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Though aboveground biomass (AGB) has an important contribution to the global carbon cycle, the information about storage and climatic effects of AGB is scare in Three-River Source Region (TRSR) shrub ecosystems. This study investigated AGB storage and its climatic controls in the TRSR alpine shrub ecosystems using data collected from 23 sites on the Tibetan Plateau from 2011 to 2013. We estimated the AGB storage (both shrub layer biomass and grass layer biomass) in the alpine shrubs as 37.49 Tg, with an average density of 1447.31 g m<sup>-2</sup>. Biomass was primarily accumulated in the shrub layer, which accounted for 92% of AGB, while the grass layer accounted for only 8%. AGB significantly increased with the mean annual temperature (<i>P</i> < 0.05). The effects of the mean annual precipitation on AGB were not significant. These results suggest that temperature, rather than precipitation, has significantly effects on of aboveground vegetation growth in the TRSR alpine shrub ecosystems. The actual and potential increase in AGB density was different due to global warming varies among different regions of the TRSR. We conclude that long-term monitoring of dynamic changes is necessary to improve the accuracy estimations of potential AGB carbon sequestration across the TRSR alpine shrub ecosystems.

This study presents an efficient strategy based on liquid-liquid extraction, high-speed counter-current chromatography, and preparative HPLC for the rapid enrichment, separation, and purification of four anthraquinones from Rheum tanguticum. A new solvent system composed of petroleum ether/ethyl acetate/water (4:2:1, v/v/v) was developed for the liquid-liquid extraction of the crude extract from R. tanguticum. As a result, emodin, aloe-emodin, physcion, and chrysophanol were greatly enriched in the organic layer. In addition, an efficient method was successfully established to separate and purify the above anthraquinones by high-speed counter-current chromatography and preparative HPLC. This study supplies a new alternative method for the rapid enrichment, separation, and purification of emodin, aloe-emodin, physcione, and chrysophanol.

In plants with infrequent pollinator services, the benefits of reproductive assurance could be eroded by severe ovule discounting and inbreeding depression (ID). However, it remains unclear how selfing evolves under complete pollinator failure and strong ID. We examined the mating system and ID under netting and robbing conditions in <i>Comastoma pulmonarium</i> (Turcz.) Toyok. (Gentianaceae), an alpine annual experiencing a high ratio of nectar robbery on the Qinghai-Tibet Plateau. <i>Comastoma pulmonarium</i> produced seeds via selfing at the study site. No pollinator was observed and thus the nectar was consumed exclusively by robbers. Inbreeding depression occurred in the life stages of seed mass and germination, and the cumulative ID was much higher than 0.5 under netting and robbing conditions. Overall, in comparison with netting, the magnitude of ID under robbing conditions tended to decrease. Our results suggested that selfing could assure reproduction for plants under complete pollinator failure and strong ID, supporting the “better than nothing” role of selfing and providing one of the few cases of the evolution of selfing under strong ID.

High throughput sequencing technology is also called Next Generation Sequencing (NGS), which can sequence hundreds and thousands sequences in different samples at the same time. In the present study, the culture-independent high throughput sequencing technology was applied to sequence the fungi metagenomic DNA of the fungal internal transcribed spacer 1(ITS 1) in the root of Sinopodophyllum hexandrum. Sequencing data suggested that after the quality control, 22 565 reads were remained. Cluster similarity analysis was done based on 97% sequence similarity, which obtained 517 OTUs for the three samples (LD1, LD2 and LD3). All the fungi which identified from all the reads of OTUs based on 0.8 classification thresholds using the software of RDP classifier were classified as 13 classes, 35 orders, 44 family, 55 genera. Among these genera, the genus of Tetracladium was the dominant genera in all samples(35.49%, 68.55% and 12.96%).The Shannon's diversity indices and the Simpson indices of the endophytic fungi in the samples ranged from 1.75-2.92, 0.11-0.32, respectively.This is the first time for applying high through put sequencing technol-ogyto analyze the community composition and diversity of endophytic fungi in the medicinal plant, and the results showed that there were hyper diver sity and high community composition complexity of endophytic fungi in the root of S. hexandrum. It is also proved that the high through put sequencing technology has great advantage for analyzing ecommunity composition and diversity of endophtye in the plant.

