Integrated metabolome and transcriptome analysis of differences in quality of ripe Lycium barbarum L. fruits harvested at different periods.

BACKGROUND: Wolfberry is well-known for its high nutritional value and medicinal benefits. Due to the continuous ripening nature of Goji berries and the fact that they can be commercially harvested within a few weeks, their phytochemical composition may change during the harvesting process at different periods. RESULT: The involved molecular mechanisms of difference in fruit quality of ripe Lycium barbarum L. harvested at four different periods were investigated by transcriptomic and metabolomics analyses for the first time. According to the results we obtained, it was found that the appearance quality of L. barbarum fruits picked at the beginning of the harvesting season was superior, while the accumulation of sugar substances in L. barbarum fruits picked at the end of the harvesting season was better. At the same time the vitamin C and carotenoids content of wolfberry fruits picked during the summer harvesting season were richer. Ascorbic acid, succinic acid, glutamic acid, and phenolic acids have significant changes in transcription and metabolism levels. Through the network metabolic map, we found that ascorbic acid, glutamic acid, glutamine and related enzyme genes were differentially accumulated and expressed in wolfberry fruits at different harvesting periods. Nevertheless, these metabolites played important roles in the ascorbate-glutathione recycling system. Ascorbic acid, phenolic substances and the ascorbate-glutathione recycling system have antioxidant effects, which makes the L. barbarum fruits harvested in the summer more in line with market demand and health care concepts. CONCLUSION: This study laid the foundation for understanding the molecular regulatory mechanisms of quality differences of ripe wolfberry fruits harvested at different periods, and provides a theoretical basis for enhancing the quality of L. barbarum fruits.

Metabolite-based genome-wide association studies enable the dissection of the genetic bases of flavonoids, betaine and spermidine in wolfberry (Lycium).

Wolfberry is a plant with medicinal and food values. However, its bioactive ingredients and the corresponding genetic bases have not been determined. Here, we de novo generated a chromosome-level genome assembly for wolfberry, yielding a genome sequence of ~1.77 Gb with contig N50 of 50.55 Mb and 39 224 predicted gene models. A variation map, using 307 re-sequenced accessions, was called based on this genome assembly. Furthermore, the fruit metabolome of these accessions was profiled using 563 annotated metabolites, which separated Lycium barbarum L. and non-L. barbarum L. The flavonoids, coumarins, alkaloids and nicotinic acid contents were higher in the former than in the latter. A metabolite-based genome-wide association study mapped 156 164 significant single nucleotide polymorphisms corresponding to 340 metabolites. This included 19 219 unique lead single nucleotide polymorphisms in 1517 significant association loci, of which three metabolites, flavonoids, betaine and spermidine, were highlighted. Two candidate genes, LbUGT (evm.TU.chr07.2692) and LbCHS (evm.TU.chr07.2738), with non-synonymous mutations, were associated with the flavonoids content. LbCHS is a structural gene that interacts with a nearby MYB transcription factor (evm.TU.chr07.2726) both in L. barbarum and L. ruthenicum. Thus, these three genes might be involved in the biosynthesis/metabolism of flavonoids. LbSSADH (evm.TU.chr09.627) was identified as possibly participating in betaine biosynthesis/metabolism. Four lycibarbarspermidines (E-G and O) were identified, and only the lycibarbarspermidines O content was higher in L. barbarum varieties than in non-L. barbarum varieties. The evm.TU.chr07.2680 gene associated with lycibarbarspermidines O was annotated as an acetyl-CoA-benzylalcohol acetyltransferase, suggesting that it is a candidate gene for spermidine biosynthesis. These results provide novel insights into the specific metabolite profile of non-L. barbarum L. and the genetic bases of flavonoids, betaine and spermidine biosynthesis/metabolism.

Wolfberry enhanced the abundance of Akkermansia muciniphila by YAP1 in mice with acetaminophen-induced liver injury.

