Effects of different nitrogen application rates and picking batches on the nutritional components of Lycium barbarum L. fruits.

Lycium barbarum L., commonly known as wolfberry, is not only a traditional Chinese medicine but also a highly nutritious food. Its main nutrients include L. barbarum polysaccharide, flavonoid polyphenols, carotenoids, alkaloids, and other compounds, demonstrating its wide application value. This study investigated the effects of nitrogen application on the accumulation of the main nutrients and metabolites in wolfberry fruits under three different nitrogen application rates, namely, N1 (20% nitrogen (N) reduction, 540 kg·ha-2), N2 (medium N, 675 kg·ha-2), and N3 (20% nitrogen increase, 810 kg·ha-2,which is a local conventional nitrogen application amount.). Additionally, due to continuous branching, blossoming, and fruiting of wolfberry plants during the annual growth period, this research also explored the variation in nutritional composition among different harvesting batches. The contents of total sugar and polysaccharide in wolfberry fruit were determined by Fehling reagent method and phenol-sulfuric acid method, respectively;The content of betaine in fruit was determined by high-performance liquid chromatography,and the flavonoids and carotene in the wolfberry fruits were determined by spectrophotometry. Analysis of data over three consecutive years revealed that as nitrogen application increased, the total sugar content in wolfberry fruits initially decreased and then increased. The levels of L. barbarum polysaccharides, total flavonoids, and total carotenoids initially increased and then decreased, while the betaine content consistently increased. Different picking batches significantly impacted the nutrient content of wolfberry fruits. Generally, the first batch of summer wolfberry fruits had greater amounts of total sugar and flavonoids, whereas other nutrients peaked in the third batch. By employing a broadly targeted metabolomics approach, 926 different metabolites were identified. The top 20 differentially abundant metabolites were selected for heatmap generation, revealing that the contents of L-citrulline, 2-methylglutaric acid, and adipic acid increased proportionally to the nitrogen gradient. Conversely, the dibutyl phthalate and 2, 4-dihydroxyquinoline contents significantly decreased under high-nitrogen conditions. The remaining 15 differentially abundant metabolites, kaempferol-3-O-sophorosid-7-O-rhamnoside, trigonelline, and isorhamnosid-3-O-sophoroside, initially increased and then decreased with increasing nitrogen levels. Isofraxidin, a common differentially abundant metabolite across all treatments, is a coumarin that may serve as a potential biomarker for wolfberry fruit response to nitrogen. Differentially abundant metabolites were analyzed for GO pathway involvement, revealing significant enrichment in metabolic pathways and biosynthesis of secondary metabolites under different nitrogen treatments. In conclusion, a nitrogen application of 675 kg·ha-2, 20% less than the local farmers' actual application, was most beneficial for the quality of four-year-old Ningqi 7 wolfberry fruits. Consumers who purchase wolfberry-dried fruit for health benefits should not consider only the first batch of summer wolfberry fruits. These results offer a broader perspective for enhancing the quality and efficiency of the wolfberry industry.

From phyllosphere to insect cuticles: silkworms gather antifungal bacteria from mulberry leaves to battle fungal parasite attacks.

BACKGROUND: Bacterial transfers from plants to insect herbivore guts have been well investigated. However, bacterial exchanges between plant phyllospheres and insect cuticles remain unclear, as does their related biological function. RESULTS: Here, we report that the cuticular bacterial loads of silkworm larvae quickly increased after molting and feeding on the white mulberry (Morus alba) leaves. The isolation and examination of silkworm cuticular bacteria identified one bacterium Mammaliicoccus sciuri that could completely inhibit the spore germination of fungal entomopathogens Metarhizium robertsii and Beauveria bassiana. Interestingly, Ma. sciuri was evident originally from mulberry leaves, which could produce a secreted chitinolytic lysozyme (termed Msp1) to damage fungal cell walls. In consistency, the deletion of Msp1 substantially impaired bacterial antifungal activity. Pretreating silkworm larvae with Ma. sciuri cells followed by fungal topical infections revealed that this bacterium could help defend silkworms against fungal infections. Unsurprisingly, the protective efficacy of ΔMsp1 was considerably reduced when compared with that of wild-type bacterium. Administration of bacterium-treated diets had no negative effect on silkworm development; instead, bacterial supplementation could protect the artificial diet from Aspergillus contamination. CONCLUSIONS: The results of this study evidence that the cross-kingdom transfer of bacteria from plant phyllospheres to insect herbivore cuticles can help protect insects against fungal parasite attacks. Video Abstract.

Eucalyptol-loaded microcapsules combined with Cynanchum komarovii extracts provide long-term and low-risk management of Chinese wolfberry (Lycium barbarum L.).

Realizing eco-friendly, long-term, and low-risk aphid control on Lycium barbarum (medicinal cash crop) using a Cynanchum komarovii extracts and eucalyptus oil-loaded microcapsules (EOMCs) formulation compositions is viable. In this study, the aim is to optimize the composition of Cynanchum komarovii extracts and EOMCs formulation for effective control of aphids, the release of EOMCs was controlled by changing the cross-linking degree of the shell to match the aphid control characteristics of Cynanchum komarovii extracts. Four types of polyamines were used as cross-linking agents for the preparation of EOMCs by interfacial polymerization. The bioactivity, wettability, and field application efficacy of Cynanchum komarovii extracts and different EOMCs formulation compositions were evaluated. These EOMCs exhibited an encapsulation efficiency exceeding 85 %. The control efficiency of the formulation compositions of microcapsules with a moderate release rate and Cynanchum komarovii extracts on aphids remained at 62.86 %, while the control efficiency of the combination of microcapsules with the fastest and slowest rates with Cynanchum komarovii extracts was only 48.62 % and 57.11 %, respectively. The formulation compositions of Cynanchum komarovii extracts with all four types of EOMCs were found to be safe for Chinese wolfberry plants. Overall, by selecting appropriate polyamines during fabrication, the release rate can be effectively controlled to achieve sustainable and low-risk aphid control in Lycium barbarum through compounding with selected microcapsules.

Achieving High Efficacy and Low Safety Risk by Balancing Pesticide Deposition on Leaves and Fruits of Chinese Wolfberry (Lycium barbarum L.).

Pesticide residue has become the main technical barrier that restricts the export of Chinese wolfberry. Can we achieve high efficacy and low safety risk by balancing pesticide deposition on the leaves and fruits of Chinese wolfberry? In this research, the structural characteristics and wettability of leaves and fruits of Chinese wolfberry at different growth stages were studied. The adaxial and abaxial surfaces of leaves were hydrophobic, whereas the fruit surfaces were hydrophilic. Adding spray adjuvant could increase the retention of droplets on the leaf surfaces of Chinese wolfberry by 52.28-97.89% and reduce the retention on the fruit surfaces by 21.68-42.14%. A structural equation model analysis showed that the adhesion tension was the key factor affecting the retention of the solutions among various interface behaviors. When the concentrations of Silwet618, AEO-5, Gemini 31551, and 1227 were 2-5 times higher than their CMCs, the retention of pesticide solutions (pyraclostrobin and tylophorine) on Chinese wolfberry leaves significantly increased, and the control efficacies on aphids and powdery mildew also dramatically improved (65.90-105.15 and 41.18-133.06%, respectively). Meanwhile, the retention of pesticides on the fruit of Chinese wolfberry was reduced. This study provides new insights into increasing the utilization of pesticides in controlling pests and improving food safety.

Dissipation Kinetics and Safety Evaluation of Flonicamid in Four Various Types of Crops.

The chemical insecticide flonicamid is widely used to control aphids on crops. Differences among crops make the universality of detection methods a particularly important consideration. The aim of this study was to establish a universal, sensitive, accurate and efficient method for the determination of flonicamid residues in peach, cucumber, cabbage and cotton. QuEChERS pretreatment was combined with ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). A satisfactory recovery rate of 84.3-99.3% was achieved at three spiking levels, and the relative standard deviation (RSD) was 0.41-5.95%. The limit of quantification (LOQ) of flonicamid in the four matrices was 0.01 mg/kg. The residue and dissipation kinetics of flonicamid in four types of crops in various locations were determined by using the optimized method. The results showed that flonicamid had a high dissipation rate in the four different types of crops and a half-life in the different matrices and locations of 2.28-9.74 days. The terminal residue of flonicamid was lower than the maximum residue limit (MRL). The risk quotient (RQ) of flonicamid was 4.4%, which is significantly lower than 100%. This result shows that the dietary risk presented by using flonicamid at the maximum recommended dose is low and acceptable. The comprehensive long-term dietary risk assessment of flonicamid performed in this study provides a reference for the protection of consumer health and safe insecticide use.

Identifying Common Genes and Pathways Associated with Periodontitis and Aging by Bioinformatics Analysis.

Background: This work used bioinformatic analysis to identify the relationship between periodontitis (PD) and aging, which could lead to new treatments for periodontal disease in the elderly. Method: Four microarray datasets were obtained from the Gene Expression Omnibus (GEO) database and analyzed in R language to identify differentially expressed genes (DEGs). The common DEGs of PD and aging were evaluated as key genes in this investigation by a Venn diagram. These common DEGs were analyzed through additional experiments and analysis, such as pathway analysis and enrichment analysis, and a network of protein-protein interactions (PPIs) was constructed. Cytoscape was used to visualize hub genes and critical modules based on the PPI network. Interaction of TF-genes and miRNAs with hub genes is identified. Result: 84 common DEGs were found between PD and aging. Cytohubba was performed on the PPI network obtained from STRING tool, and the top 10 genes (MMP2, PDGFRB, CTGF, CD34, CXCL12, VIM, IL2RG, ACTA2, COL4A2, and TAGLN) were selected as hub genes. VIM may be a potential biomarker in the analysis of linked hub gene regulatory networks, and hsa-mir-21 and hsa-mir-125b are predicted to be associated in PD and aging. Conclusion: This study investigated the key genes and pathways interactions between PD and aging, which may help reveal the correlation between PD and aging. The current research results are obtained by prediction, and follow-up biological experiments are required for further verification.

Robust route to H2O2 and H2 via intermediate water splitting enabled by capitalizing on minimum vanadium-doped piezocatalysts.

H2O2 is an environmentally friendly chemical for a wide range of water treatments. The industrial production of H2O2 is an anthraquinone oxidation process, which, however, consumes extensive energy and produces pollution. Here we report a green and sustainable piezocatalytic intermediate water splitting process to simultaneously obtain H2O2 and H2 using single crystal vanadium (V)-doped NaNbO3 (V-NaNbO3) nanocubes as catalysts. The introduction of V improves the specific surface area and active sites of NaNbO3. Notably, V-NaNbO3 piezocatalysts of 10 mg exhibit 3.1-fold higher piezocatalytic efficiency than the same catalysts of 50 mg, as more piezocatalysts lead to higher probability of aggregation. The aggregation causes reducing active sites and decreased built-in electric field due to the neutralization between different nano-catalysts. Remarkably, piezocatalytic H2O2 and H2 production rates of V-NaNbO3 (10 mol%) nanocubes (102.6 and 346.2 µmol·g-1·h-1, respectively) are increased by 2.2 and 4.6 times compared to the as-prepared pristine NaNbO3 counterparts, respectively. This improved catalytic efficiency is attributed to the promoted piezo-response and more active sites of NaNbO3 catalysts after V doping, as uncovered by piezo-response force microscopy (PFM) and density functional theory (DFT) simulation. More importantly, our DFT results illustrate that inducing V could reduce the dynamic barrier of water dissociation over NaNbO3, thus enhancing the yield of H2O2 and H2. This facile yet robust piezocatalytic route using minimal amounts of catalysts to obtain H2O2 and H2 may stand out as a promising candidate for environmental applications and water splitting. Electronic Supplementary Material: Supplementary material (typical Raman spectra of NaNbO3 and V-NaNbO3 with various doping concentrations (Fig. S1). XPS spectra of Na 1s (Fig. S2). PL spectra of solution obtained from the piezocatalytic system using NaNbO3 and V-NaNbO3 (10 mol%) as the catalysts after 1 h (Fig. S3). The length of NaNbO3 and V-NaNbO3 nanocubes calculated from XRD data of their (101) planes (Table S1)) is available in the online version of this article at 10.1007/s12274-022-4506-0.

Direct synthesis of ultrathin FER zeolite nanosheets via a dual-template approach.

The synthesis of zeolites with nanosheet morphology has been attracting extensive attention. Despite the steady progress, the direct synthesis of ultrathin nanosheets of FER zeolite with thickness of less than 10 nm is still a great challenge. Herein we report a facile synthesis of FER zeolite nanosheets (named SCM-37) by using octyltrimethylammonium chloride (OTMAC) and 4-dimethylaminopyridine (4-DMAP) as dual organic templates. The effects of synthesis parameters, including initial molar ratio of SiO2/Al2O3, crystallization temperature and time were investigated. It turned out that the crystallization field of SCM-37 was narrow. A two-step crystallization method was utilized to obtain pure and completely crystallized SCM-37 zeolite. SCM-37 was characterized by multiple techniques, including X-ray powder diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), nitrogen physisorption, Fourier transform infrared (FTIR), ammonia-temperature programmed desorption (NH3-TPD) and nuclear magnetic resonance (NMR), and compared with the conventional FER zeolite (C-FER). The two most significant features of SCM-37 are the ultrathin crystal nanosheet and extremely high external surface area. The thickness of SCM-37 along the a-axis is 4∼7 nm, while that of C-FER is over 20 nm. The external surface area reaches 198 m2 g-1, which is over ten times larger than that of C-FER. The catalytic performances of the FER zeolites were evaluated by the cracking of 1,3,5-triisopropylbenzene (TiPB). Although possessing a lower amount of total acid sites, SCM-37 showed higher conversion of TiPB, as well as selectivity to the deep cracking products. The superior performance of SCM-37 was attributed to the higher external surface area arising from the ultrathin nanosheets.

