Perspectives on the adaptation of Japanese plum-type cultivars to reduced winter chilling in two regions of Spain.

Japanese plum, like other temperate fruit tree species, has cultivar-specific temperature requirements during dormancy for proper flowering. Knowing the temperature requirements of this species is of increasing interest due to the great genetic variability that exists among the available Japanese plum-type cultivars, since most of them are interspecific hybrids. The reduction of winter chilling caused by climate change is threatening their cultivation in many regions. In this work, the adaptation perspectives of 21 Japanese plum-type cultivars were analyzed in two of the main plum-growing regions in Spain, Badajoz and Zaragoza, to future climate conditions. Endodormancy release for subsequent estimation of chilling and heat requirements was determined through empirical experiments conducted during dormancy at least over two years. Chill requirements were calculated using three models [chilling hours (CH), chilling units (CU) and chilling portions (CP)] and heat requirements using growing degree hours (GDH). Chilling requirements ranged 277-851 CH, 412-1,030 CU and 26-51 CP, and heat requirements ranged from 4,343 to 9,525 GDH. The potential adaption of the cultivars to future warmer conditions in both regions was assessed using climate projections under two Representative Concentration Pathways (RCP), RCP4.5 (effective reduction of greenhouse gas emissions) and RCP8.5 (continuous increase in greenhouse gas emissions), in two time horizons, from the middle to the end of 21st century, with temperature projections from 15 Global Climate Models. The probability of satisfying the estimated cultivar-specific chilling requirements in Badajoz was lower than in Zaragoza, because of the lower chill availability predicted. In this region, the cultivars analyzed herein may have limited cultivation because the predicted reduction in winter chill may result in the chilling requirements not being successfully fulfilled.

The dynamic changes in volatile metabolites provide a new understanding for the special flavor formation in z. Mioga flower buds during the growth stages.

Although Z. mioga flower buds are popular among consumers for its unique spicy flavor, high nutritional and medicinal value, there are few reports on the formation and changes of the flavor during its growth and maturation process. The understanding of the profile of volatile compounds would help to unravel the flavor formation for Z. mioga flower buds during growth. The volatile changes in Z. mioga flower buds were analyzed by GC-MS and a total of 182 volatile compounds identified, and the terpenoids accounted for the most abundant volatile substances. Almost all the identified volatiles presented an intuitive upward trend throughout the growth period and reached the maximum at the later stage of development (DS3 or DS4). Regarding the PCA and HCA results, there were significant differences found among the four stages, and the DS3 was the critical node. The top 50 differential volatiles screened by OPLS-DA and PLS-DA were all up-regulated, and the correlation analysis indicated that terpenoids might synergize with other chemical types of volatiles to jointly affect the flavor formation of Z. mioga flower buds during growth. The association network for flavor omics revealed that the most important sensory flavor for Z. mioga flower buds were woody and sweet, and the main contribution compounds for the unique flavor contained ß-guaiene, ß-farnesene, δ-cadinene and citronellyl isobutanoate. Taken together, the results of this study provided a reference for flavor quality evaluation of flower buds and determination of the best harvest period.

DIA-Based Quantitative Proteomics in the Flower Buds of Two Malus sieversii (Ledeb.) M. Roem Subtypes at Different Overwintering Stages.

Malus sieversii is considered the ancestor of the modern cultivated apple, with a high value for apple tolerance breeding. Despite studies on the temperature adaptability of M. sieversii carried out at a physiological response and the genome level, information on the proteome changes of M. sieversii during dormancy is limited, especially about the M. sieversii subtypes. In this study, a DIA-based approach was employed to screen and identify differential proteins involved in three overwintering periods of flower buds in two M. sieversii subtypes (Malus sieversii f. luteolus, GL; Malus sieversii f. aromaticus, HC) with different overwintering adaptabilities. The proteomic analysis revealed that the number of the down-regulated differential expression proteins (DEPs) was obviously higher than that of the up-regulated DEPs in the HC vs. GL groups, especially at the dormancy stage and dormancy-release stage. Through functional classification of those DEPs, the majority of the DEPs in the HC vs. GL groups were associated with protein processing in the endoplasmic reticulum, oxidative phosphorylation, starch and sucrose metabolism and ribosomes. Through WGCNA analysis, tricarboxylic acid cycle and pyruvate metabolism were highly correlated with the overwintering stages; oxidative phosphorylation and starch and sucrose metabolism were highly correlated with the Malus sieversii subtypes. This result suggests that the down-regulation of DEPs, which are predominantly enriched in these pathways, could potentially contribute to the lower cold tolerance observed in HC during overwintering stage.

