Himalaya-Hengduan Mountains Research Articles

Westwards and northwards dispersal of Triosteum himalayanum (Caprifoliaceae) from the Hengduan Mountains region based on chloroplast DNA phylogeography.

The varying topography and environment that resulted from paleoorogeny and climate fluctuations of the Himalaya–Hengduan Mountains (HHM) areas had a considerable impact on the evolution of biota during the Quaternary. To understand the phylogeographic pattern and historical dynamics of Triosteum himalayanum (Caprifoliaceae), we sequenced three chloroplast DNA fragments (rbcL-accD, rps15-ycf1, and trnH-psbA) from 238 individuals representing 20 populations. Nineteen haplotypes (H1–H19) were identified based on 23 single-site mutations and eight indels. Most haplotypes were restricted to a single population or neighboring populations. Analysis of molecular variance revealed that variations among populations were much higher than that within populations for the overall gene pool, as well as for the East Himalayan group (EH group) and the North Hengduan group (NHM group), but not for the Hengduan Mountains group (HM group). Ecoregions representing relatively high genetic diversity or high frequencies of private haplotypes were discovered, suggesting that this alpine herbaceous plant underwent enhanced allopatric divergence in isolated and fragmented locations during the Quaternary glaciations. The current phylogeographic structure of T. himalayanum might be due to heterogeneous habitats and Quaternary climatic oscillations. Based on the phylogeographic structure of T. himalayanum populations, the phylogenetic relationship of identified haplotypes and palaeodistributional reconstruction, we postulated both westwards and northwards expansion from the HM group for this species. The westwards dispersal corridor could be long, narrow mountain areas and/or the Yarlung Zangbo Valley, while the northwards movement path could be south–north oriented mountains and low-elevation valleys.

Long-distance dispersal or postglacial contraction? Insights into disjunction between Himalaya-Hengduan Mountains and Taiwan in a cold-adapted herbaceous genus, Triplostegia.

Current disjunct patterns can result from long‐distance dispersal or postglacial contraction. We herein investigate the evolutionary history of Triplostegia to elucidate the disjunction between the Himalaya–Hengduan Mountain region (HHM) and Taiwan (TW). Genetic structure of Triplostegia was investigated for 48 populations using sequences from five chloroplast loci and the ribosomal nuclear internal transcribed spacer. Divergence time estimation, ancestral area reconstruction, and species distribution modeling (SDM) were employed to examine the biogeographic history of Triplostegia. Substantial genetic differentiation among populations from southwestern China (SW), Central China (CC), and TW was detected. Triplostegia was inferred to have originated in SW, and diversification began during the late Miocene; CC was colonized in the mid‐Pliocene, and TW was finally colonized in the early Pleistocene. SDM suggested an expansion of climatically suitable areas during the Last Glacial Maximum and range contraction during the Last interglacial in Triplostegia. Disjunction between HHM and TW in Triplostegia is most likely the consequence of topographic isolation and postglacial contraction. The potential climatic suitability areas for Triplostegia by 2070s (2061–2080) are predicted to slightly shrink and move northward. With continued global warming and human‐induced deforestation, extinction risk may increase for the cold‐adapted species, and appropriate strategies should be employed for ecosystem conservation.

The ‘Ward Line–Mekong–Salween Divide’ is an important floristic boundary between the eastern Himalaya and Hengduan Mountains: evidence from the phylogeographical structure of subnival herbs Marmoritis complanatum (Lamiaceae)

The Ward Line-Mekong-Salween Divide (MSD), a classic geographical barrier, is considered the main driver for population differentiation of species in the Himalaya-Hengduan Mountains region. To identify the role of geological factors in shaping the population history of subnival plant species, we investigated the phylogeography of Marmoritis complanatum based on plastid and nuclear ribosomal DNA. The phylogeographical differentiation of M. complanatum was identified between two geographical regions (central-eastern Himalaya vs. Hengduan Mountains) across the MSD, and the differentiation of two lineages began in the early Pliocene (4.49 Mya), coinciding with the formation of the MSD. These results indicate the MSD was the main driver of phylogeographical differentiation in M. complanatum. High genetic diversity and inter-population differentiation were detected in M. complanatum. Mismatch distribution analyses and species distribution modelling indicated that the broad-scale species distributions remained relatively stable with signs of a slight shrinkage of populations after the Last Glacial Maximum, suggesting Quaternary glaciations had little impact on the large-scale distributional range shift of M. complanatum. This study provides the first evidence for the effects of the MSD on the phylogeographical structure of this subnival plant species, and verifies the importance of the floristic boundary represented by the MSD.

Seed dormancy and germination characteristics of two Rheum species in the Himalaya-Hengduan Mountains

Seed dormancy and germination characteristics are important factors determining plant reproductive success, and may be expected to have a major influence on plant distribution. In this study, we aimed to explore the characteristics of seed dormancy and germination in two endemic Rheum species (Rheum nobile and Rheum alexandrae) in the Himalaya-Hengduan Mountains. To determine the type of dormancy, fresh seeds of the two species (one population each) were incubated in light at 25/15 and 15/5 °C, and then dry after-ripening (DAR) seeds were incubated on water agar substrate with or without GA3. To determine the effect of temperature and light on germination, DAR seeds of the two species (two populations each) were incubated both in the light and in the dark at several temperatures, including constant and alternating temperatures. Base temperature (Tb) and thermal times for 50% germination (θ50) were calculated. DAR released physiological dormancy (PD), increasing final germination at 15/5 °C and widening the range of germination temperatures from higher to lower, indicative of type 2 non-deep PD for the two Rheum species. Light had no significant effect on germination of seeds from the two species (two populations each). Seeds of the two species germinated significantly better (>80%) at medium temperatures (10–25 °C) than at extreme low (5 °C) or high (35 °C) temperatures. Alternating temperatures (25/15 and 15/5 °C) did not significantly increase the final germination of the two species either in the light and in the dark, but it promoted seed germination more quickly than corresponding constant temperatures in the light in both Rh. alexandrae populations, especially at 15/5 °C. Germination in response to temperature was well described by the thermal-time model at suboptimal temperatures. The estimated Tb values were 1 and 0.9 °C, respectively, in two Rh. nobile populations; 4 and 4.1 °C, respectively, in two Rh. alexandrae populations; θ50 (thermal time) were 100 and 125 °Cd, respectively in two Rh. nobile populations; 76.92 and 83.33 °Cd, respectively in two Rh. alexandrae populations. The dormancy type, and germination responses to temperature and light condition does not explain why the two Rheum species are distributed in contrasting habitats. However, these findings reflect an advantageous germination strategy of these two Rheum species to adapt to the same alpine environments.

Seed dormancy, germination and soil seed bank of Lamiophlomis rotata and Marmoritis complanatum (Labiatae), two endemic species from Himalaya–Hengduan Mountains

Seed dormancy and germination characteristics are important factors determining plant reproductive success. In this study, we aimed to explore the characteristics of seed dormancy and germination of two endemic Labiatae species (Lamiophlomis rotata and Marmoritis complanatum) in the Himalaya–Hengduan Mountains. Germination was first tested in the light using freshly matured seeds at 25/15 and 15/5°C, and then again after dry after-ripening. Dried seeds were incubated in the light at a range of constant temperatures (1–35°C). The effects of dark and GA3 on germination were tested at several different temperatures. Base temperature (Tb) and thermal times for 50% final germination (θ50) were calculated. Seeds were also buried at the collection site to test seed persistence in the soil. Increased final germination after dry after-ripening indicated that the seeds of the two species exhibited non-deep physiological dormancy; however, they exhibited different germination characteristics and soil seed bank types. In L. rotata, GA3 only promoted germination at 5°C, producing no significant effect at other temperatures. Dark conditions decreased germination significantly at all temperatures. Tb and θ50 values were 0.6 and 82.7°C d. The soil seed bank of this species was classified as persistent. In M. complanatum, GA3 significantly promoted germination at all temperatures except 15°C. Dark conditions depressed germination significantly at warmer temperatures (20 and 25°C) but had no effect at lower temperatures. Tb and θ50 values were 0.1 and 92.3°C d. The soil seed bank was classified as transient. Our results suggest that the seed dormancy and germination of the two co-existing species share some commonalities but there are also species-specific adaptations to the harsh alpine environment.

Size-Mediated Interaction between a Cushion Species and Other Non-cushion Species at High Elevations of the Hengduan Mountains, SW China.

Arenaria polytrichoides (Caryophyllaceae) is a common cushion plant occurring at high elevations in the Himalaya-Hengduan Mountains, SW China. It frequently has other non-cushion species growing within its canopy, forming a contrast with the surrounding areas because it creates patches of higher diversity and greater biomass. In this study, we examined the relationship between the cushions and associated non-cushion species along a gradient of cushion size. A total of 200 A. polytrichoides individuals were selected to fit four size classes. Field measurements were carried out to assess canopy structure, functional traits relevant to growth and reproduction, and soil quality below cushions along the size gradient. Furthermore, the size effect of cushions on the richness and abundance of species and biomass production was also examined. All the morphological variables examined exhibited a positive correlation with cushion size, as did the nutrients under cushions. Large and compact cushions were associated with higher soil nutrient contents compared with small and loose cushions. As a result of these biogenic environmental changes, there was a stronger facilitation effect performed by large cushions. Data pertaining to functional traits revealed that large cushions benefit from the enhanced resources within their compact structure and exhibit greater fitness and a higher reproductive output than small cushions. Our data indicated that interactions occur between cushion species and other plants depending on the size of the cushions, probably because of the greater heterogeneity of conditions beneath larger cushions. These findings provide a clear demonstration of the generally overlooked importance of the traits of nurse plants, such as size and age, in terms of their facilitative effects.

Warm-cold colonization: response of oaks to uplift of the Himalaya-Hengduan Mountains.

Clarifying the relationship between distribution patterns of organisms and geological events is critical to understanding the impact of environmental changes on organismal evolution. Quercus sect. Heterobalanus is now distributed across the Himalaya-Hengduan Mountains (HHM) and warm lowland in East China, yet how the distribution patterns of this group changed in response to the HHM uplift remains largely unknown. This study examines the effect of tectonic events in the HHM region on the oaks, providing a biological perspective on the geological history of this region. Fifty-six populations of Quercus sect. Heterobalanus were genotyped using four chloroplast DNA regions and nine nuclear simple sequence repeat loci to assess population structure and diversity, supplemented by molecular dating and ancestral area reconstructions. The underlying demographic dynamics were compared using ecological niche models of the species distributions during the last glacial maximum and the present. These analyses illustrate that Quercus sect. Heterobalanus diversified as the HHM uplifted and climatic cooling during the mid-Miocene, colonizing the cold habitats from warm broadleaf mixed forests. Lineages in cold highlands and warm lowlands have diverged as a consequence of local adaptation to diverging climates since the late Miocene. Our results suggest that continuous uplift of the HHM in the late Miocene to early Pliocene accompanied by simultaneous cooling triggered the differentiation of oaks. The biogeography of Quercus sect. Heterobalanus illuminates the geological events responsible for the modern-day HHM.

