中草药枸杞(Lycium chinense)愈伤组织分化的最佳培养基

枸杞分布广泛,果实和根皮是贵重药材,果实滋补肝肾,治糖尿病;根自治咳嗽、吐血及肺结核等病。枸杞种子繁殖3年后才能开花结实,为了加快无性繁殖。

Drought stress,mercuric chloride,andβ-mercaptoethanol effects on hydraulic characteristics of three cultivars of wolfberry(Lycium chinense)

In order to further understand the effects of drought stress on hydraulic characteristics and the relationship between hydraulic conductivity and aquaporins or water channels of root systems of three wolfberry cultivars(Lycium chinense Mill.),hydraulic conductivity of 2-yearold pot-grown seedlings was measured under drought stress,rewatering,and treatment with exogenous mercuric chloride andβ-mercaptoethanol.Under moderate and severe drought stress levels,the most significant decrease of hydraulic conductivity was 37.3%and 24.0%,respectively,in the‘Ningqi 5’cultivar compared with the nonstressed control.After rewatering,the rate of recovery in specific conductivity was most rapid in the‘Mengqi 1’cultivar,at 0.058 and 0.072 kg MPa^-1 m^-2 s^-1 h^-1 under moderate and severe drought stress levels,respectively.The‘Mengqi 1’cultivar had the highest recovery degree of hydraulic conductivity under two concentrations ofβ-ME(500 or 1000 lmol L^-1),reaching 82.4%and 88.5%,respectively,of the initial conductivity.The adaptive capacity of hydraulic conductivity in the‘Ningqi 5’cultivar was weaker than in the‘Ningqi 1’and‘Mengqi 1’cultivars under drought stress.The recovery capacity of hydraulic conductivity in‘Mengqi 1’cultivar was stronger than the‘Ningqi 1’and‘Ningqi 5’cultivars after rewatering.Aquaporins of the‘Ningqi 1’cultivar root systems had the highest binding affinity with mercuric chloride,which was the most likely cause in the decrease in hydraulic conductivity,whereas aquaporins of‘Mengqi 1’root systems had the weakest binding affinity.The inhibitory effect of mercuric chloride was readily eliminated byβ-mercaptoethanol in the‘Mengqi 1’cultivar.The hydraulic characteristics of this cultivar were more sensitive to drought,mercuric chloride andβ-mercaptoethanol than the other cultivars.

Cloning and Characterization of a Lycium chinense Carotenoid Isomerase Gene Enhancing Carotenoid Accumulation in Transgenic Tobacco

Carotenoid isomerase （CRTISO）is a key enzyme that catalyzes the conversion of cis-lycopene to all- trans tycopene. In this study, we isolated and characterized the CRTISO gene from Lycium chinense （LcCRTISO） for the first time. The open reading flame of LcCRTISO was 1 815 bp encoding a protein of 604 amino acids with a molecular mass of 66.24 kDa. Amino acid sequence analysis revealed that the LcCRTISO had a high level of simi- larity to other CRTISO. Phylogenetic analysis displayed that LcCRTISO kept a closer relationship with the CRTISO of plants than with those of other species. Semi-quantitative PCR analysis indicated that LcCRTISO gene was expressed in all tissues tested with the highest expression in maturing fruits. The overexpression of LcCRTISO gene in transgenic tobacco resulted in an increase of total carotenoids in the leaves with [3-carotene and lutein being the predominants. The results obtained here clearly suggested that the LcCRTISO gene was a promising candidate for carotenoid production.

Lycium barbarum glycopeptide(wolfberry extract)slows N-methyl-N-nitrosourea-induced degradation of photoreceptors

Photoreceptor cell degeneration leads to blindness, for which there is currently no effective treatment. Our previous studies have shown that Lycium barbarum(L. barbarum) polysaccharide(LBP) protects degenerated photoreceptors in rd1, a transgenic mouse model of retinitis pigmentosa. L. barbarum glycopeptide(Lb GP) is an immunoreactive glycoprotein extracted from LBP. In this study, we investigated the potential protective effect of Lb GP on a chemically induced photoreceptor-degenerative mouse model. Wild-type mice received the following: oral administration of Lb GP as a protective pre-treatment on days 1–7;intraperitoneal administration of 40 mg/kg N-methylN-nitrosourea to induce photoreceptor injury on day 7;and continuation of orally administered Lb GP on days 8–14. Treatment with Lb GP increased photoreceptor survival and improved the structure of photoreceptors, retinal photoresponse, and visual behaviors of mice with photoreceptor degeneration. Lb GP was also found to partially inhibit the activation of microglia in N-methyl-N-nitrosourea-injured retinas and significantly decreased the expression of two pro-inflammatory cytokines. In conclusion, Lb GP effectively slowed the rate of photoreceptor degeneration in N-methyl-N-nitrosourea-injured mice, possibly through an anti-inflammatory mechanism, and has potential as a candidate drug for the clinical treatment of photoreceptor degeneration.

