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Cancer represents one of the leading causes of mortality worldwide. Conventional clinical treatments include radiation therapy, chemotherapy, immunotherapy, and targeted therapy. However, these treatments have inherent limitations, such as multidrug resistance and the induction of short- and long-term multiple organ damage, ultimately leading to a significant decrease in cancer survivors' quality of life and life expectancy. Paeonol, a nature active compound derived from the root bark of the medicinal plant Paeonia suffruticosa, exhibits various pharmacological activities. Extensive research has demonstrated that paeonol exhibits substantial anticancer effects in various cancer, both in vitro and in vivo. Its underlying mechanisms involve the induction of apoptosis, the inhibition of cell proliferation, invasion and migration, angiogenesis, cell cycle arrest, autophagy, regulating tumor immunity and enhanced radiosensitivity, as well as the modulation of multiple signaling pathways, such as the PI3K/AKT and NF-κB signaling pathways. Additionally, paeonol can prevent adverse effects on the heart, liver, and kidneys induced by anticancer therapy. Despite numerous studies exploring paeonol's therapeutic potential in cancer, no specific reviews have been conducted. Therefore, this review provides a systematic summary and analysis of paeonol's anticancer effects, prevention of side effects, and the underlying mechanisms involved. This review aims to establish a theoretical basis for the adjunctive strategy of paeonol in cancer treatment, ultimately improving the survival rate and enhancing the quality of life for cancer patients. [ABSTRACT FROM AUTHOR]

A long juvenile period limits the breeding process of many woody plants including tree peony. To investigate the physiological and transcriptomic differences between juvenile and adult plants of tree peony and to explore the key SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) genes, which are vital in age-dependent pathways, 1-year-old and 3-year-old Paeonia delavayi plants were used to compare the relevant physiological parameters and transcriptomic profiles of the leaves in two phases of plants. The results of the physiological parameters showed that the starch content in the leaves of adult plants remained unchanged and that the soluble sugar content significantly increased compared with those in the juvenile plants. In terms of plant hormones, the contents of cytokinin-like hormone (N6-isopentenyladenine (iP)) and jasmonic acid (JA) significantly decreased, whereas the contents of auxin (indole-3-acetic acid, IAA), abscisic acid (ABA), cytokinin-like hormone (N6-isopentenyladenenosine (iPR)), and ethylene precursor (1-aminocyclopropane-1-carboxylic acid, ACC) showed no statistic difference. Transcriptome sequencing results showed that there were 194 differentially expressed genes (DEGs) between juvenile and adult plants, including 171 up-regulated DEGs and 23 down-regulated DEGs. Circadian rhythm, plant hormone signal transduction, and sugar metabolism were closely related to the juvenile-to-adult transition in P. delavayi, involving a total of 12 DEGs. In addition, a total of 13 SPL genes were identified in the transcriptome data, but only PdSPL10 (c71307.graph_c0) was differentially expressed. It was further validated via qRT-PCR analysis, indicating that PdSPL10 might be a key gene regulating the process of juvenile-to-adult in P. delavayi. Based on the above results, a hypothetical transcriptional network regulating juvenile-to-adult transition and flowering in P. delavayi was proposed. These findings provide a reference for understanding the mechanism of juvenile-to-adult transition in tree peony. [ABSTRACT FROM AUTHOR]

Tree peonies (Paeonia Section Moutan)—including nine wild species, which belong to subsections Vaginatae and Delavayanae—are economically important plants with ornamental, nutritional, and medicinal applications. In this study, for the first time, we determined the bioactive components and antioxidant activities and antibacterial activities of the newly grown leaves of nine wild tree peony species (WTPS). A total of 276 bioactive components were identified through non-targeted metabolomics; more than 80% of the 276 metabolites identified are terpenoids and flavonoids. A total of 42 differential metabolites were quantitatively determined. The main differential metabolites were Paeoniflorin, Luteoloside, Hyperin, Apigenin-7-glucoside, Rhoifolin, and Cantharidin. Such a high terpenoid and flavonoid content of the leaf extracts renders them as species with strong antibacterial capacities, and most of the bacteria tested showed greater sensitivity derived from the members of subsection Vaginatae than those of subsection Delavayanae. All WTPS have significant antioxidant activity; this activity is attributed to high levels of the total phenolic content (TPC) and total flavonoid content (TFC), of which, among the nine WTPS, P. lutea has the strongest antioxidant capacity. Our results provided a theoretical basis for the in-deep application of tree peony leaves for food, medical, and pharmaceutical industries. [ABSTRACT FROM AUTHOR]

In order to explore the release rule of floral volatile substances and the diurnal variation of different flower development stages of Paeonia ostii 'Fengdan' in potted and ground-planted conditions, dynamic headspace adsorption combined with gas chromatography-mass spectrometry(GC-MS) was used to analyze the dynamic changes in floral volatile components and contents. Quantitative real-time PCR (qRT-PCR) was used to analyze changes in flower fragrance-regulating genes PsPAL, PsTPSs, and PsbHLH at different flower development stages and a daily change process at the full-blooming stage. The results show that there were differences in aroma components and contents of Paeonia ostii 'Fengdan' at different flower development stages and different time quantum of every day. There were 25 and 28 aroma components identified in 7 flower development stages of tree peonies planted in pots and in the field, respectively, and 23 and 22 aroma components identified at different time quantum of the day, of which the largest and highest content was alkanes. The main characteristic aroma substances were (E)-β-ocimene, 1,3,5-trimethoxybenzene, 2,4-di-tert-butylphenol, methyl jasmonate, nerol, and cinnamyl alcohol; released amounts of the abovementioned substances varied depending on the development stage and the time of the day. The expression of flower fragrance-controlling genes (PsPAL, PsTPSs, and PsbHLH) in tree peonies varied greatly in different conditions. The results of this study provide a valuable resource to investigate floral fragrance formation in tree peonies. [ABSTRACT FROM AUTHOR]

The flower bud differentiation process, morphological features and endogenous hormone changes during single-flowered Paeonia suffruticosa cultivar ‘Lihua Fen’ and lateral-flowered cultivars ‘Zimei Youchun’ and ‘Yun’e Fen’ were compared. The flower bud differentiation process of ‘Lihua Fen’, ‘Zimei Youchun’ and ‘Yun’e Fen’ were observed by anatomy and paraffin sections. The contents of auxin (IAA), abscisic acid (ABA), gibberellin (GA3) and zeatin nucleoside (ZR) in ‘Lihua Fen’ and ‘Zimei Youchun’ at different differentiation stages were determined by enzyme-linked immunosorbent assay (ELISA). The results were as follows: (1)The flower bud differentiation of ‘Lihua Fen’ was only 6 stages and that of ‘Zimei Youchun’ and ‘Yun’e Fen’ were 11 stages. The top flower primordium differentiated earlier than the lateral flower primordium. (2) The contents of ABA, GA3 and ZR of ‘Zimei Youchun’ and ‘Lihua Fen’ increased during the leaf primordium differentiating to the bract primordium, while the content of IAA decreased. As for ‘Zimei Youchun’, ABA and GA3 contents decreased at the initial stage of lateral flower primordium, while ZR and IAA increased. (3) The ABA/IAA of ‘Lihua Fen’ was the lowest at morphological differentiation stage. The study reveals that the lateral-flowered cultivars have earlier and longer flower bud differentiation compared with the single-flowered cultivar, parts of the floral organ differentiation stage of apical and lateral flowers overlaps. Higher ZR and IAA levels and lower ABA and GA3 levels are beneficial to initiate lateral flower primordium differentiation. The results provide a reference for the long florescence breeding of tree peony.

The HSP20 family is the major member of heat shock proteins that are essential components involved in plant growth, development, and stress response, but little is known about tree peony (Paeonia suffruticosa). In this study, genome-wide analysis combining HSP20 gene family expression analysis of tree peony transcriptome was conducted, and the association between SNPs of HSP20s and flower organ number-related traits was analyzed. A total of 149 members were identified in the P. ostii genome, and divided into 10 subfamilies, most of which were classified into cytoplasm or nucleus. Interestingly, their protein sequences were highly conserved, mainly containing motif 1 or 3. In addition, a total of 38 deferentially expressed HSP20s were identified from transcriptome of flower buds with 5-carpels and polycarpels of P. ostii plants at three developmental stages, among which PoHSP89 and PoHSP133 were further cloned from 271 cultivars for association analysis. It indicated that seven or 19 pairs of associated combinations were obtained with the number of carpel, petal, stamen in PoHSP89 and PoHSP133, which could explain the phenotypic variation by 1.79% to 4.06%, and 1.92% to 12.37%, respectively. It will provide a valuable basis for clarifying the phylogenetic relationship to understand their biological function within the HSP20 gene family, and the identified candidate genes and their corresponding associated SNP loci would reveal the genetic basis on floral organ number variation and be useful for molecular marker-assisted breeding of tree peony in view to improving ornamental traits and yields.

