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Differentiation of Lycium barbarum from its related Lycium species using random amplified polymorphic DNA.

The RAPD (random amplified polymorphic DNA) technique was applied for the first time to distinguish Lycium barbarum from other closely related species of the same genus. In this study, eight samples were collected, including five species, two varieties and one cultivated variety. A total of fifty arbitrary primers were used in the RAPD analysis. Distinctive DNA fingerprints corresponding to different Lycium species were successfully obtained from ten primers. Similarity index (S.I.) analysis revealed that the values are higher between intraspecies than interspecies. These results confirmed that the RAPD technique can be employed for distinguishing closely related species of Lycium.

Base Sequence↗

Effect of the Lycium barbarum polysaccharides on age-related oxidative stress in aged mice.

Oxidative damage of biomolecules increases with age and is postulated to be a major causal factor of various physiological function disorders. Consequently, the concept of anti-age by antioxidants has been developed. Lycium barbarum fruits have been used as a traditional Chinese herbal medicine and the data obtained in in vitro models have clearly established the antioxidant potency of the polysaccharides isolated from the fruits. In the present study, the age-dependent changes in the antioxidant enzyme activity, immune function and lipid peroxidation product were investigated and effect of Lycium barbarum polysaccharides on age-induced oxidative stress in different organs of aged mice was checked. Lycium barbarum polysaccharides (200, 350 and 500 mg/kg b.w. in physiological saline) were orally administrated to aged mice over a period of 30 days. Aged mice receiving vitamin C served as positive control. Enzymatic and non-enzymatic antioxidants, lipid peroxides in serum and tested organs, and immune function were measured. Result showed that increased endogenous lipid peroxidation, and decreased antioxidant activities, as assessed by superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and total antioxidant capacity (TAOC), and immune function were observed in aged mice and restored to normal levels in the polysaccharides-treated groups. Antioxidant activities of Lycium barbarum polysaccharides can be compable with normal antioxidant, vitamin C. Moreover, addition of vitamin C to the polysaccharides further increased the in vivo antioxidant activity of the latter. It is concluded that the Lycium barbarum polysaccharides can be used in compensating the decline in TAOC, immune function and the activities of antioxidant enzymes and thereby reduces the risks of lipid peroxidation accelerated by age-induced free radical.

Aging↗

Hypoglycemic and hypolipidemic effects and antioxidant activity of fruit extracts from Lycium barbarum.

The hypoglycemic and hypolipidemic effects of Lycium barbarum fruit water decoction, crude polysaccharide extracts (crude LBP), and purified polysaccharide fractions (LBP-X) in alloxan-induced diabetic or hyperlipidemic rabbits were investigated through designed sequential trials and by measuring blood glucose and serum lipid parameters. Total antioxidant capacity was also assessed using trolox equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assay. It was found that the three Lycium barbarum fruit extracts/fractions could significantly reduce blood glucose levels and serum total cholesterol (TC) and triglyceride (TG) concentrations and at same time markedly increase high density lipoprotein cholesterol (HDL-c) levels after 10 days treatment in tested rabbits, indicating that there were substantial hypoglycemic and hypolipidemic effects. Hypoglycemic effect of LBP-X was more significant than those of water decoction and crude LBP, but its hypolipidemic effect seemed to be weaker. Total antioxidant capacity assay showed that all three Lycium barbarum extracts/fractions possessed antioxidant activity. However, water and methanolc fruit extracts and crude polysaccharide extracts exhibited stronger antioxidant activity than purified polysaccharide fractions because crude extracts were identified to be rich in antioxidants (e.g., carotenoids, riboflavin, ascorbic acid, thiamine, nicotinic acid). Lycium barbarum polysaccharides (glycocojugates), containing several monosaccharides and 17 amino acids, were major bioactive constituents of hypoglycemic effect. Both polysaccharides and vitamin antioxidants from Lycium barbarum fruits were possible active principles of hypolipidemic effect.

Animals↗

Assessing phylogenetic relationships of Lycium samples using RAPD and entropy theory.

