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Objective: To optimize the culture methods and conditions for the rapid propagation of Lycium ruthenicum in vitro, explore effective proliferation methods, and screen suitable plant regeneration pathways, so as to establish an artificial high-efficiency breeding technology system. Methods: Stem segments with one or two nodes of aseptic seedlings were used as materials. MS, modified MS1 and modified MS2 were used as basic media. The effects of different plant hormones and their concentrations on callus induction, axillary bud germination, basal stem adventitious cluster bud induction and plant regeneration were studied by single factor, complete combination and L9(34) orthogonal experiments. Results: A large number of callus were induced in MS + NAA 1.0 mg/L + 6-BA 0.1 mg/L medium, but the ability of redifferentiation was weak, and the highest multiplication coefficient was only 4.36 after 35 d of culture; While in MS + NAA 0.1 mg/L + 6-BA 0.05 mg/L + KT 0.5 mg/L medium, with axillary buds starting to germinate, the node of stem segment contacted with culture medium began to swell and appeared adventitious bud points, which sprouted basal stem cluster budswith the incidence rate of 100%: And the highest multiplication coefficient could up to 42.84 after 45 d of culture. The suitable medium (MS1 + NAA 1.0 mg/L) for rooting of test-tube seedlings was modified. After 40 d of culture, the rooting rate reached 98.9%; And the survival rate of transplanted tube seedlings after refining was over 90%. Conclusion: In this study, the basal stem cluster buds were used as a new way for proliferation, and an efficient propagation system of L. barbarum in vitro was successfully established, which not only greatly improved the yield and quality of test-tube seedlings, but also provided another idea for the rapid propagation of other Lycium plants in vitro.
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Objective: Based on the transcriptome sequencing results of Lycium barbarum under different concentrations of NaCl stress, the bHLH transcription factor family members of Lycium barbarum were identified by bioinformatics method. Methods: The bHLH family genes were screened by transcriptome sequencing of leaf and root samples of Lycium ruthenicum Murr. under NaCl stress. Results: The physicochemical properties, conserved domain, gene structure, cell location and phylogenetic development of these genes were analyzed by bioinformatics method. The results showed there were 89 bHLH family of Lycium ruthenicum. under NaCl stress. Their physicochemical properties were different, of which 71.90% proteins were weakly acidic and they were hydrophilic protein. The bHLH family of Lycium ruthenicum contained two conserved domains, which were located in the alkaline amino acid region of N and the helical ring and spiral region of C, respectively. The subcellular localization prediction of these bHLHs were mainly in the nucleus and extracellular. Phylogenetic analysis showed that 89 bHLH genes were divided into 20 subgroups, among which the 3 subgroup contained the most abundant bHLH members,including 11 bHLH proteins; the 7, 11, 13, 22 subgroups had only 1 member, respectively. In the other subgroups, the range was between 2 and 8. Conclusion: There are 89 members of bHLH transcription factor family in Lycium barbarum under NaCl stress, and most of the bHLH family proteins are weakly acidic, belonging to hydrophilic proteins, which can be divided into 20 subgroups.
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Lycium ruthenicum is a kind of medicinal and edible plant with excellent health-care effect, which is a unique medicinal plant in the desert region of northwest China. Phytochemical investigations have identified that the fruit of this herb contains a variety of bioactive ingredients, including anthocyanins, flavonoids, alkaloids, and polysaccharides, as well as of fatty acids, amino acids, and some trace elements (such as manganese, selenium, and zinc, etc). Modern pharmacological researches have demonstrated that both the extract of L. ruthenicum and its constituents exhibit a wide range of pharmacological activities, such as anti-oxidation, anti-fatigue, hypoglycemic and hypolipidemic activity, cardiovascular and liver protection, as well as immune-regulatory activity. The chemical constituents of L. ruthenicum and their pharmacological effects are systematically summarized in this paper, and all information presented here may strongly facilitate further investigations on the pharmacological activities of this herb and the development and applications of the related products of this herb.
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Objective: Both Lycium barbarum L. (LB) and Lycium ruthenicum Murr. (LR) belong to the Lycium genus of Solanaceae family but their fruits have significant phenotypic differences in terms of color and shapes. This study is aimed to investigate the inter-species difference of their fruit polyphenol composition. Methods: The polyphenol composition of fresh and dried fruits of two Lycium species were comprehensively analyzed using ultra-high performance liquid chromatography-electrospray ionization-quadrupole-time of flight mass spectrometry (UHPLC-ESIQ- TOF-MS). Results: 35 polyphenolic compounds were detected and quantified in the fresh and dried Lycium fruits including phenolic acids, anthocyanins and phenolamides (i.e., the amide-adducts of polyphenolic acids and polyamines). For both species, the dried fruits had more polyphenolic compounds than their fresh fruits with significant inter-species difference; for both fresh and dried fruits, LR had more polyphenolic compounds than LB. Conclusion: There are significant inter-species differences between these two Lycium in both fresh and dried fruits. These results offer essential information for further understanding of the biological functions of these two Lycium fruits as phytomedicines and functional food.
