Targeting the AMPK/Nrf2 Pathway: A Novel Therapeutic Approach for Acute Lung Injury.

ALI(acute lung injury) is a severe respiratory dysfunction caused by various intrapulmonary and extrapulmonary factors. It is primarily characterized by oxidative stress and affects the integrity of the pulmonary barrier. In severe cases, ALI can progress to ARDS(acute respiratory distress syndrome), a condition that poses a serious threat to the lives of affected patients. To date, the etiological mechanisms underlying ALI remain elusive, and available therapeutic options are quite limited. AMPK(AMP-activated protein kinase), an essential serine/threonine protein kinase, performs a pivotal function in the regulation of cellular energy levels and cellular regulatory mechanisms, including the detection of redox signals and mitigating oxidative stress. Meanwhile, Nrf2(nuclear factor erythroid 2-related factor 2), a critical transcription factor, alleviates inflammation and oxidative responses by interacting with multiple signaling pathways and contributing to the modulation of oxidative enzymes associated with inflammation and programmed cell death. Indeed, AMPK induces the dissociation of Nrf2 from Keap1(kelch-like ECH-associated protein-1) and facilitates its translocation into the nucleus to trigger the transcription of downstream antioxidant genes, ultimately suppressing the expression of inflammatory cells in the lungs. Given their roles, AMPK and Nrf2 hold promise as novel treatment targets for ALI. This study aimed to summarise the current status of research on the AMPK/Nrf2 signaling pathway in ALI, encompassing recently reported natural compounds and drugs that can activate the AMPK/Nrf2 signaling pathway to alleviate lung injury, and provide a theoretical reference for early intervention in lung injury and future research on lung protection.

Effects of Differential Shading on Summer Tea Quality and Tea Garden Microenvironment.

Shading is an effective agronomic technique to protect tea plants from intense sunlight. However, there are currently very few studies on more effective shading methods to improve the quality of summer tea. In this study, 'Longjing43' plants were grown under four different shading treatments for 14 days, with no shading as the control. Among the four shading treatments, double-layer-net shadings had the most positive impact on the tea quality, resulting in higher levels of amino acids but lower levels of tea polyphenols. Additionally, double-layer-net shadings provided more suitable microenvironments for tea plants. The tea leaves in T4 (double nets 50 cm above the plant canopy) contained 16.13 mg∙g-1 of umami and sweet amino acids, which was significantly higher than in other treatments. T4 had the lowest air temperature and the most suitable and stable soil water content. Interestingly, the ratio of red light to far-red light in T4 was only 1.65, much lower than other treatments, which warrants further study. In conclusion, the microenvironment induced by shading can greatly affect the tea quality, and double-layer-net shading is better for improving the quality of summer tea.

Patterns and abiotic drivers of soil organic carbon in perennial tea (Camellia sinensis L.) plantation system of China.

Understanding soil organic carbon (SOC), the largest carbon (C) pool of a terrestrial ecosystem, is essential for mitigating climate change. Currently, the spatial patterns and drivers of SOC in the plantations of tea, a perennial leaf crop, remain unclear. Therefore, the present study surveyed SOC across the main tea-producing areas of China, which is the largest tea producer in the world. We analyzed the soil samples from tea plantations under different scenarios, such as provinces, regions [southwest China (SW), south China (SC), south Yangtze (SY), and north Yangtze (NY)], climatic zones (temperate, subtropical, and tropical), and cultivars [large-leaf (LL) and middle or small-leaf (ML) cultivars]. Preliminary analysis revealed that most tea-producing areas (45%) had SOC content ranging from 10 to 20 g kg-1. The highest SOC was recorded for Yunnan among the various provinces, the SW tea-producing area among the four regions, the tropical region among the different climatic zones, and the areas with LL cultivars compared to those with ML cultivars. Further Pearson correlation analysis demonstrated significant associations between SOC and soil variables and random forest modeling (RF) identified that total nitrogen (TN) and available aluminum [Ava(Al)] of soil explained the maximum differences in SOC. Besides, a large indirect effect of geography (latitude and altitude) on SOC was detected through partial least squares path modeling (PLS-PM) analysis. Thus, the study revealed a high spatial heterogeneity in SOC across the major tea-producing areas of China. The findings also serve as a basis for planning fertilization strategies and C sequestration policies for tea plantations.

Nitrogen addition reduces phosphorus availability and induces a shift in soil phosphorus cycling microbial community in a tea (Camellia sinensis L.) plantation.

Nitrogen (N) and phosphorus (P) are two important nutrient elements that limit the growth of plants and microorganisms. The effect of the N supply on soil P cycling and its mechanism remain poorly known. Here, we characterized the effects of different N application rates on soil P availability, the abundances of P-cycling functional genes, and microbial communities involved in P-cycling following the application of N for 13 years in a tea plantation. Soil available P (AP) decreased significantly under N application. The opposite pattern was observed for the activity of soil phosphatases including alkaline (ALP) and acid phosphatase (ACP). Furthermore, N addition increased the abundance of ppa but decreased the abundance of phoD in soil. Both ppa- and phoD-harboring communities varied with N application levels. Redundancy analysis (RDA) showed that soil pH was a key variable modulating ppa-harboring and phoD-harboring microbial communities. Partial least squares path modeling (PLS-PM) revealed that long-term N application indirectly reduced soil P availability by altering the abundances of phoD-harboring biomarker taxa. Overall, our findings indicated that N-induced reductions in AP increased microbial competition for P by selecting microbes with P uptake and starvation response genes or those with phosphatases in tea plantation system. This suggests that tea plantations should be periodically supplemented with P under N application, especially under high N application levels.

Magnesium promotes tea plant growth via enhanced glutamine synthetase-mediated nitrogen assimilation.

Acidic tea (Camellia sinensis) plantation soil usually suffers from magnesium (Mg) deficiency, and as such, application of fertilizer containing Mg can substantially increase tea quality by enhancing the accumulation of nitrogen (N)-containing chemicals such as amino acids in young tea shoots. However, the molecular mechanisms underlying the promoting effects of Mg on N assimilation in tea plants remain unclear. Here, both hydroponic and field experiments were conducted to analyze N, Mg, metabolite contents, and gene expression patterns in tea plants. We found that N and amino acids accumulated in tea plant roots under Mg deficiency, while metabolism of N was enhanced by Mg supplementation, especially under a low N fertilizer regime. 15N tracing experiments demonstrated that assimilation of N was induced in tea roots following Mg application. Furthermore, weighted gene correlation network analysis (WGCNA) analysis of RNA-seq data suggested that genes encoding glutamine synthetase isozymes (CsGSs), key enzymes regulating N assimilation, were markedly regulated by Mg treatment. Overexpression of CsGS1.1 in Arabidopsis (Arabidopsis thaliana) resulted in a more tolerant phenotype under Mg deficiency and increased N assimilation. These results validate our suggestion that Mg transcriptionally regulates CsGS1.1 during the enhanced assimilation of N in tea plant. Moreover, results of a field experiment demonstrated that high Mg and low N had positive effects on tea quality. This study deepens our understanding of the molecular mechanisms underlying the interactive effects of Mg and N in tea plants while also providing both genetic and agronomic tools for future improvement of tea production.

[CH4 Fluxes and Their Comprehensive Greenhouse Effects with CO2 Fluxes in Direct-seeded Rice in Poyang Lake Plain].

Paddy fields are complex ecosystems that both emit CH4 and absorb CO2, which plays an important role in the global water-carbon cycle and carbon budget. In this study, the CH4 fluxes and CO2 fluxes of double-cropping direct-seeded rice fields in 2020 in the Poyang Lake Plain were obtained using the eddy covariance method, and the variation characteristics, accumulation in the whole growth period, and comprehensive greenhouse effects of two greenhouse gases were quantitatively revealed. The results showed that, the double-cropping direct-seeded rice field in Poyang Lake Plain was the source of CH4 emission, and the emission during the whole growth period was 52.6 g·m-2, with an average daily emission of 0.208 g·(m2·d)-1. CH4 emission and daily average emission in the early rice season were 20.7 g·m-2 and 0.188 g·(m2·d)-1, respectively, which were lower than the emissions of 31.9 g·m-2 and 0.255 g·(m2·d)-1 in the late rice season. CH4 flux had significant seasonal variation characteristics. The strong emission period (emission peak) of CH4 was concentrated in the middle growth stage of early rice and the early growth stage of late rice. A total of 85.5% of CH4 in the early rice season and 92.1% of CH4 in the late rice season were released during the strong emission periods, and seasonal peak values were 0.638 g·(m2·d)-1 and 1.282 g·(m2·d)-1, respectively. The diurnal variation characteristics of CH4 flux showed three types:obvious unimodal type, non-obvious unimodal type, and irregular type. The strong emission period was mainly the unimodal type, and the peak values of 0.453 µmol·(m2·s)-1 in the early rice season and 0.977 µmol·(m2·s)-1 in the late rice season appeared at 14:00-15:00 and maintained a high emission rate at 12:30-16:00. The CO2 accumulation in the whole growth period of early rice and late rice was -990.4 g·m-2 and -1156.6 g·m-2, respectively, and the total was -2147.0 g·m-2. The comprehensive greenhouse effect of CH4 emission and CO2 exchange in the double-cropping paddy field was -673.6 g·m-2 (calculated using the CO2 equivalent), which showed a cooling effect. Excluding CH4 emissions when evaluating the greenhouse effect of the paddy field, the CO2 equivalent emission of 1473.4 g·m-2 would be underestimated, accounting for 68.6% of the net CO2 absorption. Considering CH4 emissions, CO2 exchanges, and carbon emissions caused by rice harvest, the two-season direct seeding paddy field in Poyang Lake Plain was the source of greenhouse gas emissions.

Metagenomics reveals N-induced changes in carbon-degrading genes and microbial communities of tea (Camellia sinensis L.) plantation soil under long-term fertilization.

Soil organic carbon (SOC) is an important C pool of the global ecosystem and is affected by various agricultural practices including fertilization. Excessive nitrogen (N) application is an important field management measure in tea plantation systems. However, the mechanism underlying the impact of N fertilization on SOC, especially the microscopic mechanism remain unclear. The present study explored the effects of N fertilization on C-cycling genes, SOC-degrading enzymes and microbes expressing these enzymes by using a metagenomic approach in a tea plantation under long-term fertilization with different N rates. Results showed that N application significantly changed the abundance of C-cycling genes, SOC-degrading enzymes, especially those associated with labile and recalcitrant C degradation. In addition, the beta-glucosidase and chitinase-expressing microbial communities showed a significant difference under different N rates. At the phylum level, microbial taxa involved in C degradation were highly similar and abundant, while at the genus level, only specific taxa performed labile and recalcitrant C degradation; these SOC-degrading microbes were significantly enriched under N application. Redundancy analysis (RDA) revealed that the soil and pruned litter properties greatly influenced the SOC-degrading communities; pH and DOC of the soil and biomass and total polyphenol (TP) of the pruned litter exerted significant effects. Additionally, the random forest (RF) algorithm revealed that soil pH and dominant taxa efficiently predicted the beta-glucosidase abundance, while soil pH and DOC, pruned litter TP, and the highly abundant microbial taxa efficiently predicted chitinase abundance. Our study indicated that long-term N fertilization exerted a significant positive effect on SOC-degrading enzymes and microbes expressing these enzymes, resulting in potential impact on soil C storage in a perennial tea plantation ecosystem.

Long-term nitrogen addition increases denitrification potential and functional gene abundance and changes denitrifying communities in acidic tea plantation soil.

The response of soil denitrification to nitrogen (N) addition in the acidic and perennial agriculture systems and its underlying mechanisms remain poorly understood. Therefore, a long-term (12 years) field trial was conducted to explore the effects of different N application rates on the soil denitrification potential (DP), functional genes, and denitrifying microbial communities of a tea plantation. The study found that N application to the soil significantly increased the DP and the absolute abundance of denitrifying genes, such as narG, nirK, norB, and nosZ. The diversity of denitrifying communities (genus level) significantly decreased with increasing N rates. Moreover, the denitrifying communities composition significantly differed among the soils with different rates of N fertilization. Further variance partitioning analysis (VPA) revealed that the soil (39.04%) and pruned litter (32.53%) properties largely contributed to the variation in the denitrifying communities. Dissolved organic carbon (DOC) and soil pH, pruned litter's total crude fiber (TCF) content and total polyphenols to total N ratio (TP/TN), and narG and nirK abundance significantly (VIP >1.0) influenced the DP. Finally, partial least squares path modeling (PLS-PM) revealed that N addition indirectly affected the DP by changing specific soil and pruned litter properties and functional gene abundance. Thus, the findings suggest that tea plantation is a major source of N2O emissions that significantly enhance under N application and provide theoretical support for N fertilizer management in an acidic tea plantation system.

[Status of Heavy Metal in Organic Fertilizers in Main Tea Growing Regions of China].

To provide guidance for the safe use of organic fertilizers and improve soil quality and tea safety, it is necessary to conduct systematic analyses of the heavy metal content of organic fertilizers applied in the main tea producing areas of China. In this study, we analyzed the heavy metal contents in organic fertilizer samples collected from 2017 to 2019. The risks of collected organic fertilizers from different areas and sources were calculated. The results showed that the average concentrations of ω(As), ω(Hg), ω(Pb), ω(Cd), ω(Cr), ω(Cu), ω(Zn), and ω(Ni) in the collected organic fertilizers were 4.60, 0.22, 27.1, 0.78, 27.9, 58.3, 250.1, and 16.3 mg·kg-1, respectively. According to the assessment standard in NY/T 525- 2021, the over-limit rates of As, Hg, Pb, Cd, and Cr were 6.19%, 1.33%, 4.42%, 4.42%, and 1.33%, respectively. With respect to the area, the qualified rates were 100% in Shaanxi, Jiangsu, Anhui, Fujian, and Guangxi; 80%-90% in Shandong, Zhejiang, Hubei, Sichuan, Yunnan, and Guangdong; and only 54.5% in Jiangxi. The qualified rates of sources were 100% in rapeseed cake, soybean cake, and pig manure; 95.8% in sheep manure; 91.7% in cow manure; 90.7% in chicken manure; 87.2% in manure of other animals; 82.4% in the mixture of plant and animal sources; 65.2% in other plant sources; and 63.6% in other sources. According to the recommended application rate, the accumulation rate of heavy metals in soil with pig manure, cow manure, chicken manure, and sheep manure would be much higher than that with rapeseed cake and soybean cake. The average accumulation rate of organic fertilizer from animal sources was 7-30 times higher than that from plant sources. Therefore, it is recommended to use rapeseed cake or soybean cake fertilizer in tea plantation and to increase the supervision of heavy metal accumulation in soil and tea in those high-risk areas.

Ecological management model for the improvement of soil fertility through the regulation of rare microbial taxa in tea (Camellia sinensis L.) plantation soils.

