Establishment of an in vitro Tissue Culture and Rapid Propagation System using Lonicera hypoglauca and Content Assessment of Total Flavonoids and Chlorogenic Acid

Lonicera hypoglauca is a traditional Chinese medicinal plant.In this study,the tender young leaves of L.hypoglauca were used for the first time as the explants to establish a rapid in vitro propagation and regeneration system.The results revealed that the optimal time for disinfection of the explants was 8 min and the optimal medium for callus induction was MS+2,4-D 4.0 mg·L^(-1)+sucrose 30 g·L^(-1),with an average callus induction rate of 86.67%.The optimal medium to induce differentiation of callus to bud was MS+6-BA 1.0 mg·L^(-1)+NAA 0.10 mg·L^(-1)+sucrose 30 g·L^(-1),with an average germination rate of 83.33%.The optimal medium to induce multiplication was MS+6-BA 1.5 mg·L^(-1)+NAA 0.05 mg·L^(-1)+sucrose 30 g·L^(-1),with a multiplication coefficient of 5.42.The optimal medium for root induction was 1/2 MS+NAA 0.15 mg·L^(-1)+activated carbon 0.3 g·L^(-1)+sucrose 15 g·L^(-1),with an average rooting rate of 91.11%.The survival rate of tissue-cultured seedlings in nutrient soil cultivation medium was as high as 100%.The total flavonoid content and chlorogenic acid content in the explant,callus tissue and regenerated plant were 1.83%,2.27%,1.33%and 2.77%,1.83%,1.74%respectively.This study provides novel insights into the rapid propagation and mass production of L.hypoglauca seedlings at an industrial scale and that it exhibits important application value and future prospects.

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.

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.

Lonicera caerulea polyphenols inhibit fat absorption by regulating Nrf2-ARE pathway mediated epithelial barrier dysfunction and special microbiota

Scope:High-fat diet(HFD)induces imbalance in the small intestine environment,where fat digestion and absorption mainly take place.This study aimed to elucidate the mechanisms by which Lonicera caerulea polyphenols(LCP)might inhibit fat absorption,from the perspective of small intestine microbiota and epithelial barrier integrity.Methods and results:Male Sprague-Dawley rats were given HFD with or without co-administration of LCP for 8 weeks.The results showed that LCP supplementation significantly decreased the levels of serum triglycerides(TG),total cholesterol(TC),and low-density lipoprotein cholesterol(LDL-C),and increased the contents of fecal sterols,in HFD rats.LCP also inhibited the dysfunction of the small intestine epithelial barrier,via alleviating the oxidative stress activated by Nrf2-ARE pathway,and by modulating the expressions of pro-inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),cyclooxygenase-2(COX-2),nuclear factor kappa-B p65(NF-κB p65)and inducible nitric oxide synthase(iNOS)in the small intestine.Additionally,LCP administration restored the balance in small intestine microbiota and increased the abundance of the specific bacteria,such as Lactobacillus,involved in fat absorption.Conclusion:Our results demonstrated that LCP may be beneficial to inhibit fat absorption.The mechanism seems to be associated with the protection of the epithelial barrier integrity and the modulation of specific bacteria in the small intestine.

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.

Chemical compositions and biological activities of Flos lonicerae

Flos lonicerae is the dry flower bud of Lonicera japonica Thunb,and it is a very important Chinese herbal medicine in China.In ancient times,it was widely used to treat the exopathogenic wind-heat,epidemic febrile diseases,sores,carbuncles and some infectious diseases.Modern pharmacological research has indicated that it has anti-inflammatory,anti-bacterial,anti-viral and anti-oxidation activities.This paper reviews the chemical compositions and biological activities of Flos lonicerae in order to provide reference for its application in clinical practice in the future.

