Analysis of phenotype formation mechanism of a new variety of Lonicera japonica Flos "Huajin 6" at long bud stage / 药学学报

Based on the long bud stage phenotype of a new Lonicera japonica Flos variety "Huajin 6", using "Huajin 6" and "Da Mao Hua" as materials, probing the mechanism of its phenotype formation. Detection of endogenous Jasmonic acid hormones (JAs) content; the genes related to jasmonic acid (JA) synthesis were identified by transcriptome analysis of Lonicera japonica; flower buds and flowers of "Huajin 6" and "Da Mao Hua" were collected at different periods, and the qRT-PCR (quantitative real-time PCR) technique was used to analyze the trend of the expression of synthesis-related enzyme genes in Lonicera japonica Flos during the bud stage. The study found that the content of JAs in "Huajin 6" Lonicera japonica Flos was significantly lower than that in "Da Mao Hua"; applying exogenous methyl-jasmonate (MeJA) to "Huajin 6" can restore its flowering phenotype, making it close to wild type Lonicera japonica Flos; there are significant differences in the expression of two allene oxide synthase genes (AOS), three lipoxygenase genes (LOX), and two allene oxide cyclase genes (AOC) in the flowers and buds of "Huajin 6" and "Da Mao Hua" at different periods. It is hypothesized that the low expression of JA synthesis-related enzyme genes in " Huajin 6" leads to the blockage of JA synthesis, which causes the formation of the long bud phenotype. This study laid a certain foundation for the genetic breeding of Lonicera japonica, provided a new idea for the improvement of Lonicera japonica varieties, and provided a reference for the study of JAs in plant flower organs.

Cloning and expression analysis of ANR genes from different species of Lonicera japonica Thunb / 药学学报

Anthocyanidin reductase (ANR) is one of the key enzyme in the flavonoid biosynthetic pathway, and its catalytic activity is important for the synthesis of plant anthocyanin. In this study, specific primers were designed according to the transcriptome data of Lonicera japonica Thunb., and the CDS, gDNA and promoter sequences of ANR genes from Lonicera japonica Thunb. and Lonicera japonica Thunb. var. chinensis (Wats.) Bak. were cloned. The results showed that the CDS sequences of LjANR and rLjANR were 1 002 bp, the gDNA sequences were 2 017 and 2 026 bp respectively, and the promoter sequences were 1 170 and 1 164 bp respectively. LjANR and rLjANR both contain 6 exons and 5 introns, which have the same length of exons and large differences in introns. The promoter sequences both contain a large number of light response, hormone response and abiotic stress response elements. Bioinformatics analysis showed that both LjANR and rLjANR encoded 333 amino acids and were predicted to be stable hydrophobic proteins without transmembrane segments and signal peptides. The secondary structures of LjANR and rLjANR were predicted to be mainly consisted of α-helix and random coil. Sequence alignment and phylogenetic analysis showed that LjANR and rLjANR had high homology with Actinidia chinensis var. chinensis, Camellia sinensis and Camellia oleifera, and were closely related to them. The expression levels of LjANR and rLjANR were the highest in flower buds and the lowest in roots. The expression patterns at different flowering stages were similar, with higher expression levels in S1 and S2 stages and then gradually decreased until reaching the lowest level in S4 stage, after a slow increase in S5 stage, the expression levels decreased again. The expression levels of ANR genes in the two varieties showed significant differences in roots, S2 and S5 stages, while the differences in stems, flower buds, S1, S3 and S6 stages were extremely significant. The prokaryotic expression vector pET-32a-LjANR was constructed for protein expression. The target protein was successfully expressed of about 59 kD. This study lays a foundation for further study on the function of ANR gene and provides theoretical guidance for breeding new varieties of Lonicera japonica Thunb.