Effect of Glutamic Acid and 6-benzylaminopurine on Flower Bud Biostimulation, Fruit Quality and Antioxidant Activity in Blueberry.

Blueberry is a highly demanded and consumed fruit due to its beneficial effects on human health, because of its bioactive compounds with a high antioxidant capacity. The interest in increasing the yield and quality of blueberries has led to the application of some innovative techniques such as biostimulation. The objective of this research was to assess the effect of the exogenous application of glutamic acid (GLU) and 6-benzylaminopurine (6-BAP) as biostimulants on flower bud sprouting, fruit quality, and antioxidant compounds in blueberry cv. Biloxi. The application of GLU and 6-BAP positively affected bud sprouting, fruit quality, and antioxidant content. The application of 500 and 10 mg L-1 GLU and 6-BAP, respectively, increased the number of flower buds, while 500 and 20 mg L-1 generated fruits with higher content of flavonoids, vitamin C, and anthocyanins and higher enzymatic activity of catalase and ascorbate peroxidase enzymes. Hence, the application of these biostimulants is an effective way to enhance the yield and fruit quality of blueberries.

Preparation of Male Meiotic Chromosomes for Fluorescence In Situ Hybridization and Immunodetection with Major Focus on Dogroses.

Meiosis is a very essential cell division resulting in the formation of four haploid gametes in plants. The preparation of meiotic chromosomes is a key step in plant meiotic research. Well-spread chromosomes, low background signal, and effective cell wall elimination give the best hybridization results. Dogroses (Rosa, section Caninae) are allopolyploids and frequently pentaploids (2n = 5x = 35) with asymmetrical meiosis. Their cytoplasm is enriched with organic compounds such as vitamins, tannins, phenols, essential oils, and many more. The cytoplasm is often a huge problem, avoiding successful cytogenetic experiments using fluorescence staining techniques. Here, we present a protocol with modifications for the preparation of male meiotic chromosomes suitable for fluorescence in situ hybridization (FISH) and immunolabeling with a major focus on dogroses.

Phytochemical Analysis, GC-MS Profiling, and In Vitro Evaluation of Biological Applications of Different Solvent Extracts of Leonotis nepetifolia (L.) R.Br. Flower Buds.

Leonotis nepetifolia (L.) R.Br. is a medicinally important herb belonging to the family Lamiaceae. The plant is typically found in tropical regions, and its leaf and root extracts are renowned for their ethno-botanical and therapeutic applications. This study was designed to determine the presence of various bioactive components, and to evaluate antibacterial, antifungal, antioxidant, and anti-proliferative activities. The preliminary phytochemical screening and gas chromatography-mass spectrometry (GC-MS) analysis of different solvent extracts revealed the presence of various bioactive compounds, of which methanol extract showed 24 compounds, petroleum ether extract revealed 26 compounds, and 24 compounds in hexane extracts. The major bioactive components including λ-sitosterol (16.20 %) in methanol extract, 1-nonadecanol (15.48 %) in petroleum extract, and eicosane (13.22 %) in hexane extract have been reported with various bio-therapeutic applications. In addition, the flower bud methanolic extract of L. nepetifolia exhibited inhibitory potential against all tested bacterial and fungal pathogens. The DPPH radical scavenging assay revealed that methanolic extract possessed the highest antioxidant activity. The scavenging activity increased in a concentration-dependent manner, as indicated by a 74 % inhibition rate at 1000 µg/ml. Furthermore, the in vitro cytotoxic effects of the methanolic extract on the HepG2 cell line were evaluated. The IC50 value of methanolic extract against HepG2 cells was determined to be 83.28 µg/ml. The findings reveal that different solvent extracts of L. nepetifolia flower buds contain a significant amount of various bioactive phytochemicals with antioxidant and anticancer activities; and thus, the plant could serve as a potential source of pharmacological applications.

Genome-Wide Identification of AP2/ERF Superfamily Genes in Juglans mandshurica and Expression Analysis under Cold Stress.

Juglans mandshurica has strong freezing resistance, surviving temperatures as low as -40 °C, making it an important freeze tolerant germplasm resource of the genus Juglans. APETALA2/ethylene responsive factor (AP2/ERF) is a plant-specific superfamily of transcription factors that regulates plant development, growth, and the response to biotic and abiotic stress. In this study, phylogenetic analysis was used to identify 184 AP2/ERF genes in the J. mandshurica genome, which were classified into five subfamilies (JmAP2, JmRAV, JmSoloist, JmDREB, and JmERF). A significant amount of discordance was observed in the 184 AP2/ERF genes distribution of J. mandshurica throughout its 16 chromosomes. Duplication was found in 14 tandem and 122 segmental gene pairs, which indicated that duplications may be the main reason for JmAP2/ERF family expansion. Gene structural analysis revealed that 64 JmAP2/ERF genes contained introns. Gene evolution analysis among Juglandaceae revealed that J. mandshurica is separated by 14.23 and 15 Mya from Juglans regia and Carya cathayensis, respectively. Based on promoter analysis in J. mandshurica, many cis-acting elements were discovered that are related to light, hormones, tissues, and stress response processes. Proteins that may contribute to cold resistance were selected for further analysis and were used to construct a cold regulatory network based on GO annotation and JmAP2/ERF protein interaction network analysis. Expression profiling using qRT-PCR showed that 14 JmAP2/ERF genes were involved in cold resistance, and that seven and five genes were significantly upregulated under cold stress in female flower buds and phloem tissues, respectively. This study provides new light on the role of the JmAP2/ERF gene in cold stress response, paving the way for further functional validation of JmAP2/ERF TFs and their application in the genetic improvement of Juglans and other tree species.

Leonotis nepetifolia Flower Bud Extract Mediated Green Synthesis of Silver Nanoparticles, Their Characterization, and In Vitro Evaluation of Biological Applications.

Biosynthesis of silver nanoparticles (AgNPs) using the green matrix is an emerging trend and is considered green nanotechnology because it involves a simple, low-cost, and environmentally friendly process. The present research aimed to synthesize silver nanoparticles from a Leonotis nepetifolia (L.) R.Br. flower bud aqueous extract, characterize these nanoparticles, and perform in vitro determination of their biological applications. UV-Vis spectra were used to study the characterization of biosynthesized L. nepetifolia-flower-bud-mediated AgNPs (LnFb-AgNPs); an SPR absorption maximum at 418 nm confirmed the formation of LnFb-AgNPs. The presumed phytoconstituents subjected to reduction in the silver ions were revealed by FTIR analysis. XRD, TEM, EDS, TGA, and zeta potential with DLS analysis revealed the crystalline nature, particle size, elemental details, surface charge, thermal stability, and spherical shape, with an average size of 24.50 nm. In addition, the LnFb-AgNPs were also tested for antimicrobial activity and exhibited a moderate zone of inhibition against the selected pathogens. Concentration-dependent antioxidant activity was observed in the DPPH assay. Further, the cytotoxicity increased proportionate to the increasing concentration of the biosynthesized LnFb-AgNPs with a maximum effect at 200 µg/mL by showing the inhibition cell viability percentages and an IC50 of 35.84 µg/mL. Subsequently, the apoptotic/necrotic potential was determined using Annexin V/Propidium Iodide staining by the flow cytometry method. Significant early and late apoptosis cell populations were observed in response to the pancreatic ductal adenocarcinoma (PANC-1) cell line, as demonstrated by the obtained results. In conclusion, the study's findings suggest that the LnFb-AgNPs could serve as remedial agents in a wide range of biomedical applications.

