Gentiana Straminea Research Articles

Spatial Distribution Characteristics of Plant Community and the Identification of Driving Factors in Riparian Zone of the Three-River Headwaters Region, China

The unique geographical and climatic conditions in the Three-River Headwaters Region gave birth to distinctive plant species and vegetation types. To reveal the spatial distribution of plant communities and soil habitats along the riparian zone of the Sanjiangyuan Region and their influencing mechanisms, 14 survey plots were set up （ten from the Yangtze River source, two from the Lancang River source, and two from the Yellow River source）, and the effects of soil nutrient characteristics （especially soil phosphorus morphology）, climate factors, and river topography on plant community characteristics were quantitatively analyzed. The results showed that the plant community composition in the riparian zone of the source of the three rivers was dominated by perennial herbs （72.2%）, followed by annual herbs （20.4%） and shrubs （7.4%）. The dominant plants were Stipa purpurea, Polygonum orbiculatum, Carex parvula, Potentilla anserina, and Gentiana straminea. The average plant coverage, Shannon-Wiener index, and Pielou index were （64.4% ±23.6%）, （1.31 ±0.42）, and （0.84 ±0.08）, respectively. The plant community diversity index was the highest in the Yangtze River source, followed by that in the Lancang River source, and the lowest in the Yellow River source. The soil pH of the riparian zone of the Yangtze River source was significantly higher than that of the Lancang River source, whereas the mean contents of organic matter, total nitrogen, and Fe-Al combined phosphorus were significantly lower than those of the Lancang River source. The calcium and magnesium-combined phosphorus was the main form of phosphorus in riparian soil （63.89%）. Temperature, soil organic phosphorus content, and pH had significant effects on plant composition in the riparian zone of the Three-River Headwaters Region, whereas soil calcium and magnesium-combined phosphorus content had significant effects on plant community diversities. These results may deepen the scientific understanding of the evolution trend and genetic mechanism of plant communities in the riparian zone of the Three-River Headwaters Region.

Insights from the first chromosome-level genome assembly of the alpine gentian Gentiana straminea Maxim.

Gentiana straminea Maxim. is a perennial herb and mainly distributed in the Qinghai-Tibetan Plateau. To adapt to the extreme environment, it has developed particular morphological, physiological, and genetic structures. Also, rich in iridoids, it is one of the original plants of traditional Chinese herb 'Qinjiao'. Herein, we present its first chromosome-level genome sequence assembly and compare it with the genomes of other Gentiana species to facilitate the analysis of genomic characteristics. The assembled genome size of G. straminea was 1.25 Gb, with a contig N50 of 7.5Mb. A total of 96.08% of the genome sequences was anchored on 13 pseudochromosomes, with a scaffold N50 of 92.70Mb. A total of 54,310 protein-coding genes were predicted, 80.25% of which were functionally annotated. Comparative genomic analyses indicated that G. straminea experienced two whole-genome duplication events after the γ whole-genome triplication with other eudicots, and it diverged from other Gentiana species at ~3.2 Mya. A total of 142 enzyme-coding genes related to iridoid biosynthesis were identified in its genome. Additionally, we identified differences in the number and expression patterns of iridoid biosynthetic pathway genes in G. straminea compared with two other Gentiana species by integrating whole-genome sequence and transcriptomic analyses.

Development and characterization of novel EST-SSR markers for Gentiana straminea Maxim., a traditional Tibetan herb in China and cross-amplification in related species

