Phylogenetic origins of the Himalayan endemic Dolomiaea, Diplazoptilon and Xanthopappus (Asteraceae : Cardueae) based on three DNA regions

Background and Aims It is an enduring question as to the mechanisms leading to the high diversity and the processes producing endemics with unusual morphologies in the Himalayan alpine region. In the present study, the phylogenetic relationships and origins of three such endemic genera were analysed, Dolomiaea, Diplazoptilon and Xanthopappus, all in the tribe Cardueae of Asteraceae.Methods The nuclear rDNA internal transcribed spacer (ITS) and plastid trnL-F and psbA-trnH regions of these three genera were sequenced. The same regions for other related genera in Cardueae were also sequenced or downloaded from GenBank. Phylogenetic trees were constructed from individual and combined data sets of the three types of sequences using maximum parsimony, maximum likelihood and Bayesian analyses.Key Results The phylogenetic tree obtained allowed earlier hypotheses concerning the relationships of these three endemic genera based on gross morphology to be rejected. Frolovia and Saussurea costus were deeply nested within Dolomiaea, and the strong statistical support for the Dolomiaea-Frolovia clade suggested that circumscription of Dolomiaea should be more broadly redefined. Diplazoptilon was resolved as sister to Himalaiella, and these two together are sister to Lipschitziella. The clade comprising these three genera is sister to Jurinea, and together these four genera are sister to the Dolomiaea-Frolovia clade. Xanthopappus, previously hypothesized to be closely related to Carduus, was found to be nested within a well-supported but not fully resolved Onopordum group with Alfredia, Ancathia, Lamyropappus, Olgaea, Synurus and Syreitschikovia, rather than the Cardinis group. The crude dating based on ITS sequence divergence revealed that the divergence time of Dolomiaea-Frolovia from its sister group probably occurred 13.6-12.2 million years ago (Ma), and the divergence times of the other two genera, Xanthopappus and Diplazoptilon, from their close relatives around 5.7-4.7 Ma and 2.0-1.6 Ma, respectively.Conclusions The findings provide an improved understanding of the intergeneric relationships in Cardueae. The crude calibration of lineages indicates that the uplifts of the Qiinghai -Tibetan Plateau since the Miocene might have served as a continuous stimulus for the production of these morphologically aberrant endemic elements of the Himalayan flora.

Enrichment of stable carbon and nitrogen isotopes of plant populations and plateau pikas along altitudes

Stable carbon and nitrogen isotope ratios were measured in plant populations and plateau pikas to determine enrichment in stable isotopes of three alpine meadow ecosystems at different elevations in the Qinghai-Tibet Plateau. The results indicated that stable carbon isotope signatures of plant populations at the three locations showed significant variations, delta C-13 of plant populations showed an enrichment of 0.86 parts per thousand per 1000 in over the linear proportion of the altitudinal response, while stable nitrogen isotopes showed no apparent difference. The stable nitrogen isotopes of plateau pikas became significantly isotopically heavier along altitudinal gradients and showed an enrichment of 3.17 parts per thousand/km. Stable carbon isotopes showed no significance, however, and the enrichment of 0.448 parts per thousand/km. delta C-13 and delta N-15 in plateau pikas were not significantly correlated. There appeared to be segregation between the metabolism of stable carbon and nitrogen isotopes of plateau pikas. Variances in metabolic rate at lower water availability and temperatures are presumed to be the main cause of enrichment of stable nitrogen isotopes along altitudinal gradients. Attention should be paid to construct trophic positions and to trace food chain information based oil an isotopic enrichment model.