Widely-Targeted Metabolic Profiling in Lyciumbarbarum Fruits under Salt-Alkaline Stress Uncovers Mechanism of Salinity Tolerance.

Wolfberry (Lycium barbarum L.) is an important economic crop widely grown in China. The effects of salt-alkaline stress on metabolites accumulation in the salt-tolerant Ningqi1 wolfberry fruits were evaluated across 12 salt-alkaline stress gradients. The soil pH, Na+, K+, Ca2+, Mg2+, and HCO3- contents decreased at a gradient across the salt-alkaline stress gradients. Based on the widely-targeted metabolomics approach, we identified 457 diverse metabolites, 53% of which were affected by salt-alkaline stress. Remarkably, soil salt-alkaline stress enhanced metabolites accumulation in wolfberry fruits. Amino acids, alkaloids, organic acids, and polyphenols contents increased proportionally across the salt-alkaline stress gradients. In contrast, nucleic acids, lipids, hydroxycinnamoyl derivatives, organic acids and derivatives and vitamins were significantly reduced by high salt-alkaline stress. A total of 13 salt-responsive metabolites represent potential biomarkers for salt-alkaline stress tolerance in wolfberry. Specifically, we found that constant reductions of lipids and chlorogenic acids; up-regulation of abscisic acid and accumulation of polyamines are essential mechanisms for salt-alkaline stress tolerance in Ningqi1. Overall, we provide for the first time some extensive metabolic insights into salt-alkaline stress tolerance and key metabolite biomarkers which may be useful for improving wolfberry tolerance to salt-alkaline stress.

Illumina MiSeq Sequencing Reveals Correlations among Fruit Ingredients, Environmental Factors, and AMF Communities in Three Lycium Barbarum Producing Regions of China.

The symbiotic relationship of arbuscular mycorrhizal fungi (AMF) is important for Lycium barbarum, a highly nutritious and medicinal crop. However, the influence of environmental factors on AMF communities remains largely elusive. Based on MiSeq sequencing, we analyzed AMF communities in rhizosphere soils of L. barbarum with growth synchronization in three typical L. barbarum cultivation sites in China. The Zhongning region has poor soils with a high richness of AMF communities. Geographical environmental variances lead to differences in AMF communities which in turn affects the active ingredients of L. barbarum fruit. Furthermore, different genera of AMF showed significant correlations with environmental factors and fruit ingredients. The three genera, Claroideoglomus, Dominikia, and Funneliformis correlated to environmental factors and fruits ingredients in a similar manner affecting the whole sugar (TS) and flavonoids (FLA) contents in the fruits of L. barbarum. Also, these showed a significantly positive correlation with soil pH. This fact was unknown so far due to different soil acidity/alkalinity in different studies. IMPORTANCE The climatic and ecological environment is a complex phenomenon, involving various environmental factors that regulate the diversity and population distribution structure of AMF communities affecting plant growth, crop composition, and yield. Current studies on the effects of environmental factors on AMF communities have mainly focused on soil conditions and host plants. Fewer studies have been conducted on the correlation between temperature, enzyme activity, plant fruiting, and AMF communities. The present study investigated the diversity of AMF communities and the influence of environmental factors on their distribution patterns, which showed similar effects on some AMF species. The results suggest that screening AMF fungicides that meet the target may significantly help soil restoration reducing the use of chemical fertilizers and a large amount of human and material resources.

Diversity and spatiotemporal dynamics of fungal communities in the rhizosphere soil of Lycium barbarum L.: a new insight into the mechanism of geoherb formation.