Plastid phylogenomics and adaptive evolution of Gaultheria series Trichophyllae (Ericaceae), a clade from sky islands of the Himalaya-Hengduan Mountains

Gaultheria series Trichophyllae Airy Shaw is an angiosperm clade of high-alpine shrublets endemic to the Himalaya-Hengduan Mountains and characterized by recent species divergence and convergent character evolution that has until recently caused much confusion in species circumscription. Although multiple DNA sequence regions have been employed previously, phylogenetic relationships among species in the group have remained largely unresolved. Here we examined the effectiveness of the plastid genome for improving phylogenetic resolution within the G. series Trichophyllae clade. Plastid genomes of 31 samples representing all 19 recognized species of the series and three outgroup species were sequenced with Illumina Sequencing technology. Maximum likelihood (ML), maximum parsimony (MP) and Bayesian inference (BI) phylogenetic analyses were performed with various datasets, i.e., that from the whole plastid genome, coding regions, noncoding regions, large single-copy region (LSC) and inverted-repeat region a (IRa). The partitioned whole plastid genome with inverted-repeat region b (IRb) excluded was also analyzed with ML and BI. Tree topologies based on the whole plastid genome, noncoding regions, and LSC region datasets across all analyses, and that based on the partitioned dataset with ML and BI analyses, are identical and generally strongly supported. Gaultheria series Trichophyllae form a clade with three species and one variety that is sister to a clade of the remaining 16 species; the latter comprises seven main subclades. Interspecific relationships within the series are strongly supported except for those based on the coding-region and IRa-region datasets. Eight divergence hotspot regions, each possessing >5% percent variable sites, were screened across the whole plastid genome of the 28 individuals sampled in the series. Results of morphological character evolution reconstruction diagnose several clades, and a hypothesis of adaptive evolution for plant habit is postulated.

Differentiation in drought tolerance mirrors the geographic distributions of alpine plants on the Qinghai-Tibet Plateau and adjacent highlands

Climatic tolerance, especially drought tolerance, is one of the major factors shaping the geographic distributions of plant species. Thus, the general decline in rainfall from the Himalaya-Hengduan Mountains (HHM) to the inner Qinghai-Tibet Plateau (QTP) might account for the significant differences in species distributions and richness between the two regions. To test this hypothesis, we conducted a water stress experiment using four Anisodus species (A. tanguticus, A. luridus, A. carniolicoides and A. acutangulus), which were treated with different levels of water stress in a glasshouse, and examined their differences in physiological responses. The results suggest that A. tanguticus, which inhabits the inner QTP, generally has higher fitness under severe water stress than the other species based on its high root:shoot ratio, long-term water use efficiency and photosynthetic rate, indicating that it possesses a genetically based drought tolerance mechanism. Our results suggest that plant species inhabiting the inner QTP may be more drought tolerant than those inhabiting the HHM regions. This provides a new example supporting the hypothesis that climatic tolerance plays a major role in shaping plant distributions on the QTP and its adjacent highlands and presents new insights into the patterns of geographic distribution and diversity of the plants inhabiting these areas.

Seed quality of the Sino–Himalayan endemic genus Cyananthus (Campanulaceae) increases with elevation and varies with life histories

Background: Seed quality can directly determine the survival and population dynamics of alpine plant species. However, how the quality of seeds is affected by elevation-related stress in the Hengduan mountains in the South-western China remains unclear.Aims: To determine how the seed quality of species of Cyananthus varies with elevation, life history and adaptive strategies.Methods: Nine species, including three annuals and six perennials, in the Himalaya-Hengduan Mountains from ca. 2500 m to ca. 4700 m a.s.l. were selected. For each population, seeds from at least 25 individuals were collected. After air-drying, we related seed mass and per cent germination with elevation and compared these parameters between life histories.Results: Results showed that seed mass (annual species) and germinability (annual and perennial species) were positively correlated with elevation. In addition, seeds from higher elevation exhibited explosive germination within a short period. No correlation was found between seed germination and seed mass.Conclusions: These findings indicate that selective pressure (e.g. low temperature and short growing season) might favour the formation of larger seeds in species from higher elevation, as well as a higher germination percentage and more rapid seed germination. These traits further confer high seed quality at higher elevation. We suggest that elevation is the main determinant of seed quality for Cyananthus species, but seed quality does differ between life histories.

Genetic Structure and Evolutionary History of Three Alpine Sclerophyllous Oaks in East Himalaya-Hengduan Mountains and Adjacent Regions.

The East Himalaya-Hengduan Mountains (EH-HM) region has a high biodiversity and harbors numerous endemic alpine plants. This is probably the result of combined orographic and climate oscillations occurring since late Tertiary. Here, we determined the genetic structure and evolutionary history of alpine oak species (including Quercus spinosa, Quercus aquifolioides, and Quercus rehderiana) using both cytoplasmic-nuclear markers and ecological niche models (ENMs), and elucidated the impacts of climate oscillations and environmental heterogeneity on their population demography. Our results indicate there were mixed genetic structure and asymmetric contemporary gene flow within them. The ENMs revealed a similar demographic history for the three species expanded their ranges from the last interglacial (LIG) to the last glacial maximum (LGM), which was consistent with effective population sizes changes. Effects of genetic drift and fragmentation of habitats were responsible for the high differentiation and the lack of phylogeographic structure. Our results support that geological and climatic factors since Miocene triggered the differentiation, evolutionary origin and range shifts of the three oak species in the studied area and also emphasize that a multidisciplinary approach combining molecular markers, ENMs and population genetics can yield deep insights into diversification and evolutionary dynamics of species.

Overlapping Leaves Covering Flowers in the Alpine Species Eriophyton wallichii (Lamiaceae): Key Driving Factors and Their Potential Impact on Pollination

Extrafloral structures are supposed to have evolved to protect flowers from harsh physical environments but might have effects on pollination. Overlapping leaves cover flowers in Eriophyton wallichii, an alpine perennial endemic to the Himalaya-Hengduan Mountains. In previous study, it has showed that these extrafloral leaves can protect interior flowers from temperature fluctuations caused by drastic solar radiation fluctuations, but these leaves may also protect interior flowers from rain wash and UVB damage, and we do not know which one is the main function. In this study, we investigated whether rain and UVB protection are the main functions of overlapping leaves covering flowers and their potential impact on pollination. We first measured the intensities of UVB radiation in open air, beneath leaves and corollas, and then examined pollen susceptibility to different intensities of UVB and rain in the laboratory to estimate whether corollas per se protect interior pollen from UVB and rain damage. We also carried out pollination treatments and observed pollinator visitation of flowers with and without leaves in the field to assess whether the overlapping leaves covering flowers impair pollinator attraction. Our results showed that (1) water and strong UVB significantly decreased pollen germinability, but corollas per se could protect pollen from UVB and rain damage; (2) no autonomous self-pollination and apomixis occurred, and pollinators were essential for the reproduction of E. wallichii; however, flower coverage by overlapping leaves did not limit pollination. We suggested that rain and UVB protection was not the main function of overlapping leaves covered flowers, given that this protection can be provided by corollas per se. Alternatively, this extrafloral structure in E. wallichii may have evolved in response to extreme high temperatures associated with the strong solar radiation fluctuations. This indicates that, even in alpine plants, extreme high temperature may affect the evolution of plant extrafloral structures.

Divergence in Eco-Physiological Responses to Drought Mirrors the Distinct Distribution of Chamerion angustifolium Cytotypes in the Himalaya-Hengduan Mountains Region.

Polyploid species generally occupy harsher habitats (characterized by cold, drought and/or high altitude) than diploids, but the converse was observed for Chamerion angustifolium, in which diploid plants generally inhabit higher altitudes than their polyploid derivatives. Plants at high altitudes may experience cold-induced water stress, and we therefore examined the physiological responses of diploid and hexaploid C. angustifolium to water stress to better understand the ecological differentiation of plants with different ploidy levels. We conducted a common garden experiment by subjecting seedlings of different ploidy levels to low, moderate, and severe water stress. Fourteen indicators of physiological fitness were measured, and the anatomical characteristics of the leaves of each cytotype were determined. Both cytotypes were influenced by drought, and diploids exhibited higher fitness in terms of constant root:shoot ratio (R:S ratio) and maximum quantum yield of PS II (Fv/Fm), less reduced maximal photosynthetic rate (Amax), transpiration rate (E), intercellular CO2 concentration (Ci) and stomatal conductance (gs), and higher long-term water use efficiency (WUEL) under severe water stress than did hexaploids. Analysis of leaf anatomy revealed morphological adjustments for tolerating water deficiency in diploids, in the form of closely packed mesophyll cells and small conduits in the midvein. Our results indicate that diploid C. angustifolium is more tolerant of drought than hexaploid plants, ensuring the successful survival of the diploid at high altitudes. This eco-physiological divergence may facilitate the species with different cytotypes to colonize new and large geographic ranges with heterogeneous environmental conditions.

Genetic diversity of Meconopsis integrifolia (Maxim.) Franch. In the East Himalaya–Hengduan Mountains inferred from fluorescent amplified fragment length polymorphism analysis

Meconopsis integrifolia (Maxim.) Franch. is a representative species of the genus found in the East Himalaya-Hengduan Mountain. As a species of medical and horticultural significance, M. integrifolia is threatened by over-exploitation and habitat fragmentation. In this study, the genetic diversity and structure of M. integrifolia, represented by 183 individuals from 10 populations, were studied using 6 pairs of fluorescent Amplified Fragment Length Polymorphisms primers. Our results showed a relatively high genetic variation at the species level (PPB = 82.0%, HE = 0.2356 and I = 0.3695). In contrast, population diversity was extremely low, as measured by Nei's genetic diversity index and Shannon's diversity index (HE = 0.0317, I = 0.0480). Analyses of molecular variance (AMOVA) showed that among and within populations, the genetic variation accounted for 88.9% and 11.1% of the total genetic variation, respectively. In addition, Nei's coefficient of differentiation (GST) was found to be high (0.8636), confirming the significant high level of genetic differentiation as well as low gene flow among populations of this species. The results of Neighbor-joining cluster, PCoA, and Bayesian assignment revealed similar genetic differentiation patterns, suggesting three main genetic groups that are concordant with their geographical distribution. Three main barriers—the Mekong River, the Yalong River, and a branch of the Yalong River—were detected using BARRIER software. The possible mechanisms and implications of these findings for conservation are discussed.

Floral traits influence pollen vectors' choices in higher elevation communities in the Himalaya-Hengduan Mountains.

BackgroundHow floral traits and community composition influence plant specialization is poorly understood and the existing evidence is restricted to regions where plant diversity is low. Here, we assessed whether plant specialization varied among four species-rich subalpine/alpine communities on the Yulong Mountain, SW China (elevation from 2725 to 3910 m). We analyzed two factors (floral traits and pollen vector community composition: richness and density) to determine the degree of plant specialization across 101 plant species in all four communities. Floral visitors were collected and pollen load analyses were conducted to identify and define pollen vectors. Plant specialization of each species was described by using both pollen vector diversity (Shannon’s diversity index) and plant selectiveness (d’ index), which reflected how selective a given species was relative to available pollen vectors.ResultsPollen vector diversity tended to be higher in communities at lower elevations, while plant selectiveness was significantly lower in a community with the highest proportion of unspecialized flowers (open flowers and clusters of flowers in open inflorescences). In particular, we found that plant species with large and unspecialized flowers attracted a greater diversity of pollen vectors and showed higher selectiveness in their use of pollen vectors. Plant species with large floral displays and high flower abundance were more selective in their exploitation of pollen vectors. Moreover, there was a negative relationship between plant selectiveness and pollen vector density.ConclusionsThese findings suggest that flower shape and flower size can increase pollen vector diversity but they also increased plant selectiveness. This indicated that those floral traits that were more attractive to insects increased the diversity of pollen vectors to plants while decreasing overlap among co-blooming plant species for the same pollen vectors. Furthermore, floral traits had a more important impact on the diversity of pollen vectors than the composition of anthophilous insect communities. Plant selectiveness of pollen vectors was strongly influenced by both floral traits and insect community composition. These findings provide a basis for a better understanding of how floral traits and community context shape interactions between flowers and their pollen vectors in species-rich communities.Electronic supplementary materialThe online version of this article (doi:10.1186/s12898-016-0080-1) contains supplementary material, which is available to authorized users.

The Use of DNA Barcoding on Recently Diverged Species in the Genus Gentiana (Gentianaceae) in China.

DNA barcoding of plants poses particular challenges, especially in differentiating, recently diverged taxa. The genus Gentiana (Gentianaceae) is a species-rich plant group which rapidly radiated in the Himalaya-Hengduan Mountains in China. In this study, we tested the core plant barcode (rbcL + matK) and three promising complementary barcodes (trnH-psbA, ITS and ITS2) in 30 Gentiana species across 6 sections using three methods (the genetic distance-based method, Best Close Match and tree-based method). rbcL had the highest PCR efficiency and sequencing success (100%), while the lowest sequence recoverability was from ITS (68.35%). The presence of indels and inversions in trnH-psbA in Gentiana led to difficulties in sequence alignment. When using a single region for analysis, ITS exhibited the highest discriminatory power (60%-74.42%). Of the combinations, matK + ITS provided the highest discrimination success (71.43%-88.24%) and is recommended as the DNA barcode for the genus Gentiana. DNA barcoding proved effective in assigning most species to sections, though it performed poorly in some closely related species in sect. Cruciata because of hybridization events. Our analysis suggests that the status of G. pseudosquarrosa needs to be studied further. The utility of DNA barcoding was also verified in authenticating ‘Qin-Jiao’ Gentiana medicinal plants (G. macrophylla, G. crassicaulis, G. straminea, and G. dahurica), which can help ensure safe and correct usage of these well-known Chinese traditional medicinal herbs.

Towards a comprehensive phylogeny of the large temperate genus Pedicularis (Orobanchaceae), with an emphasis on species from the Himalaya-Hengduan Mountains.

BackgroundStriking interspecific variations in floral traits of the large temperate genus Pedicularis have given rise to controversies concerning infra-generic classifications. To date, phylogenetic relationships within the genus have not been well resolved. The main goal of this study is to construct a backbone phylogeny of Pedicularis, with extensive sampling of species from the Himalaya-Hengduan Mountains. Phylogenetic analyses included 257 species, representing all 13 informal groups and 104 out of 130 series in the classification system of Tsoong, using sequences of the nuclear ribosomal internal transcribed spacer (nrITS) and three plastid regions (matK, rbcL and trnL-F). Bayesian inference and maximum likelihood methods were applied in separate and combined analyses of these datasets.ResultsThirteen major clades are resolved with strong support, although the backbone of the tree is poorly resolved. There is little consensus between the phylogenetic tree and Tsoong’s classification of Pedicularis. Only two of the 13 groups (15.4 %), and 19 of the 56 series (33.9 %) with more than one sampled species were found to be strictly monophyletic. Most opposite-/whorled-leaved species fall into a single clade, i.e. clade 1, while alternate leaves species occur in the remaining 12 clades. Excluding the widespread P. verticillata in clade 1, species from Europe and North America fall into clades 6–8.ConclusionsOur results suggest that combinations of morphological and geographic characters associated with strongly supported clades are needed to elucidate a comprehensive global phylogeny of Pedicularis. Alternate leaves are inferred to be plesiomorphic in Pedicularis, with multiple transitions to opposite/whorled phyllotaxy. Alternate-leaved species show high diversity in plant habit and floral forms. In the Himalaya-Hengduan Mountains, geographical barriers may have facilitated diversification of species with long corolla tubes, and the reproductive advantages of beakless galeas in opposite-/whorled-leaved species may boost speciation at high altitude.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-015-0547-9) contains supplementary material, which is available to authorized users.