Agricultural management is essential to enhance soil ecosystem service function through optimizing soil physical conditions and improving nutrient supply, which is predominantly regulated by soil microorganisms. Several studies have focused on soil biodiversity and function in tea plantation systems. However, the effects of different agriculture managements on soil fertility and microbes remain poorly characterized, especially for what concerns perennial agroecosystems. In this study, 40 soil samples were collected from 10 tea plantation sites in China to explore the effects of ecological and conventional managements on soil fertility, as well as on microbial diversity, community composition, and co-occurrence network. Compared with conventional management, ecological management was found to significantly enhance soil fertility, microbial diversity, and microbial network complexity. Additionally, a significant difference in community composition was clearly observed under the two agriculture managements, especially for rare microbial taxa, whose relative abundance significantly increased under ecological management. Random forest modeling revealed that rare taxa (e.g., Rokubacteria and Mortierellomycota), rather than dominant microbial taxa (e.g., Proteobacteria and Ascomycota), were key variables for predicting soil fertility. This indicates that rare taxa might play a fundamental role in biological processes. Overall, our results suggest that ecological management is more efficient than conventional management in regulating rare microbial taxa and maintaining a good soil fertility in tea plantation systems.

Foliar N Application on Tea Plant at Its Dormancy Stage Increases the N Concentration of Mature Leaves and Improves the Quality and Yield of Spring Tea.

Over 30% of the Chinese tea plantation is supplied with excess fertilizer, especially nitrogen (N) fertilizer. Whether or not foliar N application on tea plants at the dormancy stage could improve the quality of spring tea and be a complementary strategy to reduce soil fertilization level remains unclear. In this study, the effects of foliar N application on tea plants were investigated by testing the types of fertilizers and their application times, and by applying foliar N under a reduced soil fertilization level using field and 15N-labeling pot experiments. Results showed that the foliar N application of amino acid liquid fertilizer two times at the winter dormancy stage was enough to significantly increase the N concentration of the mature leaves and improved the quality of spring tea. The foliar application of 2% urea or liquid amino acid fertilizer two times at the winter dormancy stage and two times at the spring dormancy stage showed the best performance in tea plants among the other foliar N fertilization methods, as it reduced the soil fertilization levels in tea plantations without decreasing the total N concentration of the mature leaves or deteriorating the quality of spring tea. Therefore, foliar N application on tea plants at its dormancy stage increases the N concentration of the mature leaves, improves the quality and yield of spring tea, and could be a complementary strategy to reduce soil fertilization levels.

Heavy nitrogen application increases soil nitrification through ammonia-oxidizing bacteria rather than archaea in acidic tea (Camellia sinensis L.) plantation soil.

Nitrogen (N) fertilizer is widely used in agricultural ecosystems and influences N transformation processes in the soil such as nitrification. However, whether nitrification is primarily dominated by ammonia-oxidizing bacteria (AOB) or archaea (AOA) under heavy N application is still under debate. In the present work, the effect of long-term (12 years) N fertilization on soil nitrification and the key influencing factors were investigated in acidic tea plantation soil that received four different rates of N application (0, 119, 285, and 569 kg N ha-1 yr-1). Nitrification potential was measured and partitioned using chemical inhibitors. The abundance of functional genes involved in ammonia oxidation was quantified using quantitative polymerase chain reaction (qPCR). Ammonia-oxidizing communities were identified by shotgun metagenome sequencing. Potential nitrification rate in tea plantation soil was mainly dominated by autotrophic nitrification (PNRA) (71-79%). PNRA and heterotrophic nitrification (PNRH) were both significantly increased by heavy N (569 kg ha-1) application. Moreover, PNRA was mainly due to the contribution of AOB (52-66%) in N-treated soils, and N569 significantly increased the AOB contribution without affecting the AOA contribution. N569 increased the functional gene abundance of AOB and TAO100 (a non-halophilic γ-AOB) but decreased that of AOA. The dominant AOB (Nitrosomonas, Nitrosospira, and Nitrosococcus), AOA (Nitrososphaera and Nitrosopumilus) and commamox (Nitrospira) groups were profoundly altered by long-term N application rates. Partial least squares regression showed that total nitrification (PNRT), PNRA, and PNRAOB were primarily explained by the functional gene abundance of nitrifiers whereas PNRH and PNRAOA were closely associated with soil and pruned litter properties. Moreover, structural equation modeling (SEM) revealed that long-term N application significantly and indirectly affected nitrification potential by directly influencing soil properties, pruned litter properties, and functional gene abundance. Understanding the relative contribution of AOA and AOB to nitrification may help to better regulate N fertilizer use in agricultural ecosystems.