Comparative Analysis of Lonicera japonica Flower from Different Harvesting Periods Based on UV Fingerprint and Chemometrics / 中国实验方剂学杂志

ObjectiveTo investigate the quality variation of Lonicera japonica flower from different harvesting periods by ultraviolet visible（UV-Vis） fingerprint combined with chemometrics. MethodTwenty-five L. japonica flower samples from five harvesting periods， including young bud stage，green bud stage，white bud stage，silver and golden flower stages， were collected， with five samples for each stage. UV-Vis fingerprints of L. japonica flower from different harvesting periods were established in the context of the optimum extraction method based on the single factor experiment. The results showed that the absorption values at 209，216，226，250，280，303，318， and 350 nm were significantly different. Moreover，after data pretreatment and normalization，multivariate statistical analyses， such as principal component analysis （PCA），partial least squares discriminant analysis （PLS-DA），and orthogonal PLS-DA （OPLS-DA）were performed by SIMCA-P+ to establish the quality variation model of L. japonicas flower from harvesting periods. ResultAs revealed by PCA and PLS-DA， L. japonicas flower samples from five harvesting periods were clustered separately and closely in a harvesting time-dependent manner， suggesting that the content of components contained in samples from different harvesting periods was highly distinct and correlated with harvesting periods. The pairwise comparison of OPLS-DA indicated that triterpenoids or volatile oils were the main components causing the changes from the young bud stage to the green bud stage，and the content of them decreased. The main components from the green bud stage to the white bud stage were triterpenoids （or iridoids），volatile oils，phenolic acids， or flavonoids，and the content of them decreased， which was consistent with the HPLC result of chlorogenic acid. From the white bud stage to the silver flower stage， the main components were iridoids （increasing in content） and triterpenoids （or volatile oils） （decreasing in content）. The main altered components from the silver flower stage to the golden flower stage were triterpenoids （or volatile oils） whose content increased. ConclusionThis method is simple and feasible， which can provide references for the quality control of Chinese medicine.

Simultaneous determination of 11 active components in Lonicera japonica flowers and leaves at different development stages by HPLC-DAD / 中国中药杂志

This study aims to develop an HPLC-DAD method for simultaneous determination of 11 components(6 phenolic acids and 5 iridoids) in Lonicera japonica flowers(LjF) and leaves(LjL), and compare the content differences of LjF at different development stages, LjL at different maturity levels, and between LjF and LjL. One-way ANOVA, principal component analysis(PCA), and orthogonal partial least-squares discriminant analysis(OPLS-DA) were employed to compare the content of the 11 components. The content of total phenolic acids, total iridoid glycosides, and total 11 components in LjF showed an overall downward trend with the development of flowers. The content of total phenolic acids, total iridoid glycosides, and total 11 components in young leaves were higher than those in mature leaves. The results of PCA showed that the samples at different flowering stages had distinguishable differences in component content. The VIP value of OPLS-DA showed that isochlorogenic acid A, chlorogenic acid, and secologanic acid were the main differential components of LjF at different development stages or LjL with different maturity levels. LjF and LjL have certain similarities in chemical composition while significant differences in component content. The content of total phenolic acids in young leaves was significantly higher than that in LjF at various development stages. The content of total iridoid glycosides in young leaves was similar to that in LjF before white flower bud stage. The total content of 11 components in young leaves was significantly higher than that in LjF at green flower bud stage, before and during completely white flower bud stage. LjL have great potential for development. Follow-up research on the pharmacodynamic equivalence of LjF and LjL(especially young leaves) should be carried out to speed up the development and application of LjL.

Research progress on pesticide residues of Lonicera Japonica Flos / 中国中药杂志

Lonicera Japonica Flos is the dried bud or nascent flower of Lonicera japonica(Caprifoliaceae). The plant suffers from various diseases and pests in the growth period and thus pesticides are often used. As a result, the resultant pesticide residues in Lonicera Japonica Flos have aroused great concern. This review summarized the investigation, detection methods, content analysis, and risk assessment of pesticide residues in Lonicera Japonica Flos since 1996, and compared the maximum residue limits among different countries and regions. The results showed that the pesticide residues were detected in Lonicera Japonica Flos from different production areas, and only some exceeded the limits. The residual pesticides have changed from organochlorines to new types such as tebuconazole and nitenpyram. The detection method has upgraded from chromatography to chromatography-mass spectrometry. Most pesticide residues will not cause health risks, except carbofuran. Pesticide residues limit the development of Lonicera Japonica Flos industry in China. In practice, we should improve the drug registration of Lonicera Japonica Flos, promote ecological prevention and control technology, and formulate and promote pesticide residue limit standard of Lonicera Japonica Flos.

Improving compactibility properties of Lonicera Japonica Flos by Plasdone S-630 / 药学学报

To improve the fluidity and compactibility properties of raw powders of traditional Chinese medicine by particle modification technology, Lonicera Japonica Flos was used as a model drug, fluidized bed bottom spray technology was used, and Plasdone S-630 was used as a modifier to prepare modified particles. The powder properties, tablet compactibility parameters, disintegration time and dissolution were measured. The surface morphology of the powder particles before and after modification and compressed tablets were characterized by combining with scanning electron microscopy technology. The results showed that the particle size of Lonicera Japonica powder has been increased after particle modification, the fluidity, compressibility and compactibility of the powder have been improved to some extent, the disintegration time has also been reduced, and the dissolution in vitro is not affected. Therefore, this study can provide reference and ideas for the common problem that raw powder of traditional Chinese medicine that cannot meet the needs of preparation production due to poor powder properties such as fluidity and compressibility.

Research progress on chemical constituents and pharmacological activities of iridoid glycosides in Lonicera japonica / 中国中药杂志

As a traditional Chinese medicinal material, Lonicera japonica has a long medicinal history. The chemical constituents of Lonicera japonica are complex, mainly including iridoid glycosides, flavonoids, triterpenes, organic acids and volatile oil. Iridoid glycosides account for a higher proportion. In addition, modern pharmacological studies have shown that the iridoid glycosides have many pharmacological activities such as antivirus, anti-inflammation, anti-tumor, liver protection and lowering blood sugar. This review intends to systematically summarize the iridoid glycosides identified from Lonicera japonica and their pharmacological activities by searc-hing Chinese and English databases, in order to provide a reference for the further development and utilization of Lonicera japonica and for the improvement of quality standards of medicinal materials.

Three new secoiridoid glycosides from the flower buds of Lonicera japonica / 中国天然药物

Three new secoiridoid glycosides, named lonijapoglycol A (1), aldosecolohanin C (2) and aldosecolohanin B (3), together with three known ones (4-6), have been isolated from the flower the buds of Lonicera japonica. All the structures were identified by spectroscopic analyses. Lonijapoglycol A (1) expressed significant anti-inflammatory activity to inhibit the release of β-glu-curonidase induced by platelet-activating factor in rat polymorphonuclear leukocytes with an IC value of 3.76 μmol·L.

Screening novel inhibitors targeting SARS-CoV-2 S protein-ACE2 interaction based on molecular docking / 中草药

Objective: To screen inhibitors targeting SARS-CoV-2 S protein-ACE2 interaction by molecular docking. Methods: Candidate natural products were collected from Selleck China natural product library (Catalog No. L1400, 2 054 natural products). The structure of SARS-CoV-2 S protein-ACE2 had been determined by Qiang Zhou team (PDB: 6M17). The molecular docking was performed by Discovery Studio. Results: Based on the virtual amino acid mutation experiment which determined the key amino acids, the binding cavity was created. Then, 11 compounds were screened out from the natural compound library: digitonin, Lonicera grisea saponin A, forsythiaside B, L. grisea saponin B, Dipsacus asperges saponin B, hederacoside D, platycodon D, echinacoside, ginsenoside Rb2, ginsenoside Rc, and chlorogenic acid C. Conclusion: The 11 potential inhibitors targeting SARS-CoV-2 S protein-ACE2 interaction were screened out from natural products library, which provides a reference for the research of new anti SARS-CoV-2 drugs.

