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The root of Paeonia lactiflora pall. is a significant component of traditional Chinese medicine, with terpenoids and their glycosides, such as paeoniflorins, serving as key active ingredients known for their anti-inflammatory, hepatoprotective, and analgesic properties. By generating a transcriptome and functionally characterizing 32 terpene synthases (TPSs) from P. lactiflora, we successfully constructed 24 pESC-Trp-PlTPS expression vectors. Through expression in Saccharomyces cerevisiae engineered strains, we identified four mono-TPSs and five sesqui-TPSs that produce 18 compounds, including eight monoterpenes and ten sesquiterpenes in vitro. This includes a bifunctional enzyme (PlTPS22). Additionally, PlTPS21 was characterized as a pinene synthase with α-pinene as its main product. The expression pattern of PlTPS21 aligns closely with the accumulation patterns of paeoniflorins and α-pinene in the plant, suggesting that PlTPS21 is a key enzyme in the biosynthesis of paeoniflorin. [ABSTRACT FROM AUTHOR]

Powdery mildew poses a substantial threat to Chinese herbaceous peony (Paeonia lactiflora) cultivation, yet efficient identification methods for the pathogen are currently lacking, hampering early disease control. This study successfully identified Erysiphe paeoniae as the causative agent of powdery mildew in P. lactiflora in Beijing, leveraging morphological characteristics and rDNA ITS region sequencing. Furthermore, a high specificity and sensitivity specific PCR detection primers targeting E. paeoniae were developed based on ITS region sequences. Notably, these primers effectively circumvent contamination by host plant or other fungal DNA, enabling the detection of E. paeoniae DNA at concentrations as low as 1 × 10−15 ng/μL. This technological advancement provides a valuable tool for the early and rapid detection of E. paeoniae. Moreover, host range of E. paeoniae strain in our study was tested by inoculation combined with specific primers we developed. The isolated strain BJ20210618 can infect four species of Paeonia plants, including P. lactiflora, P. anomala, P. obovata, and P. veitchii. Among them, P. veitchii is a new host of E. paeoniae, reported for the first time in this study. Strikingly, this strain exhibited no infectivity towards Chinese tree peony (P. suffruticosa), diverging from prior reports suggesting E. paeoniae infects Chinese tree peony. We speculate that there may be different physiological races in E. paeoniae. We aligned the E. paeoniae strain BJ20210618 isolated from P. lactiflora in Beijing, China, E. paeoniae strain YN20220506 isolated from P. suffruticosa in Yunnan, China, along with the ITS sequences of 15 E. paeoniae strains and 13 closely related powdery mildews available on NCBI, and constructed a phylogenetic tree. Phylogenetic analysis of ITS sequences further supports these observations, distinguishing E. paeoniae from both P. lactiflora and P. suffruticosa into discrete clusters, highlighting the presence of distinct strains of E. paeoniae with different host specificities. This study not only enhances our understanding of the pathogen's behavior and specificity but also provides invaluable tools and insights for effective disease management strategies. [ABSTRACT FROM AUTHOR]

Background: This study investigates the hypolipidemic effects of a mixed extract of Salvia miltiorrhiza and Paeonia lactiflora (USCP119) in HFD-fed hamsters and in vitro cellular models. Methods: Over an 8-week period, HFD-fed hamsters were assigned to one of six groups: normal diet, HFD control, HFD with 50 mg/kg USCP119, HFD with 100 mg/kg USCP119, HFD with 50 mg/kg USCP119 twice daily (BID), and HFD with omega-3 fatty acids. Key outcomes assessed included body weight, serum triglycerides (TG), total cholesterol (TC), liver weight, hepatic TG levels, and epididymal fat. In cellular models, the impact of USCP119 on lipid accumulation and adipogenic markers was evaluated. Results: USCP119 treatment at 50 mg/kg BID resulted in the lowest weight gain (15.5%) and the most significant reductions in serum TG and hepatic TG levels compared to the HFD control. The 100 mg/kg dose also led to substantial reductions in serum TG and TC levels and notable decreases in low-density lipoprotein cholesterol. USCP119 at 50 mg/kg once daily reduced TG and TC levels but was less effective than the higher doses. In cellular models, USCP119 was non-toxic up to 400 µg/mL and effectively reduced lipid accumulation, modulated adipogenic markers, and enhanced AMPK signaling, improving lipid metabolism and insulin sensitivity. Conclusions: All USCP119 treatments demonstrated effectiveness in managing hyperlipidemia and related metabolic disorders, with variations in impact depending on the dosage. The ability of USCP119 to reduce fat accumulation, improve lipid profiles, and enhance insulin sensitivity highlights its potential as a valuable dietary supplement for addressing high-fat diet-induced hyperlipidemia and metabolic disturbances. [ABSTRACT FROM AUTHOR]

Background and research purpose: Paeoniflorin and albiflorin are monoterpene glycosides that exhibit various medicinal properties in Paeonia species. This study explored the terpene biosynthesis pathway and analyzed the distribution of these compounds in different tissues of two Korean landraces of Paeonia lactiflora to gain insights into the biosynthesis of monoterpene glycosides in P. lactiflora and their potential applications. Materials and methods: Two Korean landraces, Hongcheon var. and Hwacheon var, of P. lactiflora were used for the analyses. Contents of the paeoniflorin and albiflorin were analyzed using HPLC. RNA was extracted, sequenced, and subjected to transcriptome analysis. Differential gene expression, KEGG, and GO analyses were performed. Paeoniflorin biosynthesis genes were isolated from the transcriptomes using the genes in Euphorbia maculata with the NBLAST program. Phylogenetic analysis of of 1-Deoxy-D-xylulose 5-phosphate synthase (DOXPS), geranyl pyrophosphate synthase (GPPS), and pinene synthase (PS) was carried out with ClustalW and MEGA v5.0. Results and discussion: Analysis of paeoniflorin and albiflorin content in different tissues of the two P. lactiflora landraces revealed significant variation. Transcriptome analysis yielded 36,602 unigenes, most of which were involved in metabolic processes. The DEG analysis revealed tissue-specific expression patterns with correlations between landraces. The isolation of biosynthetic genes identified 173 candidates. Phylogenetic analysis of the key enzymes in these pathways provides insights into their evolutionary relationships. The sequencing and analysis of DOXPS, GPPS, PS revealed distinct clades and subclades, highlighting their evolutionary divergence and functional conservation. Our findings highlight the roots as the primary sites of paeoniflorin and albiflorin accumulation in P. lactiflora, underscoring the importance of tissue-specific gene expression in their biosynthesis. Conclusion: this study advances our understanding of monoterpene glycoside production and distribution in Paeonia, thereby guiding further plant biochemistry investigations. [ABSTRACT FROM AUTHOR]

Herbaceous peony (Paeonia lactiflora Pall.) is a widely used famous traditional flower in China. It prefers cold and cool climate, but is not resistant to high temperature during summer in the middle and lower reaches of the Yangtze River. Previously, we found peroxidase (POD) is an important antioxidant enzyme that played an important role in high-temperature tolerance of P. lactiflora. The present study isolated the candidate gene PlPOD45 and verified its function in resisting high-temperature stress. And the results showed that PlPOD45 had an open reading frame of 978 bp that encoded 325 amino acids. Its protein was localized to the cell membrane and cytoplasm. High-temperature stress induced PlPOD45 expression. Heterologous overexpression of PlPOD45 improved plant tolerance to high-temperature stress, decreased reactive oxygen species (ROS) accumulation, relative electrical conductivity and malondialdehyde content, and increased the ratio of variable fluorescence to highest fluorescence and POD activity. Conversely, silencing PlPOD45 in P. lactiflora could decrease POD activity, ROS scavenging capability and cell membrane stability when these plants were exposed to high-temperature stress. These results suggest that PlPOD45 positively regulates high-temperature tolerance through ROS scavenging, which would provide a theoretical basis for improving high-temperature tolerance in P. lactiflora. [ABSTRACT FROM AUTHOR]

