Effect of potassium on the agronomic traits and fruit quality of Goji (Lycium barbarum L.).

To investgate the effects of potassium (K) application on the agronomic traits and fruit quality of Lycium barbarum L. (Goji), three levels of K fertilizer, namely LK (25 g/plant), CK (50 g/plant), and HK (75 g/plant), were applied to plants in phytotron for observing and measuring relevant indicators. The investigation involved seven agronomic traits: plant height, plant stem diameter, new branch increment, yield of fresh fruits per plant, dry fruit quantity within 50 g, ratio of different grade fruits, and ratio of longitudinal diameter to transverse diameter of Goji fruits. The results showed that K application level had significant effect on ratio of the longitudinal diameter to the transverse diameter of fresh Goji fruits, and that the influence on other agronomic traits was slight. In the meanwhile, the concentrations of amino acids, betaine, polysaccharides and flavonoids of Goji fruits in different levels of K fertilizer were tested. The K treatment increased the content of glutamic acid, and decreased that of flavonoids (P < 0.05), whereas the content of other amino acids, polysaccharides and betaine were unaffected. A total of 132 flavonoid metabolites was identified. Among them, K treatment up-regulated 36 metabolites and down-regulated 30 metabolites (P < 0.05). The results provided a basis for balanced K supply to regulate the agronomic traits and nutrients of Goji fruits.

Development of a pre-column derivatization ultra-high-performance liquid chromatography method for polysaccharide and monosaccharide quantification in Lycium barbarum.

Given the limited specificity and accuracy observed in the current official colorimetric quantification of polysaccharide in Lycium barbarum, our study aims to establish a novel, specific, accurate, and economic pre-column derivatization ultra-high-performance liquid chromatography (UHPLC) method for determining the monosaccharide and polysaccharide content in L. barbarum. The optimization of extraction, hydrolysis, and derivatization (using 1-phenyl-3-methyl-5-pyrazolone) processes for polysaccharide from L. barbarum was conducted initially, followed by separation of nine monosaccharides within 20 min using UHPLC with a C18 column. Subsequently, a novel method known as quantitative analysis of multiple components by single marker was developed, utilizing either additive 2-deoxy-D-ribose or any monosaccharide present in the sample as a single reference standard to simultaneously detect the contents of polysaccharide and nine monosaccharides in L. barbarum. To validate the accuracy of the established method, the quantitative results of our approach were compared to both external and internal standard method methods. The minimal relative errors in the quantitative determination of monosaccharides among the three methods confirmed the dependability of the method. By analyzing 20 batches of L. barbarum samples, D-galacturonic acid exhibited the highest content and the polysaccharide levels ranged from 3.02 to 13.04 mg/g. All data implied the specificity and accuracy of the method.

[Comparison on effects of Lycii Fructus from different origins on NaIO_3-induced retinal degenerative diseases in mice by multivariate statistical analysis].

The multivariate statistical analysis was performed to compare the therapeutic effects of Lycii Fructus from different origins on the retinal degenerative diseases(RDD) in mice. The mouse model of RDD was established by intraperitoneal injection of NaIO_3, and the visual function and retinal apoptosis were assessed by dark-light transition and TUNEL assay. Retinal thickness was measured by fundus optical coherence tomography(OCT), and the levels of antioxidant, inflammatory, and angiogenic markers in the serum and eyeball were determined. The therapeutic effects were compared by hierarchical cluster analysis, principal component analysis, and partial least squares-discriminant analysis. The results showed that the extracts of Lycii Fructus from different origins reversed NaIO_3-induced visual damage and retinal apoptosis, reduced oxidative stress, and restored the expression of inflammatory mediators and angiogenic markers in mice. The multivariate statistical analysis based on 17 pharmacodynamic indices suggested that the extract of Lycii Fructus from Ningxia demonstrated better therapeutic effects on RDD than the samples from the other four origins. The results of this study provide a scientific basis for the selection of the advantageous production region of Lycii Fructus for the prevention and treatment of RDD.

Effects of brackish water irrigation with different exogenous salt concentrations on the growth and rhizosphere salinity of Lycium barbarum.

To investigate the effects of different typical exogenous salt concentrations on total soil salinity and the growth of Lycium barbarum under brackish water irrigation, and to determine the salinity threshold of irrigated brackish water that is conducive to the normal growth of Lycium barbarum while mitigating soil salinity accumulation. Four typical exogenous salts (NaCl, CaCl2, NaHCO3, Na2SO4) were selected and set at four concentrations (0.1, 0.5, 2.0, 4.0 g L-1) to conduct a field crossover experiments in the downstream region of the Hetao Irrigation District. The results showed that in the same fertility period, the growth rates of new branches, ground diameter, and crown width first increased and then decreased with rising concentrations of NaCl, CaCl2, and Na2SO4, but showed an inverse relationship with NaHCO3 concentrations. Furthermore, increasing salt concentrations linearly reduced the yield of dry fruits from Lycium barbarum and led to a notable accumulation of total soil salts. Utilizing an experimental research approach, a comprehensive analysis of involving multiple growth indices, stable yield, and soil salinity control of Lycium barbarum revealed that optimal growth occurs at salt concentrations of 0.1-0.5 g L-1 for different water quality areas within the irrigation area; using the method of path analysis identified the total soil salt and crown width as the primary direct and indirect factors influencing the yield of Lycium barbarum. The results of this study provide scientific basis and significant theoretical support for the safe and rational utilization of brackish water and cultivation of Lycium barbarum in typical regions with varying saline water qualities of Hetao irrigation area.

[Effects of Continuous Cropping on the Physiochemical Properties, Pesticide Residues, and Microbial Community in the Root Zone Soil of Lycium barbarum].

Continuous cropping is a common obstacle limiting the high quality and yield of Lycium barbarum （wolfberry）. To clarify the response of soil characteristics of the wolfberry root zone to continuous cropping years, we systematically determined the physicochemical properties and pesticide residues of soils in the wolfberry root zone with different continuous cropping years. In addition, soil bacterial and fungal communities were characterized using high-throughput sequencing technology. The results were as follows： The content of total salt and imidacloprid in the root zone of wolfberry increased with increasing years of continuous cropping. Compared to that with 2 and 9 years, the total salt content in the root zone of wolfberry with 15 years of continuous cropping increased by 51.97% and 54.33%, respectively, and the imidacloprid content increased by 39.58% and 36.61%, respectively. Alkaline nitrogen and available potassium showed an increasing and then decreasing trend. Compared to that with 2 and 15 years, alkaline nitrogen and available potassium in the root-soil of wolfberry with 9 years of continuous cropping increased by 16.94%-28.09% and 18.31%-18.34%, respectively. The diversity and abundance of bacterial communities and the abundance of fungal communities were higher in the root-soil of wolfberry with 9 years of continuous cropping compared to that with 15 years of continuous cropping. In addition, the increase in continuous cropping years also increased the accumulation of harmful plant pathogens such as Pseudomonas, Arthrobacter, Actinomucor, and Trichoderma in the root zone of L. barbarum. Soil total salinity, organic matter, alkaline hydrolyzable nitrogen, and available potassium were the main factors influencing the distribution of bacterial communities. Soil alkaline hydrolyzable nitrogen, available potassium, and ammonium nitrogen were the main factors influencing the distribution of fungal communities. In addition, the soil bacteria in the root zone of L. barbarum were dominated by metabolic functions； in particular, amino acid metabolism, energy metabolism, and nucleotide metabolism were most abundant in the root soil of wolfberry with 9 years of continuous cropping, whereas the highest abundance of functional genes related to membrane translocation was found in the root-soil of wolfberry with 15 years of continuous cropping. The soil fungi were all dominated by saprophytic trophic types, followed by pathogenic cross-nutrients in the root zone of L. barbarum. In conclusion, long-term continuous cropping induced changes in the soil microenvironment in the root zone of L. barbarum, increased soil residues of harmful pesticides and the enrichment of plant pathogens, and reduced the diversity of soil bacterial and fungal communities. Therefore, it is necessary to control the rate of application of soil nutrients and pesticides in the management of L. barbarum and to carry out deep ploughing and deep tilling in good time, and the turnover of old plants in the cultivation of L. barbarum.

Predicting leaf nitrogen content in wolfberry trees by hyperspectral transformation and machine learning for precision agriculture.