Drug-induced liver injury (DILI) by acetaminophen (APAP) was one of the most challenging liver diseases. Wolfberry (Lycium barbarum L.), a traditional Chinese medicinal material and food supplement, has a potential effect on increasing the abundance of Akkermansia muciniphila (A. muciniphila) in mice colons. However, the effect and mechanism of wolfberry remain unclear in APAP-induced DILI. In this study, wolfberry promoted the proliferation of activated-A. muciniphila in vitro and in vivo. For the first time, we detected that the activated-A. muciniphila but not the killed-A. muciniphila increased the expression level of Yes-associated protein 1 (YAP1) in the liver and alleviated liver injury in APAP-induced DILI mice. Mechanically, A. muciniphila improved the intestinal mucosal barrier and reduced lipopolysaccharide (LPS) content in the liver, leading to the increased expression level of YAP1. Furthermore, wolfberry increased the A. muciniphila abundance in the colon and YAP1 expression in the liver from APAP-induced DILI mice, which promoted the recovery of APAP-induced liver injury. Meanwhile, wolfberry combination with A. muciniphila synergistically increased AKK abundance and YAP1 expression in the liver. Our research provides an innovative strategy to improve DILI.

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.

Changes in m6A RNA methylation are associated with male sterility in wolfberry.

BACKGROUND: N6-methyladenosine (m6A) modification is the most abundant type of RNA modification in eukaryotic cells, playing pivotal roles in multiple plant growth and development processes. Yet the potential role of m6A in conferring the trait of male sterility in plants remains unknown. RESULTS: In this study, we performed RNA-sequencing (RNA-Seq) and m6A-sequencing (m6A-Seq) of RNAs obtained from the anther tissue of two wolfberry lines: 'Ningqi No.1' (LB1) and its natural male sterile mutant 'Ningqi No.5' (LB5). Based on the newly assembled transcriptome, we established transcriptome-wide m6A maps for LB1 and LB5 at the single nucleus pollen stage. We found that the gene XLOC_021201, a homolog of m6A eraser-related gene ALKBH10 in Arabidopsis thaliana, was significantly differentially expressed between LB1 and LB5. We also identified 1642 and 563 m6A-modified genes with hypermethylated and hypomethylated patterns, respectively, in LB1 compared with LB5. We found the hypermethylated genes significantly enriched in biological processes related to energy metabolism and lipid metabolism, while hypomethylation genes were mainly linked to cell cycle process, gametophyte development, and reproductive process. Among these 2205 differentially m6A methylated genes, 13.74% (303 of 2205) were differentially expressed in LB1 vis-à-vis LB5. CONCLUSIONS: This study constructs the first m6A transcriptome map of wolfberry and establishes an association between m6A and the trait of male sterility in wolfberry.

Transcriptional response of wolfberry to infestation with the endophytic Fusarium nematophilum strain NQ8GII4.

Fusarium nematophilum NQ8GII4 is an endophytic fungus isolated from the root of healthy wolfberry (Lycium barbarum). Previous studies have reported that NQ8GII4 could dwell in wolfberry roots and enhance the defense responses in wolfberry against root rot, which is caused by F. oxysporum. To further elucidate the molecular mechanism of wolfberry disease resistance induced by NQ8GII4, in the present study, we adopted RNA sequencing analysis to profile the transcriptome of wolfberry response to NQ8GII4 infestation over a time course of 3 and 7 days post-inoculation (dpi). Gene ontology (GO) enrichment analysis revealed that DEGs were enriched related to biological regulation, response to stimulus, signaling, detoxification, immune system process, transporter activity, electron carrier activity, transcription factor activity, nucleic acid binding transcription factor, and antioxidant activity. Through Kyoto encyclopedia of genes and genomes (KEGG) analysis, it was found that many of these DEGs were enriched in pathways related to plant-pathogen interactions, hormone signal transduction, and phenylpropanoid biosynthesis pathway in wolfberry. This suggests that innate immunity, phytohormone signaling, and numerous phenylpropanoid compounds, which comprise a complex defense network in wolfberry. Chloroplast 50S ribosomal proteins (50S RP) were consistently located at the core position of the response in wolfberry following infestation with NQ8GII4 analyzed by protein-protein interaction (PPI) network. This study elucidated the molecular mechanism underlying the interaction between NQ8GII4 and wolfberry, clarified the wolfberry immune response network to endophytic fungi infestation, identified candidate resistance genes in wolfberry, and provided a fundamental date for subsequent work.

Study on Ultrasonic Far-Infrared Radiation Drying and Quality Characteristics of Wolfberry (Lycium barbarum L.) under Different Pretreatments.

In order to explore the effects of different pretreatment methods on the ultrasonic far-infrared synergistic drying characteristics and quality of wolfberry, the bioactive components (polysaccharide, total phenol, total flavonoids, and antioxidants), the quality characteristics (rehydration ratio, color, vitamin C content, and betaine content), and the microstructure of the dried products were used as evaluation indices to test wolfberry treated by five different pretreatments (hot blanching; candied pretreatment; NaOH solution treatment; NaCl solution treatment; and Na2CO3 solution treatment). The results showed that hot blanching pretreatment improved the drying rate and shortened the drying time, and that the vitamin C content of dried products pretreated by hot blanching (92.56 mg/100 g) was higher than that of dried products pretreated by other methods. All five pretreatment methods increased the contents of the total phenols, vitamin C, and betaine of wolfberry. Wolfberry treated by candied pretreatment had lower color differences and higher contents of polysaccharide (0.83 g/g), total phenol (9.26 mg/g), and total flavonoids (2.61 mg/g) than wolfberry treated by the other pretreatment methods. Wolfberry pretreated by NaCl solution had the strongest antioxidant capacity (65.01%). Wolfberry pretreated by Na2CO3 solution had the highest betaine content (3.24%). The observation of the microstructure of the dried products revealed that hot blanching caused the most damage to wolfberry, while the candied pretreatment was less destructive to the tissue cells of wolfberry. On the whole, the dried wolfberry products obtained by the candied pretreatment were of a better quality than products obtained by the other pretreatment methods.

The Application of Handheld Near-Infrared Spectroscopy and Raman Spectroscopic Imaging for the Identification and Quality Control of Food Products.

The following investigations describe the potential of handheld NIR spectroscopy and Raman imaging measurements for the identification and authentication of food products. On the one hand, during the last decade, handheld NIR spectroscopy has made the greatest progress among vibrational spectroscopic methods in terms of miniaturization and price/performance ratio, and on the other hand, the Raman spectroscopic imaging method can achieve the best lateral resolution when examining the heterogeneous composition of samples. The utilization of both methods is further enhanced via the combination with chemometric evaluation methods with respect to the detection, identification, and discrimination of illegal counterfeiting of food products. To demonstrate the solution to practical problems with these two spectroscopic techniques, the results of our recent investigations obtained for various industrial processes and customer-relevant product examples have been discussed in this article. Specifically, the monitoring of food extraction processes (e.g., ethanol extraction of clove and water extraction of wolfberry) and the identification of food quality (e.g., differentiation of cocoa nibs and cocoa beans) via handheld NIR spectroscopy, and the detection and quantification of adulterations in powdered dairy products via Raman imaging were outlined in some detail. Although the present work only demonstrates exemplary product and process examples, the applications provide a balanced overview of materials with different physical properties and manufacturing processes in order to be able to derive modified applications for other products or production processes.

Metabolomic and transcriptomic analysis of Lycium chinese and L. ruthenicum under salinity stress.

BACKGROUND: High soil salinity often adversely affects plant physiology and agricultural productivity of almost all crops worldwide, such as the crude drug known as wolfberry. However, the mechanism of this action in wolfberry is not fully understood yet. RESULTS: Here in this study, we studied different mechanisms potentially in Chinese wolfberry (Lycium chinese, LC) and black wolfberry (L. ruthenicum, LR) under salinity stress, by analyzing their transcriptome, metabolome, and hormone changes. The hormone detection analysis revealed that the ABA content was significantly lower in LR than LC under normal condition, and increased sharply under salinity stress in LR but not in LC. The transcriptome analysis showed that the salinity-responsive genes in wolfberry were mainly enriched in MAPK signaling, amino sugar and nucleotide sugar metabolism, carbon metabolism, and plant hormone signal transduction pathways in LC, while mainly related to carbon metabolism and protein processing in endoplasmic reticulum in LR. Metabolome results indicated that LR harbored higher flavone and flavonoid contents than LC under normal condition. However, the flavone and flavonoid contents were hardly changed in LR, but increased substantially in LC when exposed to salinity stress. CONCLUSIONS: Our results adds ABA and flavone to mechanism understanding of salinity tolerance in wolfberry. In addition, flavone plays a positive role in resistance to salinity stress in wolfberry.

The branch-thorn occurrence of Lycium ruthenicum is associated with leaf DNA hypermethylation in response to soil water content.

BACKGROUND: Lycium ruthenicum is an eco-economic shrub which can exist in two forms, thorny and thornless under varying soil moisture conditions. The aim of this study was to determine if the two forms of L. ruthenicum were influenced by soil water content (SWC) and to test the three-way link among SWC, occurrence of branch-thorn and DNA methylation modification. METHODS AND RESULTS: Here, pot experiment was carried out to reveal the influence of SWC on the occurrence of branch-thorn and then paraffin sections, scanning electron microscope and methylation-sensitive amplification polymorphism(MSAP) analysis were used to determine the three-way link among SWC, branch-thorn occurrence and DNA methylation. The results showed that (a) soil drought promoted the development of thorn primordium into branch-thorn and (b) branch-thorn covered axillary bud to protect it against drought and other stresses; (c) the branch-thorn occurrence response to drought was correlated with hypermethylation of CCGG sites and (d) thorny and thornless plants of a clone were distinguished successfully based on the MSAP profiles of their leaves. CONCLUSIONS: Branch-thorns of the L. ruthenicum clone, which occurred in response to drought, covered axillary buds to protect them against drought and other stresses; thorn primordium of the clone did not develop into branch-thorn under the adequate soil moisture condition. The occurrence and absence of the branch-thorns were correlated with the hyper- and hypo-methylation, respectively. We proposed that the branch-thorn plasticity might be an adjustment strategy for the environment, which seems to support the theory of "Use in, waste out".

First Report of Anthracnose Disease Caused by Colletotrichum fioriniae on Barbary wolfberry (Lycium barbarum L.) in Gansu Province, China.

Barbary wolfberry (Lycium barbarum L.) is a well-known edible and medicinal plant, widely grown in northwest China (Gao et al. 2021). During the summer of 2019, typical anthracnose symptoms were observed on fruits of barbary wolfberry in Baiyin, Gansu province, China. Approximately 30% of the barbary wolfberry fruits had typical anthracnose symptoms. Lesions on barbary wolfberry fruits were dark, circular or irregular, sunken, and necrotic or wilted, with the presence of orange to pink conidial masses under high humidity. Small pieces cut from the margins of lesions were surface disinfested with 75% ethanol for 10 s, and 1% NaClO solution for 1 min, rinsed three times in sterile distilled water, dried on sterile filter paper, placed onto potato dextrose agar (PDA), and incubated at 25 ± 1℃ for 5 days in the dark. The pure cultures were obtained by single-spore isolation. All isolates produced pale gray and dense aerial mycelia, in reverse orange to red, at times showing concentric rings on PDA at 25℃ after 10 days in the dark. Conidia (n=100) were colorless, smooth-walled, aseptate, fusiform elliptical with one or both ends, and 8.3 to 17.6 × 3.7 to 6.2 µm. Appressoria (n = 100) were solitary, pale to medium brown, smooth-walled, subglobose to elliptical, sometimes clavate or irregular, and 5.7 to 11.7 × 4.1 to 8.5 µm. The internal transcribed spacer (ITS) region of ribosomal DNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene, bate-tubulin 2 (TUB2) gene, actin (ACT) gene, calmodulin (CAL) gene, chitin synthase 1 (CHS-1) gene, and histone H3 (HIS3) gene of the two representative isolates BY19LB02 and BY19LB06 were amplified and sequenced with primers ITS1/ITS4, GDF1/GDR1, T1/Bt2b, ACT-512F/ACT-783R, CL1/CL2A, CHS-79F/CHS-354R, CYLH3F/CYLH3R, respectively (Damm et al. 2012), and deposited on GenBank (ITS, MZ496816 and MZ505524; ACT, MZ557422 and MZ557417; CHS-1, MZ557423 and MZ557418; GAPDH, MZ557424 and MZ557419; HIS3, MZ557425 and MZ557420; TUB2, MZ557426 and MZ557421). BLAST analysis of the resulting for all the sequences showed 98 to 100% similarity with those of C. fioriniae. Based on the above, the isolates BY19LB02 and BY19LB06 were identified as C. fioriniae. To confirm the pathogenicity, detached heathy barbary wolfberry fruits were surface-sterilized with 75% ethanol for 30s, rinsed three times in sterile distilled water, allowed to dry on sterile filter paper, and then wounded using sterilized needles. Fruits were inoculated by pipetting 10 µL of conidial suspension (1×106 conidia/mL) onto each wound, and controls were inoculated with 10 µL sterile distilled water. Each treatment had 30 fruit replicates. These fruits were kept in a moist chamber at 28°C in the dark. The experiment was repeated three times. After 5 days, anthracnose symptoms were observed on all of the inoculated fruits and identical to those observed in the field, whereas control fruits did not develop symptoms. Theathogen was re-isolated from the lesions of inoculated fruits, fulfilling Koch's postulates. To the best of our knowledge, this is the first report of C. fioriniae causing anthracnose on barbary wolfberry in Gansu Province, China. The same disease on barbary wolfberry was reported in Jilin Province, China (Liu et al. 2016). Gansu is one of the main barbary wolfberry producing areas in northwest China and its geographical area, climate and environmental conditions are different from Jilin Province. Considering that barbary wolfberry is the main source of income for growers in Gansu, this identification can aid in the selection of appropriate management measures for this disease.

Changes in the Phytochemical and Bioactive Compounds and the Antioxidant Properties of Wolfberry during Vinegar Fermentation Processes.

Wolfberry (Lycium barbarum L.), as a kind of functional fruit, has various nutritional and bioactive components, which exhibit healthy benefits. However, wolfberry is not easy to preserve, and the intensive processing of wolfberry needs to be developed. In the present study, the changes in the phytochemical and bioactive compounds, as well as the antioxidant properties of wolfberry, were evaluated in the brewed processes. We found that the sugar contents were significantly decreased, and the total acids values were significantly increased during the fermentation processes. The sugar and fat contents were low in the wolfberry fruit vinegar after fermentation, which is of benefit to human health. In addition, amino acids were examined during the fermentation processes, and histidine, proline, and alanine were found to be the main amino acids in vinegar. The total phenolics and flavonoids contents were significantly increased by 29.4% and 65.7% after fermentation. 4-Hydroxy benzoic acid, 3-hydroxy cinnamic acid, and chlorogenic acid were the primary polyphenols in the wolfberry fruit vinegar. Moreover, the antioxidant activity of wolfberry fruit vinegar was significantly increased compared with that of wolfberry fruit after the fermentation processes. Polysaccharides and polyphenolics were strongly correlated with the antioxidant activity during the fermentation processes. The findings suggest that wolfberry fruit vinegar has a high antioxidant capability, and could be a beneficial food in the human diet.

Genome-Wide Comparative Analysis of the R2R3-MYB Gene Family in Five Solanaceae Species and Identification of Members Regulating Carotenoid Biosynthesis in Wolfberry.

The R2R3-MYB is a large gene family involved in various plant functions, including carotenoid biosynthesis. However, this gene family lacks a comprehensive analysis in wolfberry (Lycium barbarum L.) and other Solanaceae species. The recent sequencing of the wolfberry genome provides an opportunity for investigating the organization and evolutionary characteristics of R2R3-MYB genes in wolfberry and other Solanaceae species. A total of 610 R2R3-MYB genes were identified in five Solanaceae species, including 137 in wolfberry. The LbaR2R3-MYB genes were grouped into 31 subgroups based on phylogenetic analysis, conserved gene structures, and motif composition. Five groups only of Solanaceae R2R3-MYB genes were functionally divergent during evolution. Dispersed and whole duplication events are critical for expanding the R2R3-MYB gene family. There were 287 orthologous gene pairs between wolfberry and the other four selected Solanaceae species. RNA-seq analysis identified the expression level of LbaR2R3-MYB differential gene expression (DEGs) and carotenoid biosynthesis genes (CBGs) in fruit development stages. The highly expressed LbaR2R3-MYB genes are co-expressed with CBGs during fruit development. A quantitative Real-Time (qRT)-PCR verified seven selected candidate genes. Thus, Lba11g0183 and Lba02g01219 are candidate genes regulating carotenoid biosynthesis in wolfberry. This study elucidates the evolution and function of R2R3-MYB genes in wolfberry and the four Solanaceae species.

Nutrition, Bioactive Components, and Hepatoprotective Activity of Fruit Vinegar Produced from Ningxia Wolfberry.

Wolfberry (Lycium barbarum L.) is a nutritious and medicinal fruit, and deeply processed products of wolfberry needs to be improved. In this study, nutrition, bioactive compounds, and hepaprotective activity were explored in wolfberry vinegar (WFV). The contents of nutrients including total sugar and protein in WFV samples were 2.46 and 0.27 g/100 mL, respectively. Total phenolic and flavonoid contents in WFV were 2.42 mg GAE/mL and 1.67 mg RE/mL, respectively. p-Hydroxybenzoic acid and m-hydroxycinnamic acid were the main polyphenols in WFV. The antioxidant activity of WFV were 20.176 mM Trolox/L (ABTS), 8.614 mM Trolox/L (FRAP), and 26.736 mM Trolox/L (DPPH), respectively. In addition, WFV treatment effectively alleviated liver injury by improving histopathological changes and reducing liver biochemical indexes in CCl4-treated mice. WFV alleviated oxidative damage by inhibiting oxidative levels and increasing antioxidant levels. These results suggest that WFV can be utilized as a functional food to prevent oxidative liver injury.

Wolfberry-derived zeaxanthin dipalmitate delays retinal degeneration in a mouse model of retinitis pigmentosa through modulating STAT3, CCL2 and MAPK pathways.

