Network Pharmacology Study on Herb Pair Bletilla striata-Galla chinensis in the Treatment of Chronic Skin Ulcers.

BACKGROUND: Herb pair Bletilla striata-Galla chinensis (BS-GC) is a classic combination of topical traditional Chinese medicine formulae in the treatment of chronic skin ulcers (CSUs). OBJECTIVE: The aim of this study is to explore the effective active ingredients of BS-GC, as well as the core targets and signal transduction pathways of its action on CSUs. METHODS: The ingredients of BS-GC were obtained from TCMSP and HERB databases. The targets of all active ingredients were retrieved from the SwissTargetPrediction database. The targets of CSUs were obtained from OMIM, GeneCards, Drugbank, and DisGeNET databases. A drug-disease target protein-protein interaction (PPI) network was constructed to select the most core targets, and an herb-ingredient-target network was built by utilizing Cytoscape 3.7.2. Furthermore, we performed Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes database (KEGG) analysis and verified the results of network pharmacology through molecular docking. RESULTS: A total of 40 active ingredients from the herb pair BS-GC were initially screened, and a total of 528 targets were retrieved. Meanwhile, the total number of CSU targets was 1032. Then, the number of common targets between BS-GC and CSUs was 107. The 13 core targets of herb pair BS-GC with CSUs were filtered out according to the PPI network, including AKT1, TNF, EGFR, BCL2, HIF1A, MMP-9, etc. The 5 main core active ingredients were 1-(4-Hydroxybenzyl)-2-methoxy-9,10-dihydrophenanthrene-4,7-diol, 1-(4- Hydroxybenzyl)-4-methoxy-9,10-dihydrophenanthrene-2,7-diol, physcion, dihydromyricetin, and myricetin. The main biological processes were inflammation, oxidative stress, and immune response, involving the AGE-RAGE signaling pathway in diabetic complications, HIF-1 signaling pathway, NF-κB signaling pathway, and calcium signaling pathway. Molecular docking results showed good binding activity between the 5 main core active ingredients and 13 core targets. CONCLUSION: This study predicted the core targets and signal transduction pathways in the treatment of CSUs to provide a reference for further molecular mechanism research.

A dual network cross-linked hydrogel with multifunctional Bletilla striata polysaccharide/gelatin/tea polyphenol for wound healing promotion.

Wound healing is a dynamic and complex biological process, and traditional biological excipients cannot meet the needs of the wound healing process, and there is an urgent need for a biological dressing with multifunctionality and the ability to participate in all stages of wound healing. This study developed tea polyphenol (TP) incorporated multifunctional hydrogel based on oxidized Bletilla striata polysaccharide (OBSP) and adipic acid dihydrazide modified gelatin (Gel-ADH) with antimicrobial, antioxidant hemostatic, and anti-inflammatory properties to promote wound healing. The composite OBSP, Gel-ADH, TP (OBGTP) hydrogels prepared by double crosslinking between OBSP, TP and Gel-ADH via Schiff base bonding and hydrogen bonding had good rheological and swelling properties. The introduction of TP provided the composite hydrogel with excellent antioxidant antibacterial activities against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coil). In the rat liver hemorrhage model and skin injury model, the OBGTP composite hydrogel had significant (p < 0.001) hemostatic ability, and had the ability to accelerate collagen deposition, reduce the expression of inflammatory factors, and promote rapid wound healing. In addition, OBGTP hydrogels had adhesive properties and good biocompatibility. In conclusion, OBGTP multifunctional composite hydrogels have great potential for wound healing applications.

Isolation, invitro, invivo anti-inflammatory, analgesic and antioxidant potential of Habenaria plantegania Lindl.

Habenaira plantaginea belong to orchid family which is native to Asia. Members of this family are commonly famous for the cure of pain and inflammation. To date, no research was found on isolation of compounds from this plant for the treatment of inflammation and analgesia nor has been published to our knowledge. The purpose of this study was to evaluate an analgesic, anti-inflammatory and anti-oxidant activity of the isolated compound from the most potent chloroform sub-fraction and the isolated compounds form the habenaria plantaginea. Anti-inflammatory analgesic and antioxidant potential of the various chloroform sub-fractions and isolated compounds from the most potent sub-fraction (HP-1 & HP-1) were screened for their in vitro enzymatic assays. Furthermore, prior to in-vivo investigation, the isolated compounds were subjected for their toxicity study. The potent compound was then examined for acetic acid-induced writhing, hot plate test, carrageenan-induced inflammation assays. Further various phlogistic agents were used for the evaluation of mechanism. In the COX-2 inhibitory assay the chloroform sub fraction Cf-4 demonstrated excellent activity as compared to the other sub-fraction with 92.15% inhibition. The COX-2 enzyme make prostaglandins which are directly involved in inflammation. Likewise against 5-LOX the Cf-4 was the most potent sub-fraction with IC50 3.77 µg/mL. The 5-LOX catalyzes the biosynthesis of leukotrienes which is a group of lipid mediators of inflammation derived from arachidonic acid. Free radicals can induce inflammation through cellular damage while chronic inflammation generates a large number of free radicals, whose eventually lead to inflammation. In antioxidant assays the Cf-4 fraction was displayed excellent results against ABTS, DPPH and H2O2 free radical with 88.88, 77.44, and 65.52% inhibition at highest concentration. Likewise, the compound HP-1 demonstrated 88.81, 89.34 and 80.43% inhibition while compound HP-2 displayed 84.34, 91.52 and 82.34% inhibition against ABTS, DPPH and H2O2 free radical which were comparable to the standard drug ascorbic acid respectively. This study's findings validate the use of this species as traditional use.

First report of the complete mitochondrial genome of 3 beetles (Coleoptera: Scarabaeidae) harming Gastrodia elata (Asparagales: Orchidaceae).

Gastrodia elata Blume, a valuable traditional Chinese medicine with significant clinical and nutritional importance, is a fungal heterotrophic orchid. We present the first report of the mitochondrial genome structure and characteristics of 3 Scarabaeidae pests affecting G. elata: Sophrops peronosporus Gu & Zhang, Anomala rufiventris Kollar & Redtenbacher, and Callistethus plagiicollis Fairmaire. Each mitogenome contained 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), 2 ribosomal RNAs (rRNAs), and a control region, with no gene rearrangements observed. All 21 tRNAs, except trnS1 that lacks a dihydrouridine, had a stable cloverleaf secondary structure. Maximum likelihood and Bayesian inference analyses based on the 13 PCGs produced 2 topologically similar phylogenetic trees, both of with high nodal support. Larvae of these Scarabaeidae pests cause substantial damage by gnawing on the tubers and roots of G. elata, leading to reduced yield and compromised quality. These findings contribute to phylogenetic studies of Scarabaeidae, expand knowledge of G. elata pests, and offer valuable reference materials for their identification and control.

Distribution, Chemical Constituents and Biological Properties of Genus Malaxis.

The genus Malaxis (family Orchidaceae), comprises nearly 183 species available across the globe. The plants of this genus have long been employed in traditional medical practices because of their numerous biological properties, like the treatment of infertility, hemostasis, burning sensation, bleeding diathesis, fever, diarrhea, dysentery, febrifuge, tuberculosis, etc. Various reports highlight their phytochemical composition and biological activities. However, there is a lack of systematic review on the distribution, phytochemistry, and biological properties of this genus. Hence, this study aims to conduct a thorough and critical review of Malaxis species, covering data published from 1965 to 2022 with nearly 90 articles. Also, it examines different bioactive compounds, their chemistry, and pharmacotherapeutics as well as their traditional uses. A total of 189 unique compounds, including the oil constituents were recorded from Malaxis species. The highest active ingredients were obtained from Malaxis acuminata (103) followed by Malaxis muscifera (49) and Malaxis rheedei (33). In conclusion, this review offers an overview of the current state of knowledge on Malaxis species and highlights prospects for future research projects on them. Additionally, it recommends the promotion of domestication studies for rare medicinal orchids like Malaxis and the prompt implementation of conservation measures.

Gametophytic self-incompatibility in Maxillariinae orchids.

Gametophytic self-incompatibility (GSI) has been mainly described in species-rich clades within Orchidaceae. We report GSI for a genus within Maxillariinae, one of the most conspicuous and diverse subtribes of neotropical orchids. Here, we describe the reproductive system of Brasiliorchis picta, B. phoenicanthera, and B. porphyrostele. Anatomical studies of fruit development showed that pollen tubes of aborted, self-pollinated flowers degenerate half-way in the stylar channel and never reach the ovules. Spontaneous self-pollination and emasculation set no fruits for none of the sampled species, thus supporting the hypothesis that these three species studied rely on the agency of pollinators and pollinator-mediated cross-pollination to set fruit. Fruit set from cross-pollinations ranged from 33.4 to 77.5%. One self-pollinated fruit of B. porphyrostele developed to completion. All other fruits aborted between 10 and 21 days after pollination. These data support previous evidence of variable strength GSI being exhibited in orchid species. Additional studies of self-incompatibility systems are needed to evaluate their role in species diversification and evolution of reproductive strategies in Maxillariinae and to allow for effective conservation strategies of threatened orchid species.

Incomplete resupination during floral development leads to pollination failure in a slipper orchid.

In many families, plants undergo floral resupination by twisting through approximately 180° during floral development so that the flower is effectively positioned upside down. In most orchids, resupination results in the median petal (i.e., the labellum) becoming lowermost, which plays a crucial role in pollination by serving as a landing platform or as a trapping device, or both. Incomplete resupination is predicted to lead to reduced pollination, although tests of this assumption are still lacking. We investigated the effect of resupination using Phragmipedium vittatum, a rare lady's slipper orchid whose specialized labellum forms a trapping device. First, we surveyed the natural occurrence of incomplete resupination. Then we manipulated flowers into non- (≈0°), half- (≈90°), and fully resupinate (≈180°) positions to test the effect of orientation on pollen smear removal and deposition by pollinators (female hoverflies). We found that ca. 10% of flowers in the natural population were not fully resupinate, being either non- (upward, 0-60°) or half-resupinate (sideward, 60-120°). The change in orientation prevented the effectiveness of pollination by hoverflies since no pollen smear removal or deposition were found in flowers from non- and half-resupinate treatments. Although these flowers still attracted hoverflies, they were not trapped effectively. As this orchid is incapable of autonomous self-pollination, flowers that do not resupinate fail to set fruits. These results highlight the importance of correct floral orientation provided by resupination to ensure pollination in orchids and other resupinate flowers.

Utilizing epoxy Bletilla striata polysaccharide collagen sponge for hemostatic care and wound healing.

Hemostatic materials that are lightweight and possess good blood absorption performance have been widely considered for use in modern wound care. Natural hemostatic ingredients derived from traditional Chinese medicine have also received extensive attention. Bletilla polysaccharides are valued by researchers for their excellent hemostatic performance and good reactivity. Collagen is favored by researchers due to its high biocompatibility and low immunogenicity. In this study, Bletilla striata polysaccharide, the main hemostatic component of Bletilla striata, was activated by epoxy groups, and epoxidized Bletilla striata polysaccharide (EBSP) was prepared. Then, EBSP was crosslinked with collagen under alkaline conditions, and a new hemostatic material that was an epoxidized Bletilla polysaccharide crosslinked collagen hemostatic sponge was prepared. We demonstrated that endowing collagen with better hemostatic performance, cytocompatibility, and blood compatibility does not destroy its original three-stranded helical structure. Compared with the medical gauze, hemostasis time was shorter (26.75 ±â€¯2.38 s), and blood loss was lower (0.088 ±â€¯0.051 g) in the rat liver injury hemostasis model. In the rat model of severed tail hemostasis, hemostasis time was also shorter (47.33 ±â€¯2.05 s), and the amount of blood loss was lower (0.330 ±â€¯0.122 g). The sponge possessed good hemostatic and healing performance.

Bletilla Striata polysaccharides thermosensitive gel for photothermal treatment of bacterial infection.

Skin is the most important defense shield which touched external environment directly. Effectively clearing microbes in infected wound via non-antibiotic therapy is crucial for the promotion of recovery in complex biological environments, and the wound healing is a crucial process after sterilization to avoid superinfection. Herein, a kind of Prussian blue-based photothermal responsive gel, Bletilla striata polysaccharide-mingled, isatin-functionalized Prussian blue gel (PB-ISA/BSP gel) was reported for effective treatment of bacterial infection and wound healing. The introduction of effective components of traditional Chinese medicine (TCM), isatin (ISA), enhanced the efficiency of sterilization synergistically. Furthermore, the process of wound healing was promoted by Bletilla striata polysaccharides (BSP). PB-ISA@BSP had a considerable antibacterial rate with 98.5 % under an 808 nm laser for 10 min in vitro. Besides, PB-ISA/BSP gel showed an effective antibacterial efficacy in vivo and a fast wound healing rate as well. The as-prepared functional particles can invade and destroy bacteria membrane to kill microbes. This work highlights that PB-ISA/BSP gel is a promising antibacterial agent based on synergistically enhanced photothermal effect and wound healing promotion ability and provides inspiration for future therapy based on the synergy between photothermal agent and active components in TCM.

Phytochemical Analysis of Anacamptis coriophora Plant Cultivated Using Ex Vitro Symbiotic Propagation.

This study aims to investigate the functional groups and phytochemical profile of Anacamptis coriophora seeds, tubers, and flowers. Symbiotic seedlings produced using the ex vitro method were transferred to their natural habitat and grown to analyze the functional groups and phytochemical profiles of tubers and flowers. The life cycles of the transferred seedlings were monitored, and tubers and flowers were harvested for analysis. ATR-FTIR spectroscopy revealed the presence of functional groups such as polysaccharides, lignin, and proteins in both tubers and flowers. Differences in spectral frequencies between first-year and second-year tubers were observed. Fatty acid analysis identified 30 different compounds in seeds, flowers, and tubers, with linoleic acid being the most abundant (27 % in seeds, 33 % in tubers), and palmitic acid present in flowers (24 %). GC-MS analysis of ethanol extracts from these components highlighted the presence of 32 compounds, including hydroxyacetic acid, hydrazide, cytidine (Z)-7-hexadecenal, 2,2-dimethoxyethane, 2,5,6-trimethyldecane, and butanamide, 4-amino-N-hydroxy. A. coriophora's tubers, flowers, and seeds may contain active metabolites with therapeutic potential. These results are valuable for the commercial cultivation of the plant.

Neuroprotective neolignan glycosides from the pseudobulbs of Bletilla striata.

Four undescribed neolignan glycosides, bletineosides A-D (1-4) were isolated from the pseudobulbs of Bletilla striata. Their structures with absolute configurations were elucidated on the basis of spectroscopic analyses, along with acidic hydrolysis reactions and ECD experiments. All isolates were evaluated for their neuroprotective activities against glutamate-induced PC12 cell injury. Compound 3 and 4 showed significantly neuroprotective effects at the concentration of 10 µM when compared with the model group. Compounds 1-4 represented the first examples of neolignan glycosides from the genus Bletilla. This study disclosed the potency of Bletilla striata as a new source of anti-neurodegenerative agents.

Extraction, structural-activity relationships, bioactivities, and application prospects of Bletilla striata polysaccharides as ingredients for functional products: A review.

Bletilla striata is a well-known medicinal plant with high pharmaceutical and ornamental values. Polysaccharide is the most important bioactive ingredient in B. striata and has various health benefits. Recently, B. striata polysaccharides (BSPs) have attracted much attention from industries and researchers due to their remarkable immunomodulatory, antioxidant, anti-cancer, hemostatic, anti-inflammatory, anti-microbial, gastroprotective, and liver protective effects. Despite the successful isolation and characterization of BSPs, there is still limited knowledge regarding their structure-activity relationships (SARs), safety concerns, and applications, which hinders their full utilization and development. Herein, we provided an overview of the extraction, purification, and structural features, as well as the effects of different influencing factors on the components and structures of BSPs. We also highlighted and summarized the diversity of chemistry and structure, specificity of biological activity, and SARs of BSP. The challenges and opportunities of BSPs in the food, pharmaceutical, and cosmeceutical fields are discussed, and the potential development and future study direction are scrutinized. This article provides comprehensive knowledge and underpinnings for further research and application of BSPs as therapeutic agents and multifunctional biomaterials.