Abstract Gentiana straminea Maxim. (Gentianaceae) is an important traditional Tibetan herb that is mainly distributed on the Qinghai-Tibetan Plateau. Despite its agricultural and pharmacological importance, there remains a paucity of microsatellite markers, particularly expressed sequence tag-simple sequence repeat (EST-SSR) markers, available for this local endemic species. In this study, based on previous Illumina transcriptome data of G. straminea, a total of 96 EST-SSR markers were initially designed and tested. Thirty-two of 96 loci (33.33%) were successfully amplified and verified for validation. Among them, 10 were polymorphic and had clear bands. The polymorphism information content values were 0.09–0.799, the number of alleles per locus ranged from 3 to 14, and the levels of observed and expected heterozygosity were 0.078–0.722 and 0.238–0.884, respectively, which suggested a high level of information. Moreover, cross-amplification was successful for 10 loci in two other related species, Gentiana macrophylla Pallas and Gentiana dahurica Fischer. These EST-SSR markers provide a valuable tool for investigating the genetic diversity related to quantitative traits and population genetic studies on G. straminea and related species in sect. Cruciata Gaudin.

Quantitative comparison and evaluation between aerial and underground parts of Gentiana straminea through simultaneous determination of five major compounds by RP-HPLC

Herbal Gentiana straminea Maxim. (Family Gentianaceae), “Ma Hua Jiao” in Chinese, is a commonly used Chinese medicine. Secoiridoids and flavonoids have been identified as the major active components of herbal medicines used in the treatment of hepatitis, rheumatism and many other diseases. It is the overharvesting of the roots of this plant for medicinal purposes that has led to a drastic decline in its population. In the present study, the above and below ground parts of Gentian Bitter Glycine were quantitatively compared and evaluated for the determination of the major active constituents. Five major compounds, loganic acid, swertiamarin, gentiopicroside, sweorside and isoorientin, were extracted by solvent extraction technique and analyzed by Reversed-phase High Performance Liquid Chromatography (RP-HPLC). By analysing the principal components and calculating the composite scores, the results show that the aboveground component in different areas ranked higher compared to the underground component, with the former being able to substitute to some extent for the latter's underground component. Finally, based on hierarchical cluster analysis, we identified the ideal natural growing region for aerial parts of G. straminea distributed on the Qinghai-Tibetan Plateau. The significance of this work is that we can balance the demand for herbs with environmental preservation by selectively picking the aerial parts, which can regrow next year, instead of removing the whole plant. It protects the fragile ecological environment of the Tibetan Plateau and is important for sustainable development.

Subplenones A-J: Dimeric Xanthones with Antibacterial Activity from the Endophytic Fungus Subplenodomus sp. CPCC 401465.

Subplenones A-J (1-10), 10 new xanthone dimers, have been isolated and characterized from the endophytic fungus Subplenodomus sp. CPCC 401465, which resides within the Chinese medicinal plant Gentiana straminea. The isolation process was guided by antibacterial assays and molecular-networking-based analyses. The chemical structures of these compounds were elucidated through the interpretation of nuclear magnetic resonance (NMR) and high-resolution electrospray ionization mass spectrometry (HRESIMS) data. Furthermore, the relative configuration of the compounds was determined using NMR and single-crystal X-ray diffraction analyses, and the absolute configuration was established using electronic circular dichroism calculations. All of the isolated compounds exhibited significant inhibitory activity against Gram-positive bacteria. Notably, compounds 1, 5, and 7 displayed remarkable inhibitory activity against methicillin-resistant Staphylococcus aureus (MRSA) ATCC 700698, with a minimum inhibitory concentration (MIC) of 0.25 μg/mL, and against vancomycin-resistant Enterococcus faecium (VRE) ATCC 700221, with MIC values ranging from 0.5 to 1.0 μg/mL.

An integrated strategy for quality control of the multi-origins herb medicine of Gentianae Macrophyllae Radix based on UPLC-Orbitrap-MS/MS and HPLC-DAD.