Intension research on influencing factors of medicinal properties formation of Lonicera japonica and cause hypothesis in explaining medicinal properties of Chinese materia medica single herb / 中草药

The four natures are the basic properties of Chinese materia medica (CMM). At present, it is one of the research hotspots to reveal the scientific connotation of CMM property theory. Clarifying the formation process and influencing factors of the medicinal properties of each single herb in a long historical period is the basic condition for the induction and summary of common characteristics of TCM property theory. Through the textual research of ancient books and documents, combined with the research results of modern Chinese medicine science, this paper combs the influence factors in the formation of the medicinal properties of Lonicera japonica, and reveals its scientific connotation. That is to say, the formation of the medicinal properties of each single herb of TCM is the result of its efficacy material base, the efficacy reflected in the process of clinical treatment and the tendency of clinical medication in previous dynasties. Based on the above research, the cause hypothesis of single herb property is put forward for the first time. The hypothesis provides a reference for the theoretical study of medicine property of TCM.

Study on Fingerprint and in vitro Antiviral Activity of Lonicera japonica Polysaccharide / 中国药房

OBJECTIVE：To establis h the fingerprint of Lonicera japonica polysaccharide，and to investigate in vitro inhibitory effect of it on respiratory syndrome virus （RSV）. METHODS ：Polysaccharide from L. japonica was prepared by water extraction and twice alcohol precipitation method. After hydrolysis with trifluoroacetic acid ，derivatization with hydroxylamine hydrochloride and pyridine ，the fingerprint was established by GC method. The determination was performed on HP- 5 capillary column ，and the detector was flame ionization detector ；the temperature of the sample inlet was 250 ℃；the temperature of the detector was 300 ℃ （programmed temperature ）；the carrier gas was nitrogen （flow rate of 50 mL/min）；split sampling was adopted （split ratio of 60∶1）；the sample size was 2.0 μL. Using rhamnose as reference substance，GC fingerprint of 12 batches of L. japonica （S1-S12） was drawn ，and the similarity evaluation was performed with Similarity Evaluation System of TCM Fingerprint （2012 edition）. Cluster analysis and principal component analysis were conducted by using SPSS 21.0 software. Using ribavirin as positive control ， half effective concentration （EC50）and treatment index （TI）as indexes ，MTT assay was used to investigate in vitro inhibitory effect of L. japonica polysaccharide on RSV. RESULTS ：There were 12 common peaks in GC fingerprint of 12 batches of L. japonica. The similarity was greater than or equal to 0.994. Seven common peaks were identified ，such as rhamnose ，arabinose， fucose，mannose，glucose，galactose，inositol hexaacetate. According to the cluster analysis ，12 batches of samples could be divided into two categories ，i.e. S 1，S7，S10 and S 11 clustered into one category ，and others clustered into one category. In principal component analysis ，the eigen values of 3 principal components were all greater than 1 （5.659，2.745，1.724 respectively），and their cumulative contribution rate was 84.400％. The comprehensive score of S 12 was the highest ，the second was S 5，and the lowest was S 11. EC 50 of total polysaccharide ，80％ alcohol precipitated polysaccharide ，50％ alcohol precipitated polysaccharide and 20％ alcohol precipitated polysaccharide of L. japonica （No. S 12） were 0.76，0.61，1.03，3.04 g/L， respectively；TI were 15.36，18.51，11.69，4.22，respectively. EC 50 of 80％ ethanol alcohol precipitated polysaccharide was the lowest，and its TI was close to that of positive control （20.08）. CONCLUSIONS ：Established fingerprint provides reference for the quality evaluation of L. japonica . L. japonica polysaccharide has a certain inhibitory activity on RSV in vitro ，and the 80％ alcohol precipitated polysaccharide has the strongest activity.

Establishment of HPLC Fingerprint of Lonicera japonica and Study on Its Anti-inflammatory Spectrum-effect Relationship / 中国药房

OBJECTIVE：To establish fingerprint of Lonicera japonica ，and to study its anti-inflammatory spectrum-effect relationship. METHODS ：HPLC was adopted. The determination was performed on Diamonsil C 18 column with mobile phase consisted of 0.1％ formic acid solution-acetonitrile （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was 30 ℃，and detection wavelength was 238 nm. The sample size was 10 μL. Using chlorogenic acid as reference，HPLC fingerprint of 10 batches of L. japonica from different production areas was established according to TCM Chromatographic Fingerprint Similarity Evaluation System （2012 edition）. By comparing with reference substance ，chemical constituents corresponding to common peaks were identified ，and the similarity analysis was conducted. Acute and chronic inflammatory models of mice induced by xylene ，carrageenan and cotton ball were used to evaluate inhibition rate of 10 batches of L. japonica to ear，foot and granuloma swelling； the average value was calculated as the comprehensive pharmacodynamic index. The spectrum-effect relationship with HPLC fingerprint of L. japonica and anti-inflammatory effect was analyzed by grey relational analysis （GRA）and partial least squares regressiosn （PLSR）based on common peak area and comprehensive pharmacodynamic index . Chromatographic peaks with correlation＞0.7 and regression coefficient of PLSR model ＞0 were characteristic peaks. The percentage of peak areas of characteristic peaks to peak areas of common peak was calculated in 10 batches of L. japonica （e.g.“peak ratio ”）. RESULTS ： There were 25 common peaks in HPLC fingerprints of 10 batches of L. japonica ，with similarity of 0.775-0.994. Totally 9 peaks were confirmed ，i.e. rutin （peak 18），hyperoside（peak 20），isochlorogenc acid B （peak 22），galuteolin（peak 21），chlorogenc acid（peak 9），loganin（peak 10），neochlorogenic acid （peak 2），isochlorogenic acid C （peak 25），isochlorogenic acid A （peak 23）. All 10 batches of L. japonica had inhibitory effects on ear swelling ，foot swelling and granuloma ，with average inhibitory rate of 47.95％-56.52％. The correlation by GRA was peak 8＞12＞18＞16＞3＞11＞20＞22＞19＞21＞1＞9＞10＞13＞24＞14＞2＞ 17＞25＞23＞5＞4＞15，and all of correlations were greater than 0.7. The regression coefficient of PLSR for peaks 2，4，5，7，8， 10，12，13，14，15，16，17，18，20，21，22，24 were all greater than 0；those peaks were positively correlated with anti-inflammatory effect and were characteristic peaks except for peak 7； among them ，VIP values of peaks 5，8，10，16，18， 20，24 were greater than 1. The peak ratio of 10 batches of L. japonica was 58.61％-71.19％. CONCLUSIONS ：HPLC fingerprint of 10 batches of L. Japonica is successfully established. 10 batches of samples have similar components ，and the content of anti-inflammatory components is relatively high. The proportion of characteristic peaks to common peaks should not be less than 51.8％.

Research on remote sensing recognition of wild planted Lonicera japonica based on deep convolutional neural network / 中国中药杂志

Identification of Chinese medicinal materials is a fundamental part and an important premise of the modern Chinese medicinal materials industry. As for the traditional Chinese medicinal materials that imitate wild cultivation, due to their scattered, irregular, and fine-grained planting characteristics, the fine classification using traditional classification methods is not accurate. Therefore, a deep convolution neural network model is used for imitating wild planting. Identification of Chinese herbal medicines. This study takes Lonicera japonica remote sensing recognition as an example, and proposes a method for fine classification of L. japonica based on a deep convolutional neural network model. The GoogLeNet network model is used to learn a large number of training samples to extract L. japonica characteristics from drone remote sensing images. Parameters, further optimize the network structure, and obtain a L. japonica recognition model. The research results show that the deep convolutional neural network based on GoogLeNet can effectively extract the L. japonica information that is relatively fragmented in the image, and realize the fine classification of L. japonica. After training and optimization, the overall classification accuracy of L. japonica can reach 97.5%, and total area accuracy is 94.6%, which can provide a reference for the application of deep convolutional neural network method in remote sensing classification of Chinese medicinal materials.

Effect of calcium on ion contents and expression of photosynthetic related genes in honeysuckle under salt stress / 中国中药杂志

Exogenous calcium can enhance the resistance of certain plants to abiotic stress. Research have demonstrated that exogenous calcium could enhances the resistance of honeysuckle under salt stress by promoting the transmission of photosynthetic electrons.The aim of this study was to investigate the effects of exogenous calcium on the contents of Na~+,K~+,Ca~(2+),Mg~(2+)and the expression of photosynthetic related genes Cab and rbc L. In this study,we used ICP-OES to analysis ion contents and used qRT-PCR to analysis the expression patterns of Cab and rbc L. The results showed that CaCl_2 significantly enhanced the K~+-Na~+,Ca~(2+)-Na~+,Mg~(2+)-Na+ratio of honeysuckle treated with 50 and 100 mmol·L~(-1) NaCl. Meanwhile,Cab and rbc L were significantly up-regulated under short-term salt stress,and CaCl_2 promoted this trend. From the two gene expression patterns,rbc L rapidly up-regulated on the first day of stress and then decreased,and was more sensitive to environmental changes. In summary,exogenous calcium could alleviate salt stress and increase plant development by increasing intracellular K~+-Na~+,Ca~(2+)-Na~+,Mg~(2+)-Na+ratio,and the transient overexpression of Cab and rbc L.

Regulation of exogenous calcium on photosynthetic system of honeysuckle under salt stress / 中国中药杂志

Exogenous calcium can enhance the resistance of certain plants to abiotic stress. However,the role of calcium insaltstressed honeysuckle is unclear. The study is aimed to investigate the effects of exogenous calcium on the biomass,chlorophyll content,gas exchange parameters and chlorophyll fluorescence of honeysuckle under salt stress. The results showed that the calcium-treated honeysuckle had better photochemical properties than the salt-stressed honeysuckle,such as PIABS,PItotal,which represents the overall activity of photosystemⅡ(PSⅡ),and related parameters for characterizing electron transport efficiency φP0,ψE0,φE0,σR,and φR are significantly improved. At the same time,the gas exchange parameters Gs,Ci,Trare also maintained at a high level. In summary,exogenous calcium protects the activity of PSⅡ,promotes the transmission of photosynthetic electrons,and maintains a high Ci,therefore enhances the resistance of honeysuckle under salt stress.

Comparison of eleven active components in Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicero japonica by UPLC / 中国中药杂志

This present study aims to establish a UPLC method for simultaneously determining eleven components such as new chlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,artichoke,isochlorogenic acid A,isochlorogenic acid B,isochlorogenic acid C,rutin,hibisin and loganin in Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica and comparing the differences in the contents of phenolic acids,flavonoids and iridoid glycosides of Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.The method was carried out on an ACQUITY UPLC BEH C18column(2.1 mm×100 mm,1.7 μm) by a gradient elution using acetonitrile and 0.1% phosphoric acid.The flow rate was 0.3 mL·min-1.The column temperature was maintained at 30 ℃.The sample room temperature was 8 ℃.The wavelength was set at 326 nm for new chlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,artichoke,isochlorogenic acid A,isochlorogenic acid B and isochlorogenic acid C,352 nm for rutin and lignin,and 238 nm for loganin.The injection volume was 1 μL.The eleven components has good resolution and was separated to baseline.Each component had a wide linear range and a good linear relationship(r≥0.999 6),the average recovery rate(n=9) was 98.96%,100.7%,97.24%,97.06%,99.53%,96.78%,98.12%,95.20%,95.12%,100.2%,98.61%and with RSD was 2.5%,1.4%,1.9%,2.1%,1.7%,1.9%,1.6%,2.0%,1.4%,2.2%,2.0%,respectively.Based on the results of the content determination,the chemometric methods such as cluster analysis and principal component analysis were used to compare the Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.The results showed that Lonicerae Japonicae Flos and leaves of Lonicera japonica were similar in the chemical constituents,but both showed chemical constituents difference compored to Lonicerae Japonicae Caulis.The established multi-component quantitative analysis method can provide a reference for the quality control of Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.