PIN and PILS family genes analyses in Chrysanthemum seticuspe reveal their potential functions in flower bud development and drought stress.

Auxin affects almost all plant growth and developmental processes. The PIN-FORMED (PIN) and PIN-LIKES (PILS) family genes determine the direction and distribution gradient of auxin flow by polar localization on the cell membrane. However, there are no systematic studies on PIN and PILS family genes in chrysanthemum. Here, 18 PIN and 13 PILS genes were identified in Chrysanthemum seticuspe. The evolutionary relationships, physicochemical properties, conserved motifs, cis-acting elements, chromosome localization, collinearity, and expression characteristics of these genes were analyzed. CsPIN10a, CsPIN10b, and CsPIN10c are unique PIN genes in C. seticuspe. Expression pattern analysis showed that these genes had different tissue specificities, and the expression levels of CsPIN8, CsPINS1, CsPILS6, and CsPILS10 were linearly related to the developmental period of the flower buds. In situ hybridization assay showed that CsPIN1a, CsPIN1b, and CsPILS8 were expressed in floret primordia and petal tips, and CsPIN1a was specifically expressed in the middle of the bract primordia, which might regulate lateral expansion of the bracts. CsPIN and CsPILS family genes are also involved in drought stress responses. This study provides theoretical support for the cultivation of new varieties with attractive flower forms and high drought tolerance.

Male Meiosis as a Biomarker for Endo- to Ecodormancy Transition in Apricot.

Dormancy is an adaptive strategy in plants to survive under unfavorable climatic conditions during winter. In temperate regions, most fruit trees need exposure to a certain period of low temperatures to overcome endodormancy. After endodormancy release, exposure to warm temperatures is needed to flower (ecodormancy). Chilling and heat requirements are genetically determined and, therefore, are specific for each species and cultivar. The lack of sufficient winter chilling can cause failures in flowering and fruiting, thereby compromising yield. Thus, the knowledge of the chilling and heat requirements is essential to optimize cultivar selection for different edaphoclimatic conditions. However, the lack of phenological or biological markers linked to the dormant and forcing periods makes it difficult to establish the end of endodormancy. This has led to indirect estimates that are usually not valid in different agroclimatic conditions. The increasing number of milder winters caused by climatic change and the continuous release of new cultivars emphasize the necessity of a proper biological marker linked to the endo- to ecodormancy transition for an accurate estimation of the agroclimatic requirements (AR) of each cultivar. In this work, male meiosis is evaluated as a biomarker to determine endodormancy release and to estimate both chilling and heat requirements in apricot. For this purpose, pollen development was characterized histochemically in 20 cultivars over 8 years, and the developmental stages were related to dormancy. Results were compared to three approaches that indirectly estimate the breaking of dormancy: an experimental methodology by evaluating bud growth in shoots collected periodically throughout the winter months and transferred to forcing chambers over 3 years, and two statistical approaches that relate seasonal temperatures and blooming dates in a series of 11-20 years by correlation and partial least square regression. The results disclose that male meiosis is a possible biomarker to determine the end of endodormancy and estimate AR in apricot.

Preferential freezing avoidance localised in anthers and embryo sacs in wintering Daphne kamtschatica var. jezoensis flower buds visualised by magnetic resonance imaging.