A simple and sensitive high-performance liquid chromatographic (HPLC) method with fluorescence detection and mass spectrometric identification has been developed for analysis of 30 long-chain and short-chain free fatty acids (FFAs). The fatty acids were derivatized to their esters with 1-[2-(<i>p</i>-toluenesulfonate)ethyl]-2-phenylimidazole-[4,5-<i>f</i>]-9,10-phenanthrene (TSPP) in <i>N</i>,<i>N</i>-dimethylformamide (DMF) at 90 °C with anhydrous K<sub>2</sub>CO<sub>3</sub> as catalyst. A mixture of C<sub>1</sub>-C<sub>30</sub> fatty acids was completely separated within 60 min by gradient elution on a reversed-phase C<sub>8</sub> column. Qualitative identification of the acids was performed by atmospheric-pressure chemical ionization mass spectrometry (APCI-MS) in positive-ion mode. The fluorescence excitation and emission wavelengths were 260 and 380 nm, respectively. Quantitative determination of the 30 acids in two Tibetan medicines <i>Gentiana straminea</i> and <i>G. dahurica</i> was performed. The results indicated that the medicines contained many FFAs. Linear correlation coefficients for the FFA derivatives were >0.9991. Relative standard deviations (RSDs, <i>n</i> = 6) for the fatty acid derivatives were <3%. Detection limits (at a signal-to-noise ratio of 3:1) were 3.1-38 fmol. When the fatty acid derivatives were determined in the two real samples results were satisfactory and the sensitivity and reproducibility of the method were good.

Although large amounts of soil organic carbon (SOC) stored in the shrublands, information about SOC storage was little on the Tibetan Plateau. This study aims to evaluate the spatial patterns and storage of SOC in the shrublands and the relationships of climatic variables and soil pH on the Tibetan Plateau.<br>We used 177 profiles of soil samples obtained from 59 shrubland sites on the northeast Tibetan Plateau from 2011 to 2013. Ordinary least squares regressions, curve estimation, and multiple linear regressions were used to evaluate controlling factors on SOC stock. Kriging interpolation was used to upscale sit-level measurements to the whole study area.<br>We found that SOC storage in the northeast Tibetan shrublands was 1.36 Pg C in the top 1 m with an average SOC stock of 12.38 kg m<sup>−2</sup>. SOC stock decreased from east to west and south to north but generally increased significantly with the mean annual temperature (MAT) and the mean annual precipitation (MAP), and tended to decrease with soil pH. Although similar relationships were also observed in alpine shrublands, the trends among SOC stock, MAP, and MAT were not observed in desert shrublands. Our results indicate that a reduction in soil pH accelerates the C sequestration potential. Furthermore, global warming contributed to C sequestration in alpine shrublands, specifically, SOC stock increased 8.44 kg m<sup>−2</sup> with an increased unit of MAT in alpine shrublands just considering temperature effects. Meanwhile, the C sequestration was different among different regions due to the uneven increases in precipitation. However, in desert shrublands, MAP and MAT did not significantly affect SOC stock.<br>The results indicate that though a reduction in soil pH could contribute to C sequestration, MAT and MAP have different effects on SOC stock in different Tibetan Plateau shrublands. Increased MAT and MAP were 0.05 °C and 1.67 mm every year on the Tibetan Plateau, which will increase C sequestration in alpine shrublands, but might have limited impacts on desert shrublands, which help us comprehend soil C cycling in the global climate change scenario.

Background Rheum tanguticum Maxim. ex Balf is one of the plants generically known as rhubarb, a culinary vegetable that has long been used as a herbal remedy both in China and Europe. Increasing demand for rhubarb has triggered the overexploitation of Rh. tanguticum. Cultivation is therefore necessary for quality control and protection of wild resources. Nitrogen fertilizer plays an important role in cultivation. This study aimed to explore how nitrogen fertilizer affects the growth and quality of rhubarb on the Qinghai-Tibetan plateau. Results Nitrogen fertilizer promoted growth but had no significant influence on the active compounds of Rh. tanguticum. Generally, the N2 (150 kg ha(-1)) and N3 (225 kg ha(-1)) levels showed the most improved growth indexes, with no significant differences between them. The growth index and the amounts of eight of the nine studied active compounds in Rh. tanguticum increased from each year to the next and differed among growth stages. The contents of the active compounds were higher at the green stage and lower at the growth stage, which was opposite to the seasonal trends in root dry matter ratio. Gallic acid levels decreased with the growth of the plant. Conclusion The N2 level (150 kg ha(-1)) was the recommended nitrogen fertilizer level in this study. It was revealed that seasonal changes rather than nitrogen fertilizer influenced active compounds in the root of Rh. tanguticum. (c) 2018 Society of Chemical Industry