Effects of different temperature stress on cell membrane permeability,active oxygen metabolism and accumulation of effective substances in Lonicera japonicea / 中国中药杂志

The study is aimed to explore the effects of stress at different temperatures( 35,45,55 ℃) on membrane permeability,active oxygen metabolism and accumulation of effective substances in Lonicera japonica,and provide theoretical basis for reducing deterioration and revealing browning mechanism during postharvest processing of L. japonica. The cell membrane permeability( relative conductivity,MDA content),active oxygen metabolism( SOD,POD,PPO,CAT activity) and the accumulation of effective substances( chlorogenic acid,luteolin,neochlorogenic acid,cryptochlorogenic acid,3,5-dicaffeoylquinic acid,3,4-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid) of L. japonica were all studied by constant temperature drying method,and the results were analyzed by the SPSS 17. 0 statistical software. The results showed that MDA content in L. japonica was increased by 151. 14% at 35 ℃,SOD,POD,PPO and CAT activity were 29. 73%,42. 86%,105. 02% and 10. 74% higher than at 45 ℃,respectively. The order of effective substance content in L. japonica was 35 ℃ >45 ℃ >55 ℃. The changes of membrane permeability,activity of active oxygen metabolizing enzyme and accumulation of active components were significantly affected by different temperature stress. The indexes showed that physiological and active oxygen metabolizing enzyme activity of L. japonica was the highest under 35 ℃ stress,chlorogenic acid and luteolin were effectively accumulated,which provides basic data for solving browning problem in the postharvest processing of L. japonica.

Establishment of DNA identity card and analysis of genetic similarity among 58 varieties in Lonicera japonica / 中国中药杂志

A total of 58 varieties in Lonicera japonica from 20 producing areas were amplified by 22 pairs of SSR primers. Seven pairs of polymorphic primers were screened and their primers were used to establish DNA identity card and analyze genetic similarity.All the 58 varieties could be distinguished each other by the DNA identity card constituted by 7 pairs of core SSR primers.The genetic similarity coefficients of 58 varieties ranged from 0.366 7 to 0.916 7 by using PopGene32（vesion1.32）. Furthermore, all the varieties consistency were classified into 4 groups and constructed an evaluation table according to cluster analysis by an un-weighted pair-group average method with arithmetic mean. As expected, the results of cluster and evaluation table reflected 58 varieties relatives, which provide reference information for the selection of fine germplasm of L. japonica and the theoretical basis for the study of Dao-di herbs.

Expression analysis of Hsp70-related miRNA in Lonicera japonica Thunb. during different flowering stages / 药学学报

Plant flowering regulation is an important mechanism to response to environmental stress. Heat shock protein 70 family is one of the main molecular chaperones to resist stress; miRNA can be used as a negative regulator to participate in post-transcriptional gene in flowering network. In this paper, we obtained an Hsp70 gene from Lonicera japonica transcriptome and combined with Lonicera japonica miRNA library to obtain a novel miRNA that may target Hsp70 gene through bioinformatics method. Bioinformatics and expression during different flowering stages of the obtained Hsp70 gene and miRNA were analyzed. Phylogenetic tree showed that the obtained Hsp70 gene was clustered with Hsp110 subfamily in Oryza sativa and Arabidopis thaliana. The prediction of miRNA secondary structure showed its stable structure and high reliability. The binding site map showed that there were two base mismatches between sequences of miRNA and Hsp70 gene. The expression analysis showed that the expression of Hsp70 and miRNA in different flowering stages had opposite trends, indicating that miRNA might regulate Hsp70 to participate in the flowering stages of Lonicera japonica. This study provided new ideas for Lonicera japonica flowering regulation and response to environmental stress mechanisms.

Effect of exogenous GA₃ and DPC on active components of Lonicera japonica and its mechanism / 中国中药杂志

To reveal the effect of plant growth regulator GA₃ and DPC on the active components and its possible mechanism of Lonicera japonica. GA and DPC were applied at the stage of flower bud differentiation, and the content of active ingredients was measured by LC-MS-MS, the content of endogenous hormones were measured by ELISA, and the expression of key enzyme enes expression was determined by qRT-PCR. The level of endogenous hormone GA₃, IAA, ZR, DHZR and iPA in the GA treatment group was significantly improved, the expression of C4H1, C4H2, 4CL1 and HQT2 were also significantly increased. The content of chlorogenic acid, luteolin, luteoloside, isoquercetin and caffeic acid increased significantly. Spraying DPC did not affect or inhibit the accumulation of active components of L. japonica. Spraying GA can increase the content of endogenous GA₃, thus enhance the expression of C4H1, C4H2, 4CL1 and HQT2, and then increase the content of chlorogenic acid and luteolin.