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Abstract Background Anisodus tanguticus (Maxim.) Pascher (A. tanguticus) is a valuable botanical for extracting tropane alkaloids, which are widely used in the pharmaceutical industry. Implementing appropriate cultivation methods can improve both the quality and yield of A. tanguticus. A two-year field experiment was conducted from 2021 to 2023 using a single-factor randomized complete block design replicated three times. The study examined the effects of different nutrient levels (nitrogen: 0, 75, 150, 225, 300, 375 kg/ha; phosphorus: 0, 600, 750, 900, 1050, 1200 kg/ha; potassium: 0, 75, 112.5, 150, 187.5, 225 kg/ha) on the growth, primary alkaloid contents, and alkaloid yield of A. tanguticus at different growth stages (S-Greening, S-Growing, S-Wilting; T-Greening, T-Growing, and T-Wilting) in both the roots and aboveground portions. Results Our results demonstrate that nutrient levels significantly affect the growth and alkaloid accumulation in A. tanguticus. High nitrogen levels (375 kg/ha) notably increased both root and aboveground biomass, while phosphorus had a minimal effect, especially on aboveground biomass. For alkaloid content (scopolamine, anisodamine, anisodine, atropine), a moderate nitrogen level (225 kg/ha) was most effective, followed by low potassium (75 kg/ha), with phosphorus showing a limited impact. Increased phosphorus levels led to a decrease in scopolamine content. During the T-Growing period, moderate nitrogen addition (225 kg/ha) yielded the highest alkaloid levels per unit area (205.79 kg/ha). In the T-Wilting period, low potassium (75 kg/ha) and low phosphorus (750 kg/ha) resulted in alkaloid levels of 146.91 kg/ha and 142.18 kg/ha, respectively. This indicates nitrogen has the most substantial effect on alkaloid accumulation, followed by potassium and phosphorus. The Douglas production function analysis suggests focusing on root biomass and the accumulation of scopolamine and atropine in roots to maximize alkaloid yield in A. tanguticus cultivation. Conclusions Our findings show that the optimum harvesting period for A. tanguticus is the T-Wilting period, and that the optimal nitrogen addition is 225 kg/ha, the optimal potassium addition is 75 kg/ha, and the optimal phosphorus addition is 600 kg/ha or less.

Abstract Background Plant growth and quality are often affected by environmental factors, including geographical location, climate, and soil. In this study, we describe the effect of altitudinal differences on the growth and active ingredients in Rheum tanguticum Maxim. ex Balf. (R. tanguticum), a traditional Chinese medicinal herb known for its laxative properties. Results The results showed that plants grown at lower altitudes had better growth performances than those in higher altitude areas. The yield varied by 2.45–23.68 times with altitude, reaching a maximum of 102.01 t/ha. In addition, total anthraquinone and total sennoside contents decreased with increasing altitude, whereas total tannins increased with increasing altitude. The total anthraquinone content of the indicator compound reached 5.15% at five experimental sites, which exceeded the Chinese Pharmacopoeia standard by 70.87%. The content of the other two categories of active ingredients reached a maximum value of 0.94% (total sennosides) and 2.65% (total tannins). Redundancy analysis revealed that annual rainfall, annual average temperature, annual sunshine hours, and pH significantly affected growth and active ingredients. Moreover, key metabolites, such as flavonoids, amino acids and their derivatives, phenolic acids, lipids, and terpenes, were differentially expressed between samples from low- and high-altitude cultivation areas. These metabolites were enriched in the flavonoid and flavonol biosynthetic pathway and the monoterpene biosynthetic pathway. Conclusions These results suggest that high anthraquinone content was observed in the lowest-latitude cultivation area due to low rainfall and alkaline soil pH. Key metabolites were significantly upregulated in high-latitude cultivation areas. These results provide a scientific basis for quality control and the systematic cultivation of R. tanguticum.

Anisodus tanguticus (Maxim.) Pascher, a distinctive medicinal plant native to the Qinghai-Tibet Plateau, China, has garnered attention due to increasing market demand. This study explores the impact of environmental factors on the distribution and levels of active compounds namely anisodamine, anisodine, and atropine within A. tanguticus. Our goal was to identify suitable cultivation areas for this plant. This study employs the maximum entropy model to simulate the suitable area of A. tanguticus under current conditions and three climate change scenarios during the 2050s and 2070s. The finding revealed that altitude, precipitation in the warmest season (Bio 18), the average annual temperature (Bio 1) exerted significant influences on the distribution of A. tanguticus. Among the environmental factors considered, temperature difference between day and night (Bio 2) had the most substantial impact on the distribution of anisodamine, temperature seasonal variation variance (Bio 4) predominantly influenced anisodine distribution, and Bio 1 had the greatest effected on the distribution of atropine. The suitable areas primarily exist in the eastern Qinghai-Tibet Plateau in China, encompassing a total area of 30.78 × 104 km2. Under the climate scenarios for the future, the suitable areas exhibit increasing trends of approximately 30.2%, 30.3%, and 39.8% by the 2050s, and 25.1%, 48.8%, and 60.1% by the 2070s. This research would provide theoretical suggestions for the protection, and cultivation management of A. tanguticus resources to face the challenge of global climate change.

Anisodus tanguticus (Maxim.) Pascher, which is an important species used in traditional Tibetan medicine, is grown on the Qinghai–Tibet Plateau. A quantitative analysis of 18 elements in A. tanguticus and the soil was conducted using an inductively coupled plasma optical emission spectrometer. The elemental contents of A. tanguticus underwent principal component analysis, linear discriminant analysis, and orthogonal partial least-squares discriminant analysis. Furthermore, elemental accumulation and elemental transfer coefficients were calculated, and the correlation between environmental factors and elemental content was further explored. The multivariate statistical results indicated that A. tanguticus plants can be distinguished by their geographical origin. Elemental accumulation and transfer patterns differed significantly among the analysed elements. Some positive or negative correlation relationships were found between environmental factors and contents in root parts of A. tanguticus. The present study provides an efficient and practical method to discriminate the geographical origin of A. tanguticus from different regions.

Abstract Background Although it is well recognized that core root microorganisms contribute to plant health and productivity, little is known about their role to the accumulation of secondary metabolites. The roots of Anisodus tanguticus, a traditional herbal medication utilized by Tibetan medicine, are rich in tropane alkaloids. We collected wild A. tanguticus populations throughout a 1500 km transect on the Qinghai-Tibetan Plateau. Results Our results showed that despite sampling at a distance of 1500 km, the root of A. tanguticus selectively recruits core root bacteria. We obtained 102 root bacterial core OTUs, and although their number only accounted for 2.99% of the total, their relative abundance accounted for 73% of the total. Spearman correlation and random forest analyses revealed that the composition of core root microbiomes was related to anisodine contents, aboveground biomass and nitrogen contents of Anisodus tanguticus. Among them, the main role is played by Rhizobacter, Variovorax, Polaromonas, and Mycobacterium genus that are significantly enriched in roots. Functional prediction by FAPROTAX showed that nitrogen-cycling microorganisms and pathogenic bacteria are strongly associated with anisodine contents, aboveground biomass and nitrogen contents of Anisodus tanguticus. Conclusions Our findings show that the root selectively recruits core root bacteria and revealed that the core microbiomes and microbial functions potentially contributed to the anisodine contents, aboveground biomass and nitrogen contents of the plant. This work may increase our understanding of the interactions between microorganisms and plants and improve our ability to manage root microbiota to promote sustainable production of herbal medicines.

Rheum tanguticum is one of the primary rhubarb species used for food and medicinal purposes, and it has recently been gaining more attention and recognition. This research represents the first attempt to use stable isotopes and elemental analysis via IRMS to identify the geographical origin of Rheum tanguticum. A grand total of 190 rhubarb samples were gathered from 38 locations spread throughout the provinces of Gansu, Sichuan, and Qinghai in China. The carbon content showed a decreasing trend in the order of Qinghai, followed by Sichuan, and then Gansu. Nitrogen content was notably higher, with Qinghai and Sichuan displaying similar levels, while Gansu had the lowest nitrogen levels. Significant differences were noted in the δ13C (−28.9 to −26.5‰), δ15N (2.6 to 5.6‰), δ2H (−120.0 to −89.3‰), and δ18O (16.0‰ to 18.8‰) isotopes among the various rhubarb cultivation areas. A significant negative correlation was found between %C and both longitude and humidity. Additionally, δ13C and δ15N isotopes were negatively correlated with longitude, and δ15N showed a negative correlation with humidity as well. δ2H and δ18O isotopes exhibited a strong positive correlation with latitude, while significant negative correlations were observed between δ2H and δ18O isotopes and temperature, precipitation, and humidity. The LDA, PLS-DA, and k-NN models all exhibited strong classification performance in both the training and validation sets, achieving accuracy rates between 82.1% and 91.7%. The combination of stable isotopes, elemental analysis, and chemometrics provides a reliable and efficient discriminant model for accurately determining the geographical origin of R. tanguticum in different regions. In the future, the approach will aid in identifying the geographical origin and efficacy of rhubarb in other studies.

Abstract The relationship between spatial distribution and phylogeny has been widely debated in recent decades. To understand biogeographic and evolutionary history relationships and to explore the interspecific similarities and phylogenetic correlations of niche characteristics, we collected and recorded all distribution points for eight species of rhubarb on the Qinghai‐Tibet Plateau, used different methods to describe the ecological niche, and explored the relationship between phylogeny, ecological niche, and distribution range. The results reveal that: (1) the ranges of optimal environmental variables for species with close kinship are not exactly the same, ecologically similar species are not necessarily sister species, and the overlap of rhubarb has no significant correlation with phylogeny. Therefore, the impact of ecological dimensions on species formation is greater than that of geographical latitude for the eight species of rhubarb. (2) Among the eight species of rhubarb, the breadth of ecological niche is positively correlated with the current suitable habitat area and negatively correlated with fluctuations in future suitable habitat area. In the future, except for Rheum tanguticum and Rheum palmatum, the suitable planting areas for the other six species of rhubarb will decrease as greenhouse gas emissions concentrations and time increase. Therefore, species with smaller ecological niches are at a greater risk of habitat loss compared to species with larger ecological niches. (3) In both existing and future distribution prediction models of rhubarb, we observed that both the widely distributed Rheum spiciforme and the niche narrow Rheum nobile, all eight species of rhubarb are present in the Hengduan Mountains, based on our analysis, we propose that the Hengduan Mountains should be regarded as a priority conservation area for rhubarb, to preserve the species' biodiversity. Our study lays the groundwork for identifying evolutionary trends in ecological specialization.

Abstract Background Climate fluctuations during the Pleistocene and mountain uplift are vital driving forces affecting geographic distribution. Here, we ask how an annual plant responded to the Pleistocene glacial cycles. Methods In this study, we analyzed the population demographic history of the annual herb Swertia tetraptera Maxim (Gentianaceae) endemic to Qinghai-Tibetan Plateau (QTP). A total of 301 individuals from 35 populations of S. tetraptera were analyzed based on two maternally inherited chloroplast fragments (trnL-trnF and trnS-trnG). Phylogeographic analysis was combined with species distribution modeling to detect the genetic variations in S. tetraptera. Results The genetic diversity of S. tetraptera was high, likely due to its wide natural range, high proportion of endemic haplotypes and evolutionary history. Fifty-four haplotypes were identified in S. tetraptera. Only a few haplotypes were widespread (Hap_4, Hap_1, Hap_3), which were dispersed throughout the present geographical range of S. tetraptera, while many haplotypes were confined to single populations. The cpDNA dataset showed that phylogeographic structuring was lacking across the distribution range of S. tetraptera. Analyses of molecular variance showed that most genetic variation was found within populations (70.51%). In addition, the relationships of the haplotypes were almost completely unresolved by phylogenetic reconstruction. Both mismatch distribution analysis and neutrality tests showed a recent expansion across the distribution range of S. tetraptera. The MaxEnt analysis showed that S. tetraptera had a narrow distribution range during the Last Glacial Maximum (LGM) and a wide distribution range during the current time, with predictions into the future showing the distribution range of S. tetraptera expanding. Conclusion Our study implies that the current geographic and genetic distribution of S. tetraptera is likely to have been shaped by Quaternary periods. Multiple microrefugia of S. tetraptera existed during Quaternary glaciations. Rapid intraspecific diversification and hybridization and/or introgression may have played a vital role in shaping the current distribution patterns of S. tetraptera. The distribution range of S. tetraptera appeared to have experienced contraction during the LGM; in the future, when the global climate becomes warmer with rising carbon dioxide levels, the distribution of S. tetraptera will expand.

Agarwood is a resinous heartwood of Aquilaria sinensis that is formed in response to mechanical wounding. In the present study pre-treatment of Aquilaria sinensis was carried out, and then the dominant fungi were isolated and purified from the surface and electroshock holes of trees. The isolated Trichoderma sp. and Neurospora sp. were then screened for resistance against benzyl acetone and then inoculated into healthy Aquilaria sinensis trees. After six months, the agarwood was collected for analysis. The chemical composition of incense was analyzed using gas chromatography-mass spectroscopy, and 82 chemical constituents were identified. Agarwood products formed by using Trichoderma sp. and Neurospora sp. consisted of 50.22% and 48.71% ether extracts, respectively, which surpassed the 10% threshold specified by the Chinese Pharmacopoeia. Similarly, relative aromatic contents in the two agarwood products were 30.1% and 32.86%, while proportions of sesquiterpene constituents were 10.21% and 11.19%, respectively. These two agarwood-specific chemical constituents accounted for a large proportion of the total chemical composition, which showed that the generated agarwood was of good quality. The results of the study revealed that both Trichoderma sp. and Neurospora sp. were able to effectively induce agarwood production in Aquilaria sinensis trees in 6 months. This study expands the library of fungi that promote the production of agarwood from Aquilaria sinensis trees.

Interpreting biodiversity patterns and the underlying processes is crucial for evaluating the mechanisms of community assembly, but the view of multifaceted diversity patterns spanning broad spatial extents is less strengthened. We implemented an inventory of 1260 vegetation plots from shrublands across China with standardized methods and analyzed patterns of taxonomic and phylogenetic diversity with differential weighting of common and rare species, as well as phylogenetic co-occurrence structures. Taxonomic and phylogenetic diversity were linearly correlated when common and rare species were weighted equally, but had a logarithmic correlation when species were weighted with their relative abundances. While most shrubland communities were phylogenetically unstructured, the correlation between taxonomic and phylogenetic diversity covaried with phylogenetic relatedness when incorporating relative abundance, but only weakly so in phylogenetically over-dispersed communities. When we correlated patterns of taxonomic and phylogenetic diversity with different weightings for common versus rare species, we found an important role for geographic (e.g., longitude, altitude), climatic (temperature, precipitation) and soil factors. The importance of underlying variables varied between facets of diversity. We found a strong role for altitude in taxonomic, but less so for phylogenetic diversity. Furthermore, the importance of several environmental drivers varied depending on whether diversity metrics were strongly influenced by rare species or put more weight on common and/or dominant species. Overall, our assessment highlights the importance of synthetic analyses of patterns and processes of different facets of biodiversity to capture the full complexity of diversity in conservation studies.

Anisodus tanguticus is a valuable plant for extracting tropane alkaloids. However, the mechanisms by which plant microbiome mediate the accumulation of tropane alkaloids in Anisodus tanguticus are still not well understood. In this study, we collected 55 wild Anisodus tanguticus populations on the Tibetan Plateau and the tropane alkaloids content, and root-related bacteria and fungi diversity were analyzed using HPLC and 16 s rDNA and ITS sequencing. The results showed that tropane alkaloids content has obvious geographical distribution characteristics. Anisodine content had a significant positive correlation with latitude, while anisodamine and atropine content had a significant negative correlation with latitude. Variation partition analysis (VPA) showed that root endophytes play a significant role in promoting tropane alkaloid production in Anisodus tanguticus roots. The root endophytes alone explained 14% of the variation, which was the largest contributor. Soil properties variables could independently explain 5% of the variation, and climate variables could explain 1% of the variation. Of these, endophytic fungi alone accounted for 11%, while bacteria explained only 5%. Random forests and Mantel test showed that different regionally enriched endophytic fungi have a greater impact on the accumulation of tropane alkaloids than the whole endophytic fungi. Richness and relative abundance of enriched endophytic fungi in Hengduan-Qilian Mountains (HQ) group has a significant positive correlation with anisodine content, while richness and relative abundance of enriched endophytic fungi in Himalayas-Hengduan Mountains (HH) group has a significant positive correlation with anisodamine and atropine content. And, these enriched endophytic fungi have high network connectivity and distributed in separate network modules. This study further confirmed that endophytes were closely related to tropane alkaloids accumulation in Anisodus tanguticus and contribute to promote sustainable development, cultivation, and precision medicine of Anisodus tanguticus.

Camellia oleifera Abel, a woody oil plant, that is endemic to China. Tea oil, also referred to as “oriental olive oil,” is a superior quality plant-based cooking oil. The production of tea oil accounts for 8% of the total edible vegetable oil production in the country. Since 2022, the annual output value of C. oleifera industry has exceeded 100 billion yuan, making it one of the major economic contributors to China’s rural revitalization development strategy. In recent years, demand and production have grown in parallel. However, this has led to an increase in the incidence levels of pest and diseases. Pests and diseases significantly reduce the quality and yield of C. oleifera. C. oleifera diseases are mainly caused by pathogenic fungi. C. oleifera anthracnose, soft rot, leaf spot, coal stain, leaf gall disease, and root rot are the most important fungal diseases affecting the C. oleifera industry. However, the same disease may be caused by different pathogenic fungi. C. oleifera can be found in half of China and is found in several climatic zones. The geographical distribution of woody plant diseases is consistent with the distribution of the tree species and the ecology of the range, which also results in a highly complex distribution of fungal diseases of C. oleifera. The management of fungal diseases in C. oleifera is extremely challenging due to the variety of pathogenic fungal species, multiple routes of transmission, the lack of resistant plants, and the environmental safety of chemical measures. The optimal strategy for addressing fungal diseases in C. oleifera is to develop and apply an integrated disease management plan. This review provides a brief overview of the pathogenic species, pathogenesis, pathogenesis, geographical distribution, current management strategies, and potentially new methods of C. oleifera fungal diseases, to provide direction for the development of comprehensive management measures for C. oleifera fungal diseases in the future.

Changes in soil microorganisms caused by enclosure have long been an important topic in grassland ecology. However, few studies have described the responses of the soil microbial community composition, functional groups, and co-occurrence networks at different growth stages to long-term grazing exclusion or compared the indicative abilities of diversity and network parameters on soil ecosystem functions after grazing exclusion. In this study, we conducted a field experiment to evaluate the response of soil microbes to long-term grazing exclusion in an alpine steppe on the Qinghai-Tibetan Plateau. Our results showed that both soil fungal and bacterial community compositions at different growth stages showed significant differences. However, the enclosure only altered the fungal community composition. Differences in soil microbial functional groups between fungi and bacteria were also detected. The leading group in soil fungi was governed by grazing exclusion, and most bacterial functional groups showed significant differences between the growth stages. The co-occurrence network patterns of the fungi and bacteria differed significantly. Unexpectedly, the enclosure did not significantly alter the fungal or bacterial co-occurrence networks. Network parameters had a stronger ability to indicate soil ecosystem functions than the diversity index, displaying better relationships between network parameters and soil organic carbon and nitrate. These findings suggest that elucidating the responses of soil microbes at different growth stages is necessary to comprehensively understand soil ecosystems after long-term grazing exclusion in the alpine steppe.

Camellia oleifera (C. oleifera) is one of the four main, woody, edible oil tree species in the world, while C. oleifera anthracnose is mainly caused by the fungus Colletotrichum fructicola (C. fructicola), which severely affects the yield of C. oleifera and the quality of tea oil. Bacillus velezensis (B. velezensis) CSUFT-BV4 is an antagonistic endophytic bacterium isolated from healthy C. oleifera leaves. This study aimed to investigate the biocontrol potential of strain CSUFT-BV4 against C. oleifera anthracnose and its possible functional mechanism, and to determine its growth-promoting characteristics in host plants. In vitro, CSUFT-BV4 was shown to have efficient biofilm formation ability, as well as significant functions in the synthesis of metabolic substances and the secretion of probiotic substances. In addition, the CSUFT-BV4 fermentation broth also presented efficient antagonistic activities against five major C. oleifera anthracnose pathogens, including C. fructicola, C. gloeosporioides, C. siamense, C. camelliae, and C. kahawae, and the inhibition rate was up to 73.2%. In vivo, it demonstrated that the growth of C. oleifera treated with CSUFT-BV4 fermentation broth was increased in terms of stem width, plant height, and maximum leaf area, while the activities of various defense enzymes, e.g., superoxide dismutase (SOD), phenylalanine aminotransferase (PAL), and polyphenol oxidase (PPO), were effectively increased. The remarkable antagonistic activities against C. oleifera anthracnose, the growth-promoting characteristics, and the induction of host defense responses indicate that endophytic bacterium CSUFT-BV4 can be effectively used in the biological control of C. oleifera anthracnose in the future, which will have a positive impact on the development of the C. oleifera industry.

Traumatic brain injury (TBI) leads to structural damage in the brain, and is one of the major causes of disability and death in the world. Herein, we developed a composite injectable hydrogel (HA/Gel) composed of hyaluronic acid (HA) and gelatin (Gel), loaded with vascular endothelial growth factor (VEGF) and salvianolic acid B (SAB) for treatment of TBI. The HA/Gel hydrogels were formed by the coupling of phenol-rich tyramine-modified HA (HA-TA) and tyramine-modified Gel (Gel-TA) catalyzed by horseradish peroxidase (HRP) in the presence of hydrogen peroxide (H2O2). SEM results showed that HA/Gel hydrogel had a porous structure. Rheological test results showed that the hydrogel possessed appropriate rheological properties, and UV spectrophotometry results showed that the hydrogel exhibited excellent SAB release performance. The results of LIVE/DEAD staining, CCK-8 and Phalloidin/DAPI fluorescence staining showed that the HA/Gel hydrogel possessed good cell biocompatibility. Moreover, the hydrogels loaded with SAB and VEGF (HA/Gel/SAB/VEGF) could effectively promote the proliferation of bone marrow mesenchymal stem cells (BMSCs). In addition, the results of H&E staining, CD31 and α-SMA immunofluorescence staining showed that the HA/Gel/SAB/VEGF hydrogel possessed good in vivo biocompatibility and pro-angiogenic ability. Furthermore, immunohistochemical results showed that the injection of HA/Gel/SAB/VEGF hydrogel to the injury site could effectively reduce the volume of defective tissues in traumatic brain injured mice. Our results suggest that the injection of HA/Gel hydrogel loaded with SAB and VEGF might provide a new approach for therapeutic brain tissue repair after traumatic brain injury.

Ischemia reperfusion (I/R) is a common pathological process which occurs mostly in organs like the heart, brain, kidney, and lung. The injury caused by I/R gradually becomes one of the main causes of fatal diseases, which is an urgent clinical problem to be solved. Although great progress has been made in therapeutic methods, including surgical, drug, gene therapy, and transplant therapy for I/R injury, the development of effective methods to cure the injury remains a worldwide challenge. In recent years, exosomes have attracted much attention for their important roles in immune response, antigen presentation, cell migration, cell differentiation, and tumor invasion. Meanwhile, exosomes have been shown to have great potential in the treatment of I/R injury in organs. The study of the exosome-mediated signaling pathway can not only help to reveal the mechanism behind exosomes promoting reperfusion injury recovery, but also provide a theoretical basis for the clinical application of exosomes. Here, we review the research progress in utilizing various exosomes from different cell types to promote the healing of I/R injury, focusing on the classical signaling pathways such as PI3K/Akt, NF-κB, Nrf2, PTEN, Wnt, MAPK, toll-like receptor, and AMPK. The results suggest that exosomes regulate these signaling pathways to reduce oxidative stress, regulate immune responses, decrease the expression of inflammatory cytokines, and promote tissue repair, making exosomes a competitive emerging vector for treating I/R damage in organs.

Phytopathogens pose a devastating threat to the productivity and yield of crops by causing destructive plant diseases in natural and agricultural environments. Hemibiotrophic pathogens have a variable-length biotrophic phase before turning to necrosis and are among the most invasive plant pathogens. Plant resistance to hemibiotrophic pathogens relies mainly on the activation of innate immune responses. These responses are typically initiated after the plant plasma membrane and various plant immune receptors detect immunogenic signals associated with pathogen infection. Hemibiotrophic pathogens evade pathogen-triggered immunity by masking themselves in an arms race while also enhancing or manipulating other receptors to promote virulence. However, our understanding of plant immune defenses against hemibiotrophic pathogens is highly limited due to the intricate infection mechanisms. In this review, we summarize the strategies that different hemibiotrophic pathogens interact with host immune receptors to activate plant immunity. We also discuss the significant role of the plasma membrane in plant immune responses, as well as the current obstacles and potential future research directions in this field. This will enable a more comprehensive understanding of the pathogenicity of hemibiotrophic pathogens and how distinct plant immune receptors oppose them, delivering valuable data for the prevention and management of plant diseases.

Potentilla fruticosa, a major alpine shrubland type, is widely distributed across the Tibetan Plateau, and grazing is the most common disturbance in the shrublands of P. fruticosa. However, soil organic carbon (SOC), soil total nitrogen (STN), soil total phosphorus (STP), and their stoichiometry under different grazing intensities were unclear. In our study, we explored SOC, STN, STP, their stoichiometry, and their controlling factors in the grazing disturbance of heavy grazing (HG), moderate grazing (MG), light grazing (LG), and no grazing (NG) conditions in the Tibetan Plateau P. fruticosa shrublands. The grazing intensities were mainly assessed by considering the shrublands’ ground cover, the indicators of the road density, the distance between sampling sites and cowshed or sheep shed, the amounts of cow and sheep dung, and vegetation that had been gnawed and stampeded. Our results indicated that soil physical properties of soil temperature and bulk density have decreasing trends with decreasing grazing intensities from HG to NG. The SOC, STN, STP, and soil C:N and C:P ratios have increasing trends with decreasing grazing intensities from HG to NG, while the changes in soil N:P ratio were relatively stable along grazing intensities. Our results indicated that HG generally had stronger effects on SOC, STN, and soil C:N and C:P ratios than NG, indicating substantial effects of grazing disturbance on biogeochemical cycles of SOC and STN in the shrubland ecosystems. Therefore, for the alpine shrubland of P. fruticosa, the HG should be avoided for sustainable cycling of soil nutrients and the balance of soil nutrient stoichiometry. The grazing types can directly affect plant conditions, and plant conditions can directly affect soil physical and chemical properties and litter standing crops. Finally, soil physicochemical properties and litter standing crop resulting from different grazing intensities directly control SOC, STN, and STP. For the soil stoichiometry, the soil’s physical and chemical properties resulting from different grazing intensities have direct impacts on soil C:P and N:P ratios.

Impaired healing of diabetic wounds harms patients’ quality of life and even leads to disability and death, which is an urgent issue to be solved clinically. Despite the great progress that has been achieved, it remains a worldwide challenge to develop effective therapeutic treatments for diabetic wounds. Recently, exosomes have attracted special attention because they can be involved in immune response, antigen presentation, cell migration, cell differentiation, tumor invasion and other processes. Meanwhile, exosomes have been proven to hold great potential in the treatment of diabetic wounds. Mechanistic studies of exosomes based on signaling pathways could not only help to uncover the mechanisms by which exosomes promote diabetic wound healing but could also provide a theoretical basis for the clinical application of exosomes. Herein, our mini-review aims to summarize the progress of research on the use of various exosomes derived from different cell types to promote diabetic wound healing, with a focus on the classical signaling pathways, including PI3K/Akt, Wnt, NF-κB, MAPK, Notch, Nrf2, HIF-1α/VEGF and TGF-β/Smad. The results show that exosomes could regulate these signaling pathways to down-regulate inflammation, reduce oxidative stress, increase angiogenesis, promote fibroblast proliferation, induce re-epithelization and inhibit scar formation, making exosomes attractive candidates for the treatment of diabetic wounds.

Pine Wilt Disease (PWD), caused by the pathogenic nematode Bursaphelenchus xylophilus, is a systemic infectious disease commonly referred to as the “cancer” of pine trees. This devastating disease has gained this analogy due to its ability to rapidly spread within pine populations, leading to substantial losses in forest resources. The primary objective of this study is to investigate the bioprotective potential and underlying mechanisms of action exhibited by rhizosphere microorganisms associated with Masson pine (Pinus massoniana) in the context of controlling the pine wilt nematode. In this experiment, using high-throughput sequencing, significant differences were observed in the rhizosphere soil microbial communities among healthy Masson pine, standing dead trees, and diseased Masson pine. Furthermore, it was found that these microbial communities exhibited distinct community structures at different levels. This study successfully isolated and screened three strains of highly effective nematophagous bacteria from the rhizosphere soil. The identified strains were Lysinibacillus capsici, Bacillus Paramycoides, and Delftia tsuruhatensis. After applying the bacterial suspensions and fermentation extracts of these three strains to the roots of two-year-old Masson pine seedlings, followed by inoculation with pine wilt nematodes after a four-day period, distinct defense responses were observed in the Masson pine. Notably, the activities of phenylalanine ammonia-lyase (PAL) and peroxidase (POD) were significantly increased, leading to a substantial reduction in the incidence of pine wilt disease. Based on the changes in defense enzyme activities, it can be concluded that the fermentation extract of the Lysinibacillus capsici strain exhibits effective nematocidal effects and induces resistance. The significant biological control efficacy and induction of host defense activity indicate the potential application value of this strain and its metabolites as a biocontrol agent for pine wilt disease.

Abstract Background Rheum tanguticum (R. tanguticum) is an edible and medicinal plant that exhibits high antioxidant activity. The purpose of the present study was to investigate the bioactive components of its seeds and the potential mechanisms of antioxidant activity to provide a foundation for further developmental work on R. tanguticum seeds as a functional food. Methods In this study, the antioxidant activities of R. tanguticum seeds were measured using DPPH, ABTS and FRAP assays. LC-Q-TOF/MS was used to identify the active compounds in the seeds, and Swiss Target Prediction was used to identify their potential targets. The DisGENET, DrugBank, OMIM and GeneCard databases were used to search for antioxidant-related targets. Results The component–target–pathway network was constructed and included 5 compounds and 9 target genes. The hub genes included ESR1, APP, MAPK8, HSP90AA1, AKT1, MMP2, PTGS2, TGFB1 and JUN. The antioxidant activity signaling pathways of the compounds for the treatment of diseases were the cancer signaling pathway, estrogen signaling pathway, colorectal cancer signaling pathway, MAPK signaling pathway, etc. Molecular docking revealed that the compounds in R. tanguticum seeds could inhibit potential targets (AKT1, ESR1 and PTGS2). Conclusion Molecular docking studies revealed that the binding energy score between liriodenine and PTGS2 was the highest (8.16), followed by that of chrysophanol (7.10). This result supports the potential for PTGS2-targeted drug screening and design.

Hepato-pancreato-biliary (HPB) cancer is a serious category of cancer including tumors originating in the liver, pancreas, gallbladder and biliary ducts. It is limited by two-dimensional (2D) cell culture models for studying its complicated tumor microenvironment including diverse contents and dynamic nature. Recently developed three-dimensional (3D) bioprinting is a state-of-the-art technology for fabrication of biological constructs through layer-by-layer deposition of bioinks in a spatially defined manner, which is computer-aided and designed to generate viable 3D constructs. 3D bioprinting has the potential to more closely recapitulate the tumor microenvironment, dynamic and complex cell-cell and cell-matrix interactions compared to the current methods, which benefits from its precise definition of positioning of various cell types and perfusing network in a high-throughput manner. In this review, we introduce and compare multiple types of 3D bioprinting methodologies for HPB cancer and other digestive tumors. We discuss the progress and application of 3D bioprinting in HPB and gastrointestinal cancers, focusing on tumor model manufacturing. We also highlight the current challenges regarding clinical translation of 3D bioprinting and bioinks in the field of digestive tumor research. Finally, we suggest valuable perspectives for this advanced technology, including combination of 3D bioprinting with microfluidics and application of 3D bioprinting in the field of tumor immunology.

To study the reinforcement to soil by herb roots along the Qinghai-Tibet railway, an artificially restored grassland along the Qinghai-Tibet railway is selected as test site. By conducting tensile tests to a single root and direct shear tests to the root-soil composite system with its growth period of one year, such mechanical indices as roots’ tensile strength and shear strength of root-soil composite system are quantitatively assessed. The results show that the values of the mean root diameter, tensile resistance and tensile strength for the combination I (Agropyron cristatum + Puccinellia distans + Poa crymophila) are greater than those for the other combinations. Moreover, a positive power function can be introduced to describe the relationship between the root diameter and its tensile resistance for the three species, while a negative power function is introduced to describe the relationship between the root diameter and its tensile strength. In addition, the cohesion of the root-soil composite system for the three species exhibits a decreasing trend with the increasing soil depth. Of the values for cohesion for the three species, the cohesion for the combination I is 32.62 kPa, greater than that of the other two combinations by 15.3 % and 57.7%, respectively, suggesting the excellent capacity in reinforcing soil. The results may provide a theoretical basis for the prevention and control of the alpine grassland degradation, soil erosion, shallow landslides and other geo-hazards and eco-hazards along the Qinghai-Tibet railway, and are of a practical guiding significance for ecological restoration by planting vegetation along the railway.

Mannan oligosaccharides (MOSs) are excellent prebiotics that are usually obtained via the enzymatic hydrolysis of mannan. In order to reduce the cost of preparing MOSs, immobilized enzymes that demonstrate good performance, require simple preparation, and are safe, inexpensive, and reusable must be developed urgently. In this study, β-mannanase was immobilized on calcium alginate (CaAlg). Under the optimal conditions of 320 U enzyme addition, 1.6% sodium alginate, 2% CaCl2, and 1 h of immobilization time, the immobilization yield reached 68.3%. The optimum temperature and pH for the immobilized β-mannanase (Man-CaAlg) were 75 °C and 6.0, respectively. The Man-CaAlg exhibited better thermal stability, a high degree of pH stability, and less substrate affinity than free β-mannanase. The Man-CaAlg could be reused eight times and retained 70.34% of its activity; additionally, the Man-CaAlg showed 58.17% activity after 30 days of storage. A total of 7.94 mg/mL of MOSs, with 4.94 mg/mL of mannobiose and 3.00 mg/mL of mannotriose, were generated in the oligosaccharide production assay. It is believed that this convenient and safe strategy has great potential in the important field of the use of immobilized β-mannanase for the production of mannan oligosaccharides.

The treatment of skin wounds caused by trauma and pathophysiological disorders has been a growing healthcare challenge, posing a great economic burden worldwide. The use of appropriate wound dressings can help to facilitate the repair and healing rate of defective skin. Natural polymer biomaterials such as collagen and hyaluronic acid with excellent biocompatibility have been shown to promote wound healing and the restoration of skin. However, the low mechanical properties and fast degradation rate have limited their applications. Skin wound dressings based on biodegradable and biocompatible synthetic polymers can not only overcome the shortcomings of natural polymer biomaterials but also possess favorable properties for applications in the treatment of skin wounds. Herein, we listed several biodegradable and biocompatible synthetic polymers used as wound dressing materials, such as PVA, PCL, PLA, PLGA, PU, and PEO/PEG, focusing on their composition, fabrication techniques, and functions promoting wound healing. Additionally, the future development prospects of synthetic biodegradable polymer-based wound dressings are put forward. Our review aims to provide new insights for the further development of wound dressings using synthetic biodegradable polymers.

Abstract Anthropogenic‐driven global change, including changes in atmospheric nitrogen (N) deposition and precipitation patterns, is dramatically altering N cycling in soil. How long‐term N deposition, precipitation changes, and their interaction influence nitrous oxide (N2O) emissions remains unknown, especially in the alpine steppes of the Qinghai–Tibetan Plateau (QTP). To fill this knowledge gap, a platform of N addition (10 g m−2 year−1) and altered precipitation (±50% precipitation) experiments was established in an alpine steppe of the QTP in 2013. Long‐term N addition significantly increased N2O emissions. However, neither long‐term alterations in precipitation nor the co‐occurrence of N addition and altered precipitation significantly affected N2O emissions. These unexpected findings indicate that N2O emissions are particularly susceptible to N deposition in the alpine steppes. Our results further indicated that both biotic and abiotic properties had significant effects on N2O emissions. N2O emissions occurred mainly due to nitrification, which was dominated by ammonia‐oxidizing bacteria, rather than ammonia‐oxidizing archaea. Furthermore, the alterations in belowground biomass and soil temperature induced by N addition modulated N2O emissions. Overall, this study provides pivotal insights to aid the prediction of future responses of N2O emissions to long‐term N deposition and precipitation changes in alpine ecosystems. The underlying microbial pathway and key predictors of N2O emissions identified in this study may also be used for future global‐scale model studies.

Colletotrichum is widespread, and these pathogenic fungi can cause various plant diseases. Studies have shown that Colletotrichum fructicola cause oil-tea (Camellia oleifera) anthracnose and is widely distributed as a dominant fungus in all Ca. oleifera-producing regions. Real-time quantitative PCR(RT-qPCR) is considered the most reliable technique for simultaneously measuring relative gene expression levels in different tissues. Target genes are typically quantified using RT-qPCR to explore gene function, and reliable RT-qPCR results require data normalization using stable reference genes. No studies have reported a suitable reference gene in C. fructicola. This study has eight candidate reference genes (CfCk, CfRpp, CfUce, CfRrp, CfAdrh, CfDd, CfAct, and CfTub) which were selected from C. fructicola-Ca. oleifera transcriptome data and evaluated and sequenced using geNorm, NormFinder, and BestKeeper algorithms. The results showed that CfRrp had better stability in C. fructicola, both during the growth of pure pathogenic fungi and during the invasion of different oil-tea leaves. After normalization with CfRrp, the differentially expressed target genes were similar to the transcriptome. Our work provides suitable reference genes for future studies to quantify target gene expression levels in C. fructicola.

Swertia L. is a large genus in the family Gentianaceae. Different chloroplast gene segments have been used to study systematic evolutionary relationships between species of Swertia L. However, as gene fragment–based phylogenies lack sufficient resolution, the systematic evolutionary relationships between Swertia L. species have remained unclear. We sequenced and annotated the complete chloroplast genomes of four Swertia species, namely, S. bifolia, S. tetraptera, S. franchetian, and S. przewalskii, using next generation sequencing and the plastid genome annotator tool. The chloroplast genome sequences of 19 additional species of Swertia L. were downloaded from the NCBI database and also assessed. We found that all 23 Swertia L. species had a similar genetic structure, that is, a ring tetrad structure, but with some clear differences. The chloroplast genomes of the 23 Swertia L. species were 149036–153691 bp long, averaging 152385 bp; the genomes contained 134 functional genes: 38 tRNA, eight rRNA, and 88 protein-encoding genes. A comparative analysis showed that chloroplasts genome of Swertia was conserved in terms of genome structure, codon preference, and repeat sequences, but it differed in terms of genome sizes, gene contents, and SC/IR boundary. Using Swertia wolfangiana as a reference, we found clear divergences in most of the non-coding and intergenic regions of the complete chloroplast genomes of these species; we also found that rpoC1, ccsA, ndhI, ndhA, and rps15 protein-coding genes had large variations. These highly variable hotspots will be useful for future phylogenetic and population genetic studies. Phylogenetic analysis with high bootstrap support showed that Swertia L. was not monophyletic. The classification of subgen. Swertia and subgen. Ophelia was supported by molecular data, which also partly supported the division of sect. Ophelia, sect. Platynema, sect. Poephila, sect. Swertia, and sect. Macranthos. However, the systematic positions of other groups and species require further exploration. The Swertia L formed at 29.60 Ma. Speciation of 10 species occurred in succession after 12 Ma and 13 species occurred in succession after 2.5 Ma. Our analysis provides insight into the unresolved evolutionary relationships of Swertia L. species.

Anisodus tanguticus (Maxinowicz) Pascher (Solanaceae) is a traditional Chinese herb that is widely used in folklore and clinical practice. In recent years, wild populations have been severely impacted to the point of extinction due to over-harvesting and reclamation. Therefore, artificial cultivation is important to relieve the pressure of market demand and protect wild plant resources. Using a “3414” fertilization design, i.e., 3 factors (N, P, and K), 4 levels, and 14 fertilization treatments, with 3 replicates and a total of 42 experimental plots, A. tanguticus was harvested in October 2020, June 2021, August 2021, and October 2021, and the yield and alkaloid content were determined. The study aimed to provide a theoretical basis and technical reference for the standardization of A. tanguticus cultivation. Biomass accumulation and alkaloid content showed a trend of increasing and then decreasing with the application of nitrogen, phosphorus, and potassium, and the biomass accumulation was the highest at the application levels of nitrogen and phosphorus in T6 and T9 and at the application levels of medium and low potassium. The alkaloid content showed an increasing trend between October of the first year and June of the second year and a decreasing trend in the second year with the increase in the harvesting period. Yield and alkaloid yield showed a decreasing trend between October of the first year and June of the second year and an increasing trend in the second year with the increase in the harvesting period. The recommended application rates are 225–300 kg/ha2 for nitrogen, 850–960 kg/ha2 for phosphorus, and 65–85 kg/ha2 for potassium.

Abstract Background Sterile inflammation is a key pathological process in stroke. Inflammasome activation has been implicated in various inflammatory diseases, including ischemic stroke and hemorrhagic stroke. Hence, targeting inflammasomes is a promising approach for the treatment of stroke. Methods We applied bibliometric methods and techniques. The Web of Science Core Collection was searched for studies indexed from database inception to November 26, 2020. We generated various visual maps to display publications, authors, sources, countries, and keywords. Results Our literature search yielded 427 publications related to inflammasomes involved in stroke, most of which consisted of original research articles and reviews. In particular, we found that there was a substantial increase in the number of relevant publications in 2018. Furthermore, most of the publications with the highest citation rates were published in 2014. Relatively, the field about inflammasomes in stroke developed rapidly in 2014 and 2018. Many institutions contributed to these publications, including those from China, the United States, and worldwide. We found that NLR family pyrin domain containing 3 (NLRP3) was the most studied, followed by NLRP1, NLRP2, and NLRC4 among the inflammasomes associated with stroke. Analysis of keywords suggested that the most studied mechanisms involved dysregulation of extracellular pH, efflux of Ca2+ ions, dysfunction of K+/Na+ ATPases, mitochondrial dysfunction, and damage to mitochondrial DNA. Conclusions Given the potential diagnostic and therapeutic implications, the specific mechanisms of inflammasomes contributing to stroke warrant further investigation. We used bibliometric methods to objectively present the global trend of inflammasomes in stroke, and to provide important information for relevant researchers.

Oil tea (Camellia oleifera), mainly used to produce high-quality edible oil, is an important cash crop in China. Anthracnose of oil tea is a considerable factor that limits the yield of tea oil. In order to effectively control the anthracnose of oil tea, researchers have worked hard for many years, and great progress has been made in the research of oil tea anthracnose. For instance, researchers isolated a variety of Colletotrichum spp. from oil tea and found that Colletotrichum fructicola was the most popular pathogen in oil tea. At the same time, a variety of control methods have been explored, such as cultivating resistant varieties, pesticides, and biological control, etc. Furthermore, the research on the molecular pathogenesis of Colletotrichum spp. has also made good progress, such as the elaboration of the transcription factors and effector functions of Colletotrichum spp. The authors summarized the research status of the harm, pathogen types, control, and pathogenic molecular mechanism of oil tea anthracnose in order to provide theoretical support and new technical means for the green prevention and control of oil tea anthracnose.

Anisodus tanguticus (Maxim.) Pascher, has been used for the treatment of septic shock, analgesia, motion sickness, and anesthesia in traditional Tibetan medicine for 2,000 years. However, the chemical metabolites and geographical traceability and their network pharmacology are still unknown. A total of 71 samples of A. tanguticus were analyzed by Ultra–Performance Liquid Chromatography Q–Exactive Mass Spectrometer in combination with chemometrics developed for the discrimination of A. tanguticus from different geographical origins. Then, network pharmacology analysis was used to integrate the information of the differential metabolite network to explore the mechanism of pharmacological activity. In this study, 29 metabolites were identified, including tropane alkaloids, hydroxycinnamic acid amides and coumarins. Principal component analysis (PCA) explained 49.5% of the total variance, and orthogonal partial least-squares discriminant analysis (OPLS–DA) showed good discrimination (R2Y = 0.921 and Q2 = 0.839) for A. tanguticus samples. Nine differential metabolites accountable for such variations were identified through variable importance in the projection (VIP). Through network pharmacology, 19 components and 20 pathways were constructed and predicted for the pharmacological activity of A. tanguticus. These results confirmed that this method is accurate and effective for the geographic classification of A. tanguticus, and the integrated strategy of metabolomics and network pharmacology can explain well the “multicomponent—-multitarget” mechanism of A. tanguticus.

Abstract Background Rheum tanguticum (Rh. tanguticum) is an important traditional Chinese medicine plant, “Dahuang”, which contains productive metabolites and occupies wide habitats on the Qinghai-Tibet plateau. Plants occupying wide habitats usually vary in phenotypes such as in morphology and metabolism, thereby developing into different ecotypes. Under canopy and open habitats are a pair of dissimilar habitats which possess Rh. tanguticum plants. However, few studies have focused on the effect of habitats on Rh. tanguticum growth, particularly combining morphological and metabolic changes. This study focused on Rh. tanguticum plants growing in under canopy and open habitats where morphology and metabolism changes were quantified using non-target metabolism methods. Results The obtained results indicated that the two dissimilar habitats led to Rh. tanguticum developing into two distinct ecotypes where the morphology and metabolism were simultaneously changed. Under canopy habitats bred morphologically smaller Rh. tanguticum plants which had a higher level of metabolites (22 out of 31) which included five flavonoids, four isoflavonoids, and three anthracenes. On the other hand, the open habitats produced morphologically larger Rh. tanguticum plants having a higher level of metabolites (9 out of 31) including four flavonoids. 6 of the 31 metabolites were predicted to have effect targets, include 4 represent for under canopy habitats and 2 for open habitats. Totally, 208 targets were connected, among which 42 were communal targets for both under canopy and open habitats represent compounds, and 100 and 66 were unique targets for under canopy superior compounds and open habitats superior compounds, respectively. In addition, aloe-emodin, emodin, chrysophanol, physcion, sennoside A and sennoside B were all more accumulated in under canopy habitats, and among which aloe-emodin, emodin, chrysophanol and physcion were significantly higher in under canopy habitats. Conclusions This study determined that Rh. tanguticum growing in under canopy and in open habitats developed into two distinct ecotypes with morphological and metabolic differences. Results of network pharmacology study has indicated that “Dahuang” coming from different habitats, such as under canopy and open habitats, are different in effect targets and thus may have different medicinal use. According to target metabolomics, under canopy habitats may grow better “Dahuang”.

Anthracnose, caused by the fungus Collectotrichum fructicola (C. fructicola), is a major disease affecting the quality and yield of Camellia oleifera (C. oleifera); it reduces C. oleifera yield by 40%–80%. Bacillus subtilis (B. subtilis) YL13 is an antagonistic endophytic bacteria strain isolated from healthy C. oleifera leaves. This study was aimed at investigating the potential of YL13 for the biocontrol of C. oleifera anthracnose and the possible mechanisms involved. In in vitro assays, YL13 demonstrated remarkable antagonistic activity of C. fructicola. Its cell-free filtrates displayed antagonistic activity, which suggested that the metabolites of YL13 might play important roles. In vivo tests showed that the disease index of YL13-treated plants was obviously reduced under greenhouse conditions. YL13 secretes a variety of bioactive metabolites, including protease, cellulase, and siderophore, which might participate in the resistance to C. fructicola. In addition, C. oleifera treated with the fermentation broth of YL13 demonstrated different defense responses, e.g., accumulation of hydrogen peroxide (H2O2) and activation of the defense-related enzyme peroxidase (POD), which might contribute directly or indirectly to overcome external stresses. The significant biocontrol effect and host defense-induction activity of YL13 suggested that this B. subtilis strain as well as its metabolites have the potential to be exploited as microbial control agents for the efficient management of C. oleifera anthracnose.

Inflammatory dysfunction and angiogenesis inhibition are two main factors leading to the delayed healing of diabetic wounds. Hydrogels with anti-inflammatory and angiogenesis-promoting effects have been considered as promising wound care materials. Herein, a salvianolic acid B (SAB)-loaded hyaluronic acid (HA) self-healing hydrogel (HA/SAB) with anti-inflammatory and pro-angiogenesis capacities for diabetic wound healing is reported. The HA hydrogel was prepared via the covalent cross-linking of aldehyde groups in oxidized HA (OHA) and hydrazide groups in adipic dihydrazide (ADH)-modified HA (HA-ADH) with the formation of reversible acylhydrazone bonds. The obtained HA hydrogel exhibited multiple favorable properties such as porous structures, excellent self-healing properties, a sustainable release capacity of SAB, as well as excellent cytocompatibility. In addition, the effects of the SAB-loaded HA self-healing hydrogel were investigated via a full-thickness skin defect model using diabetic rats. The HA/SAB hydrogel showed enhanced skin regeneration effects with accelerated wound closure, shorter remaining dermal space length, thicker granulation tissue formation, and more collagen deposition. Furthermore, reduced inflammatory response and enhanced vascularization were found with HA/SAB2.5 hydrogel-treated wounds, indicating that the hydrogel promotes diabetic wound healing through the promotion of anti-inflammation and angiogenesis. Our results suggest that the fabricated SAB-loaded HA self-healing hydrogel is promising as a wound dressing for the treatment of diabetic wounds.

In the search for a novel and scalable skin scaffold for wound healing and tissue regeneration, we fabricated a class of fibrin/polyvinyl alcohol (PVA) scaffolds using an emulsion templating method. The fibrin/PVA scaffolds were formed by enzymatic coagulation of fibrinogen with thrombin in the presence of PVA as a bulking agent and an emulsion phase as the porogen, with glutaraldehyde as the cross-linking agent. After freeze drying, the scaffolds were characterized and evaluated for biocompatibility and efficacy of dermal reconstruction. SEM analysis showed that the formed scaffolds had interconnected porous structures (average pore size e was around 330 µm) and preserved the nano-scale fibrous architecture of the fibrin. Mechanical testing showed that the scaffolds’ ultimate tensile strength was around 0.12 MPa with an elongation of around 50%. The proteolytic degradation of scaffolds could be controlled over a wide range by varying the type or degree of cross-linking and by fibrin/PVA composition. Assessment of cytocompatibility by human mesenchymal stem cell (MSC) proliferation assays shows that MSC can attach, penetrate, and proliferate into the fibrin/PVA scaffolds with an elongated and stretched morphology. The efficacy of scaffolds for tissue reconstruction was evaluated in a murine full-thickness skin excision defect model. The scaffolds were integrated and resorbed without inflammatory infiltration and, compared to control wounds, promoted deeper neodermal formation, greater collagen fiber deposition, facilitated angiogenesis, and significantly accelerated wound healing and epithelial closure. The experimental data showed that the fabricated fibrin/PVA scaffolds are promising for skin repair and skin tissue engineering.

Dalbergia odorifera (D. odorifera), commonly named the fragrant rosewood, is one of the second-level protected wild plants in China, and one of 34 species of rosewood in five genera and eight categories in the National Standard of China. As a kind of traditional Chinese medicine (TCM), it plays an important role in the pharmaceutical industry, including the treatment of cardiovascular diseases, rheumatic pain, etc. With the continuous expansion of the planting area of D. odorifera, the diseases and pests of D. odorifera become more and more serious, among which leaf-eating pests are the most serious. In this study, ultraviolet rays and microwaves were used to mutagenize Beauveria bassiana (B. bassiana) strain HNCMBJ-P-01, and excellent mutant strains with high spore yield and high virulence were screened out, and then they were prepared into a wettable powder for forest control experiments to study their biocontrol effects. The virulence screening test showed that the virulence of strain HBWB-44 was the strongest, and the 10 day corrected mortality rate was 80.00%, and the lethal time was 5.622 days. The results of biological control test showed that the control effect of B. bassiana wettable powder 100 times solution reached 60.89%, second only to the botanical fungicide matrine. Generically, The B. bassiana that we screened and mutated showed a good killing effect on Plecoptera bilinealis (P. bilinealis), and the wettable powder produced by it showed a good control effect on the leaf-eating pests of D. odorifera. The application of fungal insecticides in plantations has a good prospect for controlling the occurrence of leaf-eating pests of D. odorifera.

Background & Aims: TIGIT is a co-inhibitory receptor, and its suitability as a target for cancer immunotherapy in HCC is unknown. PD1 blockade is clinically effective in about 20% of advanced HCC patients. Here we aim to determine whether co-blockade of TIGIT/PD1 has added value to restore functionality of HCC tumor-infiltrating T cells (TILs). Methods: Mononuclear leukocytes were isolated from tumors, paired tumor-free liver tissues (TFL) and peripheral blood of HCC patients, and used for flow cytometric phenotyping and functional assays. CD3/CD28 T-cell stimulation and antigen-specific assays were used to study the ex vivo effects of TIGIT/PD1 single or dual blockade on T-cell functions. Results: TIGIT was enriched, whereas its co-stimulatory counterpart CD226 was down-regulated on PD1high CD8+ TILs. PD1high TIGIT+ CD8+ TILs co-expressed exhaustion markers TIM3 and LAG3 and demonstrated higher TOX expression. Furthermore, this subset showed decreased capacity to produce IFN-γ and TNF-α. Expression of TIGIT-ligand CD155 was up-regulated on tumor cells compared with hepatocytes in TFL. Whereas single PD1 blockade preferentially enhanced ex vivo functions of CD8+ TILs from tumors with PD1high CD8+ TILs (high PD1 expressers), co-blockade of TIGIT and PD1 improved proliferation and cytokine production of CD8+ TILs from tumors enriched for PD1int CD8+ TILs (low PD1 expressers). Importantly, ex vivo co-blockade of TIGIT/PD1 improved proliferation, cytokine production, and cytotoxicity of CD8+ TILs compared with single PD1 blockade. Conclusions: Ex vivo, co-blockade of TIGIT/PD1 improves functionality of CD8+ TILs that do not respond to single PD1 blockade. Therefore co-blockade of TIGIT/PD1 could be a promising immune therapeutic strategy for HCC patients.

Nitrogen (N) and phosphorus (P) stoichiometry have significant effects on nutrient cycles in terrestrial ecosystems. However, our understanding of the patterns and the driving factors of soil N:P ratios in the Tibetan Plateau shrublands remains limited. Our study aimed to quantify the distribution of soil N:P ratio and its controlling factors based on soil, plant, and climate factors from 59 sites in shrublands across the northeast Tibetan Plateau. The kriging interpolation method was used to quantify the soil N:P distribution. Spatially, the soil N:P ratio was higher in the south than in the north and lower in the west than in the east. The soil N:P ratio in the northeast Tibetan Plateau shrublands was mainly explained by edaphic factors, which also played an important role in regulating the effects of plant and climate factors on soil N:P ratios. Mean annual precipitation, instead of mean annual temperature, significantly controlled the soil N:P ratios, and its effect on the pattern of soil N:P ratios differed between alpine shrublands and desert shrublands. The N:P ratios of different organs in shrublands also played different roles in shaping the soil N:P ratios in alpine and desert shrublands. These results provide support for the hypothesis that edaphic factors were the dominant drivers of spatial variation in soil N:P ratios across the northeast Tibetan Plateau shrublands, and our study contributes to a deeper understanding of biogeochemical cycling at high altitudes.

Although soil total nitrogen (STN) and soil total phosphorus (STP) play significant roles in terrestrial ecosystem function, their storage and driving factors in the alpine wetlands of the Qinghai-Tibetan Plateau remain unclear. In this study, we estimated STN and STP storage and their controlling factors, including vegetation, soil, and climate characteristics, using data collected from 50 sites across the wetlands in the Three Rivers Source Region. STN and STP storage in the top 30 cm of soil were 62.12 ± 37.55 Tg N and 9.24 ± 2.90 Tg P, respectively. Although STN density did not differ significantly for different vegetation types (i.e., alpine meadow and alpine wetland), belowground biomass showed a positive relationship with STN density. Mean annual precipitation (MAP) showed a significant positive relationship with STN density, whereas the effects of mean annual temperature on STN density were minor. Compared with the effects of vegetation and climatic factors, soil characteristics were found to not only exert a significant effect on STN density, but also influence the effects of climate and vegetation on STN density. For STP density, soil characteristics were found to be a significant controlling factor, whereas the effects of biomass and climatic factors were minor. The studied climate, soil, and vegetation characteristics jointly explained ∼54% of STN variance, whereas soil characteristics explained only 20% of STP variation. MAP indirectly affected STN density via effects on vegetation and soil, and its direct effect on STN density was minor. This indicated a strong relationship between biotic and abiotic effects and STN density. Identification of the factors influencing STN and STP variance in alpine wetlands contributes to our understanding of the biogeochemical cycle in high-altitude regions.

Abstract Periodic climatic oscillations and species dispersal during the postglacial period are two important causes of plant assemblage and distribution on the Qinghai‐Tibet Plateau (QTP). To improve our understanding of the bio‐geological histories of shrub communities on the QTP, we tested two hypotheses. First, the intensity of climatic oscillations played a filtering role during community structuring. Second, species dispersal during the postglacial period contributed to the recovery of species and phylogenetic diversity and the emergence of phylogenetic overdispersion. To test these hypotheses, we investigated and compared the shrub communities in the alpine and desert habitats of the northeastern QTP. Notably, we observed higher levels of species and phylogenetic diversity in the alpine habitat than in the desert habitat, leading to phylogenetic overdispersion in the alpine shrub communities versus phylogenetic clustering in the desert shrub communities. This phylogenetic overdispersion increased with greater climate anomalies. These results suggest that (a) although climate anomalies strongly affect shrub communities, these phenomena do not act as a filter for shrub community structuring, and (b) species dispersal increases phylogenetic diversity and overdispersion in a community. Moreover, our investigation of the phylogenetic community composition revealed a larger number of plant clades in the alpine shrub communities than in the desert shrub communities, which provided insights into plant clade‐level differences in the phylogenetic structures of alpine and desert shrub communities in the northeastern QTP.

Abstract Background Tea-oil tree (Camellia oleifera) is a unique edible-oil tree in China, and anthracnose occurs in wherever it is cultivated, causing great economic losses each year. We have previously identified the Ascomycete fungus Colletotrichum fructicola as the major pathogen of anthracnose in Ca. oleifera. The purpose of this study was to characterize the biological function of Snf1 protein, a key component of the AMPK (AMP-activated protein kinase) pathway, for the molecular pathogenic-mechanisms of C. fructicola. Results We characterized CfSnf1 as the homolog of Saccharomyces cerevisiae Snf1. Targeted CfSNF1 gene deletion revealed that CfSnf1 is involved in the utilization of specific carbon sources, conidiation, and stress responses. We further found that the ΔCfSnf1 mutant was not pathogenic to Ca. oleifera, resulting from its defect in appressorium formation. In addition, we provided evidence showing crosstalk between the AMPK and the cAMP/PKA pathways for the first time in filamentous fungi. Conclusion This study indicate that CfSnf1 is a critical factor in the development and pathogenicity of C. fructicola and, therefore, a potential fungicide target for anthracnose control.

Abstract There are two important allocation hypotheses in plant biomass allocation: allometric and isometric. We tested these two hypotheses in an alpine steppe using plant biomass allocation under nitrogen (N) addition and precipitation (Precip) changes at a community level. An in situ field manipulation experiment was conducted to examine the two hypotheses and the responses of the biomass to N addition (10 g N m−2 y−1) and altered Precip (±50% precipitation) in an alpine steppe on the Qinghai–Tibetan Plateau from 2013 to 2016. We found that the plant community biomass differed in its response to N addition and reduced Precip such that N addition significantly increased aboveground biomass (AGB), while reduced Precip significantly decreased AGB from 2014 to 2016. Moreover, reduced Precip enhanced deep soil belowground biomass (BGB). In the natural alpine steppe, the allocation between AGB and BGB was consistent with the isometric hypotheses. In contrast, N addition or altered Precip enhanced biomass allocation to aboveground, thus leading to allometric growth. More importantly, reduced Precip enhanced biomass allocation into deep soil. Our study provides insight into the responses of alpine steppes to global climate change by linking AGB and BGB allocation.

Anisodus tanguticus (Maxim.) Pascher, a medicinal plant growing in the Tibetan Plateau region with various medicinal values, is mainly used for the extraction of tropane alkaloids (TAs), and the increased demand for A. tanguticus has triggered its overexploitation. The cultivation of this plant is necessary for the quality control and conservation of wild resources. During 2020 and 2021, a split-plot experiment with three replicates was used to study different planting densities (D1: 30 × 50 cm; D2: 40 × 50 cm; D3: 50 × 50 cm; D4: 60 × 50 cm) and different growth periods (first withering period: October 2020; greening period: June 2021; growth period: August 2021; second withering period: October 2021) on the yield and alkaloid content (atropine, scopolamine, anisodamine, anisodine) of A. tanguticus. The results showed that the mass per plant of A. tanguticus was higher at low density, while the yield per unit area of the underground parts (25288.89 kg/ha) was greater at high density, and the mass of the aboveground parts (14933.33 kg/ha) was higher at low density. The anisodamine (0.0467%) and anisodine (0.1201%) content of D2 (40 cm × 50 cm) was significantly higher than that of the other densities during the green period. The content of all four alkaloids was highest during the greening period, and the scopolamine, anisodamine, and anisodine content was higher in the aboveground parts than in the underground parts. The total alkaloid accumulation per unit area of the whole plant reached its maximum value (1.08%, 139.48 kg/ha) in the growth period of D2; therefore, for economic efficiency and the selection of the best overall quality, it was concluded that the aboveground parts also had medicinal value, the growth period was the best harvesting period, and D2 (40 cm × 50 cm) was the best planting density for A. tanguticus.

Intercropping is a sustainable method for cultivating medicinal herbs since it requires lower dependence on chemical fertilizers than a sole cropping system. In this study, we compared the effects of sole cropping and intercropping on early bolting, yield, and the chemical composition of Angelica sinensis (Oliv.) Diels. Field experiments were conducted, in 2018 and in 2019, using different cropping systems including sole cropping of A. sinensis (AS), sole cropping of Vicia faba (VF), and intercropping (without fertilization) at three ratios: one row of A. sinensis + three rows of V. faba, AS/VF (1:3), two rows of A. sinensis + two rows V. faba, AS/VF (2:2), three rows of A. sinensis + one row V. faba, AS/VF (3:1). The effect of each cropping system was evaluated by measuring the dry biomass of V. faba and the dry biomass, ferulic acid content, and essential oil content and composition of A. sinensis. The early bolting rate of A. sinensis was significantly lower in the intercropping system as compared with that in a sole cropping system. The AS/VF (3:1) intercropping pattern resulted in an optimal yield and the highest ferulic acid content of A. sinensis, highest dry biomass of V. faba, and highest land equivalent ratio (LER). Additionally, the A. sinensis was more aggressive (the aggressivity value of A. sinensis was positive, and its competitive ratio was >1) under AS/VF (3:1) intercropping pattern, and it dominated over V. faba (which had negative aggressivity values and a competitive ratio of A. sinensis, regardless of the cropping system; however, the chemical component of essential oil was not influenced by intercropping patterns. Overall, the AS/VF (3:1) intercropping pattern without fertilization was the most productive, with the highest LER and ferulic acid content. These data indicate that intercropping can serve as an alternative for reducing the use of chemical fertilizers and intercropping also decreases the early bolting rate of A. sinensis, thus, enabling its sustainable production.

Astrocytes have distinctive morphological and functional characteristics, and are found throughout the central nervous system. Astrocytes are now known to be far more than just housekeeping cells in the brain. Their functions include contributing to the formation of the blood–brain barrier, physically and metabolically supporting and communicating with neurons, regulating the formation and functions of synapses, and maintaining water homeostasis and the microenvironment in the brain. Aquaporins (AQPs) are transmembrane proteins responsible for fast water movement across cell membranes. Various subtypes of AQPs (AQP1, AQP3, AQP4, AQP5, AQP8 and AQP9) have been reported to be expressed in astrocytes, and the expressions and subcellular localizations of AQPs in astrocytes are highly correlated with both their physiological and pathophysiological functions. This review describes and summarizes the recent advances in our understanding of astrocytes and AQPs in regard to controlling water homeostasis in the brain. Findings regarding the features of different AQP subtypes, such as their expression, subcellular localization, physiological functions, and the pathophysiological roles of astrocytes are presented, with brain edema and glioma serving as two representative AQP-associated pathological conditions. The aim is to provide a better insight into the elaborate “water distribution” system in cells, exemplified by astrocytes, under normal and pathological conditions.

Bupleurum species are well-known for their antipyretic, analgesic, gallbladder and other functions so that they are very popular in traditional Chinese medicine. However, to our knowledge, there is no completed chloroplast genome of Bupleurum genus in China. In the present study, we determined the complete chloroplast genome sequences for Bupleurum longicaule var. strictum using IIumina sequencing. Bupleurum longicaule var. strictum is 155,578 bp which is composed of two inverted repeats (IR: 26,257bp), a large single copy region (LSC: 86,977 bp), and a small single-copy region (SSC: 16,087 bp). The overall AT content is 65.32%. The chloroplast genome includes a total of 51 functional genes including 15 protein-coding genes and 36 tRNA. A total of 10 genes were duplicated in the IR regions including seven tRNA and three protein-coding genes. Phylogenetic analysis suggested that B. longicaule var. strictum formed a monophyletic clade.

Endophytes represent a ubiquitous and magical world in plants. Almost all plant species studied by different researchers have been found to harbor one or more endophytes, which protect host plants from pathogen invasion and from adverse environmental conditions. They produce various metabolites that can directly inhibit the growth of pathogens and even promote the growth and development of the host plants. In this review, we focus on the biological control of plant diseases, aiming to elucidate the contribution and key roles of endophytes and their metabolites in this field with the latest research information. Metabolites synthesized by endophytes are part of plant disease management, and the application of endophyte metabolites to induce plant resistance is very promising. Furthermore, multi-omics should be more fully utilized in plant–microbe research, especially in mining novel bioactive metabolites. We believe that the utilization of endophytes and their metabolites for plant disease management is a meaningful and promising research direction that can lead to new breakthroughs in the development of more effective and ecosystem-friendly insecticides and fungicides in modern agriculture.

As an important Tibetan medicine and a secondary protected plant in China, Pomatosace filicula is endemic to the country and is mainly distributed in the Qinghai–Tibet Plateau (QTP). However, global climate change caused by greenhouse gas emissions might lead to the extinction of P. filicula. To understand the potential spatial distribution of P. filicula in future global warming scenarios, we used the MaxEnt model to simulate changes in its suitable habitat that would occur by 2050 and 2070 using four representative concentration pathway (RCP) scenarios and five global climate models. The results showed that the QTP currently contains a suitable habitat for P. filicula and will continue to do so in the future. Under the RCP2.6 scenario, the suitable habitat area would increase by 2050 but shrink slightly by 2070, with an average reduction of 2.7%. However, under the RCP8.5 scenario, the area of unsuitable habitat would expand by an average of 54.65% and 68.20% by 2050 and 2070, respectively. The changes in the area of suitable habitat under the RCP4.5 and RCP6.0 scenarios were similar, with the unsuitable area increasing by approximately 20% by 2050 and 2070. Under these two moderate RCPs, the total suitable area in 2070 would be greater than that in 2050. The top three environmental factors impacting the habitat distribution were altitude, annual precipitation (BIO12) and annual temperature range (BIO7). The cumulative contribution rate of these three factors was as high as 82.8%, indicating that they were the key factors affecting the distribution and adaptability of P. filicula, P. filicula grows well in damp and cold environments. Due to global warming, the QTP will become warmer and drier; thus, the growing area of P. filicula will move toward higher elevations and areas that are humid and cold. These areas are mainly found near the Three-River Region. Future climate change will aggravate the deterioration of the P. filicula habitat and increase the species’ survival risk. This study describes the distribution of P. filicula and provides a basis for the protection of endangered plants in the QTP.

Global climate change and nitrogen deposition have been having broad impacts on microorganisms. On the Qinghai-Tibetan Plateau (QTP), the responses of soil microbial community assemblage and diversity to nitrogen deposition and changes in precipitation are poorly understood, especially in the alpine steppe. In this study, we conducted a field manipulative experiment of nitrogen deposition and precipitation amount in an alpine steppe on the northeastern QTP and investigated the responses of community composition, diversity, and community assemblage of soil fungi. Soil fungal community compositions were significantly altered under nitrogen addition, precipitation change, and their interaction, and positively related with soil moisture, soil pH, and plant species richness. However, they were negatively related to soil mineralizable N and soil available P content. Operational taxonomic units (OTU) richness and Chao 1 index decreased under nitrogen addition combined with precipitation reduction treatment, whereas the Shannon–Wiener index declined only under precipitation increment treatment. Convergent fungal community assembly processes were not acutely altered by both nitrogen addition and precipitation changes, indicating that environmental filtering was a dominant ecological process controlling fungal community assemblage. By elucidating the above questions, the study enhanced our ability to predict the responses of soil fungal communities to nitrogen deposition and precipitation changes at alpine steppes on the QTP in the future.