Stem strength plays a crucial role in the growth and development of plants, as well as in their flowering and fruiting. It not only impacts the lodging resistance of crops, but also influences the ornamental value of ornamental plants. Stem development is closely linked to stem strength; however, the roles of the SPL transcription factors in the stem development of herbaceous peony (Paeonia lactiflora Pall.) are not yet fully elucidated. In this study, we obtained and cloned the full-length sequence of PlSPL14, encoding 1085 amino acids. Quantitative real-time PCR (qRT-PCR) analysis revealed that the expression level of PlSPL14 gradually increased with the stem development of P. lactiflora and was significantly expressed in vascular bundles. Subsequently, utilizing the techniques of virus-induced gene silencing (VIGS) and heterologous overexpression in tobacco (Nicotiana tabacum L.), it was determined that PlSPL14-silenced P. lactiflora had a thinner xylem thickness, a decreased stem diameter, and weakened stem strength, while PlSPL14-overexpressing tobacco resulted in a thicker xylem thickness, an increased stem diameter, and enhanced stem strength. Further screening of the interacting proteins of PlSPL14 using a yeast two-hybrid (Y2H) assay revealed an interactive relationship between PlSPL14 and PlSLR1 protein, which acts as a negative regulator of gibberellin (GA). Additionally, the expression level of PlSLR1 gradually decreased during the stem development of P. lactiflora. The above results suggest that PlSPL14 may play a positive regulatory role in stem development and act in the xylem, making it a potential candidate gene for enhancing stem straightness in plants. [ABSTRACT FROM AUTHOR]

HT074, a multiherbal mixture containing extracts from Inula britannica flowers and Paeonia lactiflora roots, is used in Korean medicine for gastric disorders. This study investigated the protective mechanisms of HT074 against acute reflux esophagitis (RE) in rats. Nitric oxide (NO) production and mRNA expression of antioxidant-related genes (Nrf2, HO-1, SOD, CAT, and GPx2) were evaluated in LPS-induced RAW 264.7 cells. Gastroesophageal reflux (GER) was induced in rats, followed by HT074 (100, 300 mg/kg) or ranitidine (50 mg/kg) administration. Esophageal damage and histological changes were assessed. Gastric pH and protein expression levels of Nrf2, HO-1, SOD, CAT, and GPx-1/2 were measured. HT074 pretreatment reduced NO production and increased the expression of HO-1, CAT, and GPx2 in LPS-induced RAW 264.7 cells. In GER-induced rats, HT074 significantly decreased esophageal lesions and increased the expression of HO-1, SOD, GPx-1/2, and Nrf2. HT074 did not affect gastric pH. These findings suggest that HT074 protects against GER-induced esophagitis by inhibiting NO production and enhancing antioxidant activity. Therefore, HT074 could be a promising therapeutic agent for GER disease. [ABSTRACT FROM AUTHOR]

Rapid wilting after harvest greatly decreases the ornamental and economic value of fresh-cut flowers. To determine how chlorine dioxide (ClO2) affects postharvest flower quality, Paeonia lactiflora 'Hushui Dangxia' cut flowers in bottles were treated with different concentrations of ClO2 (0, 25, 50, 75, and 100 mg L−1). Among the treatments, ClO2 75 (i.e., 75 mg L−1) decreased the bacterial growth and lignin content at the P. lactiflora flower stem base, while also decreasing the degree of flower stem vessel blockage. Additionally, the ClO2 75 treatment increased the relative fresh weight, water balance, soluble sugar content, soluble protein content, and antioxidant enzyme activities (superoxide dismutase, peroxidase, and catalase), but decreased the malondialdehyde content and ethylene release rate of P. lactiflora cut flowers. Thus, the aging of P. lactiflora flowers was delayed, thereby maintaining the cut flower quality. Furthermore, the vase life (i.e., ornamental period) increased by three days. [ABSTRACT FROM AUTHOR]

Objective: To explore the mechanism of paeoniflorin (PF) on osteoarthritis (OA) synovial inflammation from network pharmacology to experimental pharmacology. Methods: Targets of OA were constructed by detecting the database of network database platforms (Therapeutic Target database, DrugBank and GeneCards), and the targets of PF were constructed by PubChem and Herbal Ingredients' Targets database. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of these co-targeted genes were conducted via Database for Annotation, Visualization, and Integrated Discovery (DAVID) database, and protein-protein interaction (PPI) networks were conducted via the search tool for the retrieval of interacting genes (STRING) database. Cell counting kit-8 (CCK-8) assay was performed to assess the potential toxicity of PF on human OA fibroblast-like synoviocytes (FLS), quantitative real-time polymerase chain reaction (qPCR), enzyme-linked immunosorbent assay (ELISA) and Western blot were used to verify the potential mechanism of PF in synovial inflammation. Results: Twenty-six co-targeted genes were identified. GO enrichment results showed that these co-targeted genes were most likely localized in the cytoplasm, and the biological processes mainly involved 'cellular response to hypoxia' 'lipopolysaccharide (LPS)-mediated signaling pathway' and 'positive regulation of gene expression'. KEGG pathway analysis indicated that these co-targeted genes may function through pathways associated with 'hypoxia-inducible factor-1 (HIF-1) signaling pathway' and 'tumor-necrosis factor (TNF) signaling pathway'. The PPI network showed that the top 3 hub genes were TP53, TNF, and CASP3. Molecular docking results showed that PF was well docking with TNF. CCK-8 showed no potential toxicity of 10, 20 and 50 µmol/L PF on human OA FLS. And PF significantly decreased the expression levels of interleukin-1 β, interleukin-6, TNF-α matrix metalloproteinase 13 (MMP13), and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5) and TNF-α in LPS-induced OA FLS. Conclusion: PF exhibited potent anti-inflammatory effect in OA synovial inflammation. [ABSTRACT FROM AUTHOR]

Gene function verification is a crucial step in studying the molecular mechanisms regulating various plant life activities. However, a stable and efficient homologous genetic transgenic system for herbaceous peonies has not been established. In this study, using virus-induced gene silencing technology (VIGS), a highly efficient homologous transient verification system with distinctive advantages was proposed, which not only achieves true "intact-plant" infiltration but also minimizes the operation. One-year-old roots of the representative species, Paeonia lactiflora Pall., were used as the materials; prechilling (4 °C) treatment for 3–5 weeks was applied as a critical precondition for P. lactiflora to acquire a certain chilling accumulation. A dormancy-related gene named HOMEOBOX PROTEIN 31 (PlHB31), believed to negatively regulate bud endodormancy release (BER), was chosen as the target gene in this study. GFP fluorescence was detected in directly infiltrated and newly developed roots and buds; the transgenic plantlets exhibited remarkably earlier budbreak, and PlHB31 was significantly downregulated in silenced plantlets. This study established a homologous transient silencing system featuring intact-plant infiltration and minimized manipulation for gene function research, and also offers technical support and serves as a theoretical basis for gene function discovery in numerous other geophytes. [ABSTRACT FROM AUTHOR]

Vascular wall aging has been strongly associated with cardiovascular diseases. Thus, this study aimed to investigate the efficacy of USCP-GVH-014, a mixed extract of Salvia miltiorrhiza Bunge and Paeonia lactiflora Pall. in inhibiting vascular wall aging through in vitro and in vivo experiments. The results revealed that USCP-GVH-014 inhibited abnormal cell proliferation, collagen overproduction, and MMP-2 and MMP-9 overexpression caused by various stimuli and recovered the antioxidant enzyme superoxide dismutase on human aortic smooth muscle cells. In addition, it inhibited the increase in ICAM-1 and VCAM-1 expression induced by tumor necrosis factor alpha on human aortic endothelial cells and prevented the aging of the vascular wall by regulating related proteins such as epidermal growth factor and interleukin-1ß. Furthermore, it reduced vascular aging in in vivo studies. These results demonstrate that USCP-GVH-014 effectively reduces vascular aging, thereby rendering it a potential therapeutic candidate for cardiovascular diseases. [ABSTRACT FROM AUTHOR]

Global warming causes frequent high temperatures in summer; which negatively impacts herbaceous peonies (Paeonia lactiflora Pall.) by accelerating leaf senescence and reducing biomass accumulation, leading to reduced flower quality in the subsequent year. Our findings revealed that as heat stress progressed, the high-temperature-sensitive cultivar 'Meigui Zi' (MGZ) exhibited a higher rate of chlorophyll content reduction and more pronounced premature aging symptoms than the high-temperature-tolerant cultivar 'Chi Fen' (CF). To investigate gene expression differences between CF and MGZ under high-temperature stress, we combined PacBio Iso-Seq sequencing (Iso-Seq) with next-generation sequencing (RNA-seq). Iso-seq yielded 352,891 full-length transcripts ranging from 61 bp to 49,022 bp in length. RNA-seq generated 257,562 transcripts across all samples. Further analysis revealed that differentially expressed genes (DEGs) between CF and MGZ were primarily enriched in "Photosynthesis", with most photosynthesis-related DEGs highly expressed in CF. This indicates that CF has higher stability in its photosystem compared with MGZ, which is crucial for mitigating leaf senescence caused by high temperatures. Additionally, the highly expressed chlorophyll degradation genes stay-green (SGR) and stay-green-like (SGRL) in MGZ may be involved in chlorophyll content reduction induced by high temperature. This study preliminarily revealed the molecular mechanism of high-temperature-induced leaf senescence of in herbaceous peony and provided candidate genes for further studies of the regulation mechanism of high -temperature-induced leaf senescence. [ABSTRACT FROM AUTHOR]

Our representative studies to achieve sustainable use of crude drugs and ensure their stable quality are introduced: comprehensive studies on genetic, chemical, and sometimes pharmacological diversity of Asian medicinal plants including Paeonia lactiflora, Glycyrrhiza uralensis, Ephedra spp., Saposhnikovia divaricata, and Curcuma spp., as well as their related crude drugs. (1) For peony root, after genetic and chemical diversity analysis of crude drug samples including white and red peony root in China, the value-added resources with quality similar to red peony root were explored among 61 horticultural P. lactiflora varieties, and two varieties were identified. In addition, an optimized post-harvest processing method, which resulted in high contents of the main active components in the produced root, was developed to promote cultivation and production of brand peony root. (2) Alternative resources of glycyrrhiza, ephedra herb and saposhnikovia root and rhizome of Japanese Pharmacopoeia grade were discovered in eastern Mongolia after field investigation and quality assessment comparing Mongolian plants with Chinese crude drugs. Simultaneously, suitable specimens and prospective regions for cultivation were proposed. (3) Because of the wide distribution and morphological similarities of Curcuma species, classification of some species is debated, which leads to confusion in the use of Curcuma crude drugs. Molecular analyses of the intron length polymorphism (ILP) markers in genes encoding diketide-CoA synthase (DCS) and curcumin synthase (CURS) and trnK sequences, combined with essential oils analysis, were demonstrated as useful for standardization of Curcuma crude drugs. The above studies, representing various facets, can be applied to other crude drugs. [ABSTRACT FROM AUTHOR]

Albiflorin, a principal monoterpene glycoside isolated from Paeonia lactiflora Pall., is known for its extensive pharmacological properties and medicinal value in a number of traditional Chinese medicine recipes. Herein, we consolidate current research on the biological profile, anti-inflammatory capabilities, pharmacokinetics, tissue distribution, and toxicity of albiflorin with the, aim of delineating its therapeutic potential and safety profile. Albiflorin exhibits remarkable anti-inflammatory effects in various disease models, including respiratory, urinary, gastrointestinal, and rheumatic conditions, as well as metabolic and nutritional diseases, blood system disorders, mental illnesses, and pain. Pharmacokinetic studies have revealed its efficient absorption, distribution, and elimination, indicating favorable bioavailability with minimal toxicity, as supported by traditional toxicity assessments and next-generation metabolomics analyses. Despite its proven efficacy in a broad spectrum of health conditions, comprehensive toxicological studies are scarce, highlighting the need for further research to ascertain its safety in clinical applications. This review underscores the significant therapeutic promise of albiflorin, rooted in its efficacy as the key active ingredient in relevant traditional Chinese medicine formulas and reinforced by modern scientific inquiry. Albiflorin stands at the intersection of historical wisdom and contemporary pharmacological research, warranting further exploration to harness its full medical potential, especially for the treatment of inflammation-related diseases.

The root of Paeonia lactiflora pall. is a significant component of traditional Chinese medicine, with terpenoids and their glycosides, such as paeoniflorins, serving as key active ingredients known for their anti-inflammatory, hepatoprotective, and analgesic properties. By generating a transcriptome and functionally characterizing 32 terpene synthases (TPSs) from P. lactiflora, we successfully constructed 24 pESC-Trp-PlTPS expression vectors. Through expression in Saccharomyces cerevisiae engineered strains, we identified four mono-TPSs and five sesqui-TPSs that produce 18 compounds, including eight monoterpenes and ten sesquiterpenes in vitro. This includes a bifunctional enzyme (PlTPS22). Additionally, PlTPS21 was characterized as a pinene synthase with α-pinene as its main product. The expression pattern of PlTPS21 aligns closely with the accumulation patterns of paeoniflorins and α-pinene in the plant, suggesting that PlTPS21 is a key enzyme in the biosynthesis of paeoniflorin.

Seven monoterpenoids were isolated and elucidated from the roots of Paeonia lactiflora , including two new derivatives (1 and 2), along with five known compounds (3 - 7). The structures of the new compounds (1 and 2) were determined by HR-ESI-MS, UV, and NMR spectroscopic data analyses. Moreover, all isolates were tested for their neuroprotective activities against H 2 O 2 -induced injury in PC12 cells. As a result, compound 5 displayed moderate neuroprotective activity and increased PC12 cell viability from 50 ± 2.52 % to 73 ± 1.54 % at 100 μ M. [Display omitted] • Two new derivatives were obtained from the roots of Paeonia lactiflora Pall. • The two new derivatives along with the five known ones are all monoterpenoids. • Compound 5 (galloylpaeoniflorin) showed better neuroprotective activity with cell viability which was 73 ± 1.54 % at 100 μM. [ABSTRACT FROM AUTHOR]

[Objective] Analyzing the changes of microbial community and enzyme activities in the rhizospheric soil of Paeonia lactiflora with root rot and understanding the relationship between soil environmental factors, microbial community and root rot of P. lactiflora would provide the scientific basis for controlling root rot of P. lactiflora. [Method] Illumina high throughput sequencing technology and soil physicochemical property method were employed to investigate soil bacterial and fungal communities, soil nutrients and enzyme activities of rhizosphere soil of healthy and root rot diseased plants of P. lactiflora. [Result] The content of soil organic matter in the rhizosphere of diseased plants significantly increased, the content of available potassium significantly decreased, and the content of soil alkaline phosphatase and peroxidase significantly increased, compared with healthy plants of P. lactiflora. The richness and diversity of bacteria decreased, but the richness and diversity of fungi increased in the rhizosphere soil of root rot diseased plants of P. lactiflora. The relative abundance of Proteobacteria, Bacteroidota and Sumerlaeota in the rhizosphere soil of root rot diseased plants of P. lactiflora was significantly higher than that of healthy plants, but Chloroflexi, Actinobaciota, Ethylomirabilota and Firmicutes decreased significantly. Sphingomonas, Chtoniobacter, Subgroup7 and Saccharimondales in the rhizosphere soil of root rot diseased plants of P. lactiflora was higher than that of healthy plants. The relative abundance of Ascomycota increased and Basidiomycota decreased in the rhizosphere soil of root rot diseased plants of P. lactiflora, and Zygomycota was significantly higher than that of healthy plants. The pH, organic matter, available phosphorus, available potassium, water-soluble salt, alkaline phosphatase, peroxidase, urease and sucrase were important factors that affected the changes of microbial community in the rhizosphere soil of P. lactiflora, which were mainly related to the changes of dominant bacterial phyla, Chloroflexi, Metromirabilota, Actinobacteriota, and dominant fungal phyla, Chytridiomycota, Basidiomycota, Ascomycota. [Conclusion] The relative abundance of harmful fungi such as Ilyonia, Erysiphe and Fusarium increases significantly. The accumulation of pathogenic bacteria, the changes of enzyme activities, and the nutrient imbalance are the important reasons for the occurrence of root rot of P. lactiflora. [ABSTRACT FROM AUTHOR]

In about one-quarter of childhood epilepsies, there is no complete response to the routine, available antiepileptic drugs (AEDs). In traditional Persian medicine (TPM), Paeonia lactiflora and Anacyclus pyrethrum have been known for their anticonvulsant effects. The aim of this study was to evaluate the therapeutic efficacy of a combined herbal formula consisting of the dried root of P. lactiflora and A. pyrethrum in childhood drug-resistant epilepsy (DRE). This single-arm before/after clinical trial was conducted on 46 children with DRE. The patients were observed for three months before as well as thirteen weeks after the administration of this traditional herbal preparation at a dose of 15 mg/kg/day. The frequency and duration of seizures were evaluated using the Hague seizure severity scale (HASS). There was a statistically significant reduction in both the frequency and duration of seizures after treatment with the herbal preparation (p-values were <0.001 and <0.05, respectively). A statistically significant response to the herbal formula started after eight weeks of treatment. The traditional formula of P. lactiflora and A. pyrethrum could be considered as a complementary medicine for treating childhood DRE. However, further studies are needed to provide more evidence of its benefits and possible harms and also determine the precise mechanisms of action of this formula as a medicinal remedy for epilepsy. [ABSTRACT FROM AUTHOR]

Paeonia lactiflora (P. lactiflora) has been recently reported to increase the production of proinflammatory mediators and activates phagocytosis in macrophages. Thus, in this study, we tried to verify the macrophage activation of Paeoniae Radix Rubra (PRR, also known as red peony root) and elucidate its mechanism of action. PRR upregulated the production of proinflammatory mediators and activated phagocytosis in RAW264.7 cells. However, these effects were reversed by inhibition of TLR2/4. In addition, the inhibition of p38, JNK, and ERK1/2 reduced the PRRmediated production of proinflammatory mediators, and the SPLmediated activation of p38, JNK, and ERK1/2 was blocked by the TLR4 inhibition. These findings indicate that PRR may activate macrophages through TLR4-dependent activation of p38, JNK, and ERK1/2. These indicate that PRR has immunostimulatory activity. Thus, it is believed that PRR can be used as a functional food agent that enhances the immune system. [ABSTRACT FROM AUTHOR]

Owing to its high ornamental, medicinal and horticultural values, herbaceous peony (Paeonia lactiflora Pall.) has been widely used as a landscaping and economical plant around the world. However, the lack of an efficient and stable regeneration system in P. lactiflora restricts its rapid propagation and large-scale production. By testing the key factors affecting callus formation, proliferation, adventitious bud induction and rooting, here, we developed an in vitro system for callus induction and regeneration in P. lactiflora. Our results show that callus formation was affected by explant types, culture environment, basal medium and plant growth regulators. Using cotyledons as explants, we established good conditions for P. lactiflora callus induction and callus proliferation. We effectively obtained adventitious buds differentiated from callus in Murashige and Skoog (MS) medium containing kinetin (KT) and thidiazuron (TDZ). Adventitious bud growth can be further promoted by adding gibberellin 3 (GA3), 1-naphthaleneacetic acid (NAA) and 6-benzyleaminopurine (6-BA) into the MS medium. A high percentage of rooting can be achieved by adding indolebutyric acid (IBA) and activated carbon (AC) to ½ MS medium. Overall, our system promotes callus induction and adventitious bud regeneration for P. lactiflora through improved culture conditions and plant growth regulators in the culture media, and lays a foundation for subsequent genetic engineering research. [ABSTRACT FROM AUTHOR]

The petals of Paeonia lactiflora 'Coral Sunset' change color from coral pink to pale yellow after flower opening. Pigment-targeted metabolomic analysis showed that the carotenoid and anthocyanin contents rapidly decreased after petal fading. SMART-sequencing and next-generation-sequencing analyses were performed to identify differentially expressed transcripts to characterize the candidate genes involved in petal fading. The expression of certain genes associated with anthocyanin and carotenoid synthesis and degradation was correlated with the petal-fading phenotype. The anthocyanin synthesis (AS) structural genes, CHS, F3H, F3′H, DFR, and ANS, and the carotenoid synthesis genes, LCYB and LCYE, were strongly expressed before fading, but their expression significantly declined after fading. In contrast, the expression of certain genes associated with oxidase activity and light signaling significantly increased after fading. Therefore, inhibition of pigment synthesis and accelerated pigment degradation may be crucial for petal fading. A R2R3-MYB family member of subgroup 4 (MYBs-SG4) showed the same expression pattern as the AS structural genes and functioned in the positive regulation of anthocyanin synthesis by forming the MBW protein complex. This is the first report of a SG4 member with a positive regulatory function. This study provides a foundation for elucidation of the mechanisms of pigment synthesis and metabolism, and a theoretical basis for flower-color-directed breeding. [ABSTRACT FROM AUTHOR]

HT074, a multiherbal mixture containing extracts from Inula britannica flowers and Paeonia lactiflora roots, is used in Korean medicine for gastric disorders. This study investigated the protective mechanisms of HT074 against acute reflux esophagitis (RE) in rats. Nitric oxide (NO) production and mRNA expression of antioxidant-related genes (Nrf2, HO-1, SOD, CAT, and GPx2) were evaluated in LPS-induced RAW 264.7 cells. Gastroesophageal reflux (GER) was induced in rats, followed by HT074 (100, 300 mg/kg) or ranitidine (50 mg/kg) administration. Esophageal damage and histological changes were assessed. Gastric pH and protein expression levels of Nrf2, HO-1, SOD, CAT, and GPx-1/2 were measured. HT074 pretreatment reduced NO production and increased the expression of HO-1, CAT, and GPx2 in LPS-induced RAW 264.7 cells. In GER-induced rats, HT074 significantly decreased esophageal lesions and increased the expression of HO-1, SOD, GPx-1/2, and Nrf2. HT074 did not affect gastric pH. These findings suggest that HT074 protects against GER-induced esophagitis by inhibiting NO production and enhancing antioxidant activity. Therefore, HT074 could be a promising therapeutic agent for GER disease.

Rapid wilting after harvest greatly decreases the ornamental and economic value of fresh-cut flowers. To determine how chlorine dioxide (ClO2) affects postharvest flower quality, Paeonia lactiflora ‘Hushui Dangxia’ cut flowers in bottles were treated with different concentrations of ClO2 (0, 25, 50, 75, and 100 mg L−1). Among the treatments, ClO2 75 (i.e., 75 mg L−1) decreased the bacterial growth and lignin content at the P. lactiflora flower stem base, while also decreasing the degree of flower stem vessel blockage. Additionally, the ClO2 75 treatment increased the relative fresh weight, water balance, soluble sugar content, soluble protein content, and antioxidant enzyme activities (superoxide dismutase, peroxidase, and catalase), but decreased the malondialdehyde content and ethylene release rate of P. lactiflora cut flowers. Thus, the aging of P. lactiflora flowers was delayed, thereby maintaining the cut flower quality. Furthermore, the vase life (i.e., ornamental period) increased by three days.

Simple Summary: Rhizosphere microbial composition was affected by plant cultivars, soil properties, climate and other conditions. However, the effects of cultivars on the structure and the function of rhizosphere microbial communities have not been reported completely. In this study, the rhizosphere bacterial community composition of eight cultivars of Paeonia lactiflora was revealed by high-throughput sequencing technology. The differences and similarities of rhizosphere bacterial community composition and rhizosphere bacterial interaction among different cultivars were analyzed. In addition, cultivation-related microbial resources that can be used for bioremediation, organic degradation and disease resistance have been identified. This research will contribute to the development of rhizosphere microbial resources of P. lactiflora and provide guidance for agricultural practice in the future. The composition and diversity of the rhizosphere microbial community maintain the stability of the root microclimate, and several studies have focused on this aspect of rhizosphere microorganisms. However, how these communities vary with cultivars of a species is not completely understood. Paeonia lactiflora—a perennial herb species of the family Paeoniaceae—includes a wide variety of cultivars, with rich rhizosphere microbial resources. Hence, we studied the differences in rhizosphere bacterial communities associated with eight P. lactiflora cultivars. We noted that Actinobacteria, Proteobacteria, Acidobacteria, Bacteroidetes, Firmicutes, Verrucomicrobia, Planctomycetes and Chloroflexi were the dominant phyla associated with the cultivars. The composition of rhizosphere bacterial community of different cultivars was highly similar at taxonomic levels, but there were slightly differences in the relative abundance. LEfSe analysis showed that the cultivars "Sheng Tao Hua" and "Zi Lou Xian Jin" exhibited the most biomarkers. Differential ASV analysis revealed the maximum difference in ASV abundance between "Lian Tai" and "Zi Hong Zheng Hui", as well as between "Sheng Tao Hua" and "Tao Hua Fei Xue", and the maximum similarity between "Duo Ye Zi" and "Xue Feng". Co-occurrence network analysis revealed that rhizosphere bacteria in most cultivars maintain homeostasis by cooperation, wherein Actinobacteria and Proteobacteria played a vital role. In addition, microbial resources related to cultivars like bioremediation, organic degradation and resistance to diseases are found. This study revealed the structures of the rhizosphere bacterial communities associated with different cultivars of P. lactiflora and explored their stress resistance potential, which can be used to guide future agricultural practices. [ABSTRACT FROM AUTHOR]

Paeonia lactiflora Pall., a Paeoniaceae family member, is a herbaceous perennial flower of significant horticultural importance. It holds prominence in traditional Chinese medicine due to its therapeutic applications in pain relief, inflammation reduction, and immune disorder treatment. Paeoniflorin, the active monoterpene compound in P. lactiflora, exhibits diverse biological activities, such as anti-inflammatory and immune-regulatory effects. Although the pharmacological effects of paeoniflorin have been extensively studied, its biosynthetic pathway remains elusive. Here, we combined PacBio Iso-Seq and RNA-seq techniques to investigate three distinct tissues from two different P. lactiflora ecotypes with varying paeoniflorin contents. The primary objective was to identify candidate genes involved in the biosynthesis of paeoniflorin and related monoterpenes. Differentially expressed unigenes encoding cytochrome P450s, UDP-glucoronosyl and UDP-glucosyl transferases, and transcription factors, which might be associated with monoterpene diversity in P. lactiflora, were identified. In addition, we analyzed genetic differentiation between the two P. lactiflora ecotypes. Notably, gene expression in the terpene pathway within root and rhizome tissues was downregulated compared to leaf tissues. This intriguing observation led us to speculate that paeoniflorin synthesis primarily occurs in the leaves and is subsequently transported to the roots. However, further experimentation is required to provide substantial evidence and verify this hypothesis. [ABSTRACT FROM AUTHOR]

Peony root is an important herbal drug used as an antispasmodic analgesic. To evaluate peony roots with different botanical origins, producing areas, and post-harvest processing, 1H NMR-based metabolomics analysis was employed. Five types of monoterpenoids, including albiflorin (4), paeoniflorin (6), and sulfonated paeoniflorin (25), and six other compounds, including 1,2,3,4,6-penta-O-galloyl-β-d-glucose (18), benzoic acid (21), gallic acid (22), and sucrose (26) were detected in the extracts of peony root samples. Among them, compounds 4, 6, 18, and total monoterpenoids including 21 were quantified by quantitative 1H NMR (qHNMR). Compound 25 was detected in 1H NMR spectra of sulfur-fumigated white peony root (WPR) extracts indicating that 1H NMR was a fast and effective method for identifying sulfur-fumigated WPR. The content of 26, the main factor affecting extract yield, increased significantly in peony root after low-temperature storage for one month, whereas that in WPR did not increase due to the boiling treatment after harvesting. We investigated the impact of preprocessing methods to such analysis for NMR data from commercial samples, resulting that the data matrix transformed from qHNMR spectra and normalized to internal standard were optimum for multivariate analysis. The multivariate analysis demonstrated that among commercial samples derived from P. lactiflora, peony root samples in Japanese market (PR) had high contents of 18 and 22, and red peony root (RPR) samples had high content of monoterpenoids represented by 6; and among RPR samples, those derived from P. veitchii showed higher contents of 18 and 22 than those from P. lactiflora. The 1H NMR-based metabolomics method coupled with qHNMR was useful for evaluation of peony root and would be applicable for other crude drugs. [ABSTRACT FROM AUTHOR]

Global warming causes frequent high temperatures in summer; which negatively impacts herbaceous peonies (Paeonia lactiflora Pall.) by accelerating leaf senescence and reducing biomass accumulation, leading to reduced flower quality in the subsequent year. Our findings revealed that as heat stress progressed, the high-temperature-sensitive cultivar ‘Meigui Zi’ (MGZ) exhibited a higher rate of chlorophyll content reduction and more pronounced premature aging symptoms than the high-temperature-tolerant cultivar ‘Chi Fen’ (CF). To investigate gene expression differences between CF and MGZ under high-temperature stress, we combined PacBio Iso-Seq sequencing (Iso-Seq) with next-generation sequencing (RNA-seq). Iso-seq yielded 352,891 full-length transcripts ranging from 61 bp to 49,022 bp in length. RNA-seq generated 257,562 transcripts across all samples. Further analysis revealed that differentially expressed genes (DEGs) between CF and MGZ were primarily enriched in “Photosynthesis”, with most photosynthesis-related DEGs highly expressed in CF. This indicates that CF has higher stability in its photosystem compared with MGZ, which is crucial for mitigating leaf senescence caused by high temperatures. Additionally, the highly expressed chlorophyll degradation genes stay-green (SGR) and stay-green-like (SGRL) in MGZ may be involved in chlorophyll content reduction induced by high temperature. This study preliminarily revealed the molecular mechanism of high-temperature-induced leaf senescence of in herbaceous peony and provided candidate genes for further studies of the regulation mechanism of high -temperature-induced leaf senescence.

Osteoporosis is one of the most common bone diseases in the elderly. Osteoporosis related bone fracture could seriously lower the quality of the elderly’s life. Thus, in this study, natural products which can treat or prevent osteoporosis without severe side effects have been searched. The ethanol extract of Paeonia lactiflora Pall (Paeoniceae) significantly increased the differentiation of MC3T3-E1 preosteoblastic cells determined by alkaline phosphatase assay. The bioassay-guided isolation of ethyl acetate fraction allowed to isolated 6 paeoniflorin derivatives and 2 catechins such as 4-O-methyl-moudanpioside C (1), 4-O-methylbenzoyloxypaeoniflorin (2), oxypaeoniflorin (3), (+)-catechin (4), catechuic acid (5), paeoniflorin (6), benzoylpaeoniflorin (7), albiflorin (8). Among them, 4-O-methylmoudanpioside C (1), 4-O-methylbenzoyloxypaeoniflorin (2), oxypaeoniflorin (3), benzoylpaeoniflorin (7) and albiflorin (8) significantly increased the ALP activity. Particularly, 4-O-methylmoudanpioside C (1), 4-O-methylbenzoyloxypaeoniflorin (2) were the most active. Furthermore, 4-O-methylbenzoyloxypaeoniflorin (2) significantly inhibited the osteoclastic differentiation of Raw264.7 cells. Taken together, the extract of Paeonia lactiflora and its active constituents, particularly, 4-O-methylbenzoyloxypaeoniflorin (2) could have a benefical effect on bone formation and promote bone health. [ABSTRACT FROM AUTHOR]