Seed Germination and Vegetative Propagation of Promising Decorative Lonicera tatarica L. (Caprifoliaceae Juss.), Introduced into the Tashkent Botanical Garden (Uzbekistan)

Seed germination (in laboratory and field conditions) and vegetative reproduction (by cuttings) of a promising decorative species—Lonicera tatarica L. (Caprifoliaceae Juss.) was studied for the first time in the conditions of introduction of the Tashkent Botanical Garden of Uzbekistan. Thus, the optimal temperature for germination of L. tatarica seeds in laboratory conditions is +20°C + 22°C, at which germination was 73%. The germination rate of seeds sown in autumn in the field was 62%, and the germination rate of seeds sown in spring was 71%. It was noted that in greenhouse conditions at an air temperature of 20°C - 22°C and a relative humidity of 49% - 53%, the rootability of cuttings was 75%. It was revealed that the studied species adapted well to the conditions of introduction. Taking into account the effectiveness of vegetative reproduction of L. tatarica, it can be recommended for improving the aesthetic condition and landscaping of cities, landscaping and landscape design.

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.

Lonicerae Flos:A Review of Chemical Constituents and Biological Activities

Lonicerae flos,a widely used traditional Chinese medicine(TCM),has been used for several thousand years in China.As a famous traditional Chinese herbal medicine,it was used as heat-clearing drug and alexipharmic agent and was widely cultivated in Hunan province.L.flos mainly contains biologically active compounds such as caffeic acid derivatives,essential oil,flavonoids,iridoid glycosides and terpenoids.A range of biological activities has been reported from plant extracts including anti-inflammatory,antitumor,antioxidant,antiallergy,immunomodulating and antibacterial activity.In this study,the author investigated ancient books of TCM and nowadays reports,summarized the chemical constituents,biological activity of L.flos to provide a comprehensive systematic review.

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.

Research on Tissue Culture and Rapid Propagation Technology of Superior Individuals of Lonicera edulis Turcz

[Objective] This paper aimed to study the tissue culture and rapid propagation technology of superior individuals of Lonicera edulis Turcz. [Method] Several superior individuals of Lonicera edulis Turcz were used as materials for selecting the primary medium, subculture medium, rooting medium and acclimatization substrate during the tissue culture and rapid propagation. [Result] 6-BA was the optimal cytokinin for tissue culture of Lonicera edulis Turcz, compared with ZT; modified MS＋1.0 mg/L of 6-BA ＋ 0.2 mg/L of IBA was the optimal medium as primary and subculture medium, modified MS＋ 1.5 mg/L of IBA was the optimal medium for rooting of Lonicera edulis Turcz, the rooting rate had achieved 100% after cultured for 30 d. The optimal substrate for transplanting plantlets of Lonicera edulis Turcz was composed of humus and perlite （1∶ 1, V/V）, survival rate was as high as 95% after 30 d. [Conclusion] This study provided basis for the rapid propagation of superior seedlings of Lonicera edulis Turcz, as well as the establishment of industrialized breeding technical system and the implementation of scale production.

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.

Short-term Prediction of Occurrence Period of Main Diseases and Pests of Flos lonicerae using Phenoecology

The occurrence periods of Semiaphis heraclei Takahashi,Frankliniella sp.,Haptonchus luteolus and Microsphara linicerae Enchson wint.in Rabenh.causing damage on Flos lonicerae were investigated in F.lonicerae planting area in XinCheng county of Guangxi Province during 2008-2010,which were coincided with the occurrence periods of related phenology of local Prunus persica Rootstock.With P.persica Rootstock as indicator plant,the occurrence periods of three species of pests and one species of disease were predicted,respectively,and the method was simple and accurate,which could be the foundation for preventing these pests and diseases in the local field.

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.

Comparative Analysis of the Complete Chloroplast Genome Sequences of Four Origin Plants of Lonicerae Flos(Lonicera;Caprifoliaceae)

Lonicerae Flos(LF)derived from the dried flower buds or opening flowers of four Lonicera plants(Lonicera macranthoides,L.hypoglauca,L.confusa,and L.fulvotnetosa),is a popular traditional Chinese medicine.Because the four origin plants are very similar in morphology,it is difficult to control the quality of LF in actual production.Over the past decade,many reports have pointed out the differences among them,including the botanical characteristics and active ingredients.However,there is still a lack of rapid methods that can be applied to the identification of the four origins.In this study,comparative analysis of the four chloroplast genomes was performed,and they showed low diversity(Pi=0.00267),three variation hotspots regions(rbcL-accD,rps12-ndhF and rps12-trnN-trnG)were identified as potentially molecular marker of highly informative.Meanwhile,the most obvious difference in SSR comparative analysis is reverse and complement repeats were only identified in L.confusa and L.hypoglauca,respectively.Lastly,the phylogenetic tree showed that L.confusa is more closely related to L.fulvotnetosa,while L.macranthoides is closer to L.hypoglauca.This study systematically revealed the differences among the four chloroplast genomes,and it provides valuable genetic information for identifying the origin of LF.