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Objective To clone the R1-MYB transcription factor participated in the anthocyanidin metabolism, and to analyze by bioinformatics analysis. Different expression of different varieties, different organs of the same species and salt stress conditions in Lycium were analyzed. To clone the full-length cDNA encoding R1-MYB, to perform bioinformatic analysis, and to study its expression in different cultivators and different developmental stage and in response to NaCl stress in Lycium ruthenicum and L. barbarum. Methods The full-length cDNA encoding R1-MYB was cloned using homology-based cloning and rapid amplification of cDNA ends (RACE) technique in L. ruthenicum, and the homologous gene was obtained by transcriptome in L. barbarum. The bioinformatics analysis was carried out by using Prot, Param, Smart, PSORT, and SOPMA methods. And the phylogenetic tree was constructed based on software MEGA5.0. Gene expression analysis was done by method of Real-time PCR. Results We the MYB transcription factor in L. ruthenicum was cloned and named as LrMYB1R1 (GenBank accession number KY568981), and LbMYB1R1 (GenBank accession number KY568982) in L. barbarum. Bioinformatics analysis showed that the length of LrMYB1R1 was 1 496 bp and the CDS was 927 bp. The coding products contained 308 amino acids, the molecular weight of the protein was 33 400 and 33 490, the theoretical isoelectric point was 7.80 and 7.78, belonging to the R1-MYB transcription factor, and the encoded protein is predicted to be located in the nucleus. The results of phylogenetic tree analysis showed that LrMYB1R1 and LbMYB1R1 were highly similar to MYB1R1-like protein in Solanum lycopersicum, Solanum tuberosum, and Nicotiana tabacum. Real-time PCR analysis showed that LrMYB1R1 had higher expression level in leaves and young fruits in L. ruthenicum, followed by stems, young leaves, flowers, purple fruits and black fruits, only slightly expressed in roots. In addition, the relative expression levels of LrMYB1R1 decreased in response to salt stress. Conclusion The study of R1 MYB transcription factor has been enriched, which has laid the foundation for the subsequent research on gene function and for the high-yielding anthocyanin by genetic engineering method in L. ruthenicum.
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OBJECTIVE: To study the chemical constituents from the fruits of Lycium ruthenicum Murr. METHODS: The compounds were isolated by various chromatographic methods including macroporous adsorption resin, silica gel, ODS, Sephadex LH-20 column chromatography, and preparative HPLC, and their structures were elucidated on the basis of spectroscopic data and physicochemical methods. RESULTS: Eleven compounds were isolated from the 65% ethanol extract of the fruits of Lycium ruthenicun Murr., and their structures were identified as p-hydroxyphenethyl trans-ferulate(1), kaempferol 3-O-β-D-glucopyranoside(2), quercetin 3-O-β-D-glucopyranoside(3), syringin(4), quercetin(5), ethyl caffeate(6), p-coumaric acid(7), ferulic acid(8), 2, 6-bis (1-phenylethyl) phenol(9), dotriacontane(10), and daucosterol(11). CONCLUSION: Compounds 1-4, 6, 9, and 10 are for the first time isolated from Lycium ruthenicum Murr. Compounds 1-8 show moderate protective effects on OGD-induced PC12 cell lines.
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OBJECTIVE: To isolate and identify endophytic fungi from the Tibetan drug Lycium ruthenicum Murr., then to investigate their antimicrobial activities, in order to obtain the strain with strong antimicrobial activities. METHODS: Endophytic fungi were isolated from the roots, stems and leaves of Lycium ruthenicum Murr., and then morphologically identified, and the antimicrobial activities were investigated by paper sheet method. RESULTS: Eighty-one strains of endophytic fungi were isolated from Lycium ruthenicum Murr., of which 60 strains were isolated in the general condition, 21 strains in high level of salinity. The 81 strains were i-dentified as 7 orders, 10 families, and 13genuses. The experiments showed saline-alkali fungus had stronger antimicrobial activities than the strains isolated in the general condition. CONCLUSION: It is very essential to simulate the growth environment of plants when isolating endophytic fungi.