To explore diversity in cold hardiness mechanisms, high resolution magnetic resonance imaging (MRI) was used to visualise freezing behaviours in wintering Daphne kamtschatica var. jezoensis flower buds, which have naked florets and no bud scales. MRI images showed that anthers remained stably supercooled to the range from -14 to -21°C or lower while most other tissues froze by -7°C. Freezing of some anthers detected in MRI images between -14 and -21°C corresponded with numerous low temperature exotherms and also with the 'all-or-nothing' type of anther injuries. In ovules/pistils, only embryo sacs remained supercooled at -7°C or lower, but slowly dehydrated during further cooling. Cryomicroscopic observation revealed ice formation in the cavities of calyx tubes and pistils but detected no ice in embryo sacs or in anthers. The distribution of ice nucleation activity in floral tissues corroborated the tissue freezing behaviours. Filaments likely work as the ice blocking barrier that prevents ice intrusion from extracellularly frozen calyx tubes to connecting unfrozen anthers. Unique freezing behaviours were demonstrated in Daphne flower buds: preferential freezing avoidance in male and female gametophytes and their surrounding tissues (by stable supercooling in anthers and by supercooling with slow dehydration in embryo sacs) while the remaining tissues tolerate extracellular freezing.

Structurally diverse glycosides of secoiridoid, bisiridoid, and triterpene-bisiridoid conjugates from the flower buds of two Caprifoliaceae plants and their ATP-citrate lyase inhibitory activities.

The ethanolic extracts of the dried flower buds of two Caprifoliaceae plants, Lonicera japonica and Abelia × grandiflora, showed considerable inhibitory activities against adenosine triphosphate (ATP)-citrate lyase (ACL), a new promising drug target for the treatment of metabolic disorders. Bioassay-guided purification in conjunction with HPLC-PDA profiling led to the isolation and characterization of thirty-five (1-35) and fourteen (1'-14') structurally diverse compounds from the above two plant extracts, respectively. Compounds 1-9 and 1'-6' are previously undescribed glycosides. Their structures were elucidated on the basis of spectroscopic data, electronic circular dichroism (ECD), and single crystal X-ray diffraction analyses. In particular, lonicejaposide A (1) has an unprecedented skeleton generated through the coupling of C-7 in secologanin with C-2'' in phenylacetaldehyde via an aldol condensation. Abeliflorosides A (1') and B (2') are hitherto unknown glycosides of triterpene and bisiridoid conjugates constructed through the formation of a 1,3-dioxane moiety. All the isolates were evaluated for their inhibitory activities against ACL. Compounds 9, 25-28, 31, 1', 2', and 14' displayed significant inhibitory effects, with IC50 values ranging from 0.1 to 14.2 µM. The interactions of selected compounds possessing different structure features (e.g., 9, 25, 31, and 2') with ACL were thereafter performed by employing molecular docking studies. In addition, compound 2', the most complex triterpene-bisiridoid conjugate glycoside reported herein, also inhibited acetyl-CoA carboxylase 1 (ACC1), with an IC50 value of 7.9 µM. The dried material of the flower buds of L. japonica (honeysuckle) is a well-known traditional oriental medicine (i.e., Flos Lonicerae Japonicae, FLJ) and has long been used in large quantities. The above findings not only provide new insights for the development of multipurpose utilization of FLJ in healthcare community, but also provide profitable clues indicating that the flower buds of A. × grandiflora might be a potential alternative to FLJ in the traditional Chinese medicine market.

Effect of citric acid and heat treatment on the content of less-polar ginsenosides in flower buds of Panax ginseng.

Ginseng flower bud (GFB), as an inexpensive part of Panax ginseng, attracted significant attention as a beneficial functional food with medicinal potentials due to its high content of ginsenosides. A few studies focused on the utilization of heat treatment and citric acid treatment to process ginseng flowers, converting its polar ginsenosides into rare ginsenosides to improve its biological activities. Thus, in this study, we compared the changes of ginsenosides in GFB after citric acid and heat treatment by HPLC method. The results revealed that less-polar ginsenoside, Rg6 and F4, increased to 1.01 and 0.27% by heat treatment, respectively. Further, ginsenoside F2 increased to 1.13% with 1 M citric acid treatment. Furthermore, based on the combination of these two processing methods for the first time, the conversion rate of less-polar ginsenosides surged to 80%. The content of ginsenoside Rg3(s) and Rg5 increased to 1.509 and 1.871%, respectively, by simultaneous heat and citric acid treatment. Therefore, a processing approach that simultaneously performs heat and citric acid treatments has been proposed, and this considerably inexpensive and convenient processing method could be applied to the processing of GFBs and produce less-polar ginsenosides.