Preferential assimilation of NH4+ over NO3- in tea plant associated with genes involved in nitrogen transportation, utilization and catechins biosynthesis.

Physiological effects of ammonium (NH4+) and nitrate (NO3-) on tea have confirmed that tea plants prefer NH4+ as the dominant nitrogen (N) source. To investigate the possible explanations for this preference, studies of 15NH4+ and 15NO3- assimilation using hydroponically grown tea plants were conducted. During the time course of 15NH4+ and 15NO3- assimilation, the absorption of 15N from 15NH4+ was more rapid than that from 15NO3-, as there was a more efficient expression pattern of NH4+ transporters compared with that of NO3- transporters. 15NH4+-fed tea plants accumulated more 15N than 15NO3- fed plants, which was demonstrated by that genes related to primary N assimilation, like CsNR, CsNiR, CsGDH and CsGOGAT, were more affected by 15NH4+ than 15NO3-. Markedly higher NH4+ concentrations were observed in 15NH4+-fed tea roots in comparison with NO3- treatment, whereas tea plants maintained a balanced concentration of NH4+ in tea leaves under both these two N forms. This maintenance was achieved through the increased expression of genes involved in theanine biosynthesis and the inhibition of genes related to catechins derived from phenylpropanoid pathway. The current results suggest that efficient NH4+ transportation, assimilation, and reutilization enables tea plant as an ammonium preferring plant species.

Stimulated biosynthesis of delphinidin-related anthocyanins in tea shoots reducing the quality of green tea in summer.

BACKGROUND: Greater proportions of purple tea buds and leaves usually appear in the summer, which seriously affects the color and taste quality of green tea products, yet the metabolism of purple tea shoots in summer remains unclear. Here, the metabolomic profiles and gene expression of related flavonoid metabolic pathways in the purple and normal green shoots of 'Longjing 43', and the quality of green tea made with these two phenotypes, were analyzed and compared. RESULTS: Differential metabolites identified using high-performance liquid chromatography-Orbitrap/mass spectrometry indicated that anthocyanin biosynthesis in purple leaves was enriched, with higher levels of anthocyanidins (delphinidin-hexose-coumaroyl showed the greatest increase), proanthocyanidins (oligomers of catechins) and kaempferol glycoside. Expression patterns of the genes ANR, ANS, FLS, LAR, C4H, PAL, CHI, CHS and DFR revealed that the metabolism of anthocyanin is positively regulated by high temperature and/or light levels in summer. Gas chromatography-mass spectrometry results showed that, in purple tea shoots, the metabolism of carbohydrates was enriched whereas that of amino acids was diminished, while their mannose, fructose, d-galactose, sorbose and d-glucose contents were more than double those found in green leaves. A sensory evaluation confirmed that a greater quantity of purple shoots had a greater negative impact on green tea quality because of a bitter taste and dark color (leaves and infusions were tested). CONCLUSIONS: These results highlight the need for and possibility of improving commercial tea quality via cultivation that controls the temperature or light of tea gardens during the summer. © 2019 Society of Chemical Industry.

Short-term inhibition of glutamine synthetase leads to reprogramming of amino acid and lipid metabolism in roots and leaves of tea plant (Camellia sinensis L.).

BACKGROUND: Nitrogen (N) nutrition significantly affected metabolism and accumulation of quality-related compounds in tea plant (Camellia sinensis L.). Little is known about the physiological and molecular mechanisms underlying the effects of short-term repression of N metabolism on tea roots and leaves for a short time. RESULTS: In this study, we subjected tea plants to a specific inhibitor of glutamine synthetase (GS), methionine sulfoximine (MSX), for a short time (30 min) and investigated the effect of the inhibition of N metabolism on the transcriptome and metabolome of quality-related compounds. Our results showed that GS activities in tea roots and leaves were significantly inhibited upon MSX treatment, and both tissue types showed a sensitive metabolic response to GS inhibition. In tea leaves, the hydrolysis of theanine decreased with the increase in theanine and free ammonium content. The biosynthesis of all other amino acids was repressed, and the content of N-containing lipids declined, suggesting that short-term inhibition of GS reduces the level of N reutilization in tea leaves. Metabolites related to glycolysis and the tricarboxylic acid (TCA) cycle accumulated after GS repression, whereas the content of amino acids such as glycine, serine, isoleucine, threonine, leucine, and valine declined in the MXS treated group. We speculate that the biosynthesis of amino acids is affected by glycolysis and the TCA cycle in a feedback loop. CONCLUSIONS: Overall, our data suggest that GS repression in tea plant leads to the reprogramming of amino acid and lipid metabolic pathways.

Characterization of three different classes of non-fermented teas using untargeted metabolomics.

Non-fermented teas, which are widely consumed in China, Japan, Korea, and elsewhere, have refreshing flavors and valuable health benefits. Various types of non-fermented teas look and taste similar and have no obvious differences in appearance, making their classification challenging. To date, there are very few reports about characterization and discrimination of different types of non-fermented teas. To characterize non-fermented teas and build a standard model for their classification based on their chemical composition, we employed multi-platform-based metabolomics to analyze primary and secondary metabolites in three main categories of non-fermented teas (green, yellow, and white), using 96 samples collected from China. Five hundred and ninety unique tea metabolites were identified and quantified in these three types of teas. Moreover, a partial least squares discriminant analysis (PLS-DA) model was established based on metabolomics data, in order to classify non-fermented teas into these three classes. Furthermore, our results speculate that the health benefits (e.g., antioxidant content) of these three types of non-fermented tea differ primarily because of variation in their metabolic components (e.g., ascorbate, vitexin).

Effects of nitrogen supply on flavonol glycoside biosynthesis and accumulation in tea leaves (Camellia sinensis).

Widely distributed in tea plants, the flavonoid flavonol and its glycosylated derivatives have important roles in determining tea quality. However, the biosynthesis and accumulation of these compounds has not been fully studied, especially in response to nitrogen (N) supply. In the present study, 'Longjing 43' potted tea seedlings were subjected to N deficiency (0g/pot), normal N (4g/pot) or excess N (16g/pot). Quantitative analyses using Ultra Performance Liquid Chromatography-Triple Quadrupole Mass Spectrometry (UPLC-QqQ-MS/MS) revealed that most flavonol glycosides (e.g., Quercetin-3-glucoside, Kaempferol-3-rgalactoside and Kaempferol-3-glucosyl-rhamnsoyl-glucoside) accumulated to the highest levels when treated with normal N. Results from metabolomics using Gas Chromatography-Mass Spectrometer (GC-MS) suggested that the levels of carbohydrate substrates of flavonol glycosides (e.g., sucrose, sucrose-6-phosphate, D-fructose 1,6-bisphosphate and glucose-1-phosphate) were positively correlated with flavonol glycoside content in response to N availability. Furthermore, Quantitative Real-time PCR analysis of 28 genes confirmed that genes related to flavonoid (e.g., flavonol synthase 1, flavonol 3-O-galactosyltransferase) and carbohydrate (e.g., sucrose phosphate synthase, sucrose synthase and glucokinase) metabolism have important roles in regulating the biosynthesis and accumulation of flavonol glycosides. Collectively, our results suggest that normal N levels promote the biosynthesis of flavonol glycosides through gene regulation and the accumulation of substrate carbohydrates, while abnormal N availability has inhibitory effects, especially excess N.

Transcriptomic and ionomic analysis provides new insight into the beneficial effect of Al on tea roots' growth and nutrient uptake.

KEY MESSAGE: Transcriptome profiling of roots indicated that genes involved in cell wall modification, cytoskeleton, H+ exchange and K+ influx played important roles in tea root growth under Al addition. Tea (Camellia sinensis) is considered as an Al accumulator species. It can accumulate a high concentration of Al in mature leaves without any symptom of toxicity, even improve roots' growth and nutrient uptake. However, the molecular mechanisms underlying this tolerance remain unclear. Here, we investigated the accumulation of elements and transcriptional profiles in tea roots treated with various Al doses. The results showed that the growth of tea plants was improved by a low dose of Al (0.2, 0.4, 0.6, 1 mM); however, this beneficial effect disappeared when higher concentrations of Al were supplied (2, 4, 10 mM). Ionomic analysis suggested that accumulation of P and K increased under a low Al supply (< 1 mM), while Ca and Mg contents were negatively correlated with external Al doses. The RNA seq obtained 523,391 unigenes, among which 20,448 were annotated in all databases. In total, 1876 unigenes were expressed significantly different in any Al treatment. A large number of DEGs involved in cell growth and division, such as those linked to cell wall-modifying enzymes, actin cytoskeleton, cyclin and H+-ATPase were identified, suggesting that these pathways were involved in root growth under different Al supply. Furthermore, expression of transporters significantly changed in roots supplied with Al. Among them, HAK5, which is involved in K uptake by plants, had a significant positive correlation with the K content.

Improved test to determine design parameters for optimization of free surface flow constructed wetlands.

Orthogonal tests were performed to assess the effect of design parameters on hydraulic and treatment performances of constructed wetlands. The results showed that water depth, layout of in- and outlet, flow rate, and aspect ratio mainly affected hydraulic performance, and water depth, plant spacing, and layout of in- and outlet mainly affected treatment performance. Optimal integrated performance was achieved with combination of 20-30 cm water depth, five evenly distributed inlets and one middle outlet, a flow rate of 0.4-0.55 m3/h, 20-cm plant spacing, a 1.125:1 aspect ratio, and planted with Scripus tabernaemontani. The average treatment performances of 27.2%, 16.3%, and 30.7% removal rates were received for total nitrogen, total phosphorus, and total suspended solid, respectively. The design parameters that significantly influenced hydraulic performance did not significantly influence treatment performance. Various hydraulic and purification indicators displayed extremely significant correlations. There was a significant correlation between hydraulic performance and mass removal capacity.

Geographical traceability of Chinese green tea using stable isotope and multi-element chemometrics.

RATIONALE: Deliberate and fraudulent origin mislabeling of Chinese green tea motivated by large price differences often brings significant food safety risks and damages consumer trust. Currently, there is no reliable method to verify the origin of green tea produced in China. Stable isotope and multi-element analyses combined with statistical models are widely acknowledged as useful traceability techniques for many agro-products, and could be developed to confirm the geographical origin of Chinese green tea and, more importantly, combat illegal green tea mislabeling and fraud. METHODS: An analytical strategy combining elemental analyzer/isotope ratio mass spectrometry (EA/IRMS) and inductively plasma coupled mass spectrometry (ICP-MS) with chemometrics tools was used to confirm the origin of green tea grown in the main tea production provinces around China. Stable C, N, H, O isotope ratios and twenty elements were measured to build mathematical discriminant models using unsupervised principal component analysis (PCA) and supervised linear discriminant analysis (LDA). Two main problems: (i) tracing the origin of Chinese green tea from different tea growing provinces (Zhejiang, Shandong, and other provinces); (ii) authentication of high-value Westlake Longjing tea from the Westlake region and surrounding areas in Zhejiang province, were investigated and assessed. RESULTS: The results demonstrated that PCA and follow-up LDA based on stable isotope and multi-element signatures can verify the geographical origin of Chinese green tea from different provinces, and even localized zones in the same province could be distinguishable, with discrimination accuracies higher than 92.3% and 87.8%, respectively. CONCLUSIONS: Geochemical fingerprinting techniques coupled with chemometric tools offer an accurate and effective verification method for the geographical origin of Chinese green tea, providing a promising tool to combat fraudulent mislabeling of high-value green tea.