Tree peony is a famous ornamental plant, while the low propagation rate is the main hurdles hindering the industry development. Till now, the highly efficient regeneration system for tree peony is not established. In this study, using Paeonia ostii ‘Fengdan’ mature embryos, the effects of variations in inoculation method, initiating culture, adventitious shoot induction, rooting media, plant growth regulators (PGRs), and a non-conventional PGR (plant extracts) on regeneration from explants were evaluated. In embryo cultures, embryonic callus induction rate of 1/4 embryos was the highest among those of embryos with other three technical treatments (whole embryos, 1/2 embryos, and pieces of embryos). The woody plant medium (WPM) containing 1.0 mg • L − 1 6-BA, 0.5 mg • L − 1 GA3, 30.0 g • L − 1 sucrose, and 3.0 g • L − 1 phytagel significantly improved shoot induction and multiplication. 3.0 mg • L − 1 plant extracts promoted hypocotyl germination, rooting, and root growth, in direct embryo culture, and a combination of 3.0 mg • L − 1 plant extracts + 2.0 mg • L − 1 IBA + 1.5 mg • L − 1 IAA produced optimal rooting induction rate for multiple shoots in direct embryo culture and indirect somatic embryogenesis. For the three in vitro micropropagation methods, the highest shoot proliferation coefficient (5.4 ± 0.2) was obtained with indirect somatic embryogenesis. Fortunately, the propagation ability of shoots remains high, even when culture propagation was continued for more than two years. Thus, a reliable system for plant regeneration from mature embryos derived from P. ostii ‘Fengdan’ callus and two direct embryo culture systems have been established. The novel regeneration system could facilitate uniform seedling production.

The Paeonia suffruticosa, known as 'Feng Dan', has been used for thousands of years in traditional Chinese medicine. In our chemical investigation on the root bark of the plant, five new phenolic dimers, namely, paeobenzofuranones A–E (1–5), were characterized. Their structures were determined using spectroscopic analysis including 1D and 2D NMR, HRESIMS, UV, and IR, as well as ECD calculations. Compounds 2, 4, and 5 showed cytotoxicity against three human cancer cell lines, with IC50 values ranging from 6.7 to 25.1 μM. Compounds 1 and 2 showed certain inhibitory activity on NO production. To the best of our knowledge, the benzofuranone dimers and their cytotoxicity of P. suffruticosa are reported for the first time in this paper. [ABSTRACT FROM AUTHOR]

Tree peony (Paeonia suffruticosa Andr.) is a popular cut flower among ornamental plants. However, its short vase life severely hinders the production and application of cut tree peony flowers. To extend the postharvest longevity and improve the horticultural value, silver nanoparticles (Ag-NPs) was applied for reducing bacterial proliferation and xylem blockage in cut tree peony flowers in vitro and in vivo. Ag-NPs was synthesized with the leaf extract of Eucommia ulmoides and characterized. The Ag-NPs aqueous solution showed inhibitory activity against bacterial populations isolated from stem ends of cut tree peony 'Luoyang Hong' in vitro. The minimum inhibitory concentration (MIC) was 10 mg L-1. Compared with the control, pretreatments with Ag-NPs aqueous solution at 5 and 10 mg L-1 for 24 h increased flower diameter, relative fresh weight (RFW), and water balance of tree peony 'Luoyang Hong' flowers. Additionally, malondialdehyde (MDA) and H2O2 content in pretreated petals were lower than the control during the vase life. The activities of superoxide dismutase (SOD) and catalase (CAT) in pretreated petals were lower than that of the control at the early vase stage and higher at the late vase life. Furthermore, pretreatments with Ag-NPs aqueous solution at 10 mg L-1 for 24 h could reduce bacterial proliferation in the xylem vessels on the stem ends by confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM). Overall, pretreatments with green synthesized Ag-NPs aqueous solution effectively reduced bacteria-induced xylem blockage of cut tree peony, resulting in improved water uptake, extended vase life, and enhanced postharvest quality. Therefore, this technique can be used as a promising postharvest technology in the cut flower industry. [ABSTRACT FROM AUTHOR]

Paeonia decomposita, Paeonia rotundiloba, and Paeonia rockii are three closely related species of Sect. Moutan is distributed in the montane area of the Eastern Hengduan Mountain region. Understanding the population history of these three tree peony species could contribute to unraveling the evolutionary patterns of undergrowth species in this hotspot area. We used one nuclear DNA marker (internal transcribed spacer region, ITS) and two chloroplast DNA markers (matK, ycf1) to reconstruct the phylogeographic pattern of the populations. In total, 228 individuals from 17 populations of the three species were analyzed in this study. Three nuclear clades (Clade I – Clade III) and four maternal clades (Clade A – Clade D) were reconstructed. Molecular dating suggested that young lineages diverged during the late Pliocene and early Pleistocene, younger than the uplift of the Hengduan Mountains but older than the last glacial maximum (LGM). Significant population and phylogeographic structures were detected at both markers. Furthermore, the populations of these tree peonies were overall at equilibrium during the climatic oscillations of the Pleistocene. The simulated palaeoranges of the three species during the LGM period mostly overlapped, which could have led to cross‐breeding events. We propose an evolutionary scenario in which mountain orogenesis around the Hengduan Mountain area triggered parapatric isolation between maternal lineages of tree peonies. Subsequent climatic fluctuations drove migration and range recontact of these populations along the valleys. This detailed evolutionary history provides new insights into the phylogeographic pattern of species from mountain‐valley systems. [ABSTRACT FROM AUTHOR]

Paeonia suffruticosa "Shima Nishiki" is a extremely precious double-color cultivar in the world because of its unique and attractive flower color. However, the underlying molecular mechanisms of its double-color formation have not been completely unravelled until now. In the present study, firstly, the full-length cDNA sequence, genomic DNA sequence, promoter region sequence of the key structural gene (PsDFR) in the red and pink petals of the "Shima Nishiki" cultivar were cloned and analyzed, respectively. Meanwhile, the methylation level of CpG island and promoter region of this gene in the red and pink petals was also measured. Moreover, the identification of regulatory effect of PsMYB114L/PsMYB12L and PsDFR was performed. Here, we found that the full-length cDNA sequence, genomic DNA sequence, promoter region sequence of PsDFR were identical in the red and pink petals. There were some differences for the methylation level of this gene in the red and pink petals, but these differences were little and did not show obvious regularity. Therefore, the differential expression of PsDFR should be not caused by its own sequence differences. In addition, the regulatory effect of PsMYB12L and PsDFR was successfully identified. Based on these above results, we concluded that PsMYB12L regulating the differential expression of PsDFR may be a key reason for the double-color formation. These results will advance our understanding of the molecular regulatory mechanisms of double-color formation in P. suffruticosa "Shima Nishiki". [ABSTRACT FROM AUTHOR]

The lack of pollinators in the oil tree peony field was a serious problem for the seed yield. In the preliminary studies, we found that the seed rate of oil tree peonies was significantly improved by supplementing bees during the flowering stages in the pollination net room. In addition, the performance of Apis mellifera L. was better than Bombus terrestris L. To understand the interaction relationship between flowering characteristics and the managed bees, and the pollen-collecting ability of different species of bees, the flowering traits and the pollen loaded on bees were measured in this study. We found that the bees visited flowers with preference and they prefer to forage the flowers having pollen with higher viability. The B. terrestris carried more pollen (27,000.00 ± 5613.70 grains) on the body surface than that of the A. mellifera (7690.00 ± 2873.26 grains). The A. mellifera outperformed B. terrestris both in pollen deposition per visit on the stigma and on the pollen transfer efficiency. The viability of pollen on the body surface (67.77% ± 19.06%) and hind legs (92.02% ± 10.74%) of A. mellifera were both significantly higher than that of B. terrestris (31.84% ± 4.84% and 83.77% ± 6.40%). Our study indicated that the quantity and quality of pollen loaded on stigma by A. mellifera were both better than B. terrestris, which provided evidence that the A. mellifera was the effective pollinator pollinating for the oil tree peony in the pollination net room. [ABSTRACT FROM AUTHOR]

Peony (Paeonia suffruticosa Andr.) seed oil is full of nutrition and exerts positive effects on human's health. The study detected and compared the quality, lipid characteristics, and antioxidant activities of peony seed kernel and shell oil using three extraction techniques (cold pressing, supercritical fluid extraction, and ultrasonic-assisted solvent extraction). Results indicated that the peony seed oil was rich in α-linolenic acid (43.19–44.24%) and γ-tocopherol (403.31–535.15 mg/kg). The extraction methods and shells significantly affected the beneficial phytochemicals (polyphenols, phytosterols, and tocopherols) and antioxidant activities (p < 0.05). Meanwhile, the highest lipid yield was obtained using ultrasonic-assisted solvent extraction (30.60%), while pressing showed the lowest value (17.12%). Supercritical fluid extraction had higher tocopherol (492.68–576.40 mg/kg) and phytosterol (3421.18–4006.23 mg/kg) than other methods. Additionally, ten individual polyphenols were quantified, with the number of catechin being higher. Although the pressing led to a lower lipid yield, its extracts presented a significant higher polyphenol content. In general, peony seed shell oils had a significant advantage in obtaining polyphenols, while seed kernel oil showed higher sterol and tocopherol content. The above information is valuable for manufacturers to select suitable approaches to produce peony seed oil with the required quality attributes and chemical compositions for targeted end-use. [ABSTRACT FROM AUTHOR]

Backgroud: Paeonia holds considerable value in medicinal, ornamental horticultural, and edible oil industries, but the incomplete state of phylogenetic research in this genus poses a challenge to the effective conservation and development of wild germplasm, and also impedes the practical utilization of existing cultivars. Due to its uniparental inheritance and lack of recombination, the plastome (i.e., plastid genome), which is a valuable molecular marker for phylogenetic analyses, is characterized by an appropriate rate of nucleotide evolution. Methods: In this study, 10 newly assembled data and available reported data were combined to perform a comparative genomics and phylogenetics analysis of 63 plastomes of 16 Paeonia species, primarily from East Asia, which is the origin and diversity center of Paeonia. Results: Ranging between 152,153 and 154,405 bp, most plastomes displayed a conserved structure and relatively low nucleotide diversity, except for six plastomes, which showed obvious IR construction or expansion. A total of 111 genes were annotated in the Paeonia plastomes. Four genes (rpl22, rps3, rps19 and ycf1) showed different copy numbers among accessions while five genes (rpl36, petN, psbI, rpl33 and psbJ) showed strong codon usage biases (ENC < 35). Additional selection analysis revealed that no genes were under positive selection during the domestication of tree peony cultivars whereas four core photosynthesis-related genes (petA, psaA, psaB and rbcL) were under positive selection in herbaceous peony cultivars. This discovery might contribute to the wide adaption of these cultivars. Two types of molecular markers (SSR and SNP) were generated from the 63 plastomes. Even though SSR was more diverse than SNP, it had a weaker ability to delimit Paeonia species than SNP. The reconstruction of a phylogenetic backbone of Paeonia in East Asia revealed significant genetic divergence within the P. ostii groups. Evidence also indicated that the majority of P. suffruticosa cultivars had a maternal origin, from P. ostii. The results of this research also suggest that P. delavayi var. lutea, which likely resulted from hybridization with P. ludlowii, should be classified as a lineage within the broader P. delavayi group. Conclusions: Overall, this study's research findings suggest that the Paeonia plastome is highly informative for phylogenetic and comparative genomic analyses, and could be useful in future research related to taxonomy, evolution, and domestication. [ABSTRACT FROM AUTHOR]

Floral longevity (FL) is an important floral functional trait which is critical for flowering plants. FL shows great diversity among angiosperms; however, there is limited information on the mechanisms that influence differences in floral longevity, especially the relationship between petal anatomical traits and floral longevity. We aimed to examine (1) the relationships between petal anatomical traits and FL in tree peony cultivars and (2) the petal anatomical characteristics of longer FL cultivars. Eleven traits of six tree peony cultivars with different FL were investigated, including six water conservation traits (petal thickness, cuticle thickness, number of cell layers, mesophyll thickness, adaxial epidermis thickness and abaxial epidermis thickness), three water supply traits (vein density, number of xlylem vessels and xylem vessel diameter), petal fresh mass and petal dry mass across cultivars. There are significant differences in traits related to water conservation and water supply ability of tree peonies with different FL. Tree peony cultivars with long FL were characterized by the thicker Mesophyll, cuticles, adaxial and abaxial epidermis of the petals. There was a positive correlation between FL and vessel number and vessel diameter. These results suggest that the ability to retain water in flowers is associated with floral longevity. Petal traits related to water conservation and supply, including vein densities, mesophyll thickness, and epidermis thickness, are beneficial for prolonging the flower longevity in tree peonies. [ABSTRACT FROM AUTHOR]

To investigate the amelioration of salt-induced damage on Paeonia ostii 'Fengdan' by exogenous silicon, we analyzed the photosynthetic and physiological characteristics of 1.5-year-old 'Fengdan' seedlings under NaCl stress by applying exogenous silicon (0, 0.75, and 1.5 mmol/L). Our results showed that the contents of the photosynthetic pigments chlorophyll a, chlorophyll b, and carotene, the transpiration rate, stomatal conductance, and intercellular CO2 were significantly enhanced under salt stress when silicon treatment was applied, implying that the net photosynthetic rate was greatly improved. In addition, the plant height, stem thickness, and above-ground dry biomass of tree peony seedlings were effectively increased under salt stress with low-concentration silicon (0.75 mmol/L) treatment, along with osmotic substance (SS, SP, and Pro) content, total polyamine (TP) contents, and the activities of antioxidant-related enzymes (SOD, POD, and CAT) and polyamine-related synthetases (ADC, ODC, and SAMDC). In the low-concentration silicon treatment, malondialdehyde (MDA), hydrogen peroxide (H2O2), and superoxide anions (O2−) were transformed quickly, which eventually reduced cell oxidative damage and improved seedling tolerance. This is an important finding in the understanding of how exogenous low-concentration silicon can alleviate salt-induced damage and promote the growth of tree peony seedlings, thus providing a new perspective on tree peony cultivation. [ABSTRACT FROM AUTHOR]

Tree peony seed oil (TPSO) is an important plant source of n-3 polyunsaturated fatty acid (α-linolenic acid, ALA > 40%) that is receiving increasing attention for its excellent antioxidant and other activities. However, it has poor stability and bioavailability. In this study, a bilayer emulsion of TPSO was successfully prepared using a layer-by-layer self-assembly technique. Among the proteins and polysaccharides examined, whey protein isolate (WPI) and sodium alginate (SA) were found to be the most suitable wall materials. The prepared bilayer emulsion contained 5% TPSO, 0.45% whey protein isolate (WPI) and 0.5% sodium alginate (SA) under selected conditions and its zeta potential, droplet size, and polydispersity index were −31 mV, 1291 nm, and 27%, respectively. The loading capacity and encapsulation efficiency for TPSO were up to 84% and 90.2%, respectively. It was noteworthy that the bilayer emulsion showed significantly enhanced oxidative stability (peroxide value, thiobarbituric acid reactive substances content) compared to the monolayer emulsion, which was accompanied by a more ordered spatial structure caused by the electrostatic interaction of the WPI with the SA. This bilayer emulsion also exhibited markedly improved environmental stability (pH, metal ion), rheological properties, and physical stability during storage. Furthermore, the bilayer emulsion was more easily digested and absorbed, and had higher fatty acid release rate and ALA bioaccessibility than TPSO alone and the physical mixtures. These results suggest that bilayer emulsion containing WPI and SA is an effective TPSO encapsulation system and has significant potential for future functional food development. [ABSTRACT FROM AUTHOR]

Paeonia suffruticosa (P. suffruticosa) seed meal is a byproduct of P. suffruticosa seed processing, which contains bioactive substances such as monoterpene glycosides, and has not been effectively utilized at present. In this study, monoterpene glycosides were extracted from P. suffruticosa seed meal using an ultrasound-assisted ethanol extraction process. The monoterpene glycoside extract was then purified by macroporous resin and identified using HPLC-Q-TOF-MS/MS. The results indicated the following optimal extraction conditions: ethanol concentration, 33%; ultrasound temperature, 55 °C; ultrasound power, 400 W; liquid–material ratio, 33:1; and ultrasound time, 44 min. Under these conditions, the yield of monoterpene glycosides was 121.03 mg/g. The purity of the monoterpene glycosides increased from 20.5% (crude extract) to 71.2% (purified extract) when using LSA-900C macroporous resin. Six monoterpene glycosides (oxy paeoniflorin, isomaltose paeoniflorin, albiflorin, 6′-O-β-D-glucopyranoside albiflorin, paeoniflorin, and Mudanpioside i) were identified from the extract using HPLC-Q-TOF-MS/MS. The main substances were albiflorin and paeoniflorin, and the contents were 15.24 mg/g and 14.12 mg/g, respectively. The results of this study can provide a theoretical basis for the effective utilization of P. suffruticosa seed meal. [ABSTRACT FROM AUTHOR]

Tree peony is a "spring colored-leaf" plant which has red leaves in early spring, and the red color of the leaves usually fades in late spring. Flavonols are one subgroup of flavonoids, and they affect the plant organs' color as co-pigments of anthocyanins. To investigate the color variation mechanism of leaves in tree peony, PqMYBF1, one flavonol biosynthesis-related MYB gene was isolated from Paeonia qiui and characterized. PqMYBF1 contained the SG7 and SG7-2 motifs which are unique in flavonol-specific MYB regulators. Subcellular localization and transactivation assay showed that PqMYBF1 localized to the nucleus and acted as a transcriptional activator. The ectopic expression of PqMYBF1 in transgenic tobacco caused an observable increase in flavonol level and the anthocyanin accumulation was decreased significantly, resulting in pale pink flowers. Dual-luciferase reporter assays showed that PqMYBF1 could activate the promoters of PqCHS, PqF3H, and PqFLS. These results suggested that PqMYBF1 could promote flavonol biosynthesis by activating PqCHS, PqF3H, and PqFLS expression, which leads metabolic flux from anthocyanin to flavonol pathway, resulting in more flavonol accumulation. These findings provide a new train of thought for the molecular mechanism of leaf color variation in tree peony in spring, which will be helpful for the molecular breeding of tree peony with colored foliage. [ABSTRACT FROM AUTHOR]

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Background: WRINKLED1 (WRI1) encodes a transcription factor, belonging to the APETALA2 (AP2) family, and plays a key role in regulating plant oil biosynthesis. As a newly woody oil crop, tree peony (Paeonia rockii) was notable for the abundant unsaturated fatty acids in its seed oil. However, the role of WRI1 during the accumulation of P. rockii seeds oil remains largely unknown. Results: In this study, a new member of the WRI1 family was isolated from P. rockii and was named PrWRI1. The ORF of PrWRI1 consisted of 1269 nucleotides, encoding a putative protein of 422 amino acids, and was highly expressed in immature seeds. Subcellular localization analysis in onion inner epidermal cells showed that PrWRI1 was located at the nucleolus. Ectopic overexpression of PrWRI1 could significantly increase the total fatty acid content in Nicotiana benthamiana leaf tissue and even PUFAs in transgenic Arabidopsis thaliana seeds. Furthermore, the transcript levels of most genes related to fatty acids (FA) synthesis and triacylglycerol (TAG) assembly were also up-regulated in transgenic Arabidopsis seeds. Conclusions: Together, PrWRI1 could push carbon flow to FA biosynthesis and further enhance the TAG amount in seeds with a high proportion of PUFAs. [ABSTRACT FROM AUTHOR]

The yellow colour of ornamental varieties of tree peony originated from Paeonia delavayi. However, but P. delavayi and Paeonia suffruticosa belong to different subgroups, so hybridization is difficult and results in a long breeding cycle. However, no comprehensive transcriptomic profiling has focused on the colour formation mechanisms of yellow tree peony petals. Analysing the colour formation mechanism of yellow petals in P. delavayi is very important for directional molecular breeding. In this study, the transcriptional map of yellow pigment development in petals was used to analyse the mechanism of petal colour formation. We analysed the genes related to the metabolism of flavonoids and carotenoids and the transcription factors (TFs) involved in P. delavayi var. lutea (pure yellow individual) yellow pigment development using transcriptome sequence profiling. Transcriptome sequence profiles revealed three and four differentially expressed transcripts (DETs) involved in flavonoid biosynthesis and carotenoid biosynthesis, respectively. An analysis of DETs in the flavonoid pathway showed that chalcone synthase (CHS) and chalcone 2´-glucosyltransferases (THC2'GT) act in synergy to synthesize isosalipurposide (ISP). CHS and flavonol synthase (FLS) synergistically synthesize quercetin and kaempferol. DEG analysis of the carotenoid pathway revealed that phytoene synthase (PSY), carotenoid isomerase (CRTISO) and β-carotene hydroxylases (CHYB) play a key role in regulating lutein formation, and carotenoid cleavage dioxygenase (CCD) plays an important role in the degradation of carotenoids. These two pathways may be regulated by TF families such as bHLH, ARF, and MYB. The results of the transient overexpression of genes showed that CHS and CHI are regulated by PdMYB2. In this study, the molecular mechanism of ISP synthesis was analysed in depth, and the complete metabolic pathway of carotenoids in Paeonia L. was reported for the first time. By studying the formation mechanism of yellow pigment in P. delavayi petals, a breeding strategy for improving flavonol and carotenoid contents and reducing anthocyanin synthesis by genetic engineering was suggested. [ABSTRACT FROM AUTHOR]

The cold resistance of 14 cultivars of tree peony introduced in Shenyang, China was studied. For each cultivar of peony, 3 branches with the length of 15 to 20 cm growing in the middle section of each cultivar were cut and put into a sealed plastic bag, and then they were put into a low-temperature incubator with 400 (lux) light intensity for low-temperature stress. The treatment gradients were 5 °C (CK), 0 °C, -5 °C and - 10 °C. After the branches were treated for 24 h, the physiological indexes including relative conductivity (Rec), osmotic adjustment of Soluble Sugar (SS), Soluble Protein (SP), and Superoxide dismutase (SOD) of leaves were measured. Increasing the contents of SS and Pro and the activity of SOD were the effective way to improve their resistance according to the changes of the measured indexes. Using the membership function to analyze the cold resistance of different cultivars of peony, the order of cold resistance was as follows: Juanyehong > Juhesanbian > Huihe > Daojin > Xinshiji > Murun Juelun > Taohuayushuang > Fenjinyu > Huawang > Jinge > Guijiucui > Caozhouhong > Panda > Heilongtan. This study provided a theoretical basis for the selection of peony cultivars for overwintering in the open field in northern China. [ABSTRACT FROM AUTHOR]

Peony seed oil, a type of tree nut oil, has attracted the attention of nutritionists for its rich nutritional content. The aim of this study was to extract oil from the peony seed utilizing green and efficient methods. Specifically, aqueous enzymatic extraction was optimized using the Plackett–Burman design combined with the mixture design to extract the optimal enzyme ratio of peony seed oil. When the dosage of enzymes was 10 mg protein/g peony seed, the optimal ratios of the dosages of papain, cellulase, and pectinase were 16.15%, 31.33%, and 52.53%, respectively. Subsequently, central composite design was adopted to optimize supercritical CO2 extraction to identify the process parameters of extracting residual oil from the residue of the aqueous enzymatic extraction. Almost 6.30% of peony seed oil could be obtained from the residue using continuous extraction for 1.58 h at 49.41 °C and 59.75 Mpa. After mixing the peony seed oil extracted by the two processes, its physicochemical indices were measured. Compared with commercial peony seed oil extracted based on the organic solvent leaching method, the elative density and iodine value were higher based on our approach, whereas the other indices showed no significant differences. Thus, the two-step strategy combining the aqueous enzymatic method and supercritical CO2 extraction can be effectively applied to peony seed oil production. [ABSTRACT FROM AUTHOR]

Moutan Cortex, Paeonia suffruticosa root, has long been used as a medicine for the treatment of inflammatory diseases. The aim of this study was to evaluate the modulative properties of Moutan Cortex water extract (CP) on endoplasmic reticulum (ER) stress-related macrophage activation via the calcium-CHOP pathway. RAW 264.7 mouse macrophages were activated by lipopolysaccharide (LPS), and the levels of various inflammatory mediators from RAW 264.7 were evaluated. The multiplex cytokine assay was used to investigate both cytokines and growth factors, and RT-PCR was used to investigate the expressions of inflammation-related genes, such as CHOP. Data represent the levels of NO and cytosolic calcium in LPS-stimulated RAW 264.7 were significantly inhibited by CP as well as hydrogen peroxide (p < 0.05). Minutely, NO production in LPS-stimulated RAW 264.7 incubated with CP at concentrations of 25, 50, 100, and 200 µg/mL for 24 h was 97.32 ± 1.55%, 95.86 ± 2.26%, 94.64 ± 1.83%, and 92.69 ± 2.31% of the control value (LPS only), respectively (p < 0.05). Calcium release in LPS-stimulated RAW 264.7 incubated with CP at concentrations of 25, 50, 100, and 200 µg/mL for 18 h was 95.78 ± 1.64%, 95.41 ± 1.14%, 94.54 ± 2.76%, and 90.89 ± 3.34% of the control value, respectively (p < 0.05). Hydrogen peroxide production in LPS-stimulated RAW 264.7 incubated with CP at concentrations of 25, 50, 100, and 200 µg/mL for 24 h was 79.15 ± 7.16%, 63.83 ± 4.03%, 46.27 ± 4.38%, and 40.66 ± 4.03% of the control value, respectively (p < 0.05). It is interesting that the production of IL-6, TNF-α, G-CSF, MIP-1α, MIP-2, and M-CSF in LPS-stimulated RAW 264.7 were significantly inhibited by CP (p < 0.05), while the production of LIX, LIF, RANTES, and MIP-1β showed a meaningful decrease. CP at concentrations of 25, 50, 100, and 200 µg/mL significantly reduced the transcription of Chop, Camk2α, NOS, STAT1, STAT3, Ptgs2, Jak2, c-Jun, Fas, c-Fos, TLR3, and TLR9 in LPS-stimulated RAW 264.7 (p < 0.05). CP at concentrations of 25, 50, and 100 µg/mL significantly reduced the phosphorylation of STAT3, p38 MAPK, and IκB-α in LPS-stimulated RAW 264.7 (p < 0.05). These results suggest that CP might modulate macrophage activation via LPS-induced calcium signaling and the ER stress-CHOP pathway. [ABSTRACT FROM AUTHOR]

The flower bud differentiation process, morphological features and endogenous hormone changes during single-flowered Paeonia suffruticosa cultivar ‘Lihua Fen' and lateral-flowered cultivars ‘Zimei Youchun' and ‘Yun'e Fen' were compared in order to provide a reference for the long florescence breeding of tree peony. The flower bud differentiation process of ‘Lihua Fen', ‘Zimei Youchun' and ‘Yun'e Fen' were observed by anatomy and paraffin sections. The contents of auxin (IAA), abscisic acid (ABA), gibberellin (GA3) and zeatin nucleoside (ZR) in ‘Lihua Fen' and ‘Zimei Youchun' at different differentiation stages were determined by enzyme-linked immunosorbent assay (ELISA). The results were as follows: (1) The flower bud differentiation of ‘Lihua Fen' was only 6 stages and that of ‘Zimei Youchun' and ‘Yun'e Fen' were 11 stages. The apical flower primordium differentiated earlier than the lateral flower primordium. (2) The contents of ABA, GA3 and ZR of ‘Zimei Youchun' and ‘Lihua Fen' increased during the leaf primordium differentiating to the bract primordium, while the content of IAA decreased. As for ‘Zimei Youchun', ABA and GA3 contents decreased at the initial stage of lateral flower primordium, while ZR and IAA increased. (3) The ABA/IAA of ‘Lihua Fen' was the lowest at morphological differentiation stage. The study reveals that the lateral-flowered cultivars have earlier and longer flower bud differentiation compared with the single-flowered cultivar, parts of the floral organ differentiation stage of apical and lateral flowers overlaps. Higher ZR and IAA levels and lower ABA and GA3 levels are beneficial to initiate lateral flower primordium differentiation. The results provide a reference for the long florescence breeding of tree peony. [ABSTRACT FROM AUTHOR]

Background and Aims The tree peony (Paeonia suffruticosa Andr.) has been widely cultivated as a field plant, and petal blotch is one of its important traits, which not only promotes proliferation but also confers high ornamental value. However, the regulatory network controlling blotch formation remains elusive owing to the functional differences and limited conservation of transcriptional regulators in dicots. Methods We performed phylogenetic analysis to identify MYB44-like transcription factors in P. suffruticosa blotched cultivar 'High noon' petals. A candidate MYB44-like transcription factor, PsMYB44, was analysed via expression pattern analysis, subcellular localization, target gene identification, gene silencing in P. suffruticosa petals and heterologous overexpression in tobacco. Key Results A blotch formation-related MYB44-like transcription factor, PsMYB44, was cloned. The C-terminal of the PsMYB44 amino acid sequence had a complete C2 motif that affects anthocyanin biosynthesis, and PsMYB44 was clustered in the MYB44-like transcriptional repressor branch. PsMYB44 was located in the nucleus, and its spatial and temporal expression patterns were negatively correlated with blotch formation. Furthermore, a yeast one-hybrid assay showed that PsMYB44 could target the promoter of the late anthocyanin biosynthesis-related dihydroflavonol-4-reductase (DFR) gene, and a dual-luciferase assay demonstrated that PsMYB44 could repress PsDFR promoter activity. On the one hand, overexpression of PsMYB44 significantly faded the red colour of tobacco flowers and decreased the anthocyanin content by 42.3 % by downregulating the expression level of the tobacco NtDFR gene. On the other hand, PsMYB44 -silenced P. suffruticosa petals had a redder blotch colour, which was attributed to the fact that silencing PsMYB44 redirected metabolic flux to the anthocyanin biosynthesis branch, thereby promoting more anthocyanin accumulation in the petal base. Conclusion These results demonstrated that PsMYB44 negatively regulated the biosynthesis of anthocyanin by directly binding to the PsDFR promoter and subsequently inhibiting blotch formation, which helped to elucidate the molecular regulatory network of anthocyanin-mediated blotch formation in plants. [ABSTRACT FROM AUTHOR]

Tree peony is a unique traditional flower in China, with large, fragrant, and colorful flowers. However, a relatively short and concentrated flowering period limits the applications and production of tree peony. A genome-wide association study (GWAS) was conducted to accelerate molecular breeding for the improvement of flowering phenology traits and ornamental phenotypes in tree peony. A diverse panel of 451 tree peony accessions was phenotyped for 23 flowering phenology traits and 4 floral agronomic traits over 3 years. Genotyping by sequencing (GBS) was used to obtain a large number of genome-wide single-nucleotide polymorphisms (SNPs) (107 050) for the panel genotypes, and 1047 candidate genes were identified by association mapping. Eighty-two related genes were observed during at least 2 years for flowering, and seven SNPs repeatedly identified for multiple flowering phenology traits over multiple years were highly significantly associated with five genes known to regulate flowering time. We validated the temporal expression profiles of these candidate genes and highlighted their possible roles in the regulation of flower bud differentiation and flowering time in tree peony. This study shows that GWAS based on GBS can be used to identify the genetic determinants of complex traits in tree peony. The results expand our understanding of flowering time control in perennial woody plants. Identification of markers closely related to these flowering phenology traits can be used in tree peony breeding programs for important agronomic traits. [ABSTRACT FROM AUTHOR]

Floral scent is an important and genetically complex trait in horticultural plants. Tree peony (Paeonia × suffruticosa) originates in the Pan-Himalaya and has nine wild species divided into two subsections, Delavayanae and Vaginatae. Their flowers are beloved worldwide for their sweet floral fragrance, yet the flavor-related volatiles and underlying biosynthetic pathways remain unknown. Here, we characterized the volatile blends of all wild tree peony species and found that the flavor-related volatiles were highly divergent, but linalool was a unique monoterpene in subsect. Delavayanae. Further detection of volatiles in 97 cultivars with various genetic backgrounds showed that linalool was also the characteristic aroma component in Paeonia delavayi hybrid progenies, suggesting that linalool was conserved and dominant within subsect. Delavayanae and its hybrids, instead of species and cultivars from subsect. Vaginatae. Global transcriptome analysis of all wild tree peony species and 60 cultivars revealed five candidate genes that may be involved in key steps of linalool biosynthesis; especially the expressions of three TPS genes, PdTPS1 , PdTPS2 , and PdTPS4 , were significantly positively correlated with linalool emissions across tree peony cultivars. Further biochemical evidence demonstrated that PdTPS1 and PdTPS4 were the pivotal genes determining the species-specific and cultivar-specific emission of linalool. This study revealed a new insight into floral scent divergence in tree peony and would greatly facilitate our understanding of the phylogeny and evolution of Paeonia. [ABSTRACT FROM AUTHOR]

The miRNA, a kind of endogenous non-coding small RNA, plays an essential role in regulation of gene expression in plants. Quantitative real-time PCR (qRT-PCR) assay is one of the most common methods used for quantification of miRNA expression, and levels of expression are normalized by comparing with reference genes. The present study was intended to identify the appropriate reference genes for normalizing the level of miRNA expression in various developmental stages and tissues such as the bud development process, flower development stages, and different tissues of tree peony of different flowering times. Five algorithms (Delta Ct, geNorm, NormFinder, BestKeeper, and RefFinder) were used for stability analysis. The results showed that mtr-MIR160b-p3 and gma-miR394a-5p were the most stable miRNAs expressed during the bud development process of early-flowering tree peony 'Feng dan'; PC-5p-19095 was the most stable during the bud development process of late-flowering tree peony 'Lian he', followed by gma-miR394a-5p and mtr-MIR160b-p3; the mtr-miR159a was the most stable miRNA expressed in the flower development stages of different tree peony varieties. The PC-3p-871 was the most stable miRNA expressed in different tissues of early-flowering tree peony 'Feng dan', followed by PC-5p-4, and PC-5p-4 was the most stable in late-flowering tree peony 'Lian he', followed by the mtr-miR168b. The findings of this study provide a reference for studying the changes in miRNA expression, and further exploring the regulatory mechanism of miRNA in tree peony. [ABSTRACT FROM AUTHOR]

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The Paeonia suffruticosa ANDR. (P. suffruticosa) is commonly used in traditional medicine for various purposes. Suffruticosol A (Suf‐A), isolated from P. suffruticosa, is a beneficial compound with antibiofilm, antivirulence, and anti‐inflammatory properties. The aim of the present study was to investigate the biological effects of Suf‐A on osteogenic processes in pre‐osteoblasts. It was determined here in that Suf‐A (>98.02%), isolated from P. suffruticosa, showed no cytotoxicity at 0.1–30 μM; however, it induced cytotoxicity at 50–100 μM in pre‐osteoblasts. Suf‐A increased osteogenic alkaline phosphatase activity and expression levels of noncollagenous proteins. Adhesion and trans‐migration on the extracellular matrix were potentiated by Suf‐A, but not by wound‐healing migration. Suf‐A did not affect autophagy or necroptosis during osteoblast differentiation. We found that Suf‐A increased runt‐related transcription factor 2 (RUNX2) levels and mineralized matrix formation. RUNX2 expression was mediated by Suf‐A‐induced BMP2‐Smad1/5/8 and mitogen‐activated protein kinase signaling, as demonstrated by Noggin, a BMP2 inhibitor. These results suggest that Suf‐A is a potential natural osteogenic compound. [ABSTRACT FROM AUTHOR]

Rhizosphere microorganisms are instrumental in plant drought resistance. However, knowledge of how the bacterial community structure in the rhizosphere of Paeonia ostii 'Feng Dan' responds to drought stress is limited. In the current study, the effect of bacterial community structure and diversity in the rhizosphere were investigated using high-throughput sequencing technology. Diversity analysis showed that there was no significant difference in Alpha-diversity between control (Co), drought stress (De), and rehydration treatments (Re), and the bacterial community structures in De and Re treatments were more similar and markedly different from that of the control. Taxonomic analysis revealed that Acidobacteria, Proteobacteria and Actinobacteria were the dominant phyla in the rhizosphere of Paeonia ostii 'Feng Dan', while Gp6 and Gp4 were the dominant taxa at the genus level. Drought stress did not significantly affect rhizosphere bacterial diversity but did cause significant changes in the community structure, with a marked increase in the relative abundance of Actinobacteria. Findings from this study will underpin improvements in drought tolerance of tree peony via the soil microbial community. [ABSTRACT FROM AUTHOR]

To investigate the molecular mechanism of the ADVENTITIOUS ROOTING RELATED OXYGENASE 1 (ARRO-1) gene in adventitious rooting of tree peony, the PsARRO-1 gene was isolated from Paeonia ostii. Subsequently, a yeast two-hybrid (Y2H) cDNA library using the split-ubiquitin yeast membrane system was constructed, the insert fragments lengths of pPR3-N membrane Y2H library were mainly concentrated between 500 and 3,000 bp, and the library capacity reached 1.3 × 107 CFUs, with an efficiency of recombinant of 100%, which met the requirements for subsequent screening of PsARRO-1-interacting proteins. Finally, 15 candidate PsARRO-1-interacting proteins were screened out by pBT3SUC-ARRO, among them potassium transporter 2, cytochrome b5 isoform E, and high mobility group B protein 3 (HMGB3) might be involved in the formation of roots. Both direct Y2H and bimolecular fluorescence complementation assays confirmed that PsARRO-1 interacted with PsHMGB3, which shared high identity with EgHMGB3 and MiHMGB3. The expression of PsHMGB3 was significantly up-regulated in the root outgrowth stage, and the overexpression of PsHMGB3 in Arabidopsis resulted in lengthened primary roots, more lateral roots. In short, the Y2H library provides a large number of useful resources for functional genomics research on roots of tree peony. [ABSTRACT FROM AUTHOR]

Tree peony is an important plant that integrates ornamental, oil, and medicinal use. However, drought stress seriously restricts the development of the tree peony industry and the improvement of its economic value. In this study, Paeonia ostii 'Fengdan' was used as the experimental material, their biological functions were verified by bioinformatics analysis, dual-luciferase reporter assay, overexpression transformation, and momentary transformation. The results of the degradation group showed that PomiR396g-5p cut the target gene PoACO1 at 1082 bp, and the double luciferase report test showed that PomiR396g-5p reduced the fluorescence activity of the target gene PoACO1 , and there was a targeted cutting relationship between PomiR396g-5p and PoACO1. PomiR396g-5p promoter contained a variety of cis -acting elements related to abiotic stress, and the PoACO1 protein was an intracellular hydrophilic protein, did not have a transmembrane structure, and was mainly located in the nucleus and cytoplasm. PomiR396g-5p transgenic plants had enhanced drought resistance, improved the activities of POD, SOD, CAT, and other antioxidant enzymes, and reduced the accumulation of ROS. The expression of PoACO1 in PomiR396g-5p transgenic plants was inhibited. The rate and amplitude of leaf length, width, and fresh weight decrease in the miRNAs group were lower than those in the control group and target genome. Whereas instantaneous silencing of tree peony leaves showed the opposite trend to transgenic plants and homologous instantaneous overexpression. The symptoms of acute leaf atrophy and water loss were observed in the PomiR396g-5p at 9–12 h, and the decrease rates of fresh weight, leaf length, and leaf width were significantly higher than those in the target genome and control group. In conclusion, PomiR396g-5p acted as a positive regulator of tree peony in response to drought stress and improved drought resistance by regulating the target gene PoACO1 to inhibit its expression and regulation of ROS levels in plants. Meanwhile, there was a negative regulatory relationship between PomiR396g-5p and the target gene PoACO1.Our study provides a theoretical basis and technical support for the research on drought resistance molecular breeding of tree peony. • PomiR396g-5p cut the target gene PoACO1 at 1082 bp, and PomiR396g-5p reduced the fluorescence activity of PoACO1. • PomiR396g-5p promoter contained a variety of cis -acting elements related to abiotic stress. • PomiR396g-5p transgenic plants had enhanced drought resistance. The PoACO1 in PomiR396g-5p transgenic plants was inhibited. • PomiR396g-5p acted as positive regulatory factors in response to drought stress of tree peony. [ABSTRACT FROM AUTHOR]

• The new LC-MS/MS strategy detects polyphenol metabolites in edible flowers. • The pseudo-targeted screening method enables accuracy and quantification. • Using the new strategy, 43 polyphenol metabolites were detected in 27 edible flowers. • Significant variations in polyphenol types and levels among the 27 flowers. • Enriched research on polyphenols in edible flowers;model applicable to other species. Edible flowers, as a food ingredient, are rich in bioactive substance and have received a lot of attention. In this study, we have established a pseudo-targeting method based on Multi Reaction Monitoring (MRM) - Enhanced Product Ions (EPI) in the Information Dependent Acquisition (IDA) mode. The results showed that a total of 43 metabolites, including flavonoids, phenolic acids, and terpenes were detected in 27 edible flowers using integrated liquid chromatography-tandem mass spectrometry (LC-MS/MS) strategy. Rutin was measured in all edible flowers, but the levels varied significantly among the different varieties of flowers, with the highest being about 3500 times the lowest. Albiflorin and paeoniflorin are polyphenolic substance unique to Paeonia suffruticosa. The cyanidin 3-O-glucoside chloride content in reddish-purple flowers is higher than that of other colors. Dicoumarin and 4-hydroxycoumarin were discovered in edible flowers for the first time. This study established a pseudo-targeted detection method to reveal the distribution of polyphenolic metabolites and enriched the research on polyphenolic metabolites in edible flowers. It can also serve as a general strategy to detect metabolites in other agricultural products, providing valuable information about potential metabolites for consumers. [ABSTRACT FROM AUTHOR]

Zinc (Zn) deficiency is a significant nutritional limitation to crop yield globally, particularly in calcareous soil environments. Tree peony of Peaonia ostii 'Fengdan' is regarded as an oil crop due to its seeds rich in alpha-linolenic acid, a beneficial compound for health promotion. However, low seed yield remains a primary challenge in attaining sufficient seed oil from tree peony. In this study, Zn fertilization was applied to soil or foliage of P. ostii 'Fengdan' in the growth period before fruit development. Our findings reveal that foliar Zn-spraying, as opposed to soil application, proves to be a more effective method for augmenting seed yield, Zn accumulation and photosynthetic capacity in 'Fengdan'. Comparative analyses of the leaf proteome of 'Fengdan' using iTRAQ profiling under foliar Zn-spraying identified 115 differentially expressed proteins (DEPs), including 36 upregulated proteins, which likely contribute to the observed increase in seed yields of 'Fengdan' caused by foliage Zn-spraying. Specifically, Zn2+ stimulation of phosphatidylinositol signaling initiates a cascade of metabolic regulations. Firstly, ATP synthesis promotes leaf photosynthetic capacity, facilitated by improved sucrose metabolism through upregulated pullulanase and 1,4-alpha-glucan-branching enzyme. Furthermore, lipid synthesis and transport are facilitated by upregulated lipoyl synthase and plastid lipid-associated proteins. Additionally, DEPs involved in secondary metabolism are upregulated in the production of various metabolites conducive to 'Fengdan' growth. Overall, our results demonstrate that foliage Zn-spraying enhances seed yield in P. ostii 'Fengdan' by elevating Zn content and secondary metabolite synthesis in leaves, thereby augmenting leaf photosynthetic capacity and lipid synthesis. This study provides an effective way to increase seed yield of tree peony by exogenous Zn application. [Display omitted] • Foliar Zn-spraying is a more effective method for seed yield increase in 'Fengdan'. • Foliar Zn-spraying enhances the photosynthetic capacity in 'Fengdan' leaves. • Molecular mechanism of foliage Zn-spraying was studied using leaf iTRAQ proteome. • Secondary metabolites and lipid synthesis are critical for seed yield increase in 'Fengdan'. [ABSTRACT FROM AUTHOR]

Adventitious root (AR) formation is a limiting factor in the vegetative propagation of tree peony (Paeonia suffruticosa Andr.). PoARRO-1 , which encodes an auxin oxidase involved in AR formation, plays a role in the root development of P. ostii , but its associated molecular regulatory mechanisms are not yet understood. In this study, we examined the role of PoARRO-1 in AR formation in P. ostii. The overexpression of PoARRO-1 in P. ostii test-tube plantlets led to a notable enhancement in both the rooting rate and the average number of ARs in vitro, as well as increased activities of peroxidase (POD), superoxide dismutase (SOD), and indoleacetic acid oxidase (IAAO). PoARRO-1 was involved in the conversion of IAA-Asp and IAA-Glu to OxIAA and promoted IAA oxidation. RNA sequencing analysis revealed that PoARRO-1 overexpression led to upregulation of enzyme activity, auxin metabolism related genes. Further analyses showed that PoARRO-1 interacted with the 1–175 aa position of PoIAA27b to regulate the formation of ARs. We therefore propose that PoARRO-1 interacts with PoIAA27b to promote AR formation, and it may be useful targets for enhancing the in vitro propagation of P. ostii. • Overexpression of PoARRO-1 increased the ARs number in Paeonia ostii test-tube plantlets. • PoARRO-1 was involved in the conversion of IAA-Asp and IAA-Glu to OxIAA and promoted IAA oxidation. • PoARRO-1 interacted with PoIAA27b to positively regulate the formation of ARs in P. ostii test-tube plantlets. [ABSTRACT FROM AUTHOR]

Tree peonies (Paeonia suffruticosa) are important pharmaceutical and oilseed crops widely cultivated in China. Tree peony flowers, as important by-products of the industrial crops, are conventionally used as folk medicine with health-promoting effects on human. However, the in-depth understanding of biologically active compounds in tree peony flowers is scarce. In this study, an in-depth analysis of bioactive compounds in tree peony flower buds (TPFBs) from 74 cultivars was conducted using an integrated LC-MS-based metabolomic approach. Ninety-two metabolites were tentatively identified in TPFBs. The quantitative results revealed that TPFBs were abundant in bioactive compounds. Among these, paeoniflorin, albiflorin, cynaroside, and pentagalloylglucose (PGG) emerged as the primary chemical markers of TPFBs, showing significant variations based on genotype. Each extract of TPFB exhibited potent antioxidant, antibacterial, and anti-neuroinflammatory activities. Notably, TPFB extract showed the highest effectiveness in inhibiting L. monocytogenes among all tested bacteria. Cynaroside was predicted to serve as a pivotal component in TPFB for its anti-neuroinflammatory effects by interacting with toll-like receptor 4 (TRL4) and interleukin-1beta (IL-1β). This study offers novel insights into the bioactive compounds and bioactivities of TPFBs, which lays theoretical basis for the application in pharmaceutical industry. [Display omitted] • Bioactive compound profiling through integrative metabolomics. • Tree peony flower buds are abundant in monoterpenoids and polyphenols. • Flower bud extract exhibits outstanding inhibitory effect against L. monocytogenes. • TRL4 and IL-1β might be the key targets of anti-neuroinflammation. • Tree peony flower bud can be utilized in pharmaceutical industry. [ABSTRACT FROM AUTHOR]

Tree peony (Paeonia suffruticosa Andr.) is a unique aromatic plant famous for its huge flowers, bright colors and strong fragrance, having high ornamental, medicinal, and edible value. Research on tree peony's flower fragrance has mainly focused on the comparative analysis among its varieties, leaving the inheritance of aroma compounds in this plant an overlooked area of study. Here, the volatile components of flowers at three different flowering stages of the light fragrance-type cultivar P. ostii 'Fengdan' and the strong fragrance-type cultivar P. suffruticosa 'Chunguihuawu', as well as the half-opening stage flowers of 109 F1 progeny, were collected and characterized in-depth by dynamic headspace sampling technique combined with gas chromatography-mass spectrometry (GC-MS). Diverse profiles of volatiles that included alcohols, esters, aldehydes, terpenes, benzenes, and hydrocarbons were identified from the evaluated accessions. These results revealed that the volatile components and content of parents were significantly different, and that hybridization generated more complex volatile components. Most volatile compounds in the hybrids, especially the main aromatic components, existed in at least one of the parents, being characterized by intermediate or transgressive inheritance for the floral trait; this demonstrated that volatile compounds can be inherited from parents to progeny. Further, seven de novo aroma compounds—those present in progeny yet absent in their parents—were found in progeny. This study preliminarily clarified the segregation performance of aroma traits in tree peony hybrids, which might provide a theoretical basis for selecting breeding parents and the breeding of new varieties for aroma traits.

Horticulture productivity has been increasingly restricted by heat stress from growing global warming, making it far below the optimum production capacity. As a popular ornamental cultivar of tree peony, Paeonia suffruticosa 'Yu Hong' has also been suffering from heat stress not suitable for its optimal growth. To better understand the response mechanisms against heat stress of tree peony, investigations of phenotypic changes, physiological responses, and quantitative proteomics were conducted. Phenotypic and physiological changes indicated that 24 h of exposure to heat stress (40 °C) was the critical duration of heat stress in tree peony. The proteomic analyses revealed a total of 100 heat-responsive proteins (HRPs). According to bioinformatic analysis of HRPs, the heat tolerance of tree peony might be related to signal transduction, synthesis/degradation, heat kinetic proteins, antioxidants, photosynthesis, energy conversion, and metabolism. Our research will provide some new insights into the molecular mechanism under the response against the heat stress of tree peony, which will benefit the future breeding of heat-resistant ornamental plants. [ABSTRACT FROM AUTHOR]