Refugial isolation and range expansions drive the genetic structure of Oxyria sinensis (Polygonaceae) in the Himalaya-Hengduan Mountains

The formation of the Mekong-Salween Divide and climatic oscillations in Pleistocene were the main drivers for the contemporary diversity and genetic structure of plants in the Himalaya-Hengduan Mountains (HHM). To identify the relative roles of the two historical events in shaping population history of plants in HHM, we investigated the phylogeographic pattern of Oxyria sinensis, a perennial plant endemic to the HHM. Sixteen chloroplast haplotypes were identified and were clustered into three phylogenetic clades. The age of the major clades was estimated to be in the Pleistocene, falling into several Pleistocene glacial stages and postdating the formation of the Mekong-Salween Divide. Range expansions occurred at least twice in the early and middle Pleistocene, but the spatial genetic distribution rarely changed since the Last Glacial Maximum. Our results suggest that temporary mountain glaciers may act as barriers in promoting the lineage divergence in O. sinensis and that subsequential range expansions and secondary contacts might reshape the genetic distribution in geography and blur the boundary of population differentiation created in the earlier glacial stages. This study demonstrates that Pleistocene climatic change and mountain glaciers, rather than the Mekong-Salween Divide, play the primary role in shaping the spatial genetic structure of O. sinensis.

Floral nectary morphology and evolution inPedicularis(Orobanchaceae)

Intricate associations between floral morphology and pollinator foraging behaviour are common. In this context, the presence and form of floral nectaries can play a crucial role in driving floral evolution and diversity in flowering plants. However, the reconstruction of the ancestral state of nectary form is often hampered by a lack of anatomical studies and well-resolved phylogenetic trees. Here, we studied 39 differentially pollinated Pedicularis spp., a genus with pronounced interspecific variation in colour, shape and size of the corolla. Anatomical and scanning electron microscopy observations revealed two nectary forms [bulged (N=27) or elongated (N=5)] or the absence of nectaries (N=7). In a phylogenetic context, our data suggest that: (1) the bulged nectary should be the ancestral state; (2) nectaries were independently lost in some beaked species; and (3) elongated nectaries evolved independently in some clades of beakless species. Phylogenetic path analysis showed that nectary presence is indirectly correlated with beak length/pollinator behaviour through an intermediate factor, nectar production. No significant correlation was found between nectary type and nectar production, beak length or pollinator behaviour. Some beaked species had nectary structures, although they did not produce nectar. The nectary in beaked species may be a vestigial structure retained during a recent rapid radiation of Pedicularis, especially in the Himalaya-Hengduan Mountains of south-western China.(c) 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 178, 592-607.

DNA barcoding evaluation and its taxonomic implications in the species-rich genus Primula L. in China.

The genus Primula is extremely diverse in the east Himalaya-Hengduan Mountains (HHM) in China as a result of rapid radiation. In order to overcome the difficulty of morphological classification of this genus, we surveyed three plastid regions (rbcL, matK, and trnH-psbA) and two nuclear markers (ITS and ITS2) from 227 accessions representing 66 Primula species across 18 sections, to assess their discriminatory power as barcodes. We found that ITS alone or combined with plastid regions showed the best discrimination across different infrageneric ranks and at species level. We suggest rbcL + matK + ITS as the first choice at present to barcode Primula plants. Although the present barcoding combination performed poorly in many closely related species of Primula, it still provided many new insights into current Primula taxonomy, such as the underlying presence of cryptic species, and several potential improper taxonomic treatments. DNA barcoding is one useful technique in the integrative taxonomy of the genus Primula, but it still requires further efforts to improve its effectiveness in some taxonomically challenging groups.