Paeonia lactiflora Pall. is not only a traditional ornamental plant, but also an important medicinal plant. Currently, some P. lactiflora cultivars are used for ornamental purposes, but their potential medicinal value is ignored. To explore the medicinal potential of the ornamental varieties, the medicinal cultivar 'Hangbaishao' (HS) and the ornamental cultivar 'Zifengyu' (ZFY) were selected, and microbiome and metabolome analyses were performed to compare the composition of the endophytes and metabolites in the roots. The diversity and abundance of bacteria were not significantly different between HS and ZFY; however, the diversity and abundance of endophytic fungi in the ornamental cultivar ZFY were much higher than those in the medicinal cultivar HS. The flavonoids and phenolic acid contents of the ornamental cultivar ZFY were significantly higher than those of the medicinal cultivar HS, indicating that ZFY has medicinal value. The differences in root endophytes between HS and ZFY may lead to differences in phenolic acids and flavonoids. To explore the relationship between endophytes and the accumulation of phenolic acids and flavonoids, a joint analyses of the microbiome and metabolome were performed. The key bacterium, Ruminococcaceae bacterium GD7, led to the accumulation of phenolic acids and flavonoids in the ZFY. This study contributes to future research on the potential medicinal value of ornamental P. lactiflora and provides a new approach for realizing the 'dual use of medicine and appreciation' of P. lactiflora. [ABSTRACT FROM AUTHOR]

Herbaceous peony (Paeonia lactiflora Pall.) is an ancient ornamental crop and, in recent decades, an emerging popular cut flower. Straight stems are a vital criterion for cut herbaceous peony selection, while many cultivars bend as the plant develops. Pectin helps maintain the mechanical strength of the cell wall. However, little is known about its role in the stem bending of herbaceous peony. Two herbaceous peony cultivars with contrasting stem morphologies ('Dong Fang Shao Nv', upright; 'Lan Tian Piao Xiang', bending gradually) at five developmental stages were used as materials to investigate the effects of pectin content and nanostructure on straightness using the carbazole colorimetric method and atomic force microscopy observations. The contents of water-soluble pectin (WSP), CDTA-soluble pectin (CSP), and sodium carbonate-soluble pectin (SSP) differed significantly between the two cultivars, and the contents and angle of the flower and branch showed correlations. For the pectin nanostructure, WSP showed agglomerates and long chains, with a higher proportion of broad agglomerates at the later stages of the bending cultivar than the upright cultivar. CSP showed branched chains, and the proportion of broad chains was higher in the upright cultivar at later stages, while CSP shape changed from agglomerates to chains in the bending cultivar. SSP mainly consisted of short linear main chains, and side chains in the upright stem were stacked, and the bent cultivar had more broad and short chains. It can be concluded that the contents, nanometric shape, and size of the three kinds of pectin are highly likely to affect herbaceous peony stem straightness. This study provides a theoretical basis for the role of pectin in the production and breeding of herbaceous peony cut flowers. [ABSTRACT FROM AUTHOR]

In this study, two herbaceous peony cultivars with different heat tolerances ('Fenyunu' FYN low sensitivity and 'Qiaoling' QL high sensitivity) were used as research materials. An integrated view of the factors underlying the decrease in photosynthetic rate under high-temperature (HT) stress was provided by analyzing the biochemical parameters, chloroplast ultrastructure, gas-exchange parameters, chlorophyll fluorescence, and modulated 820 nm reflection of herbaceous peony leaves. The results showed that hydrogen peroxide, superoxide anion, malondialdehyde, and electrical conductivity increased significantly, while the photosynthetic pigments content and photosynthetic capacity decreased significantly in QL than in FYN under HT. The contents of soluble sugars and proline increased greatly in FYN than in QL, while the activity of SOD decreased markedly in QL than in FYN after HT. Compared with FYN, the ultrastructure of QL was more seriously disrupted under HT. Chlorophyll fluorescence analysis showed that HT changed the shapes of OJIP curve, resulting in the increase of K phase and J phase. The PSII acceptor side was more damaged than the donor side, and the electron transfer was seriously blocked. The energy flow in the process of light energy absorption, capture, and electron transfer were significantly changed after HT stress. Meanwhile, PSI was also significantly inhibited, and the coordination of both photosystems decreased. The variation of these parameters in FYN was less than that in QL. These results suggested that FYN featured a more heat-tolerance ability as evidenced by the good performances on the antioxidant system, osmoregulatory capacity, and the thermostability of membranes and photosystems. [ABSTRACT FROM AUTHOR]

The herbaceous peony (Paeonia lactiflora Pall.) has high ornamental and nutritional value, and the seeds of P. lactiflora can be used to produce high-quality edible oil. However, low seed yield limits the application of P. lactiflora. This can be mitigated by insect pollinators. Here, we evaluated the pollination services of honeybees (Apis mellifera L.) in a P. lactiflora field. We found that A. mellifera provided pollination for P. lactiflora throughout the day especially in the period from 10:00–15:00. Seed number and weight were significantly increased when P. lactiflora was pollinated by A. mellifera. Furthermore, a DNA barcode, the rbcL gene, was used to analyze pollen samples from the corbiculae (pollen baskets) of A. mellifera (bee pollen, BP) and P. lactiflora flowers (flower pollen, FP). High homology of rbcL genes in the BP and FP suggested that BP was collected from P. lactiflora. Based on our results, A. mellifera provided efficient pollination for P. lactiflora. Therefore, A. mellifera could be a good candidate pollinator for P. lactiflora and could be applied in the field. [ABSTRACT FROM AUTHOR]

The concentration of heavy metals (HMs) in soil-plant systems was an important indicator of soil heavy metal pollution, and the migration and transportation abilities of HMs from soil to plants were of great significance for evaluating food safety and reducing health risks. This study was conducted to investigate the contamination and translocation of HMs in the soil-Paeonia lactiflora system in Bozhou, Anhui Province, China. The soil pollution was assessed by calculating the Geoaccumulation index (Igeo), Potential ecological risk index (RI), and Pollution load index (PLI). The transfer ability of HMs was evaluated by using the bioconcentration factor (BCF) and translocation factor (TF). The results showed that Fe, Mn, Cr, and As were the priority pollutants in soil. Paeonia lactiflora presented high potential to translocate Zn, Cu, and Fe from soil to roots, and had strong ability to translocate HMs except for Cd from roots to stems and leaves [ABSTRACT FROM AUTHOR]

The herbaceous peony (Paeonia lactiflora Pall.) is widely cultivated as an ornamental, medicinal and edible plant in China. Drought stress can seriously affect the growth of herbaceous peony and reduce its quality. In our previous research, a significantly differentially expressed gene, PM19L, was obtained in herbaceous peony under drought stress based on transcriptome analysis, but little is known about its function. In this study, the first PM19L that was isolated in herbaceous peony was comprised of 910 bp, and was designated as PlPM19L (OP480984). It had a complete open reading frame of 537 bp and encoded a 178-amino acid protein with a molecular weight of 18.95 kDa, which was located in the membrane. When PlPM19L was transferred into tobacco, the transgenic plants had enhanced tolerance to drought stress, potentially due to the increase in the abscisic acid (ABA) content and the reduction in the level of hydrogen peroxide (H2O2). In addition, the enhanced ability to scavenge H2O2 under drought stress led to improvements in the enzyme activity and the potential photosynthetic capacity. These results combined suggest that PlPM19L is a key factor to conferring drought stress tolerance in herbaceous peony and provide a scientific theoretical basis for the following improvement in the drought resistance of herbaceous peony and other plants through genetic engineering technology. [ABSTRACT FROM AUTHOR]