Carbon dioxide dynamics and controls in a deep-water wetland on the Qinghai-Tibetan Plateau

To initially characterize the dynamics and environmental controls of CO2, ecosystem CO2 fluxes were measured for different vegetation zones in a deep-water wetland on the Qinghai-Tibetan Plateau during the growing season of 2002. Four zones of vegetation along a gradient from shallow to deep water were dominated, respectively by the emergent species Carex allivescens V. Krez., Scirpus distigmaticus L., Hippuris vulgaris L., and the submerged species Potamogeton pectinatus L. Gross primary production (GPP), ecosystem respiration (Re), and net ecosystem production (NEP) were markedly different among the vegetation zones, with lower Re and GPP in deeper water. NEP was highest in the Scirpus-dominated zone with moderate water depth, but lowest in the Potamogeton-zone that occupied approximately 75% of the total wetland area. Diurnal variation in CO2 flux was highly correlated with variation in light intensity and soil temperature. The relationship between CO2 flux and these environmental variables varied among the vegetation zones. Seasonal CO2 fluxes, including GPP, Re, and NEP, were strongly correlated with aboveground biomass, which was in turn determined by water depth. In the early growing season, temperature sensitivity (Q(10)) for Re varied from 6.0 to 8.9 depending on vegetation zone. Q(10) decreased in the late growing season. Estimated NEP for the whole deep-water wetland over the growing season was 24 g C m(-2). Our results suggest that water depth is the major environmental control of seasonal variation in CO2 flux, whereas photosynthetic photon flux density (PPFD) controls diurnal dynamics.

Distribution and species diversity of plant communities along transect on the Northeastern Tibetan plateau

The distribution and species diversity of plant communities along a 600 km transect through the northeastern Tibetan Plateau (32 degrees 42'-35 degrees 07' N, 101 degrees 02'-97 degrees 38' E) with altitudes from 3255 to 4460 m are described. The transect started from the Youyi Bridge of Banma through Dari, Maqin and Maduo to Zaling Lake. The data from 47 plots along the transect are summarized and analyzed. The mean annual temperature, the mean annual rainfall and the length of growing season decreases from 2.6 to -4.5 degrees C, from 767.2 to 240.1 mm, from 210 to 140 days, respectively, along the transect from the southeastern Banma to northwestern Zaling Lake. The number of vascular plant species recorded in 47 plots is 242 including 2 tree, 34 shrub, 206 herb species. Main vegetation types on the transect from southeast to northwest are: Sabina convallium forest, Picea likiangensis forest, Pyracantha fortuneana + Spiraea alpina shrub, Hippophae neurocarpu shrub, Sibiraea angustata + Polygonum viviparum shrub, Stellera chamaejasme herb meadow, Potentilla fruticosa + Salix obscura + Carex sp. Shrub, Kobresia capillifolia meadow, P. froticosa + Kobresia humilis shrub, Caragana jubata + S. obscura shrub, Kobresia tibetica meadow, Kobresia pygmaea meadow, K. pygmaea + Stipa purpurea steppe meadow, Stipa purpurea steppe. Plant richness and diversity index all showed a decreasing trend with increasing of elevation along transect from southeast to northwest. Detailed information on altitudinal ranges and distribution of the alpine vegetation, vascular flora and environments over the alpine zone at northeastern Tibetan Plateau provides baseline records relevant to future assessment of probable effects of global climate changes.

Characterization of photosynthetic pathway of plant species growing in the eastern Tibetan plateau using stable carbon isotope composition

The photosynthetic pathway of plant species collected at Menyuan, Henan, and Maduo sites, east of Tibetan Plateau, China, during the growing season were studied using stable carbon isotopes in leaves. The 232 samples leaves analyzed belonged to 161 species, 30 families, and 94 genera. The delta(13)C values (from -24.6 to -29.2 %o) indicated that all the considered species had a photosynthetic C-3 pathway. The absence of plant species with C-4 photosynthetic pathway might be due to the extremely low air temperature characterizing the Tibetan Plateau. The average delta(13)C value was significantly (p < 0.05) different between annuals and perennials at the three considered study sites. Hence the longer-lived species had greater water-use efficiency (WUE) than shorter-lived species, that is, longer-lived species are better adapted to the extreme environmental conditions of the Tibetan Plateau.

Phylogeography of the Qinghai-Tibetan Plateau endemic Juniperus przewalskii (Cupressaceae) inferred from chloroplast DNA sequence variation

The vegetation of the northeast Qinghai-Tibetan Plateau is dominated by alpine meadow and desert-steppe with sparse forests scattered within it. To obtain a better understanding of the phylogeography of one constituent species of the forests in this region, we examined chloroplast trnT-trnF and trnS-trnG sequence variation within Juniperus przewalskii, a key endemic tree species. Sequence data were obtained from 392 trees in 20 populations covering the entire distribution range of the species. Six cpDNA haplotypes were identified. Significant population subdivision was detected (G(ST) = 0.772, N-ST = 0.834), suggesting low levels of recurrent gene flow among populations and significant phylogeographic structure (N-ST > G(ST), P < 0.05). Eight of the nine disjunct populations surveyed on the high-elevation northeast plateau were fixed for a single haplotype (A), while the remaining, more westerly population, contained the same haplotype at high frequency together with two low frequency haplotypes (C and F). In contrast, most populations that occurred at lower altitudes at the plateau edge were fixed or nearly fixed for one of two haplotypes, A or E. However, two plateau edge populations had haplotype compositions different from the rest. In one, four haplotypes (A, B, D and E) were present at approximately equivalent frequencies, which might reflect a larger refugium in the area of this population during the last glacial period. Phylogenetic analysis indicated that the most widely distributed haplotype A is not ancestral to other haplotypes. The contrasting phylogeographic structures of the haplotype-rich plateau edge area and the almost haplotype-uniform plateau platform region indicate that the plateau platform was recolonized by J. przewalskii during the most recent postglacial period. This is supported by the findings of a nested clade analysis, which inferred that postglacial range expansion from the plateau edge followed by recent fragmentation is largely responsible for the present-day spatial distribution of cpDNA haplotypes within the species.

Strong temperature dependence and no moss photosynthesis in winter CO2 flux for a Kobresia meadow on the Qinghai-Tibetan plateau

We examined the CO2 exchange of a Kobresia meadow ecosystem on the Qinghai-Tibetan plateau using a chamber system. CO2 efflux from the ecosystem was strongly dependence on soil surface temperature. The COZ efflux-temperature relationship was identical under both light and dark conditions, indicating that no photosynthesis could be detected under light conditions during the measurement period. The temperature sensitivity (Q(10)) of the COZ efflux showed a marked transition around -1.0 degrees C; Q(10) was 2.14 at soil surface temperatures above and equal to -1.0 degrees C but was 15.3 at temperatures below -1.0 degrees C. Our findings suggest that soil surface temperature was the major factor controlling winter COZ flux for the alpine meadow ecosystem and that freeze-thaw cycles at the soil surface layer play an important role in the temperature dependence of winter CO2 flux. (c) 2005 Elsevier Ltd. All rights reserved.

Dynamic and complex microclimate responses to warming and grazing manipulations

Synthesis efforts that identify patterns of ecosystem response to a suite of warming manipulations can make important contributions to climate change science. However, cross-study comparisons are impeded by the paucity of detailed analyses of how passive warming and other manipulations affect microclimate. Here we document the independent and combined effects of a common passive warming manipulation, open-top chambers (OTCs), and a simulated widespread land use, clipping, on microclimate on the Tibetan Plateau. OTCs consistently elevated growing season averaged mean daily air temperature by 1.0-2.0 degrees C, maximum daily air temperature by 2.1-7.3 degrees C and the diurnal air temperature range by 1.9-6.5 degrees C, with mixed effects on minimum daily air temperature, and mean daily soil temperature and moisture. These OTC effects on microclimate differ from reported effects of a common active warming method, infrared heating, which has more consistent effects on soil than on air temperature. There were significant interannual and intragrowing season differences in OTC effects on microclimate. For example, while OTCs had mixed effects on growing season averaged soil temperatures, OTCs consistently elevated soil temperature by approximately 1.0 degrees C early in the growing season. Nonadditive interactions between OTCs and clipping were also present: OTCs in clipped plots generally elevated air and soil temperatures more than OTCs in nonclipped plots. Moreover, site factors dynamically interacted with microclimate and with the efficacy of the OTC manipulations.These findings highlight the need to understand differential microclimate effects between warming methods, within warming method across ecosystem sites, within warming method crossed with other treatments, and within sites over various timescales. Methods, sites and scales are potential explanatory variables and covariables in climate warming experiments. Consideration of this variability among and between experimental warming studies will lead to greater understanding and better prediction of ecosystem response to anthropogenic climate warming.