AIM: To evaluate the phylogenetic relationships among related species of Lycium samples. METHODS: Random amplified polymorphic DNA (RAPD) fingerprinting and lab-on-a-chip electrophoresis techniques were used to analyze the characteristics of Lycium species. Seven species and 3 varieties of Lycium were studied. Based on RAPD fingerprint data obtained from 11 primers, we proposed a new index, called dispersivity, using entropy theory and projection methods to depict the diversity of the DNA fingerprints. RESULTS: Using the proposed dispersivity, primers were sorted and the dendrograms of the 7 species and 3 varieties of Lycium were constructed synthetically by merging primer information. CONCLUSION: Phylogenetic relationships among Lycium samples were constructed synthetically based on RAPD fingerprint data generated from 11 primers.

DNA Fingerprinting↗

Association of ploidy and sexual system in Lycium californicum (Solanaceae).

In North American Lycium (Solanaceae), the evolution of gender dimorphism has been proposed as a means of restoring outcrossing after polyploidization causes the loss of self-incompatibility. Previous studies of this process in Lycium focused on comparisons between species that differ in ploidy. We examined intraspecific variation in floral morphology and DNA content in populations of L. californicum to determine correlations between sexual system and cytotype. We also used nuclear ITS and GBSSI sequence data to determine whether diploid and polyploid forms represent the same phylogenetic species, and the phylogeographic relationships among populations and ploidy levels. Within populations, no variation in ploidy was found, although among populations there was a perfect correspondence between sexual system and cytotype. Diploid populations were all hermaphroditic, whereas tetraploid populations were all gender dimorphic. There was no clear geographic pattern to the occurrence of diploid and tetraploid forms. Phylogenetic analysis confirms that L. californicum, regardless of ploidy, forms a monophyletic group within the genus Lycium. Sequences from diploid and polyploid individuals did not form reciprocally monophyletic clades, indicating either multiple gains of polyploidy, ongoing gene flow between cytotypes, or lack of lineage sorting since the evolution of polyploidy. The correspondence between ploidy and sex expression is consistent with the hypothesis that polyploidization triggers the evolution of gender dimorphism in this and other Lycium species.

Arizona↗

[The effects of Ag+ on the absorption of trace metal ion during the somatic embryogenesis of Lycium barbarum. L].

We used callus of medical plant, Ningxia Lycium barbarum. L as material to induce somatic embryo genesis, and used multitracer technique and gamma-ray energy spectrum analytical method to study different concentration of AgNO3 on some absorption of trace metal ions during the somatic embryogenesis of Lycium barbarum. L. The results show that (1) when the concentration of AgNO3 is less then 50 mg/L, with the increase of AgNO3 concentration, the absorption rate of some metal ions is increasing, but when the concentration is excess of such concentration, the effects on the absorption rate are different. Ag+ has the effect of coordination, resistance or competition on the absorption of metal ions. (2) AgNO3 in suitable concentration will promote cell differentiation and somatic embryogenesis. When the concentration of AgNO3 is less then 50 mg/L, along with the increase of the concentration, the frequency of somatic embryogenesis is increased, and Ag+ has the promotion on the somatic embryogenesis of Lycium barbarum. L. While the concentration is up to 50 mg/L, it will mostly raise somatic embryogenesis in calli, the amount of callus is about 3 times as much as that of contrast group (in which AgNO3 is not added). When excess of this concentration point, Ag+ has toxic effect, and the somatic embryogenesis is inhibited remarkably.

Absorption↗

Phylogeny and biogeography of the genus Lycium (Solanaceae): inferences from chloroplast DNA sequences.

Lycium comprises approximately 70 species and is disjunctly distributed in temperate to subtropical regions in South America, North America, southern Africa, Eurasia, and Australia. Among them, only Lycium sandwicense A. Gray sporadically occurs widely on oceanic islands in the Pacific Ocean. To investigate phylogenetic and biogeographic relationships of the genus with emphasis on L. sandwicense, the coding region of matK, the two intergenic spacers trnT (UGU)-trnL (UAA) and trnL (UAA)-trnF (GAA), and the trnL (UAA) intron of chloroplast DNA (cpDNA) were sequenced. A strict consensus tree resulting from the phylogenetic analysis indicates the following: (1) New World species comprise a potentially paraphyletic assemblage; (2) southern African, Australian, and Eurasian species together are monophyletic; (3) southern African species are a paraphyletic assemblage; and (4) L. sandwicense is in a clade with certain New World species. The estimated biogeographic events based on the cpDNA analysis indicate that (1) Lycium originated in the New World, (2) all southern African, Australian, and Eurasian species have a common ancestor from the New World, (3) Australian and Eurasian species originated once from a southern African progenitor, and (4) L. sandwicense differentiated from the New World species.