Paeonia lactiflora is an important ornamental and medicinal plant, but after the flowering period, the above-ground parts of ornamental cultivars are often discarded as waste. When considering the development of medicinal plant resources, attention should be paid to the comprehensive development and utilisation of the plant. In this study, 13 ornamental herbaceous peony cultivars were selected, and their stems and leaves were analysed for antioxidant and antibacterial activities. The results showed that total phenolic content (TPC), total flavonoid content (TFC), antioxidant activity and antibacterial activity varied depending on the cultivar and plant organ. In the same cultivar, the TPC and TFC of leaves were higher than those of the stems. The antioxidant activity was significantly correlated with TPC of stems and leaves. In the antibacterial assay, the extracts of stem and leaves of the cultivars 'Hong Yan Fei Shuang' and 'Chun Xiao' were the most effective against most of the tested strains. This is an innovative study that shows the great potential of ornamental herbaceous peony stems and leaves as sources of biologically active compounds. [ABSTRACT FROM AUTHOR]

• Design and synthesis of low viscosity, highly soluble and hydrophilic new ternary deep eutectic solvent. • First extraction of polyphenolic active compounds from Paeonia lactiflora. • New ternary deep eutectic solvent with higher extraction efficiency than conventional solvent. • Deep eutectic solvent extracts have higher antioxidant activity and reducing power than traditional solvent extracts. Study has demonstrated that Paeonia lactiflora contains a significant number of polyphenolic compounds; however, there is little research literature focus on phenolic acids in Paeonia lactiflora. In this study, an ultrasound-assisted extraction method based on ternary DESs was developed for the extraction of biologically active polyphenols in Paeonia lactiflora. The results indicated that the ternary solvent composed of choline chloride, DL-lactic acid, and 1,2-propanediol in a molar ratio of 1:4:2 achieved a superior extraction of polyphenols than that of conventional solvents and other ternary solvents. Subsequently, the response surface method was used to optimize the selected parameters of solid–liquid ratio, extraction time, and temperature for the extraction of phenolic by the DES, and the optimal extraction conditions were as follows: a solid–liquid ratio of 1:13.8 g/mL, an extraction time of 47.5 min, and an extraction temperature of 34.5 °C, with a total polyphenol yield of 79.77 mg/g, which is 1.09 times that of the traditional solvent. Furthermore, the extract of Paeonia lactiflora obtained through the DES method exhibited promising in vitro antioxidant activity and reducing ability. Studies have shown that DES is an efficient and environmentally friendly extraction solvent for extracting polyphenols from natural products, providing a new direction for the development and industrial production of Paeonia lactiflora. [ABSTRACT FROM AUTHOR]

Variation in floral organ number in plants is triggered by extremely complex transcriptional and post-transcriptional regulatory processes. MicroRNAs (miRNAs) play a pivotal role in the post-transcriptional regulation of floral organs. Paeonia lactiflora Pall., originating from China, is a precious herbaceous plant with ornamental, medicinal, and oil properties. Building on previous transcriptional studies, this work examined post-transcriptional regulation of peony stamen petalization using critical-stage flower buds. Using high-throughput small RNA sequencing, a miRNA expression profile was constructed. This was used to identify 10 known, 334 conserved, and 255 novel miRNAs, 155, 151, and 44 of which were differentially expressed at three developmental stages (stamen primordium, stamen primordium partly petaloid, and stamen primordium completely petaloid). Degradome analysis identified 55 miRNAs and 127 potential target genes. A miRNA-target network suggested that miR172- PlRAP2 , miR160- PlARF , miR396- PlGRF , miR319- PlTCP4 , and miR384\394- PlFBOX modules are key regulators of peony stamen petalization. The regulation of miR319 and PlTCP4 during peony flower development was validated through transient expression in tobacco. This study provides insight into miRNA-mediated regulation of petalized stamen formation in herbaceous peony and refines the post-transcriptional network at critical stages of this process. • 10 known, 334 conserved, and 255 novel miRNAs were identified during the stamen petalization process in P. lactiflora. • Expression pattern analysis of miRNA and target genes was performed. • Degradome analysis identified 55 miRNAs and 127 potential target genes. • PlTCP4 were validated as targets of miR319-y through transient expression in tobacco. • A novel regulatory model of peony stamen petalization regulated by miRNA was proposed. [ABSTRACT FROM AUTHOR]

The petals of Paeonia lactiflora ‘Coral Sunset’ change color from coral pink to pale yellow after flower opening. Pigment-targeted metabolomic analysis showed that the carotenoid and anthocyanin contents rapidly decreased after petal fading. SMART-sequencing and next-generation-sequencing analyses were performed to identify differentially expressed transcripts to characterize the candidate genes involved in petal fading. The expression of certain genes associated with anthocyanin and carotenoid synthesis and degradation was correlated with the petal-fading phenotype. The anthocyanin synthesis (AS) structural genes, CHS, F3H, F3′H, DFR, and ANS, and the carotenoid synthesis genes, LCYB and LCYE, were strongly expressed before fading, but their expression significantly declined after fading. In contrast, the expression of certain genes associated with oxidase activity and light signaling significantly increased after fading. Therefore, inhibition of pigment synthesis and accelerated pigment degradation may be crucial for petal fading. A R2R3-MYB family member of subgroup 4 (MYBs-SG4) showed the same expression pattern as the AS structural genes and functioned in the positive regulation of anthocyanin synthesis by forming the MBW protein complex. This is the first report of a SG4 member with a positive regulatory function. This study provides a foundation for elucidation of the mechanisms of pigment synthesis and metabolism, and a theoretical basis for flower-color-directed breeding.

Owing to its high ornamental, medicinal and horticultural values, herbaceous peony (Paeonia lactiflora Pall.) has been widely used as a landscaping and economical plant around the world. However, the lack of an efficient and stable regeneration system in P. lactiflora restricts its rapid propagation and large-scale production. By testing the key factors affecting callus formation, proliferation, adventitious bud induction and rooting, here, we developed an in vitro system for callus induction and regeneration in P. lactiflora. Our results show that callus formation was affected by explant types, culture environment, basal medium and plant growth regulators. Using cotyledons as explants, we established good conditions for P. lactiflora callus induction and callus proliferation. We effectively obtained adventitious buds differentiated from callus in Murashige and Skoog (MS) medium containing kinetin (KT) and thidiazuron (TDZ). Adventitious bud growth can be further promoted by adding gibberellin 3 (GA3), 1-naphthaleneacetic acid (NAA) and 6-benzyleaminopurine (6-BA) into the MS medium. A high percentage of rooting can be achieved by adding indolebutyric acid (IBA) and activated carbon (AC) to ½ MS medium. Overall, our system promotes callus induction and adventitious bud regeneration for P. lactiflora through improved culture conditions and plant growth regulators in the culture media, and lays a foundation for subsequent genetic engineering research.

The composition and diversity of the rhizosphere microbial community maintain the stability of the root microclimate, and several studies have focused on this aspect of rhizosphere microorganisms. However, how these communities vary with cultivars of a species is not completely understood. Paeonia lactiflora—a perennial herb species of the family Paeoniaceae—includes a wide variety of cultivars, with rich rhizosphere microbial resources. Hence, we studied the differences in rhizosphere bacterial communities associated with eight P. lactiflora cultivars. We noted that Actinobacteria, Proteobacteria, Acidobacteria, Bacteroidetes, Firmicutes, Verrucomicrobia, Planctomycetes and Chloroflexi were the dominant phyla associated with the cultivars. The composition of rhizosphere bacterial community of different cultivars was highly similar at taxonomic levels, but there were slightly differences in the relative abundance. LEfSe analysis showed that the cultivars “Sheng Tao Hua” and “Zi Lou Xian Jin” exhibited the most biomarkers. Differential ASV analysis revealed the maximum difference in ASV abundance between “Lian Tai” and “Zi Hong Zheng Hui”, as well as between “Sheng Tao Hua” and “Tao Hua Fei Xue”, and the maximum similarity between “Duo Ye Zi” and “Xue Feng”. Co-occurrence network analysis revealed that rhizosphere bacteria in most cultivars maintain homeostasis by cooperation, wherein Actinobacteria and Proteobacteria played a vital role. In addition, microbial resources related to cultivars like bioremediation, organic degradation and resistance to diseases are found. This study revealed the structures of the rhizosphere bacterial communities associated with different cultivars of P. lactiflora and explored their stress resistance potential, which can be used to guide future agricultural practices.

ABSTRACT Paeonia lactiflora is a commercial crop with horticultural and medicinal value. Although interactions between plants and microbes are increasingly evident and considered to be drivers of ecosystem service, the regulatory relationship between microbial communities and the growth and root metabolites of P. lactiflora is less well known. Here, soil metabolomics indicated that carbohydrates and organic acids were enriched in the rhizosphere (RS) with higher diversity. Moreover, the variation of root-associated microbiotas between the bulk soil (BS) and the RS of P. lactiflora was investigated via 16S rRNA and internally transcribed spacer (ITS) amplicon sequencing. The RS displayed a low-diversity community dominated by copiotrophs, whereas the BS showed an oligotroph-dominated, high-diversity community. Hierarchical partitioning showed that cation exchange capacity (CEC) was the main factor affecting microbial community diversity. The null model and the dispersion niche continuum index (DNCI) suggested that stochastic processes (dispersal limitation) dominated the community assembly of both the RS and BS. The bacterial-fungal interkingdom networks illustrated that the RS possessed more complex and stable co-occurrence patterns. Meanwhile, positive link numbers and positive cohesion results revealed more cooperative relationships among microbes in the RS. Additionally, random forest model prediction and two partial least-squares path model (PLS-PM) analyses showed that the P. lactiflora root secondary metabolites were comprehensively impacted by soil water content (SWC), mean annual precipitation (MAP), pH (abiotic), and Alternaria (biotic). Collectively, this study provides a theoretical basis for screening the microbiome associated with the active components of P. lactiflora. IMPORTANCE Determining the taxonomic and functional components of the rhizosphere microbiome, as well as how they differ from those of the bulk soil microbiome, is critical for manipulating them to improve plant growth performance and increase agricultural yields. Soil metabolic profiles can help enhance the understanding of rhizosphere exudates. Here, we explored the regulatory relationship across environmental variables (root-associated microbial communities and soil metabolism) in the accumulation of secondary metabolites of P. lactiflora. Overall, this work improves our knowledge of how the rhizosphere affects soil and microbial communities. These observations improve the understanding of plant-microbiome interactions and introduce new horizons for synthetic community investigations as well as the creation of microbiome technologies for agricultural sustainability.

Isorhamnetin-3-O-glucoside and astragalin, flavonol glucosides, were isolated from the petals of Paeonia lactiflora as neurite outgrowth-promoting compounds. Isoquercitrin, formed by demethylating the B ring of isorhamnetin-3-O-glucoside or by adding a hydroxyl group to the B ring of astragalin, was evaluated for neurite outgrowth-promoting activity and was compared with the activities of isorhamnetin-3-O-glucoside and astragalin. The activities of isorhamnetin, kaempferol, and quercetin, aglycones corresponding to isorhamnetin-3-O-glucoside, astragalin, and isoquercitrin, respectively, were also evaluated. Isorhamnetin-3-O-glucoside and astragalin showed much stronger neurite outgrowth-promoting activities than the activities of the other tested flavonoids. They exhibited relatively weak anti-oxidant activities and moderate AChE inhibitory activities compared to the activities of the other tested flavonoids. Isorhamnetin-3-O-glucoside and astragalin promoted morphological neurite outgrowth and the expression of neurofilaments induced by NGF in PC12 cells. Isorhamnetin-3-O-glucoside and astragalin might be candidate compounds as neural differentiation agents in peripheral nerves and functional food ingredients preventing cognitive decline. [ABSTRACT FROM AUTHOR]

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ATP-citrate lyase (ACL) gene catalyzes the formation of acetyl-CoA to provide intermediate precursors for many secondary metabolites, and also plays an important role in anthocyanin biosynthesis of plants. Herbaceous peony (Paeonia lactiflora Pall.) is an international cut flower known for its rich flower colors, however, the function of the ACL gene in flower color regulation is still unclear. Here, double-colored P. lactiflora 'Hebao Jinlian' were used to study the molecular mechanism of red petal, and acetyl-CoA and anthocyanin biosynthesis related PlACLB2, PlCHS, PlDFR, PlANS, and PlbHLH1 genes were initially found to highly expressed in the red outer-petals. The expression pattern of PlACLB2 was consistent with the spatial accumulation of anthocyanins. The correlation analysis of PlACLB2 expression pattern, acetyl-CoA content, and anthocyanin accumulation revealed that PlACLB2 was positively correlated with the acetyl-CoA and anthocyanin contents with correlation coefficients of 0.82 and 0.80. Moreover, multiple sequence alignment identified two typical conserved domains in PlACLB2, and phylogenetic analysis clustered PlACLB2 into the ACLB clade. PlACLB2 was localized in the nucleus and cytoplasm. On the one hand, silencing PlACLB2 in P. lactiflora red outer-petal resulted in lighter petal color and decreased acetyl-CoA accumulation, and quantitative analysis detected that PlACLB2-silenced petals lost more anthocyanins than the control groups with a decrease of 31.0%, and the main pigment component cyanidin-3,5-O-diglucoside was reduced by 31.9%. On the other hand, overexpression of PlACLB2 significantly promoted red coloration, acetyl-CoA content, and anthocyanin accumulation in tobacco flowers. These results demonstrated that PlACLB2 promoted anthocyanin accumulation by increasing the abundance of its precursor substrate acetyl-CoA, thereby regulating the formation of the red petals in P. lactiflora. [ABSTRACT FROM AUTHOR]

Herbaceous peony (Paeonia lactiflora Pall.) is an important ornamental plant worldwide. In its natural state, P. lactiflora often manifests traits like rapidly elongating internodal growth, loose plant types, and soft inflorescence stems. However, very little has been known about the measures for controlling these traits. This study investigated the effect of applying paclobutrazol (PBZ) on the plant growth and flower quality in P. lactiflora. The results indicated that PBZ application reduced the plant height (8.05%), plant crown width (14.72%), and leaf area (10.90%), but increased the leaf thickness (18.18%) and stem diameter (over 11%) in P. lactiflora. Meanwhile, PBZ application was also found to increase the chlorophyll (Chl) a (29.63%), Chl b (33.33%), Chl a+b (30.56%), SPAD (27.32%), relative water content (0.47%), soluble sugar (5.09%) and activities of three antioxidant enzymes (superoxide dismutase 169.66%, peroxidase 3.59%, catalase 319.30%), but decreased the relative electrical conductivity (18.52%). Additionally, the application of PBZ was found to affect the flowering quality of P. lactiflora, increasing the flower diameter and fresh weight only in the flower-bud stage. This initiates the bloom stage, where there was a decrease in the total content of the aromatic compounds except for the flower-bud stage, and faded the flower color by reducing the content of anthocyanin. These results demonstrated that the application of PBZ can regulate the P. lactiflora plant types with no significant decrease in its ornamental values. This might provide a theoretical basis for further applying PBZ in P. lactiflora for use in urban landscape spaces. [ABSTRACT FROM AUTHOR]

Background and objectives: Bacterial antibiotics have had several side effects. Therefore, interest in natural substances with less side effects is increasing these days. Paeonia lactiflora, the root of Paeonia lactiflora, is used as a raw material for medicines. In this study, we investigated the antibacterial effect and the cytotoxicity of Paeonia lactiflora extract. Materials and Methods: For cytotoxicity, MTT analysis according to ISO 10993-5 was performed. The antibacterial test of the Paeonia lactiflora was determined from bacterial viability, Inhibition zone test, CFU (colony forming unit) and SEM (scanning electron microscope). To confirm the antibacterial component of Paeonia lactiflora, the content of flavonoids and polyphenols was analyzed. Results: Our results showed that Paeonia lactiflora extract contained flavonoids and polyphenols, which exhibited antimicrobial activity against Streptococcus mutans (S. mutans) and Candida albicans (C. ablicans). Further, the cytotoxicity of Paeonia lactiflora extract was low. Conclusions: We believe that our study makes a significant contribution to the literature because it demonstrates that Paeonia lactiflora extract can be used as an antibiotic. [ABSTRACT FROM AUTHOR]