Phenotype-fitness relationships and pollen-transfer efficiency of five orchid species with different pollination strategies.

PREMISE: Deceptive pollination, a fascinating mechanism that independently originated in several plant families for benefiting from pollinators without providing any reward, is particularly widespread among orchids. Pollination efficiency is crucial in orchids due to the aggregated pollen in a pollinarium, which facilitates pollen transfer and promotes cross-pollination as pollinators leave after being deceived. METHODS: In this study, we compiled data on reproductive ecology from five orchid species with different pollination strategies: three deceptive-strategy species (shelter imitation, food deception, sexual deception), one nectar-rewarding species, and one shelter-imitation but spontaneously selfing species. We aimed to compare the reproductive success (female fitness: fruit set; male fitness: pollinarium removal) and pollination efficiency of species representing these strategies. We also investigated pollen limitation and inbreeding depression among the pollination strategies. RESULTS: Male and female fitness were strongly correlated in all species but the spontaneously selfing species, which had high fruit set and low pollinarium removal. As expected, pollination efficiency was highest for the rewarding species and the sexually deceptive species. Rewarding species had no pollen limitation but did have high cumulative inbreeding depression; deceptive species had high pollen limitation and moderate inbreeding depression; and spontaneously selfing species did not have pollen limitation or inbreeding depression. CONCLUSIONS: Pollinator response to deception is critical to maintain reproductive success and avoid inbreeding in orchid species with non-rewarding pollination strategies. Our findings contribute to a better understanding of the trade-offs associated with different pollination strategies in orchids and highlight the importance of pollination efficiency in orchids due to the pollinarium.

Comparative analyses and phylogenetic relationships of thirteen Pholidota species (Orchidaceae) inferred from complete chloroplast genomes.

BACKGROUND: The orchid genus Pholidota Lindl. ex Hook. is economically important as some species has long been used in traditional medicine. However, the systematic status of the genus and intergeneric relationships inferred from previous molecular studies are unclear due to insufficient sampling and lack of informative sites. So far, only limited genomic information has been available. The taxonomy of Pholidota remains unresolved and somewhat controversial. In this study, the complete chloroplast (cp.) genomes of thirteen Pholidota species were sequenced and analyzed to gain insight into the phylogeny of Pholidota and mutation patterns in their cp. genomes. RESULTS: All examined thirteen Pholidota cp. genomes exhibited typical quadripartite circular structures, with the size ranging from 158,786 to 159,781 bp. The annotation contained a total of 135 genes in each cp. genome, i.e., 89 protein-coding genes, 38 tRNA genes, and eight rRNA genes. The codon usage analysis indicated the preference of A/U-ending codons. Repeat sequence analysis identified 444 tandem repeats, 322 palindromic repeats and 189 dispersed repeats. A total of 525 SSRs, 13,834 SNPs and 8,630 InDels were detected. Six mutational hotspots were identified as potential molecular markers. These molecular markers and highly variable regions are expected to facilitate future genetic and genomic studies. Our phylogenetic analyses confirmed the polyphyletic status of the genus Pholidota, with species grouped into four main clades: Pholidota s.s. was resolved as the sister to a clade containing species of Coelogyne; the other two clades clustered together with species of Bulleyia and Panisea, respectively; species P. ventricosa was placed at the basal position, deviated from all other species. CONCLUSION: This is the first study to comprehensively examine the genetic variations and systematically analyze the phylogeny and evolution of Pholidota based on plastid genomic data. These findings contribute to a better understanding of plastid genome evolution of Pholidota and provide new insights into the phylogeny of Pholidota and its closely related genera within the subtribe Coelogyninae. Our research has laid the foundation for future studies on the evolutionary mechanisms and classification of this economically and medicinally important genus.

Non-species-specific pollen transfer and double-reward production in euglossine-pollinated Vanilla.

Commonly attributed to orchids, the pollen movement in Vanilla has been associated with food deception and specific plant-pollinator relationships. This study investigated the role of flower rewards and pollinator specificity in the pollen transfer of a widely distributed member to the euglossinophilous Vanilla clade, V. pompona Schiede using data collected from Brazilian populations. These included investigations on morphology, light microscopy and histochemistry, and analysis of flowers scent using GC-MS. The pollinators and the mechanisms of pollination were recorded through focal observations. The yellow flowers of V. pompona are fragrant and offer nectar as reward. The major volatile compound of the V. pompona scent, carvone oxide, shows convergent evolution in Eulaema-pollinated Angiosperms. The pollination system of V. pompona is not species-specific, but its flowers are strongly adapted to pollination by large Eulaema males. Pollination mechanism is based in a combination of perfume collection and nectar seeking. The dogma of a species-specific pollination system based on food deception in Vanilla has been broken with the increase in studies on this Pantropical orchid genus. Here, least three bee species and dual reward-offering are involved in pollen transfer in V. pompona. Visitation frequency of bees collecting perfumes, used in courtship by male euglossines, is higher than in searching for food, as short-lived young euglossine males seem to be more interested in sex than food. A pollination system based on offering both nectar and perfumes as resources is described for the first time in orchids.

Effects of Different Drying Methods on the Quality of Bletilla striata Scented Tea.

Flower tea is widely loved as a drink, especially for the beautiful and rich flowers of the orchid family, and the drying method for different flowers is also unique. GC-MS is widely used to study volatile substances to determine the quality of flower teas. The findings show that the freeze-drying method can retain the original aroma and flavor of Bletilla striata has the highest sensory evaluation score, with the key flavor substances ethyl caproate and N-heptanal containing 1.14% and 6.28%, respectively, and their ROAV values reaching 54.46 and 100.00. Additionally, the freeze-drying method can well retain flavonoids, polysaccharides, and phenolic components, while providing better antioxidant and antibacterial properties. The stove-drying method would make Bletilla striata slightly burnt and less flavorful and efficacious than freeze-drying; the air-drying method is difficult to retain the special odor and fragrance of Bletilla striata flowers and has the lowest sensory evaluation score, with the presence of volatile components with irritating and unpleasant odors such as pyrazine and 2-pentylfuran, while not showing better efficacy. In addition, steam fixation would destroy the morphology and flavor of Bletilla striata, lose polysaccharide and phenolic components, and reduce the efficacy of Bletilla striata scented tea, but could retain the flavonoid components well. In summary, direct freeze-drying without steam fixation is the best process for drying Bletilla striata scented tea, stove-drying without steam fixation is more economical and convenient in actual production and application, steam fixation and air-drying are not suitable as drying processes for Bletilla striata scented tea. This study analyzed the quality of Bletilla striata scented tea under different drying methods, promoted the further processing of Bletilla striata scented tea, and provided a reference for the comprehensive utilization of Bletilla striata scented tea.

Isolation, structural and bioactivities of polysaccharides from Anoectochilus roxburghii (Wall.) Lindl.: A review.

Anoectochilus roxburghii (Wall.) Lindl. (A. roxburghii), a valuable herbal medicine in China, has great medicinal and edible value. Polysaccharides, as one of the main active components of A. roxburghii, comprise glucose, arabinose, xylose, galactose, rhamnose, and mannose in different molar ratios and glycosidic bond types. By varying the sources and extraction methods of A. roxburghii polysaccharides (ARPS), different structural characteristics and pharmacological activities can be elucidated. ARPS has been reported to exhibit antidiabetic, hepatoprotective, anti-inflammatory, antioxidant, antitumor, and immune regulation activities. This review summarizes the available literature on the extraction and purification methods, structural features, biological activities, and applications of ARPS. The shortcomings of the current research and potential focus in future studies are also highlighted. This review provides systematic and current information on ARPS to promote their further exploitation and application.

Sugars and sucrose transporters in pollinia of Phalaenopsis aphrodite (Orchidaceae).

The pollen grains of Phalaenopsis orchids are clumped tightly together, packed in pollen dispersal units called pollinia. In this study, the morphology, cytology, biochemistry, and sucrose transporters in pollinia of Phalaenopsis orchids were investigated. Histochemical detection was used to characterize the distribution of sugars and callose at the different development stages of pollinia. Ultra-performance liquid chromatography-high resolution-tandem mass spectrometry data indicated that P. aphrodite accumulated abundant saccharides such as sucrose, galactinol, myo-inositol, and glucose, and trace amounts of raffinose and trehalose in mature pollinia. We found that galactinol synthase (PAXXG304680) and trehalose-6-phosphate phosphatase (PAXXG016120) genes were preferentially expressed in mature pollinia. The P. aphrodite genome was identified as having 11 sucrose transporters (SUTs). Our qRT-PCR confirmed that two SUTs (PAXXG030250 and PAXXG195390) were preferentially expressed in the pollinia. Pollinia germinated in pollen germination media (PGM) supplemented with 10% sucrose showed increased callose production and enhanced pollinia germination, but there was no callose or germination in PGM without sucrose. We show that P. aphrodite accumulates high levels of sugars in mature pollinia, providing nutrients and enhanced SUT gene expression for pollinia germination and tube growth.

Gymnadenia conopsea (L.) R. Br.: Comprehensive review of propagation and breeding, traditional uses, chemical composition, pharmacology, quality control, and processing.

ETHNOPHARMACOLOGICAL RELEVANCE: Gymnadenia conopsea, a perennial herbaceous flowering plant that belongs to the family of Orchidaceae, sporadic distributed in the altitudes of 200-4700 m across northern Europe and, temperate and subtropical Asia region. The dried tubers of G. conopsea have been used to treat cough, asthma, and their syndromes, and also as a tonic in China and surrounding countries for a long history. G. conopsea is often processed deeply processed before use to enhance its efficacy. In recent years, because of its remarkable pharmacological activity and health care function, G. conopsea has been used more and more widely. Due to its extensive application and bad growth environment, the wild distribution of G. conopsea is decreasing and it has been listed as an endangered plant. AIM OF THE REVIEW: This review aims to summarize the propagation and breeding, traditional uses, chemical composition, pharmacology, quality control, and processing of G. conopsea. Moreover, it also provides suggestions for the future high-value utilization of G. conopsea. MATERIALS AND METHODS: A literature search on Gymnadenia genus and G. conopsea was performed using scientific databases including SciFinder, ACS, Web of Science, Springer, ScienceDirect, PubMed, and CNKI. Information was also collected from classic books of Chinese herbal medicine, official websites, Ph.D. and M.Sc. Dissertations, and so on. Structures of chemical compounds were drawn by ChemDraw software. RESULTS: As of submission date of this manuscript, total 170 natural compounds have been isolated and characterized from G. conopsea, and all of the compounds were isolated from the tubers. The isolated compounds including benzylester glucosides, dihydrostilbenes, phenanthrenes, phenolic compounds, alkaloids, polysaccharide, lignans, flavones, triterpenoids, steroids, and other compounds. Some of these compounds and active extracts exhibited a wide range of pharmacological activities, in which, the tonifying, anti-fatigue, anti-oxidant, anti-viral, sedative and hypnotic activities are consistent with the traditional uses for the treatment of diseases. In addition, a variety of new pharmacological activities, such as preventing and treating gastric ulcers, immunoregulatory, anti-hyperlipidemia, anti-anaphylaxis, anti-silicosis, anti-cancer and neuroprotective activities have also been reported. However, the bioactive compounds responsible for most of the above pharmacological effects have not been well summarised till now. In this manuscript, analysis, speculation and summary of compounds that responsible for pharmacological effects were conducted. CONCLUSIONS: The chemical constituents and pharmacological activities studies of G. conopsea extract have been summarised in this context, the isolated compounds responsible for the pharmacological activities were also analyzed and deduced according to the publications, all above led to suggestions for the future high-value utilization of G. conopsea.