DNA, Chloroplast↗

Protective effect of Lycium barbarum polysaccharides on streptozotocin-induced oxidative stress in rats.

Fruit from Lycium barbarum L. in the family Solanaceae is well-known in traditional Chinese herbal medicine. Lycium barbarum polysaccharides (LBP) have been identified as one of the active ingredients responsible for its biological activities. We isolated polysaccharides from dried Lycium barbarum fruits by boiling water extraction. In the study, 50 animals were divided into two groups: a nondiabetic control (n=10) and a diabetic group (n=40). Diabetes was induced by a single injection of streptozotocin (50mg/kg BW; Sigma, USA) freshly dissolved in a 0.1 mol/L citrate buffer (pH 4.5) into the intraperitonium. The normal control rats and the untreated diabetic control rats were only injected with the citrate buffer. Treated diabetic rats were administrated with LBP in drinking water through oral gavage for 30 days. At the end of experiment, oxidative indice in blood, liver and kidney of all groups were examined. The results show that administration of LBP can restore abnormal oxidative indice near normal levels. Therefore, we may assume that LBP is effective in the protection of liver and kidney tissue from the damage of STZ-induced diabetic rats and that the LBP may be of use as a antihyperglycemia agent.

Animals↗

[Effects of Lycium barbarum L and Drynaria fortunei J Smith on in vitro attachment and growth of human gingival fibroblasts on root surfaces].

In this study, the effects of Lycium barbarum L and Drynarna fortunei J Smith on in vitro attachment and growth of human gingival fibroblasts to root surfaces were investigated. The results revealed that the above herbs at dose of 1.25 mg/ml could improve attachment and growth of fibroblast, on the planed diseased root surfaces to a certain extent. When exposed to Lycium brabrum L cells on the diseased root surfaces increased markedly in number, with more even distribution, better spread, and more exuberant growth. Drynaria fortunei J Smith was not as potent as Lycium brabarum L. The results suggest that the two drugs may improve the formation of new attachment of periodontal tissue.

Cell Adhesion↗

Regeneration of transgenic Lycium barbarum L.

A simple and effective system for the transformation and regeneration of Lycium barbarum L. has been developed. Young stem segments from Lycium barbarum L. were infected by Agrobacterium tumefaciens harboring a vector containing neomycin phosphotransferase II (npt-II) gene derived from non-oncogenic Ti plasmid. Calli originating from young stem segments on selective induction medium could differentiate into buds on selective differentiation medium rapidly and finally developed into whole plants. NPT-II enzyme activity assay and DNA hybridization indicated that the foreign gene had been integrated into the genome of Lycium barbarum L. and could be expressed in plants.

Kanamycin↗

Genetic transformation of Lycium barbarum L. via A. tumefaciens.

A system for transformation and regeneration of Lycium barbarum L., an important Chinese medical plant, has been established. Young stem segments from Lycium barbarum L. were infected with Agrobacterium tumefaciens C58cl(pGV3850::neo1103), and the transformed calli selected from the callus induction medium containing 50 micrograms/ml kanamycin could regenerate buds on differentiation medium containing 25 micrograms/ml kanamycin. 30% of the regenerated buds were normal in morphology. The normal buds could develop into whole plantlets after they were transferred to the rooting medium to induce roots. Nopaline detection, NPT-II enzyme activity assay and Southern blotting hybridization indicated that the foreign genes had been integrated into the genome of Lycium barbarum L. and expressed in the plant. In the processes of experiments, it was found that (i) after the pre-processes, the explants which formed callus quickly were easy to transform; (ii) the rate of normal regenerated plants from transgenic calli was higher than that from the untransgenic ones.

Agrobacterium tumefaciens↗

R2R3-MYB transcription factor MYB113 specifically regulates anthocyanin accumulation in Lycium ruthenicum.