Leaf nitrogen content (LNC) is an important indicator for scientific diagnosis of the nutrition status of crops. It plays an important role in the growth, yield and quality of wolfberry. This study aimed to develop new spectral indices (NSIs) and constructed machine learning regression (MLR) models for predicting wolfberry tree LNC. By utilizing four smoothing methods and five mathematic transformation methods, we obtained the original spectral dataset and five spectral transformation datasets for quantitative analysis and model establishment. Subsequently, published vegetation indices (PVIs) were acquired, sensitive wavelengths (SWs) were screened and NSIs were calculated based on SWs. Then MLR models were constructed by combining NSIs from six spectral datasets with three machine learning algorithms. Finally, a comparison was made among the MLR models. The study indicated that the application of mathematical transformation highlighted the differences in spectra, the square root, first-derivative and second-derivative transformation improved the prediction accuracy of MLR models constructed based on NSIs (MLR-NSIs models). However, these transformations had little impact on improving the prediction ability of MLR models constructed based on PVIs (MLR-PVIs models). Additionally, The optimal model for predicting the LNC of wolfberry tree was obtained by using the Random Forest (RF) algorithm combined with NSIs developed by first-derivative transformation spectra. The determination coefficient of validation (Rv2) and ratio of percentage deviation (RPD) was 0.71 and 1.90, respectively. In conclusion, this study has demonstrated that the combination of hyperspectral transformation and machine learning is useful for improving the accuracy of LNC estimation in wolfberry trees.

Lycium barbarum berry extract improves female fertility against aging-related oxidative stress in the ovary.

Reproductive aging in female mammals is characterized by ovarian senescence, leading to a significant fertility decline. Lycium barbarum berry, or goji berry, is a food and medicine that appears in various formulas for treating infertility in traditional Chinese medicine. We investigated the function of an aqueous extract of Lycium barbarum berry (LB extract) to improve health status, fertility, and offspring development during female aging. Aged female mice were supplemented with LB extract, and its effects on fertility, locomotor activity, and offspring development were assessed. The results demonstrated that LB extract significantly increased pregnancy and live birth rates in naturally aged female mice. It also effectively improved aged animals' locomotor activity. Moreover, LB extract promoted the growth and development of offspring delivered from the aged animals and reduced the offspring's anxiety. During aging, fertility-related hormones gradually decline. However, the decline of anti-Müllerian hormone (AMH) and estradiol (E2) in the serum of aged mice was restored by LB extract supplementation. Immunohistochemical analysis revealed that the levels of oxidation and the inflammatory IL-6 in intra-ovarian cells were reduced by LB extract, while the antioxidant-associated proteins peroxiredoxin 4 (PRDX4) and nuclear factor erythroid 2-related factor 2 (NRF2) were increased. Bioinformatics analysis revealed a decline in egg PRDX4 expression with age across various species. This suggests that the antioxidant function protected by LB extract through PRDX4 may consistently promote fertility enhancement by improving ovarian function across different species. Importantly, LB extract did not induce significant adverse effects on aged female mice and their offspring. These findings highlight the potential of LB as a protective agent against ovarian oxidative stress, which preserves ovarian function and improves fertility rates in naturally senescent females.

In vitro digestion and fermentation of the whole goji berry: Bioactive ingredients change and impacts on human gut microbiota.

Goji berry (Lycium barbarum L.) is a nutrient-rich fruit and has received enormous interest for its health benefits. The beneficial effects of goji berry are linked to the absorption of bioactive compounds within the gastrointestinal digestion process and colon fermentation. Nonetheless, how certain bioactive compounds were released, and metabolism changed of the consumption of whole goji berries were still unclear. Therefore, the present study aimed to evaluate the digestion characteristics of key bioactive compounds in whole goji berries with an in vitro digestion model, and the effects of whole goji berries on the structure of gut microbiota were also investigated. Results showed that a significant release of carbohydrates during the digestion process, peaking within the first 15 min of the intestinal phase (421.4 ± 5.82 mg GE/g, dry weight, respectively), was observed, and the phenolic release reached the highest in the first 15 min of the gastric phase. Meanwhile, the bioaccessibilities of phenolic compounds and carbohydrates were determined to be 63.87% and 80.40%, respectively, after intestinal digestion. In addition, the undigested fractions of goji berries could be further fermented to produce short-chain fatty acids, which decreased the colon pH value (from 7.38 to 6.71) as well as the Firmicutes/Bacteroidetes ratio. Moreover, the goji berries regulated the composition of gut microbiota by promoting beneficial bacteria such as Bacteroides, Parabacteroides, and Paraclostridium, whereas inhibiting the proliferation of harmful bacteria (e.g., Fusobacterium). Our results indicated that the goji berry exhibited significant bioactivity during the digestion and fermentation stage and might provide some new insights into the utilization of goji berries in healthy food processing.