重视忍冬属Lonicera中花灌木资源在园林中的利用

忍冬属Lonicera植物绝大多数花果香色宜人,为优美的观赏树种。全世界约有200种,主要分布于温带及亚热带地区,尤以中国、日本、朝鲜为最。我国忍冬属植物资源丰富,约有100多种。对这些资源的开发利用,越来越受到国外园林界的普遍关注和高度重视。但在国内除少数种类零星应用外,大多数资源未得到应有的利用和开发,有的充其量也只不过停留在资源的保存上。为了充分利用和挖掘这些资源,丰富美化环境的材料,现将本属中观赏价值较高的主要树种以及国外广为应用的园艺品种,作如下介绍,供应用时参考。

4种忍冬属(Lonicera)植物花粉的形态比较

通过扫描电子显微镜对4种忍冬属植物的花粉进行观察,发现其花粉长圆球形至圆球形,金银忍冬的花粉最小,台儿曼忍冬的花粉最大。4种植物的花粉均为三环沟型,萌发沟为子午向,金花忍冬的花粉萌发沟最长,几乎到达两极。4种植物的花粉表面均具刺,有细小穿孔,台儿曼忍冬花粉表面的刺小,等大,且有规则的脊与凹坑,其他3种花粉表面的刺大小不等;忍冬的花粉表面除具细小穿孔外,还有不规则条纹。

Separation and Purification of Macranthoidin B and Dipsacoside B from Flos Lonicerae by HP-20 and HP-SS Macroporous Resin

[Objective] This study was conducted to develop a method for rapidly separating macranthoidin B and dipsacoside B from Flos Lonicerae. [Method] HP-20 and HP-SS macroporous resin were applied to separate and purify macranthoidin B and dipsacoside B from Flos Lonicerae. The extract of Flos Lonicerae was first loaded onto an HP-20 column to enrich saponins, which were then separated by an HP-SS macroporous resin column to get pure macranthoidin B and dipsacoside B.[Result] The optimal HP-20 purification conditions included： a concentration of sample liquid at 4.8 mg/ml, a sample volume of 2 BV, an adsorption flow rate at 1.5BV/h, an ethanol concentration for desorption at 60%, a desorption volume of 3 BV,and a desorption flow rate at 1.5 BV/h. Total saponins were then separated by an HP-SS macroporous resin column which was eluted sequentially by water, 20%ethanol, 30% ethanol, 40% ethanol and 50% ethanol. Two purified compounds were obtained in fractions eluted by 40% ethanol and 50% ethanol, respectively. The two compounds were identified as macranthoidin B and dipsacoside B by13 C and1H nuclear magnetic resonance spectroscopy. [Conclusion] The combination of HP-20 and HP-SS macroporous resin could efficiently separate macranthoidin B and dipsacoside B from Flos Lonicerae.

蓝靛果忍冬(Lonicera caerulea)种质资源研究进展

简述了蓝靛果忍冬种质资源的研究进展。并对俄罗斯的品种和我国东北地区的野生植株引种 栽培的情况进行了初步观察和分析,指出了今后对其开发利用的研究方向。

Effect of chlorogenic acid on antioxidant activity of Flos Lonicerae extracts

Flos Lonicerae is a medically useful traditional Chinese medicine herb. However, little is known about the antioxidant properties of Flos Lonicerae extracts. Here the antioxidant capacity of water, methanolic and ethanolic extracts prepared from Flos Lonicerae to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and reduce Fe3+ to Fe2+ is examined. Chlorogenic acid, a major component of Flos Lonicerae, is identified and further purified from 70% ethanolic extract with high performance liquid chromatography (HPLC) and its antioxidant capacity is characterized. The total phenolic compounds and chlorogenic acid con-tents in Flos Lonicerae are determined. The present results demonstrate that the Flos Lonicerae extracts exhibit antioxidant ac-tivity and chlorogenic acid is a major contributor to this activity.