis study tests predictions of the central‐marginal (C‐M) hypothesis using geographical patterns of genetic variation in a Cenozoic relict tree species Euptelea pleiospermum. Additionally, we explore the extent to which post‐glacial recolonization and landscape barriers created by topographical heterogeneity shape spatial genetic variation.LocationSouth‐west and central subtropical China.MethodsUsing microsatellite data of 678 individuals from 21 populations, we documented geographical patterns of genetic diversity and population genetic structure, supplemented with current and past (Last Glacial Maximum, LGM) distributions based on species distribution models (SDMs). Genetic structure was determined by non‐spatial and spatially sensitive Bayesian clustering, analysis of molecular variance, barrier analysis and a multivariate population graphing approach. We also examined both historical and contemporary gene flow.ResultsGenetic diversity significantly declined with distance from centre and showed unimodal patterns along both latitude and longitude. The C‐M pattern of genetic diversity along latitudinal gradient is high‐latitude‐skewed, with the peak at higher portion of the species' latitudinal extent. The Sichuan Basin divided individuals into south and north clusters, whereas the Qinba Mountains did not. The general shape of the predicted distribution during the LGM is strongly similar with that of current distribution, except the Sichuan Basin.Main conclusionsOur study provides genetic evidence for the C‐M hypothesis in a Cenozoic relict tree species across its distribution in China. The Sichuan Basin, rather than the Qinba Mountains, acts as dispersal barrier for a mountain tree species. Our results suggest that in situ refugia were maintained across southwest and central China during the LGM. The slight post‐glacial northward expansion and asymmetrical south–north historical‐gene flow did not result in a low‐latitude‐skewed C‐M pattern of genetic diversity. Instead, complex topography in combination with the C‐M dispersal dynamics shaped the latitudinal pattern of genetic diversity.

Contributions of historical and contemporary geographic and environmental factors to phylogeographic structure in a Tertiary relict species, Emmenopterys henryi (Rubiaceae).

Examining how historical and contemporary geographic and environmental factors contribute to genetic divergence at different evolutionary scales is a central yet largely unexplored question in ecology and evolution. Here, we examine this key question by investigating how environmental and geographic factors across different epochs have driven genetic divergence at deeper (phylogeographic) and shallower (landscape genetic) evolutionary scales in the Chinese Tertiary relict tree Emmenopterys henryi. We found that geography played a predominant role at all levels – phylogeographic clades are broadly geographically structured, the deepest levels of divergence are associated with major geological or pre-Quaternary climatic events, and isolation by distance (IBD) primarily explained population genetic structure. However, environmental factors are clearly also important – climatic fluctuations since the Last Interglacial (LIG) have likely contributed to phylogeographic structure, and the population genetic structure (in our AFLP dataset) was partly explained by isolation by environment (IBE), which may have resulted from natural selection in environments with divergent climates. Thus, historical and contemporary geography and historical and contemporary environments have all shaped patterns of genetic structure in E. henryi, and, in fact, changes in the landscape through time have also been critical factors.

Simulated herbivory does not constrain phenotypic plasticity to shade through ontogeny in a relict tree.

Ecological limits to phenotypic plasticity (PP), induced by simultaneous biotic and abiotic factors, can prevent organisms from exhibiting optimal plasticity, and in turn lead to decreased fitness. Herbivory is an important biotic stressor and may limit plant functional responses to challenging environmental conditions such as shading. In this study we investigated whether plant functional responses and PP to shade are constrained by herbivory, and whether such constraints are due to direct effects based on resource limitation by considering ontogeny. We used as a model system the relict tree Prunus lusitanica and implemented an indoor experiment to quantify the response of saplings of different ages to shade and herbivory. We measured five functional traits and quantitatively calculated PP. Results showed that herbivory did not constrain functional responses or PP to shade except for shoot:root ratio (SR), which, despite showing a high PP in damaged saplings, decreased under shade instead of increasing. Damaged saplings of older age did not exhibit reduced constraints on functional responses to shade and generally presented a lower PP than damaged saplings of younger age. Our findings suggest that herbivory-mediated constraints on plant plasticity to shade may not be as widespread as previously thought. Nonetheless, the negative effect of herbivory on SR plastic expression to shade could be detrimental for plant fitness. Finally, our results suggest a secondary role of direct effects (resource-based) on P.lusitanica plasticity limitation. Further studies should quantify plant resources in order to gain a better understanding of this seldom-explored subject.

The phylogeography of Fagus hayatae (Fagaceae): genetic isolation among populations.

The beech species Fagus hayatae is an important relict tree species in subtropical China, whose biogeographical patterns may reflect floral responses to climate change in this region during the Quaternary. Previous studies have revealed phylogeography for three of the four Fagus species in China, but study on F. hayatae, the most sparsely distributed of these species, is still lacking. Here, molecular methods based on eight simple sequence repeat (SSR) loci of nuclear DNA (nDNA) and three chloroplast DNA (cpDNA) sequences were applied for analyses of genetic diversity and structure in 375 samples from 14 F. hayatae populations across its whole range. Both nDNA and cpDNA indicated a high level of genetic diversity in this species. Significant fixation indexes and departures from the Hardy–Weinberg equilibrium, with a genetic differentiation parameter of R st of 0.233, were detected in nDNA SSR loci among populations, especially those on Taiwan Island, indicating strong geographic partitioning. The populations were classified into two clusters, without a prominent signal of isolation‐by‐distance. For the 15 haplotypes detected in the cpDNA sequence fragments, there was a high genetic differentiation parameter (G st = 0.712) among populations. A high G st of 0.829 was also detected outside but not within the Sichuan Basin. Consistent with other Fagus species in China, no recent population expansion was detected from tests of neutrality and mismatch distribution analysis. Overall, genetic isolation with limited gene flow was prominent for this species and significant phylogeographic structures existed across its range except for those inside the Sichuan Basin. Our study suggested long‐term geographic isolation in F. hayatae with limited population admixture and the existence of multiple refugia in the mountainous regions of the Sichuan Basin and southeast China during the Quaternary. These results may provide useful information critical for the conservation of F. hayatae and other Chinese beech species.