Valorizing Lycii fructus waste residue into polysaccharide-rich extracts: Extraction methodologies, physicochemical characterization, in vitro activities and economic feasibility.

The high polysaccharide content of Lycii fructus agri-food waste should be reclaimed for value liberation from both environmental and economic perspectives. In this study, waste from L. fructus pigment products was valorized on a bench scale by upcycling into active polysaccharide-rich extracts. The methodological feasibility of polysaccharide recovery from L. fructus waste was evaluated using sequential extraction techniques. Three fractions LFP-30, LFP-100, and LFP-121, were obtained under stepwise increases in temperature and pressure. Highly heterogeneous xyloglucan (XG)-pectin macromolecules composed of linear homogalacturonan (HG) and alternating intra-RG-I-linkers, with topological neutral branches and XG participation, were predominant among the L. fructus polysaccharides (LFPs). Antioxidant activities in LFPs were unaffected by waste resources and severe extraction methodology conditions. Additionally, the direct investment potential of polysaccharide recovery was evaluated for full-scale implementation. This study demonstrated the necessity and feasibility of extracting bioactive polysaccharides with potential applications from L. fructus waste, and provided a sustainable strategy for transforming L. fructus waste disposal problems into value-added products using cost-effective methodologies.

Novel glucomannan-like polysaccharide from Lycium barbarum L. ameliorates renal fibrosis via blocking macrophage-to-myofibroblasts transition.

The macrophage to myofibroblasts transition (MMT) has been reported as a newly key target in renal fibrosis. Lycium barbarum L. is a traditional Chinese medicine for improving renal function, in which its polysaccharides (LBPs) are the mainly active components. However, whether the role of LBPs in treating renal fibrosis is related to MMT process remain unclear. The purpose of this study was to explore the relationship between the regulating effect on MMT process and the anti-fibrotic effect of LBPs. Initially, small molecular weight LBPs fractions (LBP-S) were firstly isolated via Sephadex G-100 column. Then, the potent inhibitory effect of LBP-S on MMT process was revealed on bone marrow-derived macrophages (BMDM) model induced by TGF-ß. Subsequently, the chemical structure of LBP-S was elucidated through monosaccharide, methylation and NMR spectrum analysis. In vivo biodistribution characteristics studies demonstrated that LBP-S exhibited effectively accumulation in kidney via intraperitoneal administration. Finally, LBP-S showed a satisfactory anti-renal fibrotic effect on unilateral ureteral obstruction operation (UUO) mice, which was significantly reduced following macrophage depletion. Overall, our findings indicated that LPB-S could alleviate renal fibrosis through regulating MMT process and providing new candidate agents for chronic kidney disease (CKD) related fibrosis treatment.

In vivo absorption and fecal excretion of polysaccharides from the fruits of Lycium barbarum L. in rats through fluorescence labeling.

In the present study, the in vivo absorption and fecal excretion of a purified fraction of polysaccharides from the fruits of Lycium barbarum L. (LBPs-4) in rats were investigated by labelling LBPs-4 with fluorescein isothiocyanate (FITC). It was found that the fluorescent labeled LBPs-4 (LBPs-4-FITC) was not detected in the plasma within 24 h following the administration of a single dose of LBPs-4-FITC (100 mg/kg of body weight) to rats, indicating that LBPs-4 was hardly absorbed in its prototype form. Instead, a smaller fragment dissociated from LBPs-4-FITC was observed in feces and was accumulated in a time-dependent manner, suggesting that LBPs-4 was excreted into the feces with a form of degradation. Meanwhile, we observed that LBPs-4-FTIC could modulate the fecal bacterial community profile via increasing the relative abundances of Bacteroides ovatus and Alistipes and promote the production of acetic acid. Furthermore, the monoculture experiment confirmed that LBPs-4 could be metabolized into smaller fragment by B. ovatus, producing acetic acid. Collectively, our study provides information on the destiny of LBPs-4 after oral administration: non-absorbed but moved to the large intestine and catabolized by gut microbiota, especially B. ovatus.