Lycium barbarum L. is a well-known traditional geoherb in Ningxia, China. The fruits of L. barbarum contain several dietary constituents, and thus, they exert many beneficial effects on human health. However, a few studies have been conducted on the geoherb L. barbarum and its rhizosphere soil fungal community. In this study, we determined the physicochemical properties and fungal community structure of rhizosphere soil of L. barbarum from three regions of China, namely Ningxia (NX), Qinghai (QH), and Xinjiang (XJ), during three development stages of L. barbarum. Soil pH varied between 7.56 and 8.60 across the three regions, indicating that alkaline soil is conducive to the growth of L. barbarum. The majority of soil properties in NX, an authentic geoherb-producing area, were substantially inferior to those in XJ and QH during all three developmental stages. Total sugar, polysaccharide (LBP), and flavonoid contents were the highest in wolfberry fruits from NX. High-throughput sequencing showed that the abundance of the soil fungal population in NX was higher than that in QH and XJ during the flowering and fruiting stage and summer dormant stage. Moreover, the soil fungal diversity increased with the development of wolfberry. Ascomycota and Mortierellomycota were the predominant phyla in the rhizosphere fungal communities in all samples. Redundancy analysis showed a significant correlation of the soil-available phosphorus and LBP of wolfberry fruits with the fungal community composition. The characteristics of rhizosphere fungal communities determined in the present study provide insights into the mechanism of geoherb formation in NX wolfberry.

Effects of nitrogen input on soil bacterial community structure and soil nitrogen cycling in the rhizosphere soil of Lycium barbarum L.

Lycium barbarum L., goji berry, is a precious traditional Chinese medicine and it is homology of medicine and food. Its growth is heavily dependent on nitrogen. The use of chemical fertilizers has significantly promoted the yield of goji berry and the development of the L. barbarum L. industry. However, crop plants are inefficient in the acquisition and utilization of applied nitrogen, it often leads to excessive application of nitrogen fertilizers by producers, which cause negatively impact to the environment ultimately. The exploration of an interaction model which deals with crops, chemical fertilizers, and rhizosphere microbes to improve nitrogen use efficiency, is, therefore, an important research objective to achieve sustainable development of agriculture greatly. In our study, we explored the effects of nitrogen input on soil microbial community structure, soil nitrogen cycling, and the contents of nutrients in L. barbarum fruits. The structure and composition of the bacterial community in the rhizosphere soil of L. barbarum were significantly different under different nitrogen supply conditions, and high nitrogen addition inhibited the diversity and stability of bacterial communities. Low nitrogen input stimulated the relative abundance of ammonia-oxidizing bacteria (AOB), such as Nitrosospira, catalyzing the first step of the ammonia oxidation process. The results of the GLMM model showed that the level of nitrogen fertilizer (urea) input, the relative abundance of AOB, the relative abundance of Bradyrhizobium, and their combinations had significant effects on the soil nitrogen cycling and contents of nutrients in L. barbarum fruits. Therefore, we believe that moderately reducing the use of urea and other nitrogen fertilizers is more conducive to improving soil nitrogen use efficiency and Goji berry fruit quality by increasing the nitrogen cycling potential of soil microorganisms.

Changes in Metabolome and Nutritional Quality of Lycium barbarum Fruits from Three Typical Growing Areas of China as Revealed by Widely Targeted Metabolomics.

This study aimed at assessing the climatic factors influencing the wolfberry fruit morphology, and the composition of its nutritious metabolites. The cultivar Ningqi1, widely grown in Northwest China was collected from three typical ecological growing counties with contrasting climatic conditions: Ningxia Zhongning (NF), Xinjiang Jinghe (XF) and Qinghai Nomuhong (QF). During the ripening period, 45 fruits from different plantations at each location were sampled. A total of 393 metabolites were detected in all samples through the widely targeted metabolomics approach and grouped into 19 known classes. Fruits from QF were the biggest followed by those from XF and NF. The altitude, relative humidity and light intensity had negative and strong correlations with most of the metabolites, suggesting that growing wolfberry in very high altitudes and under high light intensity is detrimental for the fruit nutritional quality. Soil moisture content is highly and negatively correlated with vitamins, organic acids and carbohydrates while moderately and positively correlated with other classes of metabolites. In contrast, air and soil temperatures exhibited positive correlation with majority of the metabolites. Overall, our results suggest high soil and air temperatures, low altitude and light intensity and moderate soil moisture, as the suitable conditions to produce Lycium fruits with high content of nutritious metabolites.

Construction of indazolo[3,2-a]isoquinolines via [3 + 2] cycloaddition of benzynes.

A [3 + 2] annulation protocol for the construction of N-substituted indazolo[3,2-a]isoquinolines starting from benzynes and C,N-cyclic azomethine imines was developed. A diverse range of highly functionalized products indazolo[3,2-a]isoquinolines featuring an indazole scaffold can be easily accessed via a one-step reaction under mild conditions, and they show good anti-proliferative activity on cancer cells.