Retinal ischemia-reperfusion injury and pretreatment with Lycium barbarum glycopeptide

AIM:To investigate the antioxidant protective effect of Lycium barbarum glycopeptide(LbGP)pretreatment on retinal ischemia-reperfusion(I/R)injury(RIRI)in rats.METHODS:RIRI was induced in Sprague Dawley rats through anterior chamber perfusion,and pretreatment involved administering LbGP via gavage for 7d.After 24h of reperfusion,serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),and creatinine(CREA)levels,retinal structure,expression of Caspase-3 and Caspase-8,superoxide dismutase(SOD)activity,and malondialdehyde(MDA)in the retina were measured.RESULTS:The pretreatment with LbGP effectively protected the retina and retinal tissue from edema and inflammation in the ganglion cell layer(GCL)and nerve fiber layer(NFL)of rats subjected to RIRI,as shown by light microscopy and optical coherence tomography(OCT).Serum AST was higher in the model group than in the blank group(P=0.042),but no difference was found in ALT,AST,and CREA across the LbGP groups and model group.Caspase-3 expression was higher in the model group than in the blank group(P=0.006),but no difference was found among LbGP groups and the model group.Caspase-8 expression was higher in the model group than in the blank group(P=0.000),and lower in the 400 mg/kg LbGP group than in the model group(P=0.016).SOD activity was lower in the model group than in the blank group(P=0.001),and the decrease was slower in the 400 mg/kg LbGP group than in the model group(P=0.003).MDA content was higher in the model group than in the blank group(P=0.001),and lower in the 400 mg/kg LbGP group than in the model group(P=0.016).The pretreatment with LbGP did not result in any observed liver or renal toxicity in the model.CONCLUSION:LbGP pretreatment exhibits dosedependent anti-inflammatory,and antioxidative effects by reducing Caspase-8 expression,preventing declines of SOD activity,and decreasing MDA content in the RIRI rat model.

Supplementation of Lycium barbarum residue increases the growth rate of Tan sheep by enhancing their feed intake and regulating their rumen microbiome and metabolome

Lycium barbarum residue(LBR),a by-product of L.barbarum processing,is packed with bioactive components and can be potentially utilized as a feed additive in animal husbandry.However,the fundamental understanding of its effectiveness on livestock animals is still lacking,particularly in ruminants.To explore the effects of LBR on the growth performance,rumen fermentation parameters,ruminal microbes and metabolites of Tan sheep,sixteen fattening rams(aged 4 mon)were fed a basal diet(CON,n=8)or a basal diet supplemented with 5%LBR(LBR,n=8).The experiment lasted for 70 d,with 10 d adaptation period and 60 d treatment period.The results showed that the LBR enhanced the average daily feed intake,average daily gain(P<0.05),and ruminal total volatile fatty acids(P<0.01)while decreasing ammonia-nitrogen concentration and rumen pH value(P<0.05).Additionally,the LBR improved the relative abundances of Prevotella,Succiniclasticum,Ruminococcus,Coprococcus,Selenomonas,and Butyrivibrio(P<0.05)and reduced the relative abundances of Oscillospira and Succinivibrio(P<0.05).The LBR altered the ruminal metabolome(P<0.01)by increasing the abundances of ruminal metabolites involved in amino acids(e.g.,L-proline,L-phenylalanine,L-lysine,and L-tyrosine),pyrimidine metabolism(e.g.,uridine,uracil,and thymidine),and microbial protein synthesis(e.g.,xanthine and hypoxanthine).In conclusion,LBR had positive effects on the growth rate of Tan sheep as well as on rumen fermentation parameters,rumen microbiome and rumen metabolome.

Identification and Transcriptional Regulation of CAMTA Genes in Liriodendron chinense

This study explores CAMTA genes in the rare and endangered Chinese plant species,Liriodendron chinense.Despite the completion of whole-genome sequencing,the roles of CAMTA genes in calcium regulation and stress responses in this species remain largely unexplored.Within the L.chinense genome,we identified two CAMTA genes,Lchi09764 and Lchi222536,characterized by four functional domains:CG-1,TIG,ANK repeats,and IQ motifs.Our analyses,including phylogenetic investigations,cis-regulatory element analyses,and chromosomal location studies,aim to elucidate the defining features of CAMTA genes in L.chinense.Applying Weighted Gene Co-Expression Network Analysis(WGCNA),we explored the impact of CAMTA genes on different organs and their regulation under abiotic stress conditions.The identification of significant gene modules and the prediction of promoter binding sites revealed co-expressed genes associated with CAMTA transcription factors.In summary,this study provides initial insights into CAMTA genes in L.chinense,laying the groundwork for future research on their evolution and biological roles.This knowledge enhancement contributes to a better understanding of plant responses to environmental stress—an essential aspect of plant biology.

The flavonoids from Pentorum chinense Pursh.mediates ferroptosis to alleviate sorafenib-induced liver injury in BRL-3A cells

Background:Drug-induced liver damage is a severe medical issue that affects people all over the world.Sorafenib has some side effects that cause liver injury.A dietary medicinal plant called Penthorum chinense Pursh.(PCP)has hepatoprotective properties.There are currently few reports on PCP’s protective impact and mechanism against sorafenib-induced liver injury.Methods:To create a liver injury model,sorafenib was administered to BRL-3A cells.Cell viability assays,immunofluorescence tests,Western blotting,real-time quantitative PCR,and high-content imaging systems were utilized to examine PCP’s effect and mechanism.Results:In this study,PCP treatment mitigated the liver damage caused by sorafenib by enhancing cell survival,lowering lipid reactive oxygen species and malondialdehyde levels,and elevating glutathione levels.In addition,PCP can enhance the protein expression of cystine/glutamate transporter xCT and glutathione peroxidase 4,reduce iron content and alleviate mitochondrial toxicity.Further mechanism studies revealed that PCP inhibited ferroptosis by promoting the production of nuclear factor E2-related factor 2 nuclear translocation and subsequently affecting target genes(HO-1 and NQO1).Conclusion:Together,PCP regulates the nuclear factor E2-related factor 2 pathway,which helps to lessen ferroptosis brought on by sorafenib.

Effects of Lycium barbarum polysaccharide on cytokines in adolescents with subthreshold depression:a randomized controlled study

Strong evidence has accumulated to show a correlation between depression symptoms and inflammatory responses.Moreover,anti-inflammatory treatment has shown partial effectiveness in alleviating depression symptoms.Lycium barbarum polysaccharide(LBP),derived from Goji berries,exhibits notable antioxidative and anti-inflammatory properties.In our recent double-blinded randomized placebo-controlled trial,we found that LBP significantly reduced depressive symptoms in adolescents with subthreshold depression.It is presumed that the antidepressant effect of LBP may be associated with its influence on inflammatory cytokines.In the double-blinded randomized controlled trial,we enrolled 29 adolescents with subthreshold depression and randomly divided them into an LBP group and a placebo group.In the LBP group,adolescents were given 300 mg/d LBP.A 6-week follow up was completed by 24 adolescents,comprising 14 adolescents from the LBP group(15.36±2.06 years,3 men and 11 women)and 10 adolescents from the placebo group(14.9±1.6 years,2 men and 8 women).Our results showed that after 6 weeks of treatment,the interleukin-17A level in the LBP group was lower than that in the placebo group.Network analysis showed that LBP reduced the correlations and connectivity between inflammatory factors,which were associated with the improvement in depressive symptoms.These findings suggest that 6-week administration of LBP suppresses the immune response by reducing interleukin-17A level,thereby exerting an antidepressant effect.

Optimization of Extraction Process for Total Flavonoids from Penthorum chinense Pursh and Comparison of Their Contents from Different Parts

[Objectives]This study was conducted to optimize the extraction process of total flavonoids from Penthorum chinense Pursh and compare their contents from different parts.[Methods]Single factor and orthogonal experiments were designed to optimize the extraction process of total flavonoids from P.chinense Pursh with the volume fraction of ethanol,the ratio of material to liquid,heating reflux extraction time and extraction times as factors,and the content of total flavonoids as the index.A verification test was carried out.The optimized extraction process was adopted to compare the contents of total flavonoids from different parts of P.chinense Pursh.[Results]The best extraction process was extracting the powder of P.chinense Pursh for 2.0 h with 20 times of 55%ethanol by reflux twice.Under this condition,the contents of total flavonoids were 3.63%,8.90%,11.28%,and 4.36%from stems,leaves,flowers and whole grass of P.chinense Pursh,respectively.[Conclusions]The process is reasonable,feasible and stable,and can effectively extract total flavonoids from P.chinense Pursh.The contents of total flavonoids from different parts of P.chinense Pursh were quite different,and the value was higher in the leaves and flowers,so the proportions of leaves and flowers should be paid attention to in the industrial processing of P.chinense Pursh.

Establishment of Somatic Cell Clones in Thesium chinense Turcz and Its in vitro Rooting Technique

[Objective] The aim of this study is to establish the rapid micro-propagation system in Thesium chinense Turcz.[Method]With stem fragments of wild Thesium chinense Turcz as explants,different culture media were designed to conduct induction culture,strengthening plantlet culture and in vitro rooting.[Result]The optimum medium for inducing clustered shoots was determined to be MS medium appended with 1.5 mg/L 6-BA,0.01 mg/L NAA and 0.3 mg/L 2,4-D;in addition,60 mg/kg ABT was suitable for rooting,by which the percentage of rooted plantlets reached 76.6%.[Conclusion]This study simplified the procedures of tissue culture in Thesium chinense Turcz and enhanced the proliferation rate,providing basis for artificial cultivation and resource protection of Thesium chinense Turcz.

Study on the Relationship between the Main Secondary Metabolites and Polysaccharide in Fruits of Lycium barbarum at Different Application Amounts of Nitrogen

[Objective] The aim of this study is to investigate the optimum application amount of nitrogen in Lycium barbarum based on considering the relationship between main secondary metabolites and polysaccharide.[Method]Under field conditions,the effects of different application amounts of nitrogen on main secondary metabolites of betaine,carotenoid and flavone of Lycium barbarum and the relationship between main secondary metabolites and polysaccharide.[Result] The main secondary metabolites of betaine,carotenoid and flavone of Lycium barbarum varied under different application amounts of nitrogen.The proper application amount of nitrogen(600-900 kg/hm2)was beneficial to the formation and accumulation of secondary metabolites such as carotenoids.Correlation analysis results showed that polysaccharide were negatively correlated with betaine,carotenoid and flavone at significant probability level.[Conclusion]Considering the relationship between the output and quality of the fruits of Lycium barbarum,the optimum nitrogen application amount should be 600-900 kg/hm2.

不同类型群落中濒危植物永瓣藤(Monimopetalum chinense)种群的空间分布格局

为揭示永瓣藤(Monimopetalum chinense)种群的分布格局特征及其成因,采用分形分析中的计盒维数对取自4个分布地的13个样方进行了研究。结果显示:(1)同一研究地不同群落中永瓣藤种群的分形特征差异明显,而不同研究地同一类型群落之间的差异不大,分析认为这与永瓣藤在群落中的生态地位及生境异质性有关。(2)永瓣藤种群计盒维数由高到低依次为杉木林(1.911±0.025)、杂木林(1.806±0.079)、马尾松林(1.692±0.054)和毛竹林(1.629±0.081),表明不同群落中其个体分布的非均匀性存在较大差异。(3)永瓣藤种群的拐点尺度由高到低依次为毛竹林(4.111m±0.484m)、马尾松林(3.778m±0.222m)、杂木林(2.857m)、杉木林(2.013m±0.117m),次序与计盒维数相反。(4)分析认为,不同群落中永瓣藤无性繁殖能力的不同对其种群格局的计盒维数以及拐点尺度有重要影响。

Genetic Polymorphism of Eighteen Lycium barbarum Resources Based on nrDNA ITS Sequence

[Objective] The study aimed to investigate the genetic polymorphism of eighteen Lycium barbarum resources via nrDNA ITS sequencing. [Method] The genomic DNAs from Lycium barbarum leaves were isolated by modified CTAB method for PCR amplification on the nrDNA ITS region using specifically synthesized primers; the amplified fragments were cloned and sequenced, then the sequencing results were clustered. [Result] nrDNA ITS sequences of the tested eighteen Lycium barbarum were firstly obtained in the present study. For all eighteen tested materials, the variation range of whole ITS region was 559-634 bp, with an average of 612 bp; alignment analyses showed that the whole length of internal transcribed spacer (ITS1+ITS2) was 480 bp, within which there are 194 variation sites (accounting for 40.4%) and 286 conserved sites (accounting for 59.6%). The cluster results showed that the eighteen tested materials could be grouped into three classes. [Conclusion] Analysis of nrDNA ITS sequence may avail to identify the Lycium barbarum germplasm resources.