Hypoglycaemic activity of the anthocyanin enriched fraction of Lycium ruthenicum Murr. Fruits and its ingredient identification via UPLC-triple-TOF-MS/MS.

Lycium ruthenicum Murr. is mainly distributed in the northwest region of China and its berries are rich in anthocyanin. This study evaluated the hypoglycaemic activity of the anthocyanin-enriched fraction (AEF) of L. ruthenicum Murr. on α-glucosidase in vivo and in vitro. Overall, 10 anthocyanins were identified via UPLC-Triple-TOF-MS/MS. The AEF exhibited strong inhibitory activity against α-glucosidase, with an IC50 value of 4.468 mg/mL. It behaved as a reversible, mixed-type inhibitor. Molecular docking and dynamic results indicated that the compounds in AEF interacted with enzymes primarily through van der Waals and hydrogen bond and the complex system was stable. The postprandial blood glucose and area under the curve of diabetic mice was significantly decreased by AEF in the carbohydrate tolerance experiments. The results indicate that the AEF from L. ruthenicum Murr. berries could be as a promising food supplement for managing blood sugar levels in patients with diabetes mellitus.

The combination of HSI and NMR techniques with deep learning for identification of geographical origin and GI markers of Lycium barbarum L.

Lycium barbarum L. (L. barbarum) is renowned worldwide for its nutritional and medicinal benefits. Rapid and accurate identification of L.barbarum's geographic origin is essential because its nutritional content, medicinal efficacy, and market price significantly vary by region. This study proposes an innovative method combining hyperspectral imaging (HSI), nuclear magnetic resonance (NMR), and an improved ResNet-34 deep learning model to accurately identify the geographical origin and geographical indication (GI) markers of L.barbarum. The deep learning model achieved a 95.63% accuracy, surpassed traditional methods by 6.26% and reduced runtime by 29.9% through SHapley Additive exPlanations (SHAP)-based feature selection. Pearson correlation analysis between GI markers and HSI characteristic wavelengths enhanced the interpretability of HSI data and further reduced runtime by 33.99%. This work lays the foundation for portable multispectral devices, offering a rapid, accurate, and cost-effective solution for quality assurance and market regulation of L.barbarum products.

A review of Lycium barbarum polysaccharides: Extraction, purification, structural-property relationships, and bioactive molecular mechanisms.

Lycium barbarum L. is of great significance medicinal and edible plant, which is native to N. & Central China. The extensive health benefits of L. barbarum have earned it great respect in traditional medicine for centuries. Lycium barbarum polysaccharides (LBPs) being recognized as one of the most crucial bioactive compounds found within this plant, with it exhibit a diverse range of pharmacological activities and nutritional functions, thereby generating substantial market demand and broad application prospects. To gain a more comprehensive understanding of LBPs, the review discussed the extraction, purification and structural-property relationships of these compounds. In addition, this review provides a comprehensive summary of the potential mechanisms underlying various biological activities attributed to LBPs, including immune modulation, antioxidant effects, neuroprotection, hepatoprotection, and antitumor properties. The application status and the future research directions of LBPs were subsequently presented. This review will establish a robust foundation and serve as an invaluable resource for future research and advancements in the field of LBPs.

Lycium barbarum glycopeptide promotes neuroprotection in ET-1 mediated retinal ganglion cell degeneration.

BACKGROUND: Vascular dysregulation is one of the major risk factors of glaucoma, and endothelin-1 (ET-1) may have a role in the pathogenesis of vascular-related glaucoma. Fruit extract from Lycium Barbarum (LB) exhibits anti-ageing and multitarget mechanisms in protecting retinal ganglion cells (RGC) in various animal models. To investigate the therapeutic efficacy of LB glycoproteins (LbGP) in ET-1 induced RGC degeneration, LbGP was applied under pre- and posttreatment conditions to an ET-1 mouse model. Retina structural and functional outcomes were characterised using clinical-based techniques. METHODS: Adult C57BL/6 mice were randomly allocated into four experimental groups, namely vehicle control (n = 9), LbGP-Pretreatment (n = 8), LbGP-Posttreatment (day 1) (n = 8) and LbGP-Posttreatment (day 5) (n = 7). Oral administration of LbGP 1 mg/Kg or PBS for vehicle control was given once daily. Pre- and posttreatment (day 1 or 5) were commenced at 1 week before and 1 or 5 days after intravitreal injections, respectively, and were continued until postinjection day 28. Effects of treatment on retinal structure and functions were evaluated using optical coherence tomography (OCT), doppler OCT and electroretinogram measurements at baseline, post-injection days 10 and 28. RGC survival was evaluated by using RBPMS immunostaining on retinal wholemounts. RESULTS: ET-1 injection in vehicle control induced transient reductions in arterial flow and retinal functions, leading to significant RNFL thinning and RGC loss at day 28. Although ET-1 induced a transient loss in blood flow or retinal functions in all LbGP groups, LbGP treatments facilitated better restoration of retinal flow and retinal functions as compared with the vehicle control. Also, all three LbGP treatment groups (i.e. pre- and posttreatments from days 1 or 5) significantly preserved thRNFL thickness and RGC densities. No significant difference in protective effects was observed among the three LbGP treatment groups. CONCLUSION: LbGP demonstrated neuroprotective effects in a mouse model of ET-1 induced RGC degeneration, with treatment applied either as a pretreatment, immediate or delayed posttreatment. LbGP treatment promoted a better restoration of retinal blood flow, and protected the RNFL, RGC density and retinal functions. This study showed the translational potential of LB as complementary treatment for glaucoma management.

Lycium chinense Mill Induces Anti-Obesity and Anti-Diabetic Effects In Vitro and In Vivo.

This study investigated the effects of Lycium chinense Mill (LCM) extract on obesity and diabetes, using both in vitro and high-fat diet (HFD)-induced obesity mouse models. We found that LCM notably enhanced glucagon-like peptide-1 (GLP-1) secretion in NCI-h716 cells from 411.4 ± 10.75 pg/mL to 411.4 ± 10.75 pg/mL compared to NT (78.0 ± 0.67 pg/mL) without causing cytotoxicity, implying the involvement of Protein Kinase A C (PKA C) and AMP-activated protein kinase (AMPK) in its action mechanism. LCM also decreased lipid droplets and lowered the expression of adipogenic and lipogenic indicators, such as Fatty Acid Synthase (FAS), Fatty Acid-Binding Protein 4 (FABP4), and Sterol Regulatory Element-Binding Protein 1c (SREBP1c), indicating the suppression of adipocyte differentiation and lipid accumulation. LCM administration to HFD mice resulted in significant weight loss (41.5 ± 3.3 g) compared to the HFD group (45.1 ± 1.8 g). In addition, improved glucose tolerance and serum lipid profiles demonstrated the ability to counteract obesity-related metabolic issues. Additionally, LCM exhibited hepatoprotective properties by reducing hepatic lipid accumulation and diminishing white adipose tissue mass and adipocyte size, thereby demonstrating its effectiveness against hepatic steatosis and adipocyte hypertrophy. These findings show that LCM can be efficiently used as a natural material to treat obesity and diabetes, providing a new approach for remedial and therapeutic purposes.

Three-dimensional mass transfer modeling and phenolic chemistry exploration for ultrasound-assisted and microwave drying of goji berry.

Herein, goji berries were pretreated with sodium carbonate (Na2CO3) and then dried via ultrasound-assisted air drying or microwave drying. Water migration and phenolic chemistry of goji berries were studied under drying. A three-dimensional ellipsoid water transport model, accounting for porosity and temperature fluctuations, was established to explore the intricacies of the drying mechanism. Generally, microwave drying promoted interior water transport compared to ultrasound drying. Among all the drying methods, microwave drying at 240 W (MW-240 W) exhibited the highest De (from 7.34 × 10-9 to 9.61 × 10-9 m2/s) and kc (6.78 × 10-4 m/s) values. The goji berries received a considerably high water content gradient between its surface and center within the first 2 s of all the drying treatments. Microwave drying diminished the water content gradient earlier than air drying and ultrasound-assisted air drying treatments. Furthermore, most correlations observed among phenolics, oxidase activity, and cell wall pectin did not align with the established theories, highlighting the highly nonlinear nature of phenolic chemistry during goji berry drying. This study provides a three-dimensional model to study the mass transfer mechanism of goji berries and analyzes the evolution of polyphenols during the drying process.

Development and evaluation of suitable reference genes for qRT-PCR normalization of hybrids derived from Lycium barbarum and Lycium ruthenicum.

BACKGROUND: A correct and stably expressing reference gene is prerequisite for successful quantitative real-time PCR (qRT-PCR). Investigating gene expression profiling during flower development could enhance our understanding of the molecular mechanisms of flower formation and fertility in Lycium. METHODS AND RESULTS: In this study, 11 candidate reference genes in Lycium flower development were selected from transcriptome sequence data and evaluated with five traditional housekeeping genes from previous studies based on qRT-PCR amplification. Comparing the expression stability result of 16 candidate genes using GeNorm, NormFinder, BestKeeper, and Delta Ct algorithms, Lba04g01649 and Lba12g02820 were validated as the optimal reference genes for the flower development of Lycium. CONCLUSIONS: The reference genes identified in this study would improve the accuracy of qRT-PCR quantification of target gene expression in Lycium flower development and facilitate future functional genomics studies on flower development. This research could lay the foundation for the study of the reproduction and development of the Lycium flower.

Lycium ruthenicum Murray derived exosome-like nanovesicles inhibit Aß-induced apoptosis in PC12 cells via MAPK and PI3K/AKT signaling pathways.

Plant-derived exosome-like nanovesicles (ELNs) are nano-sized vesicles extracted from edible plants. Lycium ruthenicum Murray (LRM) has been gaining increasing attention due to its nutritional and medicinal value, but the ELNs in LRM has not been reported. In this study, LRM-ELNs were obtained, and the proteins, lipids, microRNAs (miRNAs) and active components in LRM tissues and LRM-ELNs was analyzed by LC-MS/MS, LC-MS, high-throughput sequencing techniques, and physical and chemical analysis. LRM-ELNs can be uptaken by PC12 cells through macropinocytosis and caveolin-mediated endocytosis primarily. Transcriptomic and western blot experiments indicate that LRM-ELNs can inhibit Aß-induced apoptosis in PC12 cells through the MAPK and PI3K/AKT signaling pathways, with miRNAs playing a crucial role. These results indicate that LRM-ELNs have the protection effect on PC12 cells and can be considered as dietary supplements for alleviating neurodegenerative diseases.

Nutritional Value, Phytochemical Composition and Biological Activities of Lycium barbarum L. fruits from Serbia.

Cultivation of goji berries (GB), fruits of Lycium barbarum L. (Solanaceae), is expanding worldwide, including in Europe. In this study, a comparative analysis of the nutritional value, chemical composition and in vitro biological activities of GB from different locations in Serbia was performed. Proximate compositions were evaluated according to standard methods. Minerals were assessed by inductively coupled plasma techniques, while fatty acids, sterols, and phenolic profiles were analyzed by gas- and liquid chromatography-based techniques coupled with flame-ionization, mass spectrometry, or diode array detection. The total content of phenolics, flavonoids, carotenoids, and polysaccharides was assessed using spectrophotometric methods. Methanol extracts from GB were examined for their antioxidant, enzyme inhibitory (α-amylase, α-glucosidase, acetylcholinesterase and tyrosinase) and antibacterial activities. Despite significant variations among samples from different locations, the results confirmed that GB are a valuable source of dietary fiber and protein and are characterized by favorable fatty acid profiles. Phytochemical analysis revealed that ß-sitosterol, Δ5-avenasterol, and 24-methyldesmosterol are the predominant sterols and caffeic acid, gallic acid, quercetin and rutin are the main phenols. All GB samples showed both antioxidant and mild antimicrobial activity. A dose-dependent anti-enzymatic activity (IC50 ranging 1.68-6.88 mg/mL) was demonstrated. The results support further promotion of GB cultivation in Serbia and further investigations on their potential applications in various industries.