Concentrations of 20 free amino aicds (FAAs) in a famous Tibetan medicine Gentiana dahurica was first investigated using 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) as the pre-column fluorescence derivatization reagent by reversed-phase high performance liquid chromatography (RP-LC). 20 amino acid derivatives (AAD) were separated on a Hypersil BDS C<sub>18</sub> column with a good baseline resolution within 65 min. Identification of 20 AAD was by online post-column mass spectrometry with an electrospray ionization (ESI) source. The validation of the method was examined by linearity, repeatability, and detection limits. Most linear correlation coefficients for AAD were &gt;0.9990, and detection limits (at signal-to-noise of 3:1) were 6.5-178.2 fmol. There were 18 FAAs found in G. dahurica, of which seven FAAs were necessary to the people's health and related to the treatment of liver and gall disease. Variation of concentrations of the 20 FAAs showed geographical distribution difference among populations. Meanwhile a stable genetic diversity of FAAs composition of G. dahurica was also revealed at the species level. Results of the present study proved that the established method was rapid and reproducible for further separation and determination of FAAs in more medicinal plants.

Anthraquinone glycosides, such as chrysophanol 1-O-β-d-glucoside, chrysophanol 8-O-β-d-glucoside, and physion 8-O-β-d-glucoside, are the accepted important active components of Rheum tanguticum Maxim. ex Balf. due to their pharmacological properties: antifungal, antimicrobial, cytotoxic, and antioxidant activities. However, an effective method for the separation of the above-mentioned anthraquinone glycosides from this herb is not currently available. Especially, greater difficulty existed in the separation of the two isomers chrysophanol 1-O-β-d-glucoside and chrysophanol 8-O-β-d-glucoside. This study demonstrated an efficient strategy based on preparative high-performance liquid chromatography and high-speed countercurrent chromatography for the separation of the above-mentioned anthraquinone glycosides from Rheum tanguticum Maxim.ex Balf.

Emissions of nitrous oxide (N₂O) contribute to global warming and stratospheric ozone depletion. Anthropogenic N₂O emissions predominately result from the addition of synthetic nitrogen (N) fertilizers to terrestrial ecosystems. Usually, an exponential increase in N₂O emissions occurs as N addition rates increase to exceed plant demands. However, most evidence to date is from temperate areas, with little information available for alpine ecosystems. Here we examined the changes in N₂O flux under eight N addition levels and the mechanisms regulating these changes in a Tibetan alpine steppe. Our results showed that N₂O emission rate increased linearly with increasing N additions. Even when soil N availability exceeded plant N uptake, no sharp N₂O emissions were observed. The likely explanation was that decreased soil temperature limited the growth of nitrification-related microorganisms, mainly ammonia-oxidizing archaea, which further attenuated the positive response of N₂O emissions to excess N supply. These findings suggest that the N-induced changes in soil temperature regulate the growth of nitrifying microorganisms and the subsequent N₂O fluxes in this alpine steppe, and the exponential N₂O emission-N rate relationship observed in warm regions may not be simply extrapolated to alpine ecosystems.<br>N₂O emission exhibited a linear, rather than an exponential, response to increasing N additionsN₂O flux was explained by the changes in AOA along this N addition gradientDecreased soil temperature limited the growth of AOA, weakening the positive response of N₂O flux to excess N supplies

Alpine shrubland ecosystems in the Three Rivers Source Region (TRSR) store substantial soil organic carbon (SOC), but the storage, patterns and control of SOC in those ecosystems have rarely been investigated. In this study, using data from 66 soil profiles surveyed from 22 sites between 2011 and 2013, we estimated the storage and patterns of SOC, and their relationships with climatic factors, elevation, ground cover and slope. Our results showed that SOC storage in the top 100 cm across the TRSR shrubland was 0.68 ± 0.38 Pg C, with an average SOC density (soil carbon storage per area) of 26.21 ± 14.58 kg m−2. Spatially, SOC density increased with longitude and latitude. Vertically, SOC in the topsoil at 30 cm and 50 cm accounted for 56% and 75%, respectively, of the total at 100 cm. SOC density showed a decreasing trend with increasing elevation, but it was greater in regions of higher ground cover. The density had no relationship with either mean annual precipitation or slope. Increasing mean annual temperature had positive effects on SOC density, which is inconsistent with the global trend. With increasing soil depth, however, the effects of temperature on SOC density were not significant. Therefore, in a global warming scenario, increasing temperature gives shrubland considerable C sink potential on the topsoil, and the regions of C sequestration differ as a result of uneven increases in temperature. Hence, further monitoring of dynamic changes is necessary to provide a more accurate assessment of potential C sequestration in TRSR shrubland.<br>• Storage and patterns of SOC were investigated in the TRSR shrubland. • SOC has no relationship with both precipitation and slope. • SOC was decreasing with elevation, but larger with both ground cover and temperature.