中宁枸杞(Lycium barbarum L.)的核型分析

报道了中宁枸杞( Lycium barbarum L.)的核型.核型公式为K(2n)=2 X=24=2 2 m(4 SAT)+2 sm.核型类型为1A，为较原始的对称核型，并与前人的报道结果进行了对比分 析.

杀线虫真菌Simplicillium chinense菌株Snef5强毒和弱毒菌株的诱变筛选

杀线虫真菌Simplicillium chinense菌株Snef5分离自柞蚕茧蛹体内,其发酵液对多种线虫均具有较高的致死作用。为筛选对南方根结线虫强毒与弱毒的菌株,采用紫外照射、硫酸二乙酯处理及农杆菌介导的遗传转化等3种诱变方法对菌株Snef5进行诱变育种,结果表明:共筛选得到8株强毒菌株,3株弱毒菌株。复筛后获得毒力稳定的菌株共4株,包括强毒菌株UV-17和UV-18,弱毒菌株UV-15和T-25,以农杆菌介导的遗传转化法诱变处理效果最佳。诱变菌株UV-17、UV-18、UV-15和T-25的获得,为将来对杀线虫活性基因簇进行分析研究奠定了基础。

Saikosaponin q-1 from Bupleurum chinense

A new saikosaponin was isolated from Bupleurum chinense DC., and its structure was identified as 3β,16α,23,28,30_pentahydroxy_olean_11,13(18)_dien_3_O_β_D_glucopyranosyl(1→6)_[α_L_rhamnopyranosyl (1→4)]_β_D_glucopyranoside on the basis of chemical and spectral evidence, named as saikosaponin q_1. In addition, two known saikosaponins, 3″_O_acetyl_saikosaponin d and 3″_O_acetyl_saikosaponin b 2, were also isolated and identified from this plant for the first time.

辣椒C.frutescens×C.chinense种间杂种的获得与鉴定

运用有性杂交方法,以强辣灌木辣椒Capsicum frutescens H101为母本(P1)、强辣C.chinense PI439487为父本(P2)进行种间杂交,获得了C.frutescens×C.chinense种间杂种。对种间杂种F125个表型性状进行了观察比较,结果表明:F1在生长势方面具有明显的杂种优势,其他表型性状则多介于P1与P2之间。SRAP分析显示,F1与P1、P2共有带220条,占总位点数的53.92%,与P1或P2共有带143条,占35.05%,F1特异带和特异缺失带12条,点2.95%。F1与P1遗传相似系数为0.749,大于其与P2的遗传相似系数(0.740),说明杂种在DNA水平上更趋向于母本。C.frutescens×C.chinense种间杂种的获得,为C.chinense香味和多花基因的转移及新材料创制奠定了基础。

Effects of Different Culture Conditions on Vitrification of Lycium barbarum L. Plantlets in Tissue Culture

The tender stems from new Lycium barbarum L. cultivar ＂Ningqi 3＂ released by Ningxia Academy of Agricultural and Forestry Sciences were regarded as explants to investigate the vitrification of Lycium barbarum plantlets in tissue culture under different concentrations of 6-BA, sucrose, agrose, culture temperature, and illumination duration with MS as basic medium. The results show that the conditions for maximal proliferation coefficient and min- imal vitrification are as following： the basic medium with 0.2 mg/L 6-BA, 3% sucrose and 0.65% agarose; culture at 25℃; 12 h/d（ daylight lamp, 2 000 lx） illumination.

Effects of NaCl Stress on Seed Germination of Lycium ruthenicum Murr.

[Objective]To study the seeds germination of Lycium ruthenicum Murr.under different concentrations of NaCl,as well as to find the optimal concentration of NaCl for the germination of L.ruthenicum.[Method]The seeds of L.ruthenicum were treated with different concentrations of NaCl,and the state of seed germination was measured.[Result]With the increasing of concentration of NaCl,the seed germination rate of L.ruthenicum showed an obvious increasing trend.when the concentration was of 0.3%-0.4 %,the germination rate was the highest,and when the concentration of NaCl was greater than 0.4,the germination rate showed a decline trend.[Conclusion]After treated with appropriate concentrations of NaCl before sowing,the germination rate of seeds of L.ruthenicum would increase.