Gentianae Macrophyllae Radix, the dried root of Gentiana macrophylla Pall., Gentiana crassicaulis Duthie ex Burk., Gentiana straminea Maxim., or Gentiana dahurica Fisch., is a traditional Chinese medicine with multi-origins and some adulterants. Liquid chromatography coupled to electrostatic orbitrap high-resolution mass spectrometry (LC-Orbitrap-MS) was used to search the different components of Gentianae Macrophyllae Radix of the four species. High-performance liquid chromatography (HPLC) combined with fingerprint analysis, principal components analysis (PCA), and partial least-squares discrimination analysis (PLS-DA) was also utilized to distinguish them and their adulterants based on the critical components identified by LC-MS. A single standard to determine the multi-components (SSDMC) method was established for the determination of the critical markers. A total of 93 compounds were identified from Gentianae Macrophyllae Radix, including 58 common ones. Their HPLC fingerprints show a significant difference with the adulterants. In addition, PCA and PLS-DA could make a distinction among the four species. Loganic acid, 6'-O-β-d-glucosylgentiopicroside, swertiamarine, gentiopicroside, and sweroside were identified as the critical markers and then quantified by the SSDMC method. The developed strategy is powerful for the quality control and authentication of Gentianae Macrophyllae Radix.

Ethanol Extract of Gentiana straminea Maxim Displays Anti-Hypoxia Effects by Regulating Antioxidant Enzymes and Energy Metabolism

Ethanol Extract of Gentiana straminea Maxim Displays Anti-Hypoxia Effects by Regulating Antioxidant Enzymes and Energy Metabolism

Preliminary verification of the anti-hypoxia mechanism of Gentiana straminea maxim based on UPLC-triple TOF MS/MS and network pharmacology

BackgroundAnoxia is characterized by changes in the morphology, metabolism, and function of tissues and organs due to insufficient oxygen supply or oxygen dysfunction. Gentiana straminea Maxim (G.s Maxim) is a traditional Tibetan medicine. Our previous work found that G.s Maxim mediates resistance to hypoxia, and we found that the ethyl acetate extract had the best effect. Nevertheless, the primary anti-hypoxia components and mechanisms of action remain unclear.MethodsCompounds from the ethyl acetate extraction of G.s Maxim were identified using UPLC-Triple TOF MS/MS. Then Traditional Chinese Medicine Systematic Pharmacology Database was used to filtrate them. Network pharmacology was used to forecast the mechanisms of these compounds. Male specific pathogen-free Sprague Dawley rats were randomly divided into six groups: (1) Control; (2) Model; (3) 228 mg/kg body weight Rhodiola capsules; (4) 6.66 g/kg body weight the G.s Maxim’s ethyl acetate extraction; (5) 3.33 g/kg body weight the G.s Maxim’s ethyl acetate extraction; (6) 1.67 g/kg body weight the G.s Maxim’s ethyl acetate extraction. After administering intragastric ally for 15 consecutive days, an anoxia model was established using a hypobaric oxygen chamber (7000 m, 24 h). Then Histology, enzyme-linked immunosorbent assays, and western blots were performed to determine these compounds’ anti-hypoxic effects and mechanisms. Finally, we performed a molecular docking test to test these compounds using Auto Dock.ResultsEight drug-like compounds in G.s Maxim were confirmed using UPLC-Triple TOF MS/MS and Lipinski’s rule. The tumor necrosis factor (TNF) signaling pathway, the hypoxia-inducible factor 1 (HIF-1) signaling pathway, and the nuclear factor kappa-B (NF-κB) signaling pathway was signaling pathways that G.s Maxim mediated anti-anoxia effects. The critical targets were TNF, Jun proto-oncogene (JUN), tumor protein p53 (TP53), and threonine kinase 1 (AKT1). Animal experiments showed that the ethyl acetate extraction of G.s Maxim ameliorated the hypoxia-induced damage of hippocampal nerve cells in the CA1 region and reversed elevated serum expression of TNF-α, IL-6, and NF-κ B in hypoxic rats. The compound also reduced the expression of HIF-1α and p65 and increased the Bcl-2/Bax ratio in brain tissue. These findings suggest that G.s Maxim significantly protects against brain tissue damage in hypoxic rats by suppressing hypoxia-induced apoptosis and inflammation. Ccorosolic acid, oleanolic acid, and ursolic acid had a strong affinity with core targets.ConclusionsThe ethyl acetate extraction of G.s Maxim mediates anti-hypoxic effects, possibly related to inhibiting apoptosis and inflammatory responses through the HIF-1/NF-κB pathway. The primary active components might be corosolic, oleanolic, and ursolic acids.