Retinitis pigmentosa (RP) is a group of inherited photoreceptor degeneration diseases that causes blindness without effective treatment. The pathogenesis of retinal degeneration involves mainly oxidative stress and inflammatory responses. Zeaxanthin dipalmitate (ZD), a wolfberry-derived carotenoid, has anti-inflammatory and anti-oxidative stress effects. Here we investigated whether these properties of ZD can delay the retinal degeneration in rd10 mice, a model of RP, and explored its underlying mechanism. One shot of ZD or control vehicle was intravitreally injected into rd10 mice on postnatal day 16 (P16). Retinal function and structure of rd10 mice were assessed at P25, when rods degenerate substantially, using a visual behavior test, multi-electrode-array recordings and immunostaining. Retinal pathogenic gene expression and regulation of signaling pathways by ZD were explored using transcriptome sequencing and western blotting. Our results showed that ZD treatment improved the visual behavior of rd10 mice and delayed the degeneration of retinal photoreceptors. It also improved the light responses of photoreceptors, bipolar cells and retinal ganglion cells. The expression of genes that are involved in inflammation, apoptosis and oxidative stress were up-regulated in rd10 mice, and were reduced by ZD. ZD further reduced the activation of two key factors, signal transducer and activator of transcription 3 and chemokine (C-C motif) ligand 2, down-regulated the expression of the inflammatory factor GFAP, and inhibited extracellular signal regulated protein kinases and P38, but not the JNK pathways. In conclusion, ZD delays the degeneration of the rd10 retina both morphologically and functionally. Its anti-inflammatory function is mediated primarily through the signal transducer and activator of transcription 3, chemokine (C-C motif) ligand 2 and MAPK pathways. Thus, ZD may serve as a potential clinical candidate to treat RP.

Bioactive phenylpropanoid derivatives from the fruits of Lycium ruthenicum Murr.

Eight new (1-7 and 15) and 18 known (8-14 and 16-26) phenylpropanoid derivatives were isolated from the fruits of Lycium ruthenicum Murr. (black wolfberry). Their structures were determined by comprehensive spectroscopic analyses, chemical methods, and comparisons of spectroscopic data. Four known compounds (16, 17, 24, and 26) were firstly isolated from the genus Lycium. Interestingly, compounds 1/2 and 4/5 were isolated as two pairs of inseparable anomers owing to the tautomerism of the free hemiacetal at C-1'' in solution. The antioxidant, α-glucosidase inhibitory, and acetylcholinesterase (AChE) inhibitory activities of compounds 1-26 were evaluated. Some compounds possessed DPPH radical scavenging activity, and all compounds (1-26) exhibited different levels of oxygen radical absorbance capacity (ORAC). One compound displayed α-glucosidase inhibitory activity with potency close to that of the positive control (acarbose).

The inhibitory effect of volatile organic compounds produced by Bacillus subtilis CL2 on pathogenic fungi of wolfberry.

Bacillus subtilis strain CL2 is antagonistic to wolfberry postharvest pathogenic fungi. In this study, we isolated and screened this strain for in vitro experiments. The result of the two-sealed-base-plates method revealed that volatile organic compounds (VOCs) emitted from the strain CL2 inhibited the hyphal growth of four pathogenic fungi Mucor circinelloides LB1, Fusarium arcuatisporum LB5, Alternaria iridiaustralis LB7, and Colletotrichum fioriniae LB8. After exposure to VOCs for 5 days, the hyphal growth of the pathogen C. fioriniae LB8 was inhibited by 73%. Scanning electron microscopy revealed that the VOCs produced by B. subtilis CL2 caused the mycelium morphology of the pathogenic fungi to deform, twist, fold, and shrink. In the in vivo experiments, we noticed that VOCs could significantly reduce the weight loss rate of wolfberry fruits caused by the pathogenic fungus M. circinelloides LB1 and that the decay incidence rate were caused by the pathogenic fungi F. arcuatisporum LB5, A. iridiaustralis LB7, and C. fioriniae LB8. On the basis of the headspace-gas chromatography-ion mobility spectrometry analysis, seven VOCs produced by strain CL2 were identified. Among them, 2,3-butanedione and 3-methylbutyric acid are the main antifungal active substances. This study investigated the antifungal properties of VOCs produced by the strain CL2 on postharvest pathogenic fungi isolated from wolfberry fruits both in vivo and in vitro, thereby providing the theoretical basis for its future applications.

Lycium Barbarum Polysaccharides and Wolfberry Juice Prevent DEHP-Induced Hepatotoxicity via PXR-Regulated Detoxification Pathway.

Environmental di(2-Ethylhexyl) phthalate (DEHP) is widely used in various industries as a plasticizer, and has been reported to induce reproductive and developmental toxicities in organisms. The purpose of this study was to evaluate the detoxification capacity of Lycium barbarum polysaccharides (LBP) and wolfberry juice (WJ) against DEHP-induced hepatotoxicity. Two groups of rats were purchased to study two different intervention method experiments: LBP (50, 100, 200 mg/kg·bw) intervention before DEHP (2000 mg/kg·bw) exposure, and LBP (200 mg/kg·bw) or WJ (8 mL/kg·bw) intervention after DEHP (3000 mg/kg·bw) exposure. The rats were exposed to DEHP once, while the intervention lasted for seven days. At the end of the intervention, enzyme-linked immunosorbent assay (ELISA) was used to measure the related index. The LBP intervention before DEHP exposure experiment (the first experimental method) found that LBP group rats showed a strong capacity toward DEHP detoxification, evidenced by the significant upregulation of activities and concentrations of the partner retinoid, X receptor alpha (RXRα), and downstream regulators Cytochrome P4502E1 (CYP2E1), Cytochrome P4503A1 (CYP3A1), Glutathione S-Transferase Pi (GSTpi), and UDP-glucuronosyltransferase 1 (UGT1) in a dose-dependent manner. The LBP and WJ intervention after DEHP exposure experiment (the second intervention experiment) found that WJ could downregulate pregnane X receptor (PXR), and upregulate downstream regulators, CYP2E1, CYP3A1, and Glutathione S-Transferase (GST) with the extension of intervention time, to alleviate the toxicity of DEHP. However, the intervention effect of WJ was more obvious than that of LBP. These results suggested that LBP and WJ might be effective detoxification agents against DEHP-induced toxic effects, by activating PXR and PXR-related detoxifying enzymes.

The major zeaxanthin dipalmitate derivatives from wolfberry.

Zeaxanthin dipalmitate (3) and two zeaxanthin dipalmitate derivatives, including one new compound (1), were obtained from wolfberry [the fruit of Lycium barbarum L. (Solanaceae)]. Their structures were unambiguously elucidated by spectroscopic analyses. Compound 2 is isolated from the genus Lycium for the first time, and its 1D/2D NMR data are firstly reported. All the compounds belong to carotenoids which are a kind of major bioactive constituents in wolfberry and are also responsible for wolfberry's red color.

[Genetic diversity and genetic structure evaluation of cultivated Chinese wolfberry (Lycium barbarum) based on microsatellite population genetics].

A total of 178 Chinese wolfberry individuals from 17 populations were detected by 7 pairs of SSR primers to evaluate genetic diversity and structure, using software GenALEx 6.5,NTSYS,STRUCTURE, the effects of cultivation on genetic diversity and structure were clarified aiming to find the strategies for genetic management and sustainable use. The results showed that the genetic diversity of cultivated Chinese wolfberry was low. The average number of alleles N_A, expected heterozygosity H_E, observed heterozygosity H_O, and Shannon's information index H' was 3.9, 0.443 7, 0.556 6, 0.788 1, respectively. STRUCTURE, UPGMA clustering and PCA test indicated that Chinese wolfberry varieties were severely intermixed but no differentiation among varieties. Mantel test showed no significant correlation between genetic distance and geographic distance. AMOVA analysis showed that genetic variation mainly occurred among individuals within the population(84.58%, P<0.001), and there was almost no genetic differentiation between varieties(3.63%, P<0.001) and between populations(11.79%, P<0.001). The cultivation has caused a significant decline in the genetic diversity of Chinese wolfberry, which may cause inbreeding decline. New germplasm resources should be sought from the wild to improve the existing cultivars. On the other hand, there are obvious homogenization and germplasm intermixing between cultivated varieties and populations. Meanwhile, Chinese wolfberry cultivars should be purified and prevented from flowing into the wild population, in case of causing pollution of the wild germplasm.