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Study region: Alpine shrubland on the northeastern Qinghai-Tibetan Plateau. Study focus: Water provision ability is a pivotal ecological service of high-altitude alpine regions and is controlled by precipitation, evapotranspiration (ET), and soil water storage whereas the underlying ecohydrological processes remain highly unquantified. Here, we investigated continuous 19-year flux measurements to quantify the temporal patterns of ET and water budget (precipitation minus ET, P−ET), as well as 0-20 cm soil water storage change (ΔSWS). New hydrological insights for the region: At a monthly scale, ET peaked in July (96.7 ± 26.4 mm, Mean ± S.D.) and averaged 41.7 ± 31.9 mm, whose variations were determined by the slope of the saturation vapor pressure curve at air temperature, air and soil temperatures, regardless of vegetation growth stage. P−ET averaged 18.3 ± 26.3 mm in August and September while stayed deficit during the other months. The variations in P−ET were controlled by precipitation in the May-October growing season whereas by ET in the non-growing season from November to April. ΔSWS peaked in May (28.8 ± 11.2 mm) and September (3.0 ± 2.7 mm) and almost accumulated to zero over the whole season. At annual scales, none of ET, P−ET, and ΔSWS changed significantly. ET averaged 512.2 ± 68.4 mm and exceeded precipitation (459.1 ± 58.4 mm), likely due to the lateral flow supply of uphill locations. The variations in ET were regulated directly by bulk canopy resistance and indirectly by net radiation. P−ET averaged −53.2 ± 95.4 mm and demonstrated a clear water deficit (−51.6 ± 21.0 mm) during the non-growing season. The variations of P−ET were driven jointly by precipitation and ET, with opposite but equivalent effects. The dominance of thermal conditions and energy availability on ET variability manifested an energy-limited feature of water loss in the alpine shrubland. The temporal patterns in P−ET elucidated that the alpine shrubland plays the water retention rather than water provision function through transforming variable precipitation input into stable ET loss.

In recent decades, alpine meadows have experienced severe degradation owing to external disturbances. Although soil microorganisms are critical for ecosystem services, little is known about their responses to soil degradation and the potential patterns in alpine meadows. To solve this question, we collected and analyzed soil samples from three degraded alpine meadows situated on the Qinghai-Tibet Plateau. We aimed to examine the effects of degradation on soil microbial diversity and identify the ecological predictors for the diversity of bacteria and fungi. Our results showed that alpine meadow degradation significantly changed soil bacterial and fungal diversity and community composition. Specifically, the relationship between bacterial and fungal diversity and degradation intensity was a hump-shaped, with the highest diversity observed at a moderate degradation level. Additionally, alpine meadow degradation-induced changes in microbial diversity were strongly correlated with decreased plant production, with fungal diversity showing a closer link with below-ground biomass (BGB) than with bacterial diversity. Our findings offer empirical evidence that intermediate disturbance (i.e., moderate degradation) may be beneficial for supporting soil biodiversity. This has important implications for informing policy and management strategies meant to conserve soil biodiversity and ecosystem services when facing anthropogenic change.

Accurate estimation of livestock carrying capacity (LCC) and implementation of an appropriate actual stocking rate (ASR) are key to the sustainable management of grazing adapted alpine grassland ecosystems. The reliable determination of aboveground biomass is fundamental to these determinations. Peak aboveground biomass (AGBP) captured from satellite data at the peak of the growing season (POS) is widely used as a proxy for annual aboveground biomass (AGBA) to estimate LCC of grasslands. Here, we demonstrate the limitations of this approach and highlight the ability of POS in the estimation of ASR. We develop and trail new approaches that incorporate remote sensing phenology timings of grassland response to grazing activity, considering relations between biomass growth and consumption dynamics, in an effort to support more accurate and reliable estimation of LCC and ASR. The results show that based on averaged values from large-scale studies of alpine grassland on the Qinghai-Tibet Plateau (QTP), differences between AGBP and AGBA underestimate LCC by about 31%. The findings from a smaller-scale study that incorporate phenology timings into the estimation of annual aboveground biomass reveal that summer pastures in Haibei alpine meadows were overgrazed by 11.5% during the study period from 2000 to 2005. The methods proposed can be extended to map grassland grazing pressure by predicting the LCC and tracking the ASR, thereby improving sustainable resource use in alpine grasslands.