Characterizing CO2 fluxes for growing and non-growing seasons in a shrub ecosystem on the Qinghai-Tibet Plateau

To assess carbon budget for shrub ecosystems on the Qinghai-Tibet Plateau, CO2 flux was measured with an open-path eddy covariance system for an alpine shrub ecosystem during growing and non-growing seasons. CO2 flux dynamics was distinct between the two seasons. During the growing season from May to September, the ecosystem exhibited net CO2 uptake from 08:00 to 19:00 (Beijing Standard Time), but net CO2 emission from 19:00 to 08:00. Maximum CO2 uptake appeared around 12:00 with values of 0.71, 1,19, 1.46 and 0.67 g CO2 m(-2) h(-1) for June, July, August and September, respectively. Diurnal fluctuation Of CO2 flux showed higher correlation with photosynthetic photon flux density than temperature. The maximum net CO2 influx occurred in August with a value of 247 g CO2 m(-2). The total CO2 uptake by the ecosystem was up to 583 g CO2 m(-2) for the growing season. During the non-growing season from January to April and from October to December, CO2 flux showed small fluctuation with the largest net CO2 efflux of 0.30 g CO2 m(-2) h(-1) in April. The diurnal CO2 flux was close to zero during most time of the day, but showed a small net CO2 eff lux from 11:00 to 18:00. Diurnal CO2 flux, is significantly correlated to diurnal temperature in the non-growing season. The maximum monthly net CO2 eff lux appeared in April, with a value of 105 g CO2 m(-2). The total net CO2 eff lux for the whole non-growing season was 356 g CO2 m(-2).

Energy exchange between the atmosphere and a meadow ecosystem on the Qinghai-Tibetan Plateau

To reveal the potential contribution of grassland ecosystems to climate change, we examined the energy exchange over an alpine Kobresia meadow on the northeastern Qinghai-Tibetan Plateau. The annual pattern of energy exchange showed a clear distinction between periods of frozen soil with the daily mean soil temperature at 5 cm (T-s5 &LE; 0 &DEG; C) and non-frozen soil (T-s5 > 0 &DEG; C). More than 80% of net radiation was converted to sensible heat (H) during the frozen soil period, but H varied considerably with the change in vegetation during the non-frozen soil period. Three different sub-periods were further distinguished for the later period: (1) the pre-growth period with Bowen ratio (β) > 1 was characterized by a high β of 3.0 in average and the rapid increase of net radiation associated with the increases of H, latent heat (LE) and soil heat; (2) during the Growth period when β &LE; 1, the LE was high but H fluxes was low with β changing between 0.3 and 0.4; (3) the post-growth period with average β of 3.6 when H increased again and reached a second maximum around early October. The seasonal pattern suggests that the phenology of the vegetation and the soil water content were the major factors affecting the energy partitioning in the alpine meadow ecosystem. © 2005 Elsevier B.V. All rights reserved.

Experimental warming causes large and rapid species loss, dampened by simulated grazing, on the Tibetan Plateau

We investigated the independent and combined effects of experimental warming and grazing on plant species diversity on the north-eastern Tibetan Plateau, a region highly vulnerable to ongoing climate and land use changes. Experimental warming caused a 26-36% decrease in species richness, a response that was generally dampened by experimental grazing. Higher species losses occurred at the drier sites where N was less available. Moreover, we observed an indirect effect of climate change on species richness as mediated by plant-plant interactions. Heat stress and warming-induced litter accumulation are potential explanations for the species' responses to experimental warming. This is the first reported experimental evidence that climate warming could cause dramatic declines in plant species diversity in high elevation ecosystems over short time frames and supports model predictions of species losses with anthropogenic climate change.

Analysis of diets of upland buzzards using stable carbon and nitrogen isotopes

We measured the stable carbon and nitrogen isotope ratios for muscles of the upland buzzards (Buteo hemilasius) and their potential food sources, plateau pikas (Ochotona curzoniae), Qinghai voles (Lasiopodomys fuscus), plateau zokors (Myospalax fontanierii), and several passerine bird species at the alpine meadow in Maduo county, Guoluo prefecture of Qinghai province, People's Republic of China, to provide diet information of upland buzzards, highlighting different diet composition of upland buzzards exposed to different locations. The results demonstrated that stable carbon isotope ratios of upland buzzards, passerine birds, plateau pikas, plateau zokors, and Qinghai voles were -24.42 +/- 0.25parts per thousand, -22.89 +/- 1.48parts per thousand, -25.30 +/- 1.47parts per thousand, -25.78 +/- 0.22parts per thousand, and -25.41 +/- 0.01parts per thousand, respectively, and stable nitrogen isotope ratios were 7.89 +/- 0.38parts per thousand, 8.37 +/- 2.05parts per thousand, 5.83 +/- 1.10parts per thousand, 5.23 +/- 0.34parts per thousand, and 8.86 +/- 0.06parts per thousand, respectively. Fractionation of stable carbon and nitrogen isotope ratios between upland buzzards and their food were 1.03parts per thousand and 2.11parts per thousand, respectively. Based on mass balance principle of stable isotopes and the Euclidean distance mixing model, upland buzzards depended mainly on plateau pikas as food (74.56%). Plateau zokors, Qinghai voles, and passerine birds only contributed a small proportion (25.44%) to diets of upland buzzards. The results were closely accordant with analyses of stomach contents and food pellets, which firmly supported the feasibility of using stable carbon and nitrogen isotope ratios to investigate diet information of upland buzzards. Another study based on stable carbon isotopes showed that upland buzzards living in the Haibei prefecture (another prefecture located in the southeast Qinghai province) mainly preyed on passerine birds (64.96% or more) as food supply. We were alarmed by the preliminary results that widespread poisoning activities of small mammals could reshape the food composition of upland buzzards, influencing the stability and sustainability of the alpine meadow. Bio-control on rodent pests should be carried out rather than the chemical measures.

Methane emissions from different vegetation zones in a Qinghai-Tibetan Plateau wetland

We measured methane (CH4) emissions in the Luanhaizi wetland, a typical alpine wetland on the Qinghai-Tibetan Plateau, China, during the plant growth season (early July to mid-September) in 2002. Our aim was to quantify the spatial and temporal variation of CH4 flux and to elucidate key factors in this variation. Static chamber measurements of CH4 flux were made in four vegetation zones along a gradient of water depth. There were three emergent-plant zones (Hippuris-dominated; Scirpus-dominated; and Carex-dominated) and one submerged-plant zone (Potamogeton-dominated). The smallest CH4 flux (seasonal mean = 33.1 mg CH4 m(-2) d(-1)) was, observed in the Potamogeton-dominated zone, which occupied about 74% of the total area of the wetland. The greatest CH4 flux (seasonal mean = 214 mg CH4 m(-2) d(-1)) was observed in the Hippuris-dominated zone, in the second-deepest water area. CH4 flux from three zones (excluding the Carex-dominated zone) showed a marked diurnal change and decreased dramatically under dark conditions. Light intensity had a major influence on the temporal variation in CH4 flux, at least in three of the zones. Methane fluxes from all zones increased during the growing season with increasing aboveground biomass. CH4 flux from the Scirpus-dominated zone was significantly lower than in the other emergent-plant zones despite the large biomass, because the root and rhizome intake ports for CH4 transport in the dominant species were distributed in shallower and more oxidative soil than occupied in the other zones. Spatial and temporal variation in CH4 flux from the alpine wetland was determined by the vegetation zone. Among the dominant species in each zone, there were variations in the density and biomass of shoots, gas-transport system, and root-rhizome architecture. The CH4 flux from a typical alpine wetland on the Qinghai-Tibetan Plateau was as high as those of other boreal and alpine wetlands. (C) 2004 Elsevier Ltd. All rights reserved.

