Study on Tissue Culture of Japanese Honeysuckle ( Lonicera japonica )

[Objectives]This study was conducted to increase the reproduction coefficient of Japanese honeysuckle(Lonicera japonica)to keep the character of optimal benign.[Methods]The young leaves of medicinal Japanese honeysuckle were selected as explants,and MS was used as the basic culture medium.Suitable culture concentrations and conditions were screened through different concentration gradients of growth regulators and cytokinin.[Results]As the concentration of 6-BA in the culture medium increased,the browning rate increased,and the browning phenomenon occurred earlier.On the contrary,a lower concentration of 6-BA was suitable for the differentiation and growth of young leaves,and the browning response was slow.However,if the cultivation time was too long and the materials were not transferred in a timely manner,browning would also occur.The optimal combination of levels was obtained through a 3×3 orthogonal experiment(three parallel groups for each of 6-BA and NAA).The culture conditions included a constant temperature of 26℃and light intensity of 1200 lx.The optimal medium for inducing callus proliferation was MS+6-BA 0.5 mg/L+NAA 0.5 mg/L;and the optimal medium for inducing bud differentiation was MS+6-BA 1.0 mg/L+NAA 0.1 mg/L.[Conclusions]This study provides a theoretical basis for accelerating the development of the honeysuckle industry.

Analysis of Volatile Constituents in Lonicera japonica Thunb. from Different Origins by GC-MS

[Objective] This study aimed to analyze the volatile constituents in Lonicera japonica Thunb. from different origins. [Method] HP-5MS capillary columns were used and column temperature was controlled by a program. MS analysis was performed with EI and quadruple mass analyzer. The volatile constituents in L. japonica Thunb. were identified by NIST02 and Wiley275 libraries, and their relative contents were determined with chromatographic peak area normalization method. [Result] According to GC-MS total ion-current chromatograms, 35 volatile constituents were identified in L. japonica Thunb. from Guangxi Zhuang Autonomous Region, mainly including methyl linolenate, n-hexadecanoic acid and ζ-muurolene; 18 volatile constituents were identified in L. japonica Thunb. from Hunan Province, mainly including n-hexadecanoic acid, linoleic acid and α-curcumene. [Conclusion] Main volatile constituents in L. japonica Thunb. from two different origins varied significantly.

Leaf epidermal characters of Lonicera japonica and Lonicera confuse and their ecology adaptation

The leaf epidermis of Japanese honeysuckle （Lonicera japonica Thunb.） and Wild Honeysuckle （Lonicera confusa） in the genus of Flos Lonicerae were mainly observed by scanning electron microscopes （SEM） to study the characteristics of stomata, trichomes and dermal cell, etc.. The results showed that stoma exists only on the lower epidermis and its distribution is irregular, and leaf epidermis consist of epidermis cells, stoma complexes and bushy trichomes including glandular hair and non-glandular hair. On the upper epidermis, anticlinal wall caves in sinuous groove to countercheck the transpiration. Evidences from leaf morphological structures serve as another proof on drought-resistant mechanisms. Some strumaes distributing regularly are hypothesized as oxalic calcium on the lower epidermis under laser scanning confocal microscopy （LSCM） with Fluo-3/AM, which can increase their endurance to drought stress. Therefore, the above characteristics of Flos Lonicerae can reduce the loss of water and make Japanese honeysuckle and Wild Honeysuckle adapt to the droughty environment at Karst area in southwest China. However, there is some difference of the two species. From the SEM （Scanning Electron Microscopy） result, it is shown that on the upper epidermis, some glandular hair regularly present along the midrib of Japanese honeysuckle, but Wild Honeysuckle has no glandular hair on the upper epidermis, which can verify the relationships of Flos Lonicerae species and provide the significance for classification of Flos Lonicerae.

Effect of Different Factors on Rooting Characteristics and Survival of Lonicera japonica Thunb

[Objective] The aim was to explore the effects of different factors on rapid propagation and seedling survival of Lonicera japonica Thunb. [Method] The cuttings of 1, 2 or 3 years old, with different length, were soaked in different hormones at different concentrations for different time and planted at different dates. Finally, their rooting rate and survival rate in different treatments were measured and compared. [Result] The rooting rate and survival rate of one-year old cuttings with length of 10-13 cm and planted on September 20 and October 20, 2012 were relatively higher among all the treatments. Hormone treatment could significantly promote the callus formation and accelerate the rooting of the cuttings, and improve their survival rate. The cuttings treated with 120-200 mg/L ABT1 or NAA for 35-45 min had higher rooting rate and survival rate. Among them, the survival rate of cuttings treated with 160 mg/L ABT1 for 35 min was the highest, up to 92.5%. [Conclusion] The study provided basis for improving the survival rate of L. japonica by cuttage, and for optimizing the rapid propagation technique.

