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As a plant-specific gene family, class III peroxidases (PODs) play an important role in plant growth, development, and stress responses. However, the POD gene family has not been systematically studied in Paeonia ostii. In this study, a total of 57 PoPOD genes were identified in the P. ostii genome. Subsequently, phylogenetic analysis and chromosome mapping revealed that PoPODs were classified into six subgroups and were unevenly distributed across five chromosomes. The gene structure and conserved motifs indicated the potential for functional divergence among the different subgroups. Meanwhile, four PoPODs were identified as tandem duplicated genes, with no evidence of segmental duplication. Using RNA-seq data from eight different tissues, multiple PoPODs exhibited enhanced expression in apical and adventitious roots (ARs). Next, RNA-seq data from AR development combined with trend analysis showed that PoPOD30/34/43/46/47/57 are implicated in the formation of ARs in tree peony. Through WGCNA based on RNA-seq, two key genes, PoPOD5/15, might be involved in heat tolerance via ABA and MeJA signaling. In addition, real-time quantitative PCR (qRT-PCR) analysis indicated that PoPOD23 may play an important role in flower senescence. These findings deepened our understanding of POD-mediated AR development, heat tolerance, and petal senescence in tree peony. [ABSTRACT FROM AUTHOR]

SERK is a marker gene for early somatic embryogenesis. We screened and functionally verified a SERK-interacting protein to gain insights into tree-peony somatic embryogenesis. Using PoSERK as bait, we identified PorbcL (i.e., the large subunit of Rubisco) as a SERK-interacting protein from a yeast two-hybrid (Y2H) library of cDNA from developing tree-peony somatic embryos. The interaction between PorbcL and PoSERK was verified by Y2H and bimolecular fluorescence complementation analyses. PorbcL encodes a 586-amino-acid acidic non-secreted hydrophobic non-transmembrane protein that is mainly localized in the chloroplast and plasma membrane. PorbcL was highly expressed in tree-peony roots and flowers and was up-regulated during zygotic embryo development. PorbcL overexpression caused the up-regulation of PoSERK (encoding somatic embryogenesis receptor-like kinase), PoAGL15 (encoding agamous-like 15), and PoGPT1 (encoding glucose-6-phosphate translocator), while it caused the down-regulation of PoLEC1 (encoding leafy cotyledon 1) in tree-peony callus. PorbcL overexpression led to increased indole-3-acetic acid (IAA) content but decreasing contents of abscisic acid (ABA) and 6-benzyladenosine (BAPR). The changes in gene expression, high IAA levels, and increased ratio of IAA to ABA, BAPR, 1-Aminocyclopropanecarboxylic acid (ACC), 5-Deoxystrigol (5DS), and brassinolide (BL) promoted embryogenesis. These results provide a foundation for establishing a tree-peony embryogenic callus system. [ABSTRACT FROM AUTHOR]

SUMMARY: Drought is a detrimental environmental factor that restricts plant growth and threatens food security throughout the world. WRKY transcription factors play vital roles in abiotic stress response. However, the roles of IIe subgroup members from WRKY transcription factor family in soluble sugar mediated drought response are largely elusive. In this study, we identified a drought‐responsive IIe subgroup WRKY transcription factor, PoWRKY69, from Paeonia ostii. PoWRKY69 functioned as a positive regulator in response to drought stress with nucleus expression and transcriptional activation activity. Silencing of PoWRKY69 increased plants sensitivity to drought stress, whereas conversely, overexpression of PoWRKY69 enhanced drought tolerance in plants. As revealed by yeast one‐hybrid, electrophoretic mobility shift assay, and luciferase reporter assays, PoWRKY69 could directly bind to the W‐box element of fructose‐1,6‐bisphosphate aldolase 5 (PoFBA5) promoter, contributing to a cascade regulatory network to activate PoFBA5 expression. Furthermore, virus‐induced gene silencing and overexpression assays demonstrated that PoFBA5 functioned positively in response to drought stress by accumulating fructose to alleviate membrane lipid peroxidation and activate antioxidant defense system, these changes resulted in reactive oxygen species scavenging. According to yeast two‐hybrid, bimolecular fluorescence complementation, and firefly luciferase complementation imaging assays, valine‐glutamine 11 (PoVQ11) physically interacted with PoWRKY69 and led to an enhanced activation of PoWRKY69 on PoFBA5 promoter activity. This study broadens our understanding of WRKY69‐VQ11 module regulated fructose accumulation in response to drought stress and provides feasible molecular measures to create novel drought‐tolerant germplasm of P. ostii. Significance Statement: This manuscript identified a novel WRKY69‐VQ11 module that regulated fructose accumulation in response to drought stress, which provides feasible molecular measures to create novel drought‐tolerant germplasm of Paeonia ostii. [ABSTRACT FROM AUTHOR]

Fengdan (Paeonia × suffruticosa Andrews), a woody oil crop abundant in unsaturated fatty acids, notably linolenic acid, holds promise in agriculture. However, selenium deficiency in Chinese soil poses a challenge to its growth. This study investigated the impact of Selenium nanoparticles (SeNPs) on Fengdan's physiological traits and selenium enrichment. Employing a randomized complete block design, five SeNPs spraying levels were tested on Fengdan cultivar Feng Dan. Results showed substantial enhancements in photosynthetic parameters with Se application, particularly at 6.00 g/ha (S3) and 8.25 g/ha (S4). Moreover, Se application improved protein, soluble sugar, and proline solubility, along with increased activities of SOD and POD, while decreased MDA content. Se content in Fengdan organs notably increased with SeNPs concentration, especially in the testa, leaf, and kernel. In conclusion, comprehensive SeNPs fertilizer application positively affected Fengdan's growth, photosynthesis, and selenium enrichment. The most effective treatments were observed at 6.00 g/ha (S3) and 8.25 g/ha (S4). This study offers valuable insights into selenium enrichment strategies for oil crops, stressing the significance of Se application dosage and organ specificity in biological Se enrichment. [ABSTRACT FROM AUTHOR]

With persistent elevation in global temperature, water scarcity becomes a major threat to plant growth and development, yield security, agricultural sustainability, and food production. Proline, as a key osmolyte and antioxidant, plays a critical role in regulating drought tolerance in plants, especially its key biosynthetic enzyme, delta-1-pyrroline-5-carboxylate synthase (P5CS), which always positively responds to drought stress. As an important woody oil crop, the expansion of Paeonia ostii cultivation needs to address the issue of plant drought tolerance. Here, we isolated a PoP5CS gene from P. ostii, with an open reading frame of 1842 bp encoding 613 amino acids. PoP5CS expression progressively increased in response to increasing drought stress, and it was localized in the cytoplasm. Silencing of PoP5CS in P. ostii reduced drought tolerance, accompanied by decreased proline content, elevated reactive oxygen species (ROS) accumulation, and increased relative electrical conductivity (REC) and malondialdehyde (MDA) levels. Conversely, overexpression of PoP5CS in Nicotiana tabacum plants enhanced drought resistance, manifested by increased proline levels, reduced ROS accumulation, and lower REC and MDA contents. This study isolates PoP5CS from P. ostii and validates its role in regulating drought tolerance, providing valuable genetic resources and theoretical insights for the development of drought-resistant P. ostii cultivars. [ABSTRACT FROM AUTHOR]

Tree peony (Paeonia suffruticosa Andr.) is a woody plant with high ornamental, medicinal, and oil values. However, its low rooting rate and poor rooting quality are bottleneck issues in the micropropagation of P. ostii. The WUSCHEL-related homeobox (WOX) family plays a crucial role in root development. In this study, based on the screening of the genome and root transcriptome database, we identified ten WOX members in P. ostii. Phylogenetic analysis revealed that the ten PoWOX proteins clustered into three major clades, the WUS, intermediate, and ancient clade, respectively. The conserved motifs and tertiary structures of PoWOX proteins located in the same clade exhibited higher similarity. The analysis of cis-regulatory elements in the promoter indicated that PoWOX genes are involved in plant growth and development, phytohormones, and stress responses. The expression analysis revealed that PoWOX genes are expressed in distinct tissues. PoWOX4, PoWOX5, PoWOX11, and PoWOX13b are preferentially expressed in roots at the early stage of root primordium formation, suggesting their role in the initiation and development of roots. These results will provide a comprehensive reference for the evolution and potential function of the WOX family and offer guidance for further study on the root development of tree peony. [ABSTRACT FROM AUTHOR]

The oil tree peony (Paeonia ostii) is widely cultivated for their high value seed oil, and its roots also have high medicinal value, but they are often discarded and wasted as by-products. In this study, a set of steam explosion (SE) and ultrasound (US)-assisted extraction/purification methods of insoluble bound phenols (IPs) from oil peony roots (PR) were explored and developed. The IPs were obtained through SE pre-treatment (120 °C, 15 min) and US-assisted alkaline extraction (0.25 W/cm2, 4 h and 4 M NaOH). Their total phenolic content (TPC) significantly increased from 112.26 mg gallic acid/g dry weight (mg GAE/g dw) with traditional method to 185.59 mg GAE/g dw. Subsequently, the IPs were purified into PIPs using LX-83 resin. The optimized purification conditions were pH = 4, 3.0 mg GAE/mL of IPs, 0.25 W/cm2 adsorption for 120 min, as well as 60 % ethanol desorption solution, and 0.25 W/cm2 desorption for 30 min. The TPC of the PIPs increased to 254.74 mg GAE/g dw. Notably, typical compounds in IPs and PIPs were identified and quantified by HPLC-MS and their antioxidant and anti-tyrosinase activities were also evaluated by chronometry. The results showed that paeoniflorin and benzoyl-paeoniflorin increased by about 10-fold in the PIPs. Additionally, they demonstrated superior antioxidant and tyrosinase inhibitory activities compared to IPs. Therefore, the innovative extraction and purification process greatly improved the yield of bound phenols and recovered paeoniflorin from non-medicinal PR. The process may also provide a reference for the in-depth development of medicinal PR, ultimately achieving more efficient PR development and utilization. [Display omitted] • The integrated process improves the yield of insoluble phenols from peony (IP). • IP are raised by steam explosion and ultrasound-assisted alkaline extraction. • US enhances adsorption/desorption of IPs using LX-83 resins. • The sum of 5 typical compounds increases from 0.47 (IPs) to 3.24 mg/g (PIPs). • PIPs showed higher antioxidant and anti-tyrosinase activities. [ABSTRACT FROM AUTHOR]

The effects of selenite (0, 15, 30, 45 mg L-1) on physiological characteristics and medicinal components of Paeonia ostii were analyzed. The results showed that selenite application promoted the activity of superoxide dismutase and the contents of soluble sugar, proline, carotenoids, total flavonoids, and total polyphenols, and decreased the contents of reactive oxygen species, relative electrical conductivity, and malondialdehyde. In addition, selenite also increased chlorophyll content, improved electron transfer ability, PSI and PSII performance, and the coordination between PSI and PSII, which significantly improved photosynthetic capacity. Moreover, selenite treatment also greatly increased the contents of gallic acid, catechin, albiflorin, paeoniflorin, benzoic acid, and paeonol in Moutan cortex radicis (MCR). These results showed that selenite effectively protected the photosynthetic apparatus from photooxidative damage by enhancing antioxidant capacity, improving photosynthetic capacity, and increasing the content of the medicinal compounds in MCR.

Trehalose-6-phosphate synