Plant metabolomics for studying the effect of two insecticides on comprehensive constituents of Lonicerae Japonicae Flos.

Pesticides' overuse and misuse have been reported to induce ingredient variations in herbal medicine, which is now gaining attention in the medicinal field as a form of alternative medicine. To date, available studies on pesticide-induced ingredient variations of herbal medicine are limited only on a few compounds and remain most others unexamined. In this study, a plant metabolomics-based strategy was performed to systematically explore the effects of two frequently used insecticides on the comprehensive constituents of Lonicerae Japonicae Flos (LJF), the flower buds of Lonicera japonica Thunb. Field trials were designed on a cultivating plot of L. japonica with controls and treatments of imidacloprid (IMI) and compound flonicamid and acetamiprid (CFA). Unbiased metabolite profiling was conducted by ultra-high performance liquid chromatography/quadrupole-Orbitrap mass spectrometer. After data pretreatment by automatic extraction and screening, a data matrix of metabolite features was submitted for statistical analyses. Consequently, 29 metabolic markers, including chlorogenic acids, iridoids and organic acid-glucosides were obtained and characterized. The relative quantitative assay was subsequently performed to monitor their variations across flowering developments. This is the first study that systematically explored the insecticide-induced metabolite variations of LJF while taking into account the inherent variability of flowering development. The results were beneficial for holistic quality assessment of LJF and significant for guiding scientific use of pesticides in the large-scale cultivation.

Inhibitory Activity of Honeysuckle Extracts against Influenza A Virus In Vitro and In Vivo.

Honeysuckle has been used in the treatment of influenza virus infection for thousands of years in China. However, its main active components and the functional mechanisms remain to be elucidated. Here, four honeysuckle extracts, including acids extract, flavonoids extract, total extract and acids-flavonoids mixture, were prepared to clarify the main active antiviral components. The cytopathic effect reduction assay showed that all the four extracts inhibited the replication of influenza viruses H1N1, H3N2 and the oseltamivir-resistant mutant strain H1N1-H275Y. The acids-flavonoids mixture had the strongest inhibitory effects in vitro with EC50 values of 3.8, 4.1, and > 20 µg/mL against H1N1, H3N2 and H1N1-H275Y, respectively, showing competitive antiviral activity with oseltamivir and ribavirin. Honeysuckle acids extract also showed the most significant antiviral activity in vivo. Oral administration of the acids extract at a dosage of 600 mg/kg/d effectively alleviated viral pneumonia, maintained body weight and improved the survival rate to 30% of the mice infected with a lethal dose of H1N1. The results of time-of-drug addition experiment and neuraminidase (NA) inhibition assay showed that honeysuckle extracts had a broad-spectrum inhibitory effect against influenza virus NAs. The flavonoid extract showed the strongest inhibitory effect on the NA of influenza virus H7N9 with an IC50 of 24.7 µg/mL. These results suggested that these extracts might exert their antiviral activity by suppressing the release of influenza viruses. Briefly, our findings demonstrate that acids and flavonoids extracts of honeysuckle are the major antiviral active components, and the acids extract has the potential to be developed into an antiviral agent against influenza virus, especially for oseltamivir-resistant viruses.

[Regulation of exogenous calcium on photosynthetic system of honeysuckle under salt stress].

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.

Saline stress enhanced accumulation of leaf phenolics in honeysuckle (Lonicera japonica Thunb.) without induction of oxidative stress.

Honeysuckle (Lonicera japonica Thunb.) is a traditional medicinal plant in Chinese, and chlorogenic acid and luteolosid are its specific bioactive phenolic compounds. This study was to investigate leaf antioxidant responses in honeysuckle to saline stress with emphasis on phenolics through hydroponic experiments and field trials. NaCl stress did not stimulate antioxidant system including superoxide dismutase, ascorbate peroxidase, catalase and ascorbate, and had no significant effect on lipid peroxidation in the leaves. Consistently, no inhibition on photochemical capacity of photosystems suggested that reactive oxygen species (ROS) was maintained at a normal level under NaCl stress. However, leaf phenolic synthesis was activated by NaCl stress, indicated by elevated genes transcription and activity of phenylalanine ammonia-lyase and increased phenolics concentration. Specifically, leaf chlorogenic acid concentration was increased by 67.43% and 48.86% after 15 days of 150 and 300 mM NaCl stress, and the increase of luteolosid concentration was 54.26% and 39.74%. The accumulated phenolics hardly helped detoxify ROS in vivo in absence of oxidative stress, but the elevated phenolic synthesis might restrict ROS generation by consuming reduction equivalents. As with NaCl stress, soil salinity also increased concentrations of leaf phenolics including chlorogenic acid and luteolosid without exacerbated lipid peroxidation. In conclusion, leaf phenolics accumulation is a mechanism for the acclimation to saline stress probably by preventing oxidative stress in honeysuckle; leaf medicinal quality of honeysuckle can be improved by saline stress due to the accumulation of bioactive phenolic compounds.

The role of silicon in physiology of the medicinal plant (Lonicera japonica L.) under salt stress.

Silicon(Si) is the only element which can enhance the resistance to multiple stresses. However, the role of silicon in medicinal plants under salt stress is not yet understood. This experiment was conducted to study the effects of silicon addition on the growth, osmotic adjustments, photosynthetic characteristics, chloroplast ultrastructure and Chlorogenic acid (CGA) production of Honeysuckle plant (Lonicera japonica L.) under salt-stressed conditions. Salinity exerted an adverse effect on the plant fresh weight and dry weight, whilst 0.5 g L(-1) K2SiO3 · nH2O addition obviously improved the plant growth. Although Na(+) concentration in plant organs was drastically increased with increasing salinity, higher levels of K(+)/Na(+) ratio was obtained after K2SiO3 · nH2O addition. Salinity stress induced the destruction of the chloroplast envelope; however, K2SiO3 · nH2O addition counteracted the adverse effect by salinity on the structure of the photosynthetic apparatus. K2SiO3 · nH2O addition also enhanced the activities of superoxide dismutase and catalase. To sum up, exogenous Si plays a key role in enhancing its resistance to salt stresses in physiological base, thereby improving the growth and CGA production of Honeysuckle plant.

Influence of sulfur fumigation on the chemical constituents and antioxidant activity of buds of Lonicera japonica.

Lonicera japonica flos is widely used as a pharmaceutical resource and a commonly-employed ingredient in healthy food, soft beverages and cosmetics in China. Sometimes, sulfur fumigation is used during post-harvest handling. In this study, a comprehensive comparison of the chemical profile between sun-dried and sulfur-fumigated samples was conducted by HPLC fingerprints and simultaneous quantification of nine constituents, including secologanic acid, along with another eight usually-analyzed markers. Secologanic acid was destroyed, and its sulfonates were generated, whereas caffeoylquinic acids were protected from being oxidized. The residual sulfur dioxide in sulfur-fumigated samples was significantly higher than that in sun-dried samples, which might increase the potential incidence of toxicity to humans. Meanwhile, compared with sun-dried samples, sulfur-fumigated samples have significantly stronger antioxidant activity, which could be attributed to the joint effect of protected phenolic acids and flavonoids, as well as newly-generated iridoid sulfonates.

[Investigation on formation mechanism of secologanic acid sulfonates in sulfur-fumigated buds of Lonicera japonica].

To investigate formation mechanism of secologanic acid sulfonates in sulfur-fumigated buds of Lonicera japonica, secologanic acid was enriched and purified from the sun-dried buds of L. japonica by various column chromatography on macroporus resin HPD-100, silica gel and ODS. The stimulation experiments of sulfur-fumigation process were carried out using secologanic acid reacted with SO2 in the aqueous solution. The reaction mechanism could be involved in the esterification or addition reaction. The present investigation provides substantial evidences for interpreting formation pathway of secologanic acid sulfonates in sulfur-fumigated buds of L. japonica.

Profiling and characterization of volatile components from non-fumigated and sulfur-fumigated Flos Lonicerae Japonicae using comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry coupled with chemical group separation.

Flos Lonicerae Japonicae (FLJ) is a popular herb used for many centuries in Traditional Chinese Medicine as a treatment of fever and inflammation. Non-fumigated processing of FLJ has been the traditional approach used in post-harvest preparation of the commodity for commercial use. However, in recent years, natural drying processing of FLJ has been replaced by sulfur-fumigation for efficiency and pest control. Sulfur-fumigation can induce changes in the volatile compounds of the herb, altering its medicinal properties. A comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC×GC-TOF/MS) method was established for the resolution and determination of volatile components in non-fumigated and sulfur-fumigated FLJ. In this paper, analysis of the volatile oils in non-fumigated and sulfur-fumigated (including lab-prepared sulfur-fumigated and industrial sulfur-fumigated) FLJ was performed using GC×GC-TOF/MS. Seventy-three representative volatile components were identified, including furans, alkalies, acids, aldehydes, ketones, alcohols, terpenes, esters, and others, as the main components of FLJ volatile oils. The proposed method was successfully applied for rapid and accurate quality evaluation of FLJ and its related medicinal materials and preparations.

[Effects of irrigation volume on growth and quality of Lonicera japonica].

OBJECTIVE: To study the effects of irrigation volume on the growth and quality of Lonicera japonica. METHOD: Different volume of irrigation water was applied to the potted L. japonica before the first florescence of honeysuckle in order to keep the relative moisture content of the soil as 100%, 80%, 50%, 30%, and observe the growth and development of shoot and leaves, yield and quality of honeysuckle. RESULT: As the of irrigation volume was reduced, the internodal elongation of shoot and the increase of the knot were restrained, and the yield of honeysuckle dropped. But the content of chlorogen acid in the buds treated by 80% irrigation volume was not affected, while that of the other treatment all decreased. The moisture contents of the leaves treated by 30% irrigation volume was significantly lower than that of the CK, while the specific leaf weight (SLW) increased significantly comparing to the CK. The content of chlorophyl in the leaves of 50% and 30% increased, while the ratio of chlorophyl A and B fell significantly. The content of dissoluble suger in the leaves of 80% and 50% irrigation volume was much higher than that in the CK. The results also showed that, the content of dissoluble protein in the leaves of 30% irrigation volume increased comparing with that in 50%, and the content of proline in leaveas of 30% irrigation volume increased significantly. CONCLUSION: Using less irrigation volume (keeping 80% of relative moisture content of the soil) does not affect the quality of honeysuckle, but decreased dry weight of honeysuckle. It is necessary to take irrigation management during cultivation of honeysuckle.