Photosynthesis of Saussurea superba and Gentiana straminea is not reduced after long-term enhancement of UV-B radiation

Experiments were conducted in an alpine Kobresia humilis meadow near Haibei Alpine Meadow Ecosystem Research Station (37degrees29'-37degrees45'N, 101degrees12'-101degrees33'E; altitude 3200 m). Effects of enhanced ultraviolet-B (UV-B) radiation on photosynthesis of the alpine plants of Saussurea superba and Gentiana straminea were investigated. Both species were exposed to a UV-B-BE density at 15.80 kJ m(-2) per day, simulating nearly 14% ozone (O-3) reduction during the plant growing season. Neither photosynthetic CO2 uptake rate nor photosynthetic O-2 evolution rate were decreased after a long period of enhanced UV-B radiation treatment. On the contrary, there was a tendency to increase of both parameters in both species. The photosynthetic pigments were also increased, when expressed on a leaf area basis. UV-B absorbing compounds, detected by the absorbance values at 300 mm, had a tendency to increase in both species after enhanced UV-B radiation. After long-term exposure of plants to enhanced UV-B radiation, leaf morphology was also affected. Leaf thickness in both S. superba and G. straminea were increased significantly (P < 0.001). This supports our hypothesis that the increase of leaf thickness in both species after long-term exposure of enhanced UV-B radiation could compensate for the photodestruction of photosynthetic pigments when light passes through the leaf. Therefore, photosynthesis is not reduced in either species when expressed on leaf area basis. (C) 2003 Elsevier B.V. All rights reserved.

Molecular phylogeny and biogeography of the Qinghai-Tibet Plateau endemic Nannoglottis (Asteraceae)

All taxa endemic to the Qinghai-Tibet Plateau are hypothesized to have originated in situ or from immediately adjacent areas because of the relatively recent formation of the plateau since the Pliocene, followed by the large-scaled biota extinction and recession caused by the Quaternary ice sheet. However, identification of specific progenitors remains difficult for some endemics, especially some endemic genera. Nannoglottis, with about eight species endemic to this region, is one such genus. Past taxonomic treatments have suggested its relationships with four different tribes of Asteraceae. We intend to identify the closest relatives of Nannoglottis by evaluating the level of monophyly, tribal delimitation, and systematic position of the genus by using molecular data from ndhF gene, trnL-F, and ITS region sequences. We find that all sampled species of Nannoglottis are a well-defined monophyly. This supports all recent taxonomic treatments of Nannoglottis, in which all sampled species were placed in one broadly re-circumscribed genus. Nannoglottis is most closely related to the Astereae, but stands as an isolated genus as the first diverging lineage of the tribe, without close relatives. A tentative relationship was suggested for Nannoglottis and the next lineage of the tribe was based on the ITS topology, the "basal group," which consists of seven genera from the Southern Hemisphere. Such a relationship is supported by some commonly shared plesiomorphic morphological characters. Despite the very early divergence of Nannoglottis in the Astereae, the tribe must be regarded to have its origin in Southern Hemisphere rather than in Asia, because based on all morphological, molecular, biogeographical, and fossil data, the Asteraceae and its major lineages (tribes) are supposed to have originated in the former area. Long-distance dispersal using Southeast Asia as a steppingstone from Southern Hemisphere to the Qinghai-Tibet Plateau is the most likely explanation for this unusual biogeographic link of Nannoglottis. The 23-32-million-year divergence time between Nannoglottis and the other Astereae estimated by DNA sequences predated the formation of the plateau. This estimation is further favored by the fossil record of the Asteraceae and the possible time of origin of the Astereae. Nannoglottis seems to have reached the Qinghai-Tibet area in the Oligocene-Eocene and then re-diversified with the uplift of the plateau. The molecular infragenetic phylogeny of the genus identifies two distinct clades, which reject the earlier infrageneric classification based on the arrangement of the involucral bracts and the length of the ligules, but agree well with the habits and ecological preferences of its current species. The "alpine shrub" vs. "coniferous forest" divergence within Nannoglottis was estimated at about 3.4 million years ago when the plateau began its first large-scale uplifting and the coniferous vegetation began to appear. Most of the current species at the "coniferous forest" clade of the genus are estimated to have originated from 1.02 to 1.94 million years ago, when the second and third uprisings of the plateau occurred, the climate oscillated and the habitats were strongly changed. The assumed evolution, speciation diversity, and radiation of Nannoglottis based on molecular phylogeny and divergence times agree well with the known geological and paleobotanical histories of the Qinghai-Tibet Plateau. (C) 2002 Elsevier Science (USA). All rights reserved.