As a candidate national flower of China, tree peony has extremely high ornamental, medicinal and oil value. However, the short florescence and rarity of early-flowering and late-flowering varieties restrict further improvement of the economic value of tree peony. Specific miRNAs and their target genes engaged in tree peony floral florescence, development and senescence remain unknown. This report presents the integrated analysis of the miRNAome, transcriptome and degradome of tree peony petals collected from blooming, initial flowering, full blooming and decay stages in early-flowering variety Paeonia ostii 'Fengdan', an early-flowering mutant line of Paeonia ostii 'Fengdan' and late-flowering variety Paeonia suffruticosa 'Lianhe'. Transcriptome analysis revealed a transcript ('psu.G.00014095') which was annotated as a xyloglucan endotransglycosylase/hydrolase precursor XTH-25 and found to be differentially expressed across flower developmental stages in Paeonia ostii 'Fengdan' and Paeonia suffruticosa 'Lianhe'. The miRNA-mRNA modules were presented significant enrichment in various pathways such as plant hormone signal transduction, indole alkaloid biosynthesis, arachidonic acid metabolism, folate biosynthesis, fatty acid elongation, and the MAPK signaling pathway. Multiple miRNA-mRNA-TF modules demonstrated the potential functions of MYB-related, bHLH, Trihelix, NAC, GRAS and HD-ZIP TF families in floral florescence, development, and senescence of tree peony. Comparative spatio-temporal expression investigation of eight floral-favored miRNA-target modules suggested that transcript 'psu.T.00024044' and microRNA mtr-miR166g-5p are involved in the floral florescence, development and senescence associated agronomic traits of tree peony. The results might accelerate the understanding of the potential regulation mechanism in regards to floral florescence, development and abscission, and supply guidance for tree peony breeding of varieties with later and longer florescence characteristics. [ABSTRACT FROM AUTHOR]

Considering the problem of food perishability and the environmental pollution of plastic packaging, the natural active packaging prepared by incorporating plant leaf (e.g. peony leaf) extract into biodegradable food packaging materials may be a key to addressing these issues. In the study, a novel green bioactive composite film (CS-PLE) with antioxidant and antibacterial propertiese was developed by blending peony leaf extract (PLE) into chitosan (CS) film through the solution casting method. The physical properties and biological activities of a series of films, including CS film and CS-PLEs incorporated with different concentrations of PLE, were studied by appropriate experimental protocols. The results demonstrated that addition of PLE improved thermal stability, the barrier performance of UV-A and total phenolic content of CS film, and the antioxidant activities of CS-PLEs increased by 60.6 % ~ 73.2 % with the appending proportion of PLE from 0.1 wt% ~ 0.7 wt% compared to CS film. Furthermore, CS-PLEs had better inhibiting effect on bacteria. Notably, CS-PLE (0.7 wt%) wrapping apples showed the least degree of browning among all the groups. Hence, the study of CS-PLEs can provide a new strategy for the preparation of food packaging materials that are green alternatives and reduce environmental pollution. [Display omitted] • Chitosan (CS) films incorporated with peony leaf extract (PLE) were prepared for the first time. • PLE significantly improved the antioxidant and antibacterial activities of CS films. • The incorporation of PLE enhanced UV-shielding properties and water vapor barrier performance of CS films. • CS-PLE composite films were suggested as a new active packaging material. [ABSTRACT FROM AUTHOR]

Paeonia suffruticosa 'Er Qiao' is a famous double-color cultivar in the world till now. However, the regulatory mechanism of this double-color phenotype is still unknown. In this study, the P. suffruticosa cultivar 'Er Qiao' with purple and pink petals on the same flower was selected as the research materials. Based on high-throughput sequencing technology, microRNA (miRNA) sequences of purple (Ps-Z) and pink (Ps-F) petals were analyzed. 260 known and 204 novel miRNAs were identified, among which 17 known and 51 novel miRNAs were differentially expressed between the Ps-Z and Ps-F libraries. Meanwhile, their target genes of these differentially expressed miRNAs (DEMs) were successfully predicted and annotated. Based on the results of differential expression of miRNAs and annotation of the corresponding target genes, one DEM (miR156-9) was screened out, which may be related to double-color formation. Furthermore, the quantitative real-time PCR (qRT-PCR) result also showed that the expression level of miR156-9 in purple and pink petals of the double-color flowers was significantly different. Based on these above results, we concluded that miR156-9 should play a key role in the formation of the double-color flower phenotype of the 'Er Qiao' cultivar in P. suffruticosa at the post transcriptional level. The study will provide a theoretical basis and a new perspective for further research on molecular regulatory mechanism of double-color formation in P. suffruticosa 'Er Qiao'. [ABSTRACT FROM AUTHOR]

Paeonia suffruticosa Andr., a member of Paeoniaceae, is native to China. In its 1600 years' cultivation, more than 2000 cultivars for different purposes (ornamental, medicinal and oil use) have been inbred. However, there are still some controversies regarding the provenance of tree peony cultivars and the phylogenetic relationships between and within different cultivar groups. In this study, plastid genome sequencing was performed on 10 representative tree peony cultivars corresponding to 10 different flower types. Structure and comparative analyses of the plastid genomes showed that the total lengths of the chloroplast genome of the 10 cultivars ranged from 152,153 to 152,385 bp and encoded 84–88 protein-coding genes, 8 rRNAs and 31–40 tRNAs. The number of simple sequence repeats and interspersed repeat sequences of the 10 cultivars ranged from 65–68 and 40–42, respectively. Plastid phylogenetic relationships of Paeonia species/cultivars were reconstructed incorporating data from our newly sequenced plastid genomes and 15 published species, and results showed that subsect. Vaginatae was the closest relative to the central plains cultivar group with robust support, and that it may be involved in the formation of the group. Paeonia ostii was recovered as a successive sister group to this lineage. Additionally, eleven morphological characteristics of flowers were mapped to the phylogenetic skeleton to reconstruct the evolutionary trajectory of flower architecture in Paeoniaceae. [ABSTRACT FROM AUTHOR]

The aim of this review is to perform a systematic review of scientific papers and an in-depth analysis of the latest research related to Paeonia × suffruticosa Andrews as a valuable plant species, important in pharmacy and cosmetology. P. × suffruticosa bark root-Moutan cortex is a medicinal raw material formerly known from traditional Chinese medicine (TCM) but less common in official European medicine. It was introduced for the first time in the European Pharmacopoeia Supplement 9.4 in 2018. In this work, the numerous possible applications of this raw material were depicted based on modern professional pharmacological studies documenting its very valuable medicinal values, including antioxidant, cytoprotective, anti-cancer, anti-inflammatory, cardioprotective, anti-atherosclerotic, anti-diabetic and hepatoprotective activities. The scientific studies indicated that the profile of raw material activity is mainly due to paeonol, paeoniflorin and 1,2,3,4,6-penta-O-galloyl-β-D-glucopyranose. Moreover, the significance of this plant (its different organs) in the production of cosmetics was underlined. P. × suffruticosa finds increasing application in cosmetology due to research on its chronic dermatitis, anti-aging and brightening effects. Furthermore, some biotechnological research has been described aimed at developing effective in vitro micropropagation protocols for P. × suffruticosa. [ABSTRACT FROM AUTHOR]

Tree peony (Paoenia ostii) is a famous Chinese traditional flower well-known in many countries of the world. However, the short and concentrated flowering period of tree peony greatly affects the ornamental and economic value of the flowers. Exogenous brassinosteroid (BR) treatment can delay the flowering period of ostii T. Hong et J. X. Zhang var. lishizhenenii B. A. Shen for 3 days, but the underlying regulatory mechanism remains elusive. Here, full-length transcriptome and transcriptome sequencing were used to mine key genes related to BR-induced delayed flowering in tree peony. The transcriptome sequencing of the petals yielded 21.27 G clean data and 62,229 isoforms. Among them, 58,218 isoforms were annotated in NR, NT, SwissProt, KEGG, KOG, InterPro and GO databases. GO and KEGG analyses showed that 2460 DEGs were related to delayed flowering in response to BR. Additionally, a total of seven genes affecting flowering were annotated from 11 isoforms, which responded to BR through three pathways to delay the flowering of P. ostii var. lishizhenii. BR treatment increased the expression of BRASSINOSTEROID-SIGNALING KINASE3 (BSK3), potentially by promoting BRI1 ASSOCIATED KINASE RECEPTOR 1 (BAK1). Moreover, BR treatment suppressed the expression of SQUAMOSA-PROMOTER BINDING PROTEIN-LIKE 1 (SPL1), possibly to inhibit the expressions of downstream WRKY genes and APETALA 2 (AP2). Meanwhile, BR treatment promoted the expression of DELLA, which might inhibit the expressions of CONSTANS (CO) and SPL. These results suggest a theoretical basis for further analyses of the molecular mechanism of flowering regulation in tree peony. [ABSTRACT FROM AUTHOR]

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