Interplay among Antioxidant System, Hormone Profile and Carbohydrate Metabolism during Bud Dormancy Breaking in a High-Chill Peach Variety.

(1) Background: Prunus species have the ability to suspend (induce dormancy) and restart growth, in an intricate process in which environmental and physiological factors interact. (2) Methods: In this work, we studied the evolution of sugars, antioxidant metabolism, and abscisic acid (ABA) and gibberellins (GAs) levels during bud dormancy evolution in a high-chill peach variety, grown for two seasons in two different geographical areas with different annual media temperature, a cold (CA) and a temperate area (TA). (3) Results: In both areas, starch content reached a peak at ecodormancy, and then decreased at dormancy release (DR). Sorbitol and sucrose declined at DR, mainly in the CA. In contrast, glucose and fructose levels progressively rose until DR. A decline in ascorbate peroxidase, dehydroascorbate reductase, superoxide dismutase and catalase activities occurred in both seasons at DR. Moreover, the H2O2-sensitive SOD isoenzymes, Fe-SOD and Cu,Zn-SOD, and two novel peroxidase isoenzymes, were detected. Overall, these results suggest the occurrence of a controlled oxidative stress during DR. GA7 was the major bioactive GA in both areas, the evolution of its levels being different between seasons and areas. In contrast, ABA content decreased during the dormancy period in both areas, resulting in a reduction in the ABA/total GAs ratio, being more evident in the CA. (4) Conclusion: A possible interaction sugars-hormones-ROS could take place in high-chill peach buds, favoring the DR process, suggesting that, in addition to sugar metabolism, redox interactions can govern bud DR, regardless of chilling requirements.

Six new dammarane-type triterpene saponins from Panax ginseng flower buds and their cytotoxicity.

BACKGROUND: Panax ginseng has been used for a variety of medical purposes in eastern countries for more than two thousand years. From the extensive experiences accumulated in its long medication use history and the substantial strong evidence in modern research studies, we know that ginseng has various pharmacological activities, such as antitumor, antidiabetic, antioxidant, and cardiovascular system-protective effects. The active chemical constituents of ginseng, ginsenosides, are rich in structural diversity and exhibit a wide range of biological activities. METHODS: Ginsenoside constituents from P. ginseng flower buds were isolated and purified by various chromatographic methods, and their structures were identified by spectroscopic analysis and comparison with the reported data. The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H- tetrazolium bromide method was used to test their cytotoxic effects on three human cancer cell lines. RESULTS: Six ginsenosides, namely 6'-malonyl formyl ginsenoside F1 (1), 3ß-acetoxyl ginsenoside F1 (2), ginsenoside Rh24 (6), ginsenoside Rh25 (7), 7ß-hydroxyl ginsenoside Rd (8) and ginsenoside Rh26 (10) were isolated and elucidated as new compounds, together with four known compounds (3-5 and 9). In addition, the cytotoxicity of these isolated compounds was shown as half inhibitory concentration values, a tentative structure-activity relationship was also discussed based on the results of our bioassay. CONCLUSION: The study of chemical constituents was useful for the quality control of P. ginseng flower buds. The study on antitumor activities showed that new Compound 1 exhibited moderate cytotoxic activities against HL-60, MGC80-3 and Hep-G2 with half inhibitory concentration values of 16.74, 29.51 and 20.48 µM, respectively.

Rb1, the Primary Active Ingredient in Panax ginseng C.A. Meyer, Exerts Antidepressant-Like Effects via the BDNF-Trkb-CREB Pathway.

Panax ginseng C.A. Meyer (Araliaceae), a popular tonic and dietetic herbal medicine, has been traditionally prescribed in China and other countries to treat affective disorders. The medicinal parts of ginseng, the roots and flower buds, have become increasingly popular as dietary supplements due to the current holistic healthcare trend. We have investigated for the first time the antidepressive actions of the different medicinal parts, namely, the main roots, fibrous roots, and flower buds (in water extract and powder), of garden-cultivated ginseng through behavioral and drug-induced tests in mice. The water extracts, but not the powders of ginseng fibrous roots, flower buds, and main roots (1.5 g of crude drug per kilogram, p.o.), significantly reduced the immobility time in the forced swim test (FST) and tail suspension test (TST); moreover, the water extracts enhanced the 5-hydroxytryptophan (5-HTP)-induced head-twitch response and antagonized the action of reserpine in the mouse. We then explored the antidepressive mechanism of action of the ginsenoside Rb1 (Rb1) related to the brain-derived neurotrophic factor (BDNF) and its downstream proteins in mice exposed to chronic unpredictable mild stress (CUMS). Treatment with Rb1 (20 mg/kg, p.o.) for 21 days significantly attenuated the CUMS-induced decrease in the activities of BDNF, tropomyosin-related kinase B (TrkB), protein kinase B (AKT), extracellular regulatory protein kinase (ERK), and cyclic adenosine monophosphate (cAMP) response element binding protein (CREB) in the mouse hippocampal CA3 region and prefrontal cortex (PFC). Interestingly, treatment with the novel TrkB antagonist ANA-12 (0.5 mg/kg, i.p.) did not alter the level of BDNF but significantly blocked the antidepressive effects of Rb1 on proteins downstream of BDNF in CUMS-treated mice. These results suggest that BDNF-TrkB-CREB signaling may be involved in the antidepressive mechanism of the action of Rb1.

[Studies on chemical constituents of saponins from Panax ginseng flower buds].

This project is to investigate the chemical constituents of ginsenosides from the flower buds of Panax ginseng. The compounds were isolated by using a variety of chromatographic methods including Diaion HP-20,silica gel,MCI gel and semi-preparative HPLC chromatography. Their structures were identified by NMR,and MS data. As a result,32 compounds were isolated from the extract of P. ginseng flower buds,and identified as ginsenoside Rk_3( 1),ginsenoside Rh_4( 2),ginsenoside Rh_8( 3),pseudoginsenoside Rc_1( 4),ginsenoside Rc( 5),ginsenoside Rb_2( 6),ginsenoside Rg_6( 7),20( E)-ginsenoside F_4( 8),ginsenoside Rb_1( 9),vinaginsenoside R_(16)( 10),ginsenoside Rh_6( 11),vinaginsenoside R_3( 12),5,6-didehydro-ginsenoside Rd( 13),vinaginsenoside R_4( 14),vinaginsenoside R_8( 15),ginsenoside Rf( 16),notoginsenoside E( 17),ginsenoside Ⅲ( 18),3-O-ß-D-glucopyranosyl-3ß,7ß,12ß,20 S-tetrahydroxydammar-5( 6),24-diene-20-O-ß-D-glucopyranoside( 19),20( S)-ginsenoside Rg_2( 20),20( R)-ginsenoside Rg_2( 21),notoginsenoside R_2( 22),ginsenoside F_2( 23),quinquenoside I( 24),ginsenoside M_1( 25),quinquenoside L_(10)( 26),ginsenoside Rh_5( 27),ginsenoside Rg_5( 28),ginsenoside Rk_1( 29),20( R)-ginsenoside Rg_3( 30),oleanolic acid 3-O-ß-D-glucopyranosyl-( 1→2)-ß-D-( 6'-methyl ester)-glucuronopyranoside( 31) and ginsenoside MC( 32). Among them,compounds 10,12,13,15,19,22,24,31 and 32 were isolated from P. ginseng for the first time,and compound 19 was a genuine ginsenoside firstly obtained by separation and identification,with NMR data that were also reported. Compounds 1-3,7,8,23,25-30 were isolated from P. ginseng flower buds for the first time.

New Depsides and Neuroactive Phenolic Glucosides from the Flower Buds of Rugosa Rose ( Rosa rugosa).

The flower buds of Rosa rugosa Thunb. have been commonly used as a source of rose oil and as an ingredient in tea in eastern Asia, including China, Japan, and Korea. Repeated chromatography of a hot water extract from the flower buds of R. rugosa led to the isolation and characterization of three new depside glucosides, rosarugosides A-C (1-3), along with three phenolic compounds, one ionone glucoside, four flavonoids, and two tannins having known chemical structures. Linarionoside A and 2-phenylethyl-(6- O-galloyl)-ß-d-glucopyranoside were isolated from R. rugosa for the first time in this study. The structures of the new compounds 1-3 were elucidated by interpreting one- and two-dimensional nuclear magnetic resonance spectroscopic and mass spectrometric data. Among the isolates, a new depside glucoside (1) and two major phenolic glucosides (4 and 5) improved MK-801-induced sensorimotor gating deficits, which were measured via an acoustic startle response test in mice.

Protective effects of extract of Cleistocalyx operculatus flower buds and its isolated major constituent against LPS-induced endotoxic shock by activating the Nrf2/HO-1 pathway.

The flower buds of Cleistocalyx operculatus are used as an important ingredient in herbal tea and herbal products in several tropical countries. However, their protective effects and underlying mechanisms on lipopolysaccharide (LPS)-induced endotoxic shock remain unclear. The aim of this study was to investigate the anti-inflammatory effects of ethanol extract of C. operculatus flower buds (ECO) and its major constituent 2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone (DMC) in macrophages and in an experimental LPS-induced sepsis mouse model. ECO inhibited the LPS-induced production and expression of pro-inflammatory mediators in macrophages. In an endotoxic shock mouse model, the oral administration of ECO rescued LPS-induced mortality, and attenuated LPS-induced increases in the serum levels of pro-inflammatory mediators, and damage of the lung and liver tissues. ECO increased the nuclear translocation of the nuclear factor erythroid 2-related factor 2 (Nrf2), as well as the expression of Nrf2 target genes, including heme oxygenase-1 (HO-1), in macrophages. Similar to the effects of ECO, DMC also inhibited the LPS-induced inflammatory response in macrophages and endotoxic shock in mice, and activated the Nrf2/HO-1 pathway. In conclusion, our findings suggested that ECO and its major constituent, DMC, attenuated LPS-induced endotoxic shock by activating the Nrf2/HO-1 pathway.

Extraction and Separation of Eight Ginsenosides from Flower Buds of Panax Ginseng Using Aqueous Ionic Liquid-Based Ultrasonic-Assisted Extraction Coupled with an Aqueous Biphasic System.

Ionic liquids (ILs) are recognized as a possible replacement of traditional organic solvents, and ILs have been widely applied to extract various compounds. The present work aims to extract ginsenosides from Panax ginseng flower buds using aqueous ionic liquid based ultrasonic assisted extraction (IL-UAE). The extraction yields of 1-alkyl-3-methylimidazolium ionic liquids with different anions and alkyl chains were evaluated. The extraction parameters of eight ginsenosides were optimized by utilizing response surface methodology (RSM). The model demonstrated that a high yield of total ginsenosides could be obtained using IL-UAE, and the optimum extraction parameters were 0.23 M [C4mim][BF4], ultrasonic time of 23 min, temperature of extraction set to 30 °C, and liquid-solid ratio of 31:1. After that, an aqueous biphasic system (ABS) was used to separate ginsenosides further. The nature and concentration of salt, as well as the value of pH in ionic liquid were evaluated, and the optimal conditions (6.0 mL IL extract, 3 g NaH2PO4, and pH 5.0) were obtained. The preconcentration factor was 2.58, and extraction efficiency reached 64.53%. The results indicate that as a simple and efficient method, an IL-UAE-ABS can be considered as a promising method for extracting and separating the natural active compounds from medicinal herbs.