Age Structure and Self-Thinning of Four Poisonous Forbs in Alpine Meadows on the Qinghai-Tibetan Plateau

Age structure and self-thinning indicate the development of plant species and their adaptability to the environment, and the proportion of poisonous plants could reflect the status of grasslands degradation. Little is known about age structure and self-thinning of poisonous dicotyledonous forbs in the alpine meadows on the Tibetan Plateau. We selected four poisonous dicotyledonous species, namely Anaphalis lacteal, Stellera chamaejasme, Pedicularis kansuensis, and Gentiana straminea, and determined their age structures by using number of branches, crown diameters, and plant heights and their self-thinning by using dry weights and different components (branches, leaves, and inflorescences). The populations of the four species were pyramid shaped with a rich seedling and juvenile base, which demonstrated a strong ability to increase in number rapidly and adapt to the environment. The Godron index indicated that the stability of age structures of the four species were A. lacteal > S. chamaejasme > G. straminea > P. kansuensis. Significant self-thinning was exhibited by branches of A. lacteal, branches and leaves of S. chamaejasme, and inflorescences of P. kansuensis. The self-thinning exponents from the allometric relationships between biomass and density differed at the level of branches, leaves, and inflorescences in the four poisonous forbs. We concluded that the populations of the four poisonous species were stable and able to increase in number rapidly on the Qinghai-Tibetan Plateau.

Effective Parts of Gentiana straminea Maxim Attenuates Hypoxia-Induced Oxidative Stress and Apoptosis.

Hypoxia occurs in physiological situations and several pathological situations, inducing oxidative stress. G straminea Maxim (G.s Maxim) is a traditional Tibetan medicine that exerts several biological effects. This study focused on the protective effects of G.s Maxim in hypoxia-induced oxidative stress and apoptosis. We found that G.s Maxim significantly increased survival and reduced oxidative stress in hypoxic mice. Various extraction parts of G.s Maxim showed antioxidant activity and significantly improved survival in hypoxia-injured PC12 cells. G.s Maxim reduced hypoxia-induced cell apoptosis and leakage of lactate dehydrogenase. Hypoxic cells had increased malondialdehyde levels but reduced superoxide dismutase activity and G.s Maxim reversed these effects. Moreover, G.s Maxim suppressed hypoxia-induced apoptosis by inducing protein expression of B cell leukemia/lymphoma-2 and reducing the expression of hypoxia-inducible factor-1α, Bcl-2-associated X, and nuclear factor-k-gene binding. These findings suggest that G.s Maxim attenuates hypoxia-induced injury associated with oxidative stress and apoptosis.

Gentiana straminea Maxim. polysaccharide decolored via high-throughput graphene-based column and its anti-inflammatory activity

Gentiana straminea Maxim. exhibits various biological activities. However, the purification and functions of polysaccharides in Gentiana straminea Maxim. have never been reported. Herein, by proposing a flexible 3D graphene-based decoloration column (3DD column), Gentiana straminea Maxim. polysaccharide (GMP) was high-throughput obtained and its anti-inflammatory activity was investigated. Benefiting from the large macroporous network of 3D NH2-graphene oxide hydrogel with selective adsorption towards pigments, the 3DD column exhibits high decoloration ratio (96.41%). In addition, the 3DD column provides superior practical functionality as compared to the traditional approaches, which are time-consuming and need toxic solvents, and exhibiting widespread-application for the purification of polysaccharide from other common plant species. More importantly, the decolored GMP as a natural product has promising anti-inflammatory activity on RAW264.7 cells without negative impact on cell viability. Overall, this work reveals a new functional polysaccharides and provides a flexible approach for polysaccharide decoloration, exhibiting a promising prospect for natural polysaccharides in practical application of pharmaceutical.