Two Triterpenoid Saponins from Lonicera Japonica

A new triterpenoid saponin, together with a known saponin. was isolated from the flowers of Lonicera Japonica Thunb. Using chemical and spectroscopic methods, mainly 2D NMR technique, their structures were deduced to be 3-beta-D-glucopyranosyl-(1 --> 4)-beta-D-glucopyranosyl-(1 --> 3)-alpha-L-rhamnospyranosyl-(1 --> 2)-alpha-L-arabinopyranosyl hederagenin 28-O-beta-D-glucopyranosyl-(l --> 6)-beta-D-glucopyranoside and 3-alpha-L-rhamnospyranosyl-(1 --> 2)-alpha-L-arabinopyranosyl hederagenin 28-O-alpha-L-rhamnospyranosyl- (1 --> 2)-[beta-D-xylcopyranosyl-( 1 --> 6)]-beta-D-glucopyranoside, respectively.

金银花(Lonicera japonica Thunb.)愈伤组织的诱导和分化

以金银花的茎、叶片、芽为外植体,在70个不同浓度IAA、NAA、6-BA组合的MS培养基上进行愈伤组织诱导,分析了金银花的快速繁殖体系的培养条件,发现芽是金银花组织培养的最佳外植体,IAA和NAA的最佳浓度均为1mg·L-1,6-BA的最适浓度为0.1mg·L-1,确定适宜的初代培养基为:MS+6-BA0.1mg·L-1+NAA1mg·L-1,诱芽培养基为:MS+6-BA0.1mg·L-1+NAA0.02mg.L-1.关键词愈伤组织,金银花,快速繁殖。

Effects of Biochar on Physiological Characteristics of Lonicera Japonica and Accumulation of Cd and Pb in Cd and Pb Contaminated Soil

A pot experiment was conducted to study the effects of biochar on physiological characteristics and accumulation of Cd and Pb of Lonicera japonica in Cd and Pb contaminated soil.The results showed that adding 2%~16%biochar could significantly increase the net photosynthetic rate,chlorophyll content and root activity of Lonicera japonica,and significantly reduce its SOD activity.When 6%~16%biochar was added,the contents of Cd and Pb in roots and stems of Lonicera japonica decreased significantly;when 2%~16%biochar was added,the contents of Cd and Pb in the leaves and flowers of Lonicera japonica decreased significantly.Adding more than 8%biochar can reduce the content of Cd and Pb in Lonicera japonica to a safe range level.

Lonicera japonica-induced inhibition of interleukin-1 beta thermogenesis and E-type prostaglandin receptor-3 expression in the preoptic area of rabbits

BACKGROUND： It has been shown that interleukin-1β（IL-1β） can induce fever by activating vascular endothelial cells and macrophages of the supraoptic crest to generate prostaglandin E2, which binds with receptors of the thermo-sensitive hypothalamic neurons. Lonicera japonica is one of the medicinal plants used widely in Asia for its antipyretic properties. However, these mechanisms have not yet been intensively studied. OBJECTIVE： To investigate the antipyretic effect and mechanisms of Lonicera japonica on IL-1β- induced febrile New Zealand rabbits by observing expression changes of E-type prostaglandin receptor-3 （EP3） mRNA in the preoptic anterior hypothalamus （POAH）. DESIGN： A randomized controlled study. SETTING： Electrophysiological Laboratory at the Department of Pathophysiology, Medical College of Jinan University; Department of Orthopaedics, First Hospital Affiliated to Medical College of Jinan University. MATERIALS： The experiment was performed from April to December 2005, using a total of 32 New Zealand white rabbits of both sexes, weighing 1.5 2.0 kg. All the animal experiments were performed according to the internationally accepted ethical guidelines. Lonicera japonica injection was purchased from Huanghe pharmaceutical factory of Xi＇an, China. IL-1βwas purchased from Sigma, USA. METHODS： A total of 32 rabbits were divided randomly into four groups： ① Normal saline （NS） control group;② Lonicerajaponica treatment group; ③ IL-1βtreatment group; and ④Lonicerajaponica plus IL-1βtreatment group. In the first 3 groups, the rabbits were given separate intravenous （i.v.） injections of l mL NS, l mL Lonicera japonica, and 100 ng IL-l β （dissolved in 0.9% NS without pyrogen）. In the Lonicerajaponica plus IL-1βgroup each rabbit was given i.v. injections of l mL NS and, 30 minutes later, 100 ng IL-1 β. MAIN OUTCOME MEASURES： Colonic temperature of each rabbit was measured at 0, 10, 20, 30, 40, 50, 60, and 70 minutes after injection and the maximum temperature rise （ A T） and the temperature response index after l hour （TRII） was calculated. Subsequently, in situ hybridization （ISH） was done with an ISH kit, EP3 mRNA expression in the POAH of all groups was measured （number of positive cells and average gray scale value）. RESULTS： A total of 32 rabbits were included in the result analysis, without any loss, （i） A T and TRII： there was no significant difference between the Lonicera japonica group and NS group （P 〉 0.05）. The IL-1β group was significantly greater compared to NS group （P 〈 0.01）. The Lonicera japonica plus IL-1β group was significantly less than the IL-1β group （P 〈 0.05）. ② In the NS and Lonicera japonica groups, the EP3 mRNA expression was negative （no coloration） or only weakly positive （only a few brown yellow particles in the cytoplasm cells could not be identified）. The number of positive cells and the average gray scale value were not significantly different between the two groups （P 〉 0.05）. In the IL-1β group, the number of positive cells was remarkably higher and the average gray scale value was lower than the NS group （P 〈 0.0 l）. In the Lonicera japonica plus IL-1β group, the number of positive cells was significantly less than the IL-1β group （P 〈 0.05）. However, the average gray scale value was greater than the IL-1β group （P 〈 0.05）. CONCLUSION： Lonicera japonica has obvious antipyretic effects on IL-1β-induced febrile rabbits and acts by inhibiting expression of EP3 mRNA in the POAH.

Extraction Process and Content Determination of Total Flavonoids in Lonicera japonica

[Objectives]To optimize the extraction process of total flavonoids of Lonicerae japonica and establish a method for its extraction and content determination.[Methods]The total flavonoids of L.japonica were extracted by reflux extraction method.Through a single factor experiment,the effects of extraction method,extraction solvent concentration,extraction volume and extraction time on the total flavonoids content of medicinal materials were investigated to select the optimal extraction process of the total flavonoids of L.japonica.[Results]The optimal extraction process of the total flavonoids of L.japonica was 70%ethanol,1∶30 of solid-to-liquid ratio,and 1.0 h of reflux extraction time.[Conclusions]This method can effectively determine the content of total flavonoids in L.japonica and is expected to provide a certain scientific basis for the study of the quality standard of L.japonica.This method has high reproducibility.It is stable and feasible in extraction of total flavonoids from L.japonica.

Protective effect of different medicinal parts of Lonicera japonica Thunb. on myocardial ischemic injury induced by isoproterenol in rats

Background:To study the protective effect of different medicinal parts of honeysuckle on myocardial ischemia injury in rats.Method:SD mice were randomly divided into 6 groups:the normal group,model group,compound salvia miltiorrhiza group,Lonicera japonica Thunb.of flower bud group,flower group,and the stem leaf group.There were 8 rats in each group,including 4 males and 4 females.The normal group and model group were given equal amount of distilled water,whereas the other dosage groups were given the appropriate drug by intragastric administration,continuous gavage for 14 days.From the 12 days,in addition to the normal group were injected of the same amount of normal saline,the other groups were injected with 5g/kg.Injection of isoproterenol to establish the mouce mycardial ischemia model,its continuously 3d.To detect the serum of AST,CK,CK-MB,LDH and the myocardial issue of Chinese super oxideismutase and malondialdehyde level.Results:Compared with normal group,the levels of AST,CK,CK-MB,cTn-I,LDH and malondialdehyde were significantly increased in the model group(P<0.01)and the level of super oxideismutase decreased significantly(P<0.01)the levels of AST,CK,CK-MB,cTn-I,LDH and malondialdehyde in the serum of flower bud group,flower group and stem leaf group were significantly decreased(P<0.05),and the decreasing level of flower bud group,flower,leaf group of AST,CK,CK-MB,cTn-I,LDH and malondialdehyde was flower bud group>flower>leaf group,the activity of super oxideismutase was obviously increased,and the activity level of super oxideismutase of flower bud group,flower group and stem leaf group was flower bud group>flower>leaf group.Conclusion:In the myocardial ischemic injury model of rats induced by isoproterenol,Lonicera japonica Thunb.had a protective effect in all parts,and Lonicera japonica Thunb.bud had the greatest protective effect.

Anti-encystment and amoebicidal activity of Lonicera japonica Thunb. and its major constituent Chlorogenic acid in vitro

Objective:To examine the acanthamoebicidal effects of ethyl acetate,aqueous and butanol fractions of dried flower buds of Lonicera japonica(L.japonica) Thunb.(Flos Lonicerae) in vitro.Methods:Acanthamoeba triangularis isolates were obtained from environmental water samples and identified by PCR.They were exposed to ethyl acetate,water and butanol fractions of L.japonica Thunb.at concentrations ranging from 0.5 mg/m L to 1.5 mg/m L.The extracts were evaluated for growth inhibition at 24,48 and 72 h,respectively.Chlorogenic acid at a concentration of 1 mg/mL was examined for inhibition of encystment.Results:Ethyl acetate fraction at a concentration of 1.5 mg/mL evoked a significant reduction of trophozoite viability by 48.9% after 24 h,49.2% after 48 h and 33.7% after 72 h Chlorogenic acid,the major active constituent of L.japonica Thunb.at the concentration of 1 mg/mL reduced the cysts/trophozoite ratio by 100% after 24 h,84.0% after 48 h and 72.3% after 72 h.This phenolic compound at concentration of 1 mg/mL concurrent with 0.6% hydrogen peroxide inhibited hydrogen peroxide-induced encystment by 92.8% at 72 h.Conclusions:Results obtained from this study show that ethyl acetate fraction at 1.5 mg/m L is the most potent fraction of L.japonica Thunb.and its major constituent chlorogenic acid showed the remarkable inhibition of encystment at a concentration of 1 mg/mL.

Review of Factors Influencing Active Components of Lonicera japonica Thunb.

Lonicera japonica Thunb. is a typical medicinal and edible plant in China. It has antibacterial, anti-oxidant, anti-endotoxin, and anti-inflammatory effects, etc. Its pharmacological action is closely related to its active components. Based on the results of previous studies, this paper summarized the effects of variety, origin, cultivation conditions, pruning, harvesting period and processing methods on the active components of L. japonica , in the hope of providing theoretical guidance for high-yield and high-quality cultivation of L. japonica.

Chemical constituents and pharmacological effects of Lonicerae japonicae

Lonicera japonica,also known as honeysuckle,is an evergreen shrub in the family of Syzygium.By consulting Sciencedirect databases and Web of Science databases,79 related articles were found,of which 22 were related to chemical composition and pharmacological activity.These articles show that L.japonica has a wide range of pharmacological activities,including antiviral,anti-tumor,and antioxidant effects.These activities have important applications in the pharmaceutical,food,and fragrance industries.This review focuses on the chemical composition and pharmacological effects of L.japonicae,which is of great significance to the development of new drugs and therapeutic methods.

Screening hepatoprotective effective components of Lonicerae japonica Flos based on the spectrum-effect relationship and its mechanism exploring

Lonicerae japonicae Flos(LF)is a kind of healthcare food with hepatoprotective function.This study was designed to explore the spectrum-effect relationships between UPLC fingerprints and the hepatoprotective effects of LF.Fingerprints of ten batches of LF were established by UPLC-PDA.The inhibitory levels of AST and ALT were used as pharmacological indexes,and secoxyloganin,isochlorogenic acid A and isochlorogenic acid C were screened as hepatoprotective active compounds by grey relational analysis(GRA)and partial least squares regression analysis(PLSR).Caspase-3 was obtained by network pharmacology as a key target of hepatoprotective active compounds.Molecular docking is used to explore the interaction between small molecules and proteins.This work provided a general model of the combination of UPLC-PDA and hepatoprotective effect to study the spectrum-effect relationship of LF,which can be used to considerable methods and insight for the fundamental research of the material basis of similar healthcare food.

Two 1-(6'-O-acyl-β-D-glucopyranosyl)pyridinium-3-carboxylates from the flower buds of Lonicera japonica

Two new l-（6＇-O-acyl-β-D-glucopyranosyl）pyridinium-3-carboxylates,lonijaponinicotinosides A（1）and B（2）,were isolated from an aqueous extract of the flower buds of Lonicera japonica.Their structures were determined by spectroscopic data analysis,and confirmed by comparison with synthetic 1-β-Dglucopyranosylpyridinium-3-carboxylate.

Comparison of anti-bacterial activity of three types of di-O-caffeoylquinic acids in Lonicera japonica flowers based on microcalorimetry

The anti-bacterial activities of three types of di-O-caffeoylquinic acids(diCQAs) in Lonicera japonica flowers, a traditional Chinese medicine(TCM), on Bacillus shigae growth were investigated and compared by microcalorimetry. The three types of diCQAs were 3, 4-di-O-caffeoylquinic acid(3, 4-diCQA), 3, 5-di-O-caffeoylquinic acid(3, 5-diCQA), and 4, 5-di-O-caffeoylquinic acid(4, 5-diCQA). Some qualitative and quantitative information of the effects of the three diCQAs on metabolic power–time curves, growth rate constant k, maximum heat-output power Pm, and the generation time tG, total heat output Qt, and growth inhibitory ratio I of B. shigae were calculated. In accordance with a thermo-kinetic model, the corresponding quantitative relationships of k, Pm, Qt, I and c were established. Also, the half-inhibitory concentrations of the drugs(IC50) were obtained by quantitative analysis. Based on the quantity–activity relationships and the IC50 values, the sequence of inhibitory activity was 3, 5-diCQA > 4, 5-diCQA > 3, 4-diCQA. The results illustrate the possibility that the caffeoyl ester group at C-5 is the principal group that has a higher affinity for the bacterial cell, and that the intramolecular distance of the two caffeoyl ester groups also has an important influence on the anti-bacterial activities of the diCQAs.

LjaFGD:Lonicera japonica functional genomicsdatabase

Lonicera japonica Thunb.,a traditional Chinese herb,has been used for treating human diseases for thousands of years.Recently,the genome of L.japonica has been decoded,providing valuable information for research into gene function.However,no comprehensive database for gene functional analysis and mining is available for L.japonica.We therefore constructed LjaFGD(www.gzybioinformatics.cn/LjaFGD and bioinformatics.cau.edu.cn/LjaFGD),a database for analyzing and comparing gene function in L.japonica.We constructed a gene co-expression network based on 77 RNA-seq samples,and then annotated genes of L.japonica by alignment against protein sequences from public databases.We also introduced several tools for gene functional analysis,including Blast,motif analysis,gene set enrichment analysis,heatmap analysis,and JBrowse.Our co-expression network revealed that MYB and WRKY transcription factor family genes were co-expressed with genes encoding key enzymes in the biosynthesis of chlorogenic acid and luteolin in L.japonica.We used flavonol synthase 1(LjFLS1)as an example to show the reliability and applicability of our database.LjaFGD and its various associated tools will provide researchers with an accessible platform for retrieving functional information on L.japonica genes to further biological discovery.

Three new secoiridoid glycosides from the flower buds of Lonicera japonica

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 ofβ-glucuronidase induced by platelet-activating factor in rat polymorphonuclear leukocytes with an IC50 value of 3.76μmol·L^-1.

Two homosecoiridoids from the flower buds of Lonicera japonica

Two new homosecoiridoids, named loniaceticiridoside （1） and lonimalondialiridoside （2）, were isolated from an aqueous extract of the flower buds of Lonicera japonica. Their structures including the absolute configuration were determined by extensive spectroscopic studies, especially by 2D NMR and CD data analysis. A proposed biosynthetic pathway and preliminary investigations of the biological activity of compounds 1 and 2 are also discussed.

Main Diseases and Pests of Lonicerae japonica in Guangxi Province and Fluctuation Pattern of Their Natural Enemies

[ Objective ] The paper was to study the main diseases and pests in Lonicerae japonica in Guangxi Province and fluctuation pattern of their natural ene- mies. [Method] The population change of main diseases and pests, and main predatory naturaI enemies of pests in different species of Lonicerae japonica in Xincheng County of Guangxi Province were systematically investigated during 2008 -2010. [ Result ] The variation cycle of population number of Semiaphis heraclei （Takahashi） belongs to single peak type, and it has large occurrence in April. The variation cycle of population number of Franklinlella sp. and Haptonchus luteo- ius （Erichson） belongs to double peak type, and they have large occurrence at April and September. The variation cycle of disease severity of Microsphaera lonicerae （ Wint. ） belongs to double peak type, and the disease index will be larger at April and September. The variation cycle of diseased shoot rate belongs to single peak type; diseased shoot rote of Lonicera dasystyla Rehd. and Loniccra hvpolzlauca Miquel is relatively high during April to June, while the index of Lonice- ra confusa （Sweet） DC. is relatively high during February to May. The variation cycle of population number of main natural enemies such as Lemnia biplagiata （ Swartz）, Coccinella septcmpunctata （ Linnaeus ） and Harmonia axyridis （Pallas） belongs to double peak type, and they have large occurrence at April and Sep- tember. [ Conclusion] The results could provide scientific basis for determination of optimum control period against major diseases and pests in Loniceraejaponica and development of integrated control programs.