Abstract Biomass temporal stability plays a key role in maintaining sustainable ecosystem functions and services of grasslands, and climate change has exerted a profound impact on plant biomass. However, it remains unclear how the community biomass stability in alpine meadows responds to changes in some climate factors (e.g., temperature and precipitation). Long‐term field aboveground biomass monitoring was conducted in four alpine meadows (Haiyan [HY], Henan [HN], Gande [GD], and Qumalai [QML]) on the Qinghai‐Tibet Plateau. We found that climate factors and ecological factors together affected the community biomass stability and only the stability of HY had a significant decrease over the study period. The community biomass stability at each site was positively correlated with both the stability of the dominant functional group and functional groups asynchrony. The effect of dominant functional groups on community stability decreased with the increase of the effect of functional groups asynchrony on community stability and there may be a ‘trade‐off’ relationship between the effects of these two factors on community stability. Climatic factors directly or indirectly affect community biomass stability by influencing the stability of the dominant functional group or functional groups asynchrony. Air temperature and precipitation indirectly affected the community stability of HY and HN, but air temperature in the growing season and nongrowing season had direct negative and direct positive effects on the community stability of GD and QML, respectively. The underlying mechanisms varied between community composition and local climate conditions. Our findings highlighted the role of dominant functional group and functional groups asynchrony in maintaining community biomass stability in alpine meadows and we highlighted the importance of the environmental context when exploring the stability influence mechanism. Studies of community stability in alpine meadows along with different precipitation and temperature gradients are needed to improve our comprehensive understanding of the mechanisms controlling alpine meadow stability.

Alpine wetlands sequester large amounts of soil carbon, so it is vital to gain a full understanding of their land-atmospheric CO2 exchanges and how they contribute to regional carbon neutrality; such an understanding is currently lacking for the Qinghai—Tibet Plateau (QTP), which is undergoing unprecedented climate warming. We analyzed two-year (2018–2019) continuous CO2 flux data, measured by eddy covariance techniques, to quantify the carbon budgets of two alpine wetlands (Luanhaizi peatland (LHZ) and Xiaobohu swamp (XBH)) on the northeastern QTP. At an 8-day scale, boosted regression tree model-based analysis showed that variations in growing season CO2 fluxes were predominantly determined by atmospheric water vapor, having a relative contribution of more than 65%. Variations in nongrowing season CO2 fluxes were mainly controlled by site (categorical variable) and topsoil temperature (Ts), with cumulative relative contributions of 81.8%. At a monthly scale, structural equation models revealed that net ecosystem CO2 exchange (NEE) at both sites was regulated more by gross primary productivity (GPP), than by ecosystem respiration (RES), which were both in turn directly controlled by atmospheric water vapor. The general linear model showed that variations in nongrowing season CO2 fluxes were significantly (p < 0.001) driven by the main effect of site and Ts. Annually, LHZ acted as a net carbon source, and NEE, GPP, and RES were 41.5 ± 17.8, 631.5 ± 19.4, and 673.0 ± 37.2 g C/(m2 year), respectively. XBH behaved as a net carbon sink, and NEE, GPP, and RES were –40.9 ± 7.5, 595.1 ± 15.4, and 554.2 ± 7.9 g C/(m2 year), respectively. These distinctly different carbon budgets were primarily caused by the nongrowing season RES being approximately twice as large at LHZ (p < 0.001), rather than by other equivalent growing season CO2 fluxes (p > 0.10). Overall, variations in growing season CO2 fluxes were mainly controlled by atmospheric water vapor, while those of the nongrowing season were jointly determined by site attributes and soil temperatures. Our results highlight the different carbon functions of alpine peatland and alpine swampland, and show that nongrowing season CO2 emissions should be taken into full consideration when upscaling regional carbon budgets. Current and predicted marked winter warming will directly stimulate increased CO2 emissions from alpine wetlands, which will positively feedback to climate change.

Accurate approaches to aboveground biomass (AGB) estimation are required to support appraisal of the effectiveness of land use measures, which seek to protect grazing-adapted grasslands atop the Qinghai-Tibet Plateau (QTP). This methodological study assesses the effectiveness of one commonly used visible band vegetation index, Red Green Blue Vegetation Index (RGBVI), obtained from unmanned aerial vehicle (UAV), in estimating AGB timely and accurately at the local scale, seeking to improve the estimation accuracy by taking into account in situ collected information on disturbed grassland. Particular emphasis is placed upon the mapping and quantification of areas disturbed by grazing (simulated via mowing) and plateau pika (Ochotona curzoniae) that have led to the emergence of bare ground. The initial model involving only RGBVI performed poorly in AGB estimation by underestimating high AGB by around 10% and overestimating low AGB by about 10%. The estimation model was modified by the mowing intensity ratio and bare ground metrics. The former almost doubled the estimation accuracy from R2 = 0.44 to 0.81. However, this modification caused the bare ground AGB to be overestimated by about 38 and 19 g m−2 for 2018 and 2019, respectively. Although further modification of the model by bare ground metrics improved the accuracy slightly to 0.88, it markedly reduced the overestimation of low AGB values. It is recommended that grazing intensity be incorporated into the micro-scale estimation of AGB, together with the bare ground modification metrics, especially for severely disturbed meadows with a sizable portion of bare ground.

Abstract Background The yield of major crops is generally limited by sink capacity and source strength. Cucumber is a typical raffinose family oligosaccharides (RFOs)–transporting crop. Non-coding RNAs and alternative polyadenylation (APA) play important roles in the regulation of growth process in plants. However, their roles on the sink‒source regulation have not been demonstrated in RFOs–translocating species. Results Here, whole–transcriptome sequencing was applied to compare the leaves of cucumber under different sink strength, that is, no fruit-carrying leaves (NFNLs) and fruit-carrying leaves (FNLs) at 12th node from the bottom. The results show that 1101 differentially expressed (DE) mRNAs, 79 DE long non–coding RNAs (lncRNAs) and 23 DE miRNAs were identified, which were enriched in photosynthesis, energy production and conversion, plant hormone signal transduction, starch and carbohydrate metabolism and protein synthesis pathways. Potential co–expression networks like, DE lncRNAs–DE mRNAs/ DE miRNAs–DE mRNAs, and competing endogenous RNA (ceRNA) regulation models (DE lncRNAs–DE miRNAs–DE mRNAs) associated with sink‒source allocation, were constructed. Furthermore, 37 and 48 DE genes, which enriched in MAPK signaling and plant hormone signal transduction pathway, exist differentially APA, and SPS (CsaV3_2G033300), GBSS1 (CsaV3_5G001560), ERS1 (CsaV3_7G029600), PNO1 (CsaV3_3G003950) and Myb (CsaV3_3G022290) may be regulated by both ncRNAs and APA between FNLs and NFNLs, speculating that ncRNAs and APA are involved in the regulation of gene expression of cucumber sink‒source carbon partitioning. Conclusions These results reveal a comprehensive network among mRNAs, ncRNAs, and APA in cucumber sink–source relationships. Our findings also provide valuable information for further research on the molecular mechanism of ncRNA and APA to enhance cucumber yield.

Objective To evaluate the safety and efficacy of removing blood stasis (RBS) herbal medicine for the treatment of acute intracerebral haemorrhage (AICH) within a 6-hour time window. Study design: A randomised, multicentre, double-blind, placebo-controlled study performed in 14 hospitals in China. Participants: and interventions Patients with AICH were randomly assigned to receive a placebo, the ICH-1 (Intracerebral Haemorrhage) formula (eight herbs, including the RBS herbs hirudo and tabanus) or the ICH-2 formula (six herbs without the RBS herbs hirudo and tabanus) within 6 hours of ICH onset. Outcomes: The primary safety outcome was the incidence of haematoma enlargement at 24 hours and at 10 days after treatment. The secondary outcome was the incidence of poor prognosis (mortality or modified Rankin Scale score ≤5) assessed at 90 days after symptom onset. Results: A total of 324 subjects were randomised between October 2013 and May 2016: 105 patients received placebo; 108 patients received the ICH-1 formula; and 111 patients received the ICH-2 formula. The incidence of haematoma enlargement at 24 hours was 7.8% in the placebo group, 12.3% in the ICH-1 group and 7.5% in the ICH-2 group; the incidence of haematoma enlargement on day 10 was 1.1% in the placebo group, 1.1% in the ICH-1 group, and 3.1% in the ICH-2 group, with no significant differences among the groups (P>0.05). The mortality rates were 3.8% in the placebo group, 2.8% in the ICH-1 group, and 0.9% in the ICH-2 group; the incidences of poor prognosis were 7.1% in the placebo group, 6.0% in the ICH-1 group and 4.8% in the ICH-2 group at 3 months, with no significant differences among the groups (p>0.05). However, the overall frequency of treatment-emergent adverse events in the ICH-1 group (12.1%) was higher among the three groups (5.8% and 2.8%, respectively, p<0.05). All three cases of serious adverse events were in the ICH-1 group. Conclusions: Ultra-early administration of ICH-1 formula for AICH patients did not exert significant beneficial effects on clinical outcomes but increased the risk of bleeding, which probably resulted from the inclusion of RBS herbal medicines in ICH-1. [ABSTRACT FROM AUTHOR]

The optimization of the sink-source relationship is of great importance for crop yield regulation. Cucumber is a typical raffinose family oligosaccharide (RFO)-transporting crop. DNA methylation is a common epigenetic modification in plants, but its role in sink-source regulation has not been demonstrated in RFO-translocating species. Here, whole-genome bisulfite sequencing (WGBS-seq) was conducted to compare the nonfruiting-node leaves (NFNLs) and leaves of fruit setting (FNLs) at the 12th node by removing all female flowers in other nodes of the two treatments. We found considerable differentially methylated genes enriched in photosynthesis and carbohydrate metabolic processes. Comparative transcriptome analysis between FNLs and NFNLs indicated that many differentially expressed genes (DEGs) with differentially methylated regions were involved in auxin, ethylene and brassinolide metabolism; sucrose metabolism; and RFO synthesis pathways related to sink-source regulation. Moreover, DNA methylation levels of six sink-source-related genes in the pathways mentioned above decreased in leaves after 5-aza-dC-2′-deoxycytidine (5-Aza-dC, a DNA methyltransferase inhibitor) treatment on FNLs, and stachyose synthase (CsSTS) gene expression, enzyme activity and stachyose content in RFO synthesis pathway were upregulated, thereby increasing fruit length and dry weight. Taken together, our findings proposed an up-to-date inference for the potential role of DNA methylation in the sink-source relationship, which will provide important references for further exploring the molecular mechanism of DNA methylation in improving the yield of RFO transport plants.

The bZIP (basic leucine zipper) proteins play crucial roles in various biological functions. Nitrogen (N) is an essential element for plant growth, especially in cucumber (Cucumis sativus) due to its shallow roots. However, the regulation of bZIP genes in cucumber nitrogen metabolism has not been studied yet. In this study, we identified a total of 72 bZIP genes (CsbZIPs) in the cucumber genome that could be classified into 13 groups. These genes were unevenly distributed on seven chromosomes, and synteny analysis showed that the CsbZIP genes were expanded in a segmentally duplicating manner. Furthermore, our genome-wide expression analysis suggested that CsbZIP genes had different patterns and that five CsbZIP genes were regulated by nitrogen treatment in both leaves and roots. Consistent with CsNPF, CsbZIP55 and CsbZIP65 were regulated by nitrogen treatment in leaves and roots. Moreover, the subcellular localization showed that CsbZIP55 and CsbZIP65 were specifically located in the nucleus, and the transcriptional activation assay showed that CsbZIP55 and CsbZIP65 have transcriptional activation activity. Additionally, in the CsbZIP55 and CsbZIP65 overexpression plants, most nitrogen-regulated CsNPF genes were downregulated. Taken together, our comprehensive analysis of the bZIP gene family lays a foundation for understanding the molecular and physiological functions of CsbZIPs.

HD-Zip, a special class of transcription factors in high plants, has a role in plant development and responding to external environmental stress. Heat stress has always been an important factor affecting plant growth, quality, and yield. Carotenoid content is also an important factor affecting the color of the inner leaf blades of Chinese cabbage. In this study, the genomes of three Brassicaceae plants were selected: Chinese cabbage (Brassica rapa subsp. pekinensis), Brassica oleracea, and Brassica napus. We identified 93, 96, and 184 HD-Zip genes in the B. rapa, B. oleracea, and B. napus, respectively. The HD-Zip gene family was classified into four subfamilies based on phylogeny: I, II, III, and IV;. The results of cis-acting element analysis suggested that HD-Zip family genes may participate in various biological processes, such as pigment synthesis, cell cycle regulation, defense stress response, etc. Conserved motifs prediction revealed that three motifs exist among the four HD-Zip gene families and that different motifs exhibit significant effects on the structural differences in HD-Zips. Synteny, Ks, and 4DTv results displayed that genome-wide triplication events act in HD-Zip gene family expansion. Transcriptome data showed that 18 genes responded (>1.5-fold change) to heat stress in Chinese cabbage, and 14 of 18 genes were from the HD-Zip I subfamily. Three genes had up-regulation, and eight genes had down-regulation in high-carotenoid-content Chinese cabbage. The BraA09g011460.3C expression level was up-regulated after heat stress treatment and significantly reduced in varieties with high carotenoid content, indicating its potential for heat stress tolerance and carotenoid content regulation. This study provided important gene resources for the subsequent breeding of Chinese cabbage.

The NITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER family (NPF) proteins perform an essential role in regulating plant nitrate absorption and distribution and in improving plant nitrogen use efficiency. In this study, cucumber (Cucumis sativus L.) NPF genes were comprehensively analyzed at the whole genome level, and 54 NPF genes were found to be unevenly distributed on seven chromosomes in the cucumber genome. The phylogenetic analysis showed that these genes could be divided into eight subfamilies. We renamed all CsNPF genes according to the international nomenclature, based on their homology with AtNPF genes. By surveying the expression profiles of CsNPF genes in various tissues, we found that CsNPF6.4 was specifically expressed in roots, indicating that CsNPF6.4 may play a role in N absorption; CsNPF6.3 was highly expressed in petioles, which may be related to NO3− storage in petioles; and CsNPF2.8 was highly expressed in fruits, which may promote NO3− transport to the embryos. We further examined their expression patterns under different abiotic stress and nitrogen conditions, and found that CsNPF7.2 and CsNPF7.3 responded to salt, cold, and low nitrogen stress. Taken together, our study lays a foundation for further exploration of the molecular and physiological functions of cucumber nitrate transporters.