This study explores the basic characteristics and potential functions of the terpene synthase(TPS) gene family members in Lonicera japonica. The L. japonica TPS(LjTPS) gene family was identified and functionally analyzed using bioinformatics methods. The results showed that a total of 70 members of the LjTPS gene family were identified in L. japonica, with protein lengths ranging from 130 to 1 437 amino acids. Most of these proteins were hydrophilic, and they were unevenly distributed across nine chromosomes. Phylogenetic analysis showed that the LjTPS gene family members were divided into six subfamilies, mainly consisting of members from the TPS-a, TPS-b, and TPS-e subfamilies. Promoter cis-acting element analysis showed that LjTPS members contained a large number of stress-responsive cis-acting elements. Aphid inoculation experiments showed that key enzyme genes in the MVA pathway for terpenoid backbone synthesis in L. japonica, such as HMGS, HMGR, MK, MPD, and the key enzyme gene in the DXP pathway, DXS, exhibited an initial increase followed by a decrease under aphid stress. The qRT-PCR analysis showed that the expression levels of the α-farnesene synthase genes LjTPS34 and LjTPS39 were down-regulated, while the expression levels of(E)-β-caryophyllene synthase genes LjTPS15 and LjTPS17 were up-regulated 12 h before aphid feeding, then began to decline. Farnesyl pyrophosphate synthase(FPS), which interacted with these genes, also displayed a pattern of increasing followed by decreasing expression. The expression of linalool synthase genes LjTPS12 and LjTPS33 was significantly up-regulated after 72 h of aphid feeding(P<0.000 1), reaching 24.39 and 22.64 times the initial expression, respectively. This pattern was in close alignment with the trend of linalool content in L. japonica. This study provides a theoretical foundation for future research on the interaction between L. japonica and pests, as well as on the functional roles of the LjTPS gene family.
Animals ; Aphids/physiology* ; Alkyl and Aryl Transferases/chemistry* ; Lonicera/parasitology* ; Phylogeny ; Plant Proteins/chemistry* ; Gene Expression Regulation, Plant ; Multigene Family ; Terpenes/metabolism*

To explore the regulatory mechanism of drought stress on the synthesis of chlorogenic acid and luteoloside in Lonicera japonica, we designed five drought gradients (soil water contents of 30%, 24%, 17%, 14%, and 10%) and screened and verified the differentially expressed genes (DEGs) by RNA sequencing (RNA-seq) and reverse transcription quantitative polymerase chain reaction (RT-qPCR). Furthermore, we employed HPLC to systematically measure the content changes of chlorogenic acid and luteoloside. The results revealed that drought significantly reduced the accumulation of secondary metabolites, and severe drought led to more obvious reductions. Under extreme drought (soil water content of 10%), the content of chlorogenic acid and luteoloside decreased significantly to 25.73 mg/g and 11.33 mg/g (with the decrease rates of 37.85% and 9.58%, respectively). A total of 77 454 genes were identified via transcriptome analysis, among which the number of DEGs reached 1 128 under the extraordinary drought. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses revealed that the DEGs were mainly involved in flavonoid synthesis, secondary metabolite biosynthesis, plant hormone signal transduction and the plant-pathogen interaction pathways, and the expression of key genes regulating the synthesis of chlorogenic acid and luteoloside was significantly downregulated. RT-qPCR verified the accuracy of the RNA-seq data. This study revealed that drought stress reduced the content of chlorogenic acid and luteoloside, the main secondary metabolites, by inhibiting the expression of key genes in the secondary metabolism pathways. The findings provide candidate gene resources for molecular breeding of drought-tolerant Lonicera japonica.
Lonicera/physiology* ; Chlorogenic Acid/metabolism* ; Droughts ; Stress, Physiological ; Gene Expression Regulation, Plant ; Glucosides/metabolism* ; Luteolin

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.
Calcium ; pharmacology ; Chlorophyll ; analysis ; Lonicera ; drug effects ; physiology ; Photosynthesis ; Plant Leaves ; Salt Stress

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.
Cell Membrane Permeability ; Chlorogenic Acid/metabolism* ; Hot Temperature ; Lonicera/physiology* ; Luteolin/metabolism* ; Oxygen/metabolism* ; Stress, Physiological

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.
Calcium ; physiology ; Cations ; analysis ; Lonicera ; physiology ; Photosynthesis ; Salt Stress

To establish HPLC specific chromatogram and its correlation with the protection effect of Shuanghuanglian on MDCK (Madin-Darby canine kidney) cell injury induced by influenza A virus( H1N1). Nine recipes of Shuanghuanglian based on the official prescription were prepared according to orthogonal test for HPLC analysis and MDCK cells protection experiment separately (cytopathic effect (CPE) method was used for observing the virus infectivity and MTT staining results were used as the determining indexes for drug concentration selection and analyzing cell viability). The results suggested that all the other Shuang-Huang-Lian recipes except recipe1 demonstrate protecting effect on MDCK cell injury induced by influenza A virus (P < 0.01, P < 0.001). Stepwise regression analysis was used for analyzing the relationships between HPLC fingerprint and the protecting effect of Shuanghuanglian on influenza A virus induced MDCK cell injury. Peak 2, 3, 6, 8 and 12 were found to be strongly related with anti-influenza A virus efficacy. Stepwise regression analysis of recipes data and efficacy data showed that Lonicerae Japonicae Flos and Forsythiae Fructus were positively associated with the protecting effect of cells injury. From HPLC fingerprints, we found that peak 2, 3, 12 were from Lonicerae Japonicae Flos and peak 6, 8 were from Forsythiae Fructus. Four peaks were identified through comparing the retention time between the standard and Shuanghuanglian recipes, and they were chlorogenicacid, cryptochlorogenic acid, forsythoside B and 3,4-dicaffeoylquinic acid respectively. Caffeic acid derivatives in Lonicerae Japonicae Flos and Forsythiae Fructus were found to be greatly correlated with anti-influenza A virus efficacy and maybe the substance basis of Shuanghuanglian.
Animals ; Antiviral Agents ; analysis ; pharmacology ; Chromatography, High Pressure Liquid ; methods ; Dogs ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Forsythia ; chemistry ; Influenza A Virus, H1N1 Subtype ; drug effects ; physiology ; Lonicera ; chemistry ; Madin Darby Canine Kidney Cells ; Scutellaria baicalensis ; chemistry

The objective of this study is to define the population dynamics of Semiaphis heraclei in the main-producing district of Lonicera japonica in Shandong, and screen for highly efficient, safety control technique. Through fixed field investigation, we tested the toxicity of eight kinds of insecticides by using dipping methods, and carried out the field experiment. The results showed that the aphids' emergence peak appeared in May. The aphids on the Sijihua variety of L. japonica was more susceptible and the peak was also seven days earlier than Damao variety of L. japonica. The aphid populations on Sijihua were 1 fold than those on the Daomao in happened peak. Comparing the eight kinds of insecticides, the LC50 of lambda-cyhaothrin, abamectin, imidacloprid and pyrethrin to wingless aphids were 1.494, 1.690, 2.840, 2.861 mg x L(-1), respectively, whose toxicity were higher, the toxicity of matrine, pymetrozine and azadirachtin were also high. The field efficacy trials indicated that during the period of aphids occurred, 25% imidacloprid wettable powder, 1.8% abamectin missible oil, 2.5% lambda-cyhaothrin missible oil, 25% pymetrozine wettable powder, 5% pyrethrin missible oil, 1% matrine water aqua were sprayed at concentrations of 20,000, 2,000, 2,500, 5,000, 500 and 50 times, respectively,the control effect achieved 91.69%, 98.90%, 96.18%, 95.06%, 99.24%, 90.10%, respectively, after 5 days. During the growing period of L. japonica in spring, application of thiamethoxam, thiacloprid, pymetrozine and imidacloprid, all of the control effect against aphids achieved above 98.88% after 50 days. The result indicated that May was the S. heraclei Takahashi's emergence peak in Pingyi, Shandong. The efficient safety and environmentally friendly insecticides by spraying and systemic insecticide of pymetrozine and imidacloprid by root application were all efficient controlled aphids. These insecticides were long for controlling S. heraclei Takahashi and worthy of being widely applied.
Animals ; Aphids ; drug effects ; physiology ; Insect Control ; Insecticides ; toxicity ; Lonicera ; parasitology ; Plant Diseases ; parasitology ; prevention & control ; Population Dynamics

OBJECTIVETo study the occurrence of Cercospora leaf spot of Lonicera macrathoides and fungicides control method, so as to provide scientific basis for its integrated pests management (IPM).