LrMYB113 drives anthocyanin biosynthesis in Lycium ruthenicum by forming an MBW complex and directly activating LrDFR and LrANS promoters, providing a genetic target for enhancing flavonoid production. Lycium ruthenicum Murray (black goji berry), a Solanaceae medicinal plant, is valued for its high flavonoid content. However, the transcriptional regulation of flavonoid biosynthesis in L. ruthenicum remains unclear, hindering its pharmaceutical development. Here, we identified and characterized LrMYB113, an R2R3-MYB transcription factor, as a key regulator of anthocyanin biosynthesis in L. ruthenicum. Phylogenetic analysis grouped LrMYB113 into the anthocyanin-associated S6 subgroup of MYBs. Heterologous expression of LrMYB113 in tobacco induced pigment accumulation and upregulated anthocyanin pathway genes. LrMYB113 overexpression in L. ruthenicum hairy roots enhanced accumulation of four acylated anthocyanins and activated anthocyanin pathway genes. Yeast two-hybrid and bimolecular fluorescence complementation assays showed LrMYB113 interacts with bHLHs (LrJAF13/LrAN1b) and WD40 (LrAN11) to form an MBW complex. Promoter binding and transactivation assays demonstrated LrMYB113 directly binds to and activates LrDFR and LrANS promoters. Dual-luciferase assays showed LrMYB113 alone strongly activates LrDFR and LrANS promoters; MBW complexes enhanced activity compared to individual bHLH/WD40 but not to LrMYB113 alone. Our findings identify LrMYB113 as a critical regulator of anthocyanin biosynthesis in L. ruthenicum, shedding light on flavonoid molecular mechanisms and supporting genetic improvement for pharmaceutical use.

Anthocyanins↗

Experimental study on the enhancement of murine splenic lymphocyte proliferation by Lycium barbarum glycopeptide.

In order to investigate the immunoactivity of Lycium Barbarum glycopeptide (LBG), the routinely prepared murine splenic lymphocyte suspension was separately added into the samples with different concentrations (500, 100, 10, 1 microg/ml) of LBG as LBG groups. Blank control group in the absence of Lycium Barbarum glycopeptide or ConA and positive control group in the presence of 0.5 ml ConA but in the absence of LBG were created. 0.5 ml LBG samples with different concentrations in combination with 0. 5 ml ConA (10 microg/ml) into each well to observe the synergic effects of LBG and ConA as LBG+ConA groups. After incubation for 72 h at 37 degrees C, the samples were analyzed by CFSE-labeled cells combined with flow cytometry, and MTT. Flow cytometry revealed that both LBG could enhance the murine splenic lymphocyte proliferative reaction. Combined use of LBG and ConA had synergic effects. MTT demonstrated that sample A could obviously promote the murine splenic lymphocyte proliferative reaction as compared with control group (P<0. 01), while sample B could also enhance the lymphocyte proliferation at a high dose. In combination with ConA, sample A had synergic effects at high dose, while sample B showed obviously synergic effects (P<0.05). It was concluded that both samples (A and B) had strong immunocompetence.

Animals↗

Fermentation of a milk-soymilk and Lycium chinense Miller mixture using a new isolate of Lactobacillus paracasei subsp. paracasei NTU101 and Bifidobacterium longum.

A milk-soymilk mixture was fermented using Lactobacillus paracasei subsp. paracasei NTU101 and Bifidobacterium longum BCRC11847 at different inoculum ratios (1:1, 1:2, 1:5, 2:1, and 5:1). When the inoculum ratio was 1:2, the cell numbers of both strains were balanced after 12 h of cultivation. The pH and titratable acidity were very similar at the various inoculum ratios of cultivation. The milk-soymilk mixture was supplemented with 5, 10, 15, and 20% Lycium chinense Miller juice and fermented with Lactobacillus paracasei subsp. paracasei NTU101 and B. longum BCRC11847. Sensory evaluation results showed that supplementation with 5% Lycium chinense Miller juice improved the acceptability of the fermented milk-soymilk. The fermented beverage was stored at 4 degrees C for 14 days; variations in pH and titratable acidity were slight. The cell numbers of L. paracasei subsp. paracasei NTU101 and B. longum BCRC11847 in the fermented beverage were maintained at 1.2x10(9) CFU/ml and 6.3x10(8) CFU/ml, respectively, after 14 days of storage.

Animals↗

Quantification of zeaxanthin dipalmitate and total carotenoids in Lycium fruits (Fructus Lycii).

An HPLC-DAD method has been developed to quantitatively analyze for the content of zeaxanthin dipalmitate, a major carotenoid in Fructus Lycii, in different species of the genus Lycium. Determination was performed using an Alltima C18 column with the mobile phase consisting of acetonitrile and dichloromethane (42:58). The total contents of carotenoids in these samples were also determined by using UV spectrophotometric assay. Total carotenoid concentrations of different Fructus Lycii are within the range of 0.03-0.5%. Zeaxanthin dipalmitate is a predominant carotenoid, comprising 31-56% of the total carotenoids in Fructus Lycii. This study is the first systematic quantification of the carotenoids in the fruits of different Lycium species. The results demonstrated that these methods are reliable and facile techniques for rapid analysis of carotenoids for crude drug and plant-derived food supplements.

Antioxidants↗

Effect of lycium barbarum polysaccharide on human hepatoma QGY7703 cells: inhibition of proliferation and induction of apoptosis.

Lycium barbarum polysaccharide (LBP), extracted from Lycium barbarum that is a kind of traditional Chinese herb, is found to have anticancer activity. In this study, the effect of LBP on the proliferation rate, cell cycle distribution and apoptosis in the human hepatoma QGY7703 cell line were investigated. The effects of this compound were also tested on the concentration of calcium in cells. LBP treatment caused inhibition of QGY7703 cell growth with cycle arrest in S phase and apoptosis induction. The amount of RNA in cells and the concentration of intracellular Ca2+ were increased. Moreover, the distribution of calcium in cells was changed. Taken together, the study suggests that the induction of cell cycle arrest and the increase of intracellular calcium in apoptotic system may participate in the antiproliferative activity of LBP in QGY7703 cells.

Antineoplastic Agents, Phytogenic↗

2-O-(beta-D-Glucopyranosyl)ascorbic acid, a novel ascorbic acid analogue isolated from Lycium fruit.

A novel stable precursor of ascorbic acid (vitamin C), 2-O-(beta-D-glucopyranosyl)ascorbic acid, was isolated from both the ripe fresh fruit and dried fruit of Lycium barbarum L., a plant of the Solanaceae family. The chemical structure was inferred by instrumental analyses and confirmed by chemical synthesis. The dried fruit of Lycium barbarum L. contained ca. 0.5% of it, which is comparable to the ascorbic acid content of fresh lemons. It increased the blood ascorbic acid by oral administration to rats, and it was also detected in blood from the portal vein.

Animals↗

Gametophytic self-incompatibility in Lycium parishii (Solanaceae): allelic diversity, genealogical structure, and patterns of molecular evolution at the S-RNase locus.

We characterized allelic diversity at the locus controlling self-incompatibility (SI) for a population of Lycium parishii (Solanaceae) from Organ Pipe National Monument, Arizona. Twenty-four partial sequences of S-RNase alleles were recovered from 25 individuals. Estimates of allelic diversity range from 23 to 27 alleles and, consistent with expectations for SI, individuals are heterozygous. We compare S-RNase diversity, patterns of molecular evolution, and the genealogical structure of alleles from L. parishii to a previously studied population of its congener L. andersonii. Gametophytic SI is well characterized for Solanaceae and although balancing selection is hypothesized to be responsible for high levels of allelic divergence, the pattern of selection varies depending on the portion of the gene considered. Site-specific models investigating patterns of selection for L. parishii and L. andersonii indicate that positive selection occurs in those regions of the S-RNase gene hypothesized as important to the recognition response, whereas positive selection was not detected for any position within regions previously characterized as conserved. A 10-species genealogy including S-RNases from a pair of congeners from each of five genera in Solanaceae reveals extensive transgeneric evolution of L. parishii S-RNases. Further, within Lycium, the Dn/Ds ratios for pairs of closely related alleles for intraspecific versus interspecific comparisons were not significantly different, suggesting that the S-RNase diversity recovered in these two species was present prior to the speciation event separating them. Despite this, two S-RNases from L. parishii are identical to two previously reported alleles for L. andersonii, suggesting gene flow between these species.

Base Sequence↗