Gentiana straminea supplementation improves feed intake, nitrogen and energy utilization, and methane emission of Simmental calves in northwest China.

ObjectiveNative plants can be used as additives to replace antibiotics to improve ruminant feed utilization and animal health. An experiment was conducted to evaluate the effects of Gentiana straminea (GS) on nutrient digestibility, methane emissions, and energy metabolism of Simmental calves.MethodsThirty-two (5-week-old) male Simmental clves, with initial body weight (BW) of 155±12 kg were fed the same basal diet of concentrates (26%), alfalfa hay (37%), and oat hay (37%) and were randomly separated into four treatment groups according to the amount of GS that was added to their basal diet. The four different groups received different amounts of GS as a supplement to their basal diet during whole experiment: (0 GS) 0 mg/kg BW, the control; (100 GS) 100 mg/kg BW; (200 GS) 200 mg/kg BW; and (300 GS) 300 mg/kg BW.ResultsFor calves in the 200 GS and 300 GS treatment groups, there was a significant increase in dry matter (DM) intake (p<0.01), average daily gain (ADG) (p<0.05), organic matter intake (p<0.05), DM digestibility (p<0.05), neutral detergent fibre (NDF) digestibility (p<0.05), and acid detergent fibre (ADF) digestibility (p<0.05). Dietary GS supplementation result in quadratic increases of DM intake (p<0.01), ADG (p<0.05), NDF intake (p<0.05), and ADF intake (p<0.05). Supplementing the basal diet with GS significantly increased nitrogen (N) retention (p<0.001) and the ratio of retention N to N intake (p<0.001). Supplementing the basal diet with GS significantly decreased methane (CH4) emissions (p<0.01), CH4/BW0.75 (p<0.05) and CH4 energy (CH4-E) (p<0.05). Dietary GS supplementation result in quadratic increases of CH4 (p<0.01) and CH4/DM intake (p<0.01). Compared with 0 GS, GS-supplemented diets significantly improved their gross energy intake (p<0.05). The metabolizable energy and digestive energy intake were significantly greater for calves in the 100 GS and 200 GS calves than for 0 GS calves (p<0.05).ConclusionFrom this study, we conclude that supplementing calf diets with GS could improve utilization of feed, energy, and N, and may reduce CH4 emissions without having any negative effects on animal health.

Quantitative analysis of the profiles of twelve major compounds in Gentiana straminea Maxim. Roots by LC-MS/MS in an extensive germplasm survey in the Qinghai-Tibetan plateau

Ethnobotanical relevanceGentiana straminea Maxim. is a well-known Tibetan traditional herb, which has been used to treat rheumatic arthritis, iceteric hepatitis, and other diseases for thousands years. However, there is still lack of comprehensive active constituents profiling of this species throughout the Qinghai-Tibet Plateau (QTP). Aim of studyThis study was designed to provide a comprehensive quality map of G.straminea germplasm based on twelve active constituents (loganic acid, gentiopicroside, swertiamarin, sweroside, 6-O-β-D-glucosylgentiopicroside, oleanic acid, morroniside, trilobatin, isoorientin, isovite, Shanzhisidemethylester and quercetin) on the QTP. Materials and methodsG.straminea root samples collected throughout QTP in the flowering period were analyzed by the LC-MS/MS. Statistics analysis methods PCA, clustering and ecological regions analysis for G.straminea constituents differentiation was demonstrated. ResultsThe active constituents varied greatly across the QTP; the majority of constituents were secoiridoid derivatives, with gentiopicroside being the most abundant compound. Most constituents were significantly affected by the latitudes and altitudes other than longitudes. PCA and hierarchical clustering analysis showed that all samples could be separated into six distinct groups, and 15 populations showed the highest constituent abundances. Further, geographical region analysis showed that the highest quality populations mainly located near the source region of Yellow River, especially in the Qinghai and Sichuan areas. Additionally, correlation analysis showed that there were relationships among genetiopicroside, loganic acid, and other compounds, which might be related to the enzymatic pathways involved in the metabolism of these constituents. Conclusion: LC-MS/MS method allowed separation of quality profiling of G.straminea on the QTP, 15 populations showed the highest constituent abundances. In six geographical groups, the highest quality populations mainly located near the source region of Yellow River, especially in the Qinghai and Sichuan areas, which may be due to the climate caused by the westerlies and Indian Ocean monsoons in the QTP.

A novel method to rapidly evaluate the quality of the traditional Chinese medicine Gentiana straminea Maxim. using near infrared spectroscopy coupled with chemometrics

The test is to control quality of Gentiana straminea Maxim. by near infrared spectroscopy. Samples were collected throughout Qinghai-Tibet Plateau, in which the bioactive ingredients were gentiopicroside and loganic acid. Sample set was divided using concentration gradient method. Modeling optimization was processed with a designed orthogonal test L9 (33). Pretreatment of spectra was multiplicative scatter correction + second derivative, and partial least square was used to establish model. The model showed high R and low root mean square error. Moreover, residual predictive deviation was 3.14. The present quantitative model can fast and accurately determine the quality of G. straminea.

Integrated quality evaluation strategy for multi-species resourced herb medicine of Qinjiao by metabolomics analysis and genetic comparation

BackgroundQuality evaluation of multi-species resourced herb medicine (MSRHM) is a main problem for quality control of herb medicine. Current quality evaluation methodology lost consideration of species discrepancy. New quality evaluation strategy for MSRHM is in urgent need. Qinjiao, a representative MSRHM, originated from Gentiana macrophylla Pall., Gentiana straminea Maxim., Gentiana crassicaulis Duthie ex Burk. or Gentiana dahurica Fisch., has been used as an important herb medicine over 2000 years for expelling wind-dampness and relieving impediment pain. However, quality evaluation among species has never been revealed. The current work proposes an integrated quality evaluation strategy for MSRHM of Qinjiao, which may promote innovation of quality control of MSRHM.MethodsIn this work, 58 batches of Qinjiao covering 4 species were collected. Genetic comparative analysis based on ITS2 sequence was conducted. Metabolomics analysis based on TOF–MS and NMR spectrum were carried out. Compounds underlying species differences were identified and their discrepancies among species were investigated by ANOVA analysis and multivariate analysis.ResultsFour species of Qinjiao can be authenticated by ITS2 sequence comparation. Metabolomics analysis by TOF/MS and NMR revealed chemical discrepancies among species of Qinjiao. Maximum discrepancy was present between Gentiana crassicaulis Duthie ex Burk. and Gentiana dahurica Fisch. Chemical difference among species were tentative explored. For TOF–MS profiling, 28 constituents were tentative identified, 17 of which were further confirmed by standards. For 1H-NMR profiling, signals from 5 compounds were assigned. Contents discrepancies were investigated by ANOVA analysis. It seems that (seco)iridoids like loganic acid, gentiopicroside or swertiamarin were richer in specie of Gentiana crassicaulis Duthie ex Burk., while flavonoid (morroniside) and triterpenoids (roburic aicd, ursolic acid, oleanolic acid, β-sitosterone) were richer in specie of Gentiana dahurica Fisch. The current research demonstrates that metabolite profiling based on both UPLC/Q-TOF MS and 1H-NMR coupled with ITS2 sequence comparation can be a powerful tool for quality investigation of MSRHM of Qinjiao.ConclusionsA comprehensive quality evaluation strategy for MSRHM was proposed by integrating UPLC-Q-TOF–MS, NMR based metabolic analysis and ITS2 sequence genetic comparation. The proposed quality evaluation strategy shall promote innovation of quality control of traditional Chinese medicine.

Traceability of Geographical Origin in Gentiana straminea by UPLC-Q Exactive Mass and Multivariate Analyses.

The root of Gentiana straminea Maxim. (Gentianaceae), is officially listed as “Qin-Jiao” in the Chinese Pharmacopoeia for the treatment of rheumatic arthritis, icteric hepatitis, constipation, pain, and hypertension. To establish the geographical origin traceability in G. straminea, its chemical profiles were determined by a UPLC-Q exactive mass spectrometer, from which 43 compounds were identified by comparing retention times and mass spectrometry. Meanwhile, a pair of isomers (loganin and secologanol) was identified by mass spectrometry based on their fragmentation pathway. A total of 42 samples from difference habitats were determined by an UPLC-Q exactive mass spectrometer and the data were assayed with multivariate statistical analysis. Eight characteristic compounds were identified to determine the geographical origin of the herb. To estimate the key characteristic markers associated with pharmacological function, the inhibiting activities of nitric oxide (NO) production in lipopolysaccharide (LPS)-induced macrophages were examined. This finding is crucial in realizing the determination of botanical origin and evaluating the quality of G. straminea.

Food and habitat provisions jointly determine competitive and facilitative interactions among distantly related herbivores

Abstract Interactions between distantly related herbivores exert powerful influences on ecosystems, but most studies to date have only considered unidirectional effects. Few have simultaneously examined the mutual effects that vertebrate herbivores and insect herbivores have on one another. We conducted a set of manipulative experiments to evaluate the potential competition and facilitation between two pairs of distantly related herbivore taxa: an insect caterpillar (Gynaephora alpherakii) and two large vertebrate herbivores, yak (Bos grunniens) and Tibetan sheep (Ovis aries tibetica). We found that these large herbivores consistently increased the density of caterpillars likely by improving the habitat for caterpillars. The caterpillars, in turn, decreased yak but increased Tibetan sheep foraging time and intake bites by differently changing available food resources of the two large herbivores. Diet preferences of herbivores modified the habitat and food resources, thereby causing a diet‐mediated competition between yak and caterpillars, and facilitation between sheep and caterpillars. The vertebrate herbivores non‐feeding upon Lamiophlomis rotata and Gentiana straminea, the caterpillars preferred habitat, increased densities of the two plant species, thus favouring the caterpillars. In turn, the caterpillar preference for Kobresia pygmaea significantly reduced food resources for yak, while promoting food resources (multiple forbs) for sheep. Synthesis. Our study indicates that two different mechanisms (the changes in habitat and food availability) induced by herbivore foraging jointly determine competitive and facilitative interactions between distantly related herbivore species. We also suggest that examining the bi‐directional effects between herbivores offers a better understanding of competition and facilitation in terrestrial animal communities.

Effects of 8-Year Nitrogen and Phosphorus Treatments on the Ecophysiological Traits of Two Key Species on Tibetan Plateau.

Understanding how nitrogen (N) and/or phosphorus (P) addition affects plants carbon- and water- related ecophysiological characteristics is essential for predicting the global change impact on the alpine meadow ecosystem structure and function in carbon and water cycling. The Qinghai-Tibetan Plateau (QTP) with the largest alpine meadow in the world is regarded as the third pole in the earth and has been experiencing increased atmospheric N deposition. In this project, we focused on two key species (Elymus dahuricus and Gentiana straminea) of the alpine meadow on the Tibetan Plateau and investigated the variability of photosynthetic and stomatal responses to 8-year N and/or P treatments through field measurements and modeling. We measured photosynthesis- and gs-response curves to generate parameter estimates from individual leaves with two widely used stomatal models (the BWB model and MED model) for validation of growth and ecosystem models and to elucidate the physiological basis for observed differences in productivity and WUE. We assessed WUE by means of gas exchange measurements (WUEi) and stable carbon isotope composition (Δ13C) to get the intrinsic and integrated estimates of WUE of the two species. P and N+P treatments, but not N, improved the photosynthetic capacity (Anet and Vcmax) for both species. Stomatal functions including instaneous measurements of stomatal conductance, intrinsic water-use efficiency and stomatal slope parameters of the two widely used stomatal models were altered by the addition of P or N+P treatment, but the impact varied across years and species. The inconsistent responses across species suggest that an understanding of photosynthetic, stomatal functions and water-use should be evaluated on species separately. WUE estimated by Δ13C values had a positive relationship with Anet and gs and a negative relationship with WUEi. Our findings should be useful for understanding the underlying mechanisms of the response of alpine plants growth and alpine meadow ecosystem to global change.

An Intronless β-amyrin Synthase Gene is More Efficient in Oleanolic Acid Accumulation than its Paralog in Gentiana straminea.

Paralogous members of the oxidosqualene cyclase (OSC) family encode a diversity of enzymes that are important in triterpenoid biosynthesis. This report describes the isolation of the Gentiana straminea gene GsAS2 that encodes a β-amyrin synthase (βAS) enzyme. Unlike its previously isolated paralog GsAS1, GsAS2 lacks introns. Its predicted protein product was is a 759 residue polypeptide that shares high homology with other known β-amyrin synthases (βASs). Heterologously expressed GsAS2 generates more β-amyrin in yeast than does GsAS1. Constitutive over-expression of GsAS2 resulted in a 5.7 fold increase in oleanolic acid accumulation, while over-expression of GsAS1 led to a 3 fold increase. Additionally, RNAi-directed suppression of GsAS2 and GsAS1 in G. straminea decreased oleonolic acid levels by 65.9% and 21% respectively, indicating that GsAS2 plays a more important role than GsAS1 in oleanolic acid biosynthesis in G. straminea. We uses a docking model to explore the catalytic mechanism of GsAS1/2 and predicted that GsAS2, with its Y560, have higher efficiency than GsAS1 and mutated versions of GsAS2 in β-amyrin produce. When the key residue in GsAS2 was mutagenized, it produced about 41.29% and 71.15% less β-amyrin than native, while the key residue in GsAS1 was mutagenized to that in GsAS2, the mutant produced 38.02% more β-amyrin than native GsAS1.

Chloroplast genome structures in Gentiana (Gentianaceae), based on three medicinal alpine plants used in Tibetan herbal medicine.

The genus Gentiana is the largest in the Gentianaceae family with ca. 400 species. However, with most species growing on the Qinghai-Tibet plateau, the processes of adaptive evolution and speciation within the genus is not clear. Also, the genomic analyses could provide important information. So far, the complete chloroplast (cp) genome data of the genus are still deficient. As the second and third sequenced members within Gentianaceae, we report the construction of complete cp sequences of Gentiana robusta King ex Hook. f. and Gentiana crassicaulis Duthie ex Burk., and describe a comparative study of three Gentiana cp genomes, including the cp genome of Gentiana straminea Maxim. published previously. These cp genomes are highly conserved in gene size, gene content, and gene order and the rps16 pseudogene with exon2 missing was found common. Three repeat types and five SSR types were investigated, and the number and distribution are similar among the three genomes. Sixteen genome divergent hotspot regions were identified across these cp genomes that could provide potential molecular markers for further phylogenetic studies in Gentiana. The IR/SC boundary organizations in Gentianales cp genomes were compared and three different types of boundaries were observed. Six data partitions of cp genomes in Gentianales were used for phylogenetic analyses and different data partitions were largely congruent with each other. The ML phylogenetic tree was constructed based on the fragments in cp genomes commonly available in 33 species from Lamiids, including 12 species in Gentianales, 1 in Boraginaceae, 10 in Solanales, and 10 in Lamiales. The result strongly supports the position of Boraginaceae (Ehretia acuminata) as the sister of Solanales, with the bootstrap values of 97%. This study provides a platform for further research into the molecular phylogenetics of species in the order Gentianales (family Gentianaceae) notably in respect of speciation and species identification.