METHODThe field investigation and the field controlling trial were carried out for the research.

RESULTCercospora leaf spot was caused by C. rhamni. There was obvious relationship between the damage rate and the altitude, soil organic matter, available K, available P as well as variety. The controlling effect of 10% difenoconazole WG was the highest. 50% Thiram WP, 70% Mancozeb WP also had well controlling effect.

CONCLUSIONCercospora leaf spot of L. macrathoides could be controlled by suitability agricultural and chemical controls.

Ascomycota ; drug effects ; isolation & purification ; pathogenicity ; physiology ; Fungicides, Industrial ; pharmacology ; Lonicera ; microbiology ; Plant Diseases ; microbiology ; prevention & control ; Plant Leaves ; microbiology ; Virulence

AIMTo investigate the effect of Honeysuckle flower (HF) and Scutellaria Baicalensis Georgi (SBG) on contraction and electric activity of small intestine smooth muscle in rabbit and the underlying mechanisms.

METHODSUsing organ bath technique to observe the effect of HF and SBG on contractive and electric activity of small intestine smooth muscle in rabbit.

RESULTSHF and SBG significantly decreased the amplitude, frequency and area under the curve of contractive, as well as electric activity in a dose-depended manner. IC50 of the contractive amplitude was 6.30 g/L and 1.56 g/L by Logit Loglinear analysis. The inhibitive effect of HF and SBG on contractive activity could be partly decreased by beta-receptor blocker Propranolol, NO synthase inhibitor L-NAME, and K+ channel blocker Glibenclamide. Also HF and SBG inhibited acetylcholine-induced both intracellular and extracellular calcium-depended contraction significantly.

CONCLUSIONHF and SBG obviously inhibit the contractive and electric activity of small intestine smooth muscle of rabbit. The mechanisms are related to several pathways.

Action Potentials ; drug effects ; Animals ; Depression, Chemical ; Drugs, Chinese Herbal ; pharmacology ; Female ; Flowers ; chemistry ; In Vitro Techniques ; Intestine, Small ; physiology ; Lonicera ; chemistry ; Male ; Muscle Contraction ; drug effects ; Muscle, Smooth ; physiology ; Rabbits ; Scutellaria baicalensis ; chemistry

OBJECTIVETo explore the effects of traditional Tibetan medicine, Fructus Lonicerae microphyllae (FLM) on phagecytosis and cytokines production of murine macrophages.

METHODThe phagecytosis of murine macrophages was analyzed by neutral red phagecytosis assay. The activities of IL-1 and TNF-alpha were measured by biological methods. The mRNA of TNF-alpha and INF-gamma expressed by macrophages was detected by RT-PCR.

RESULTThe phagecytosis of murine macrophages was significantly enhanced by FLM at a concentration from 1 microg x mL(-1) to 100 microg x mL(-1) and the secretions of IL-1, and TNF-alpha from macrophages were markedly induced by FLM. Meanwhile, FLM also increased the expression of TNF-alpha mRNA and INF-gamma mRHA from macrophages in vitro.

CONCLUSIONFLM could promote phagecytosis and cytokines production of murine macrophages.

Animals ; Cell Line ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Fibroblasts ; cytology ; Fruit ; chemistry ; Interferon-gamma ; biosynthesis ; genetics ; Interleukin-1 ; secretion ; Lonicera ; chemistry ; Macrophages, Peritoneal ; metabolism ; physiology ; Mice ; Mice, Inbred C57BL ; Mice, Inbred ICR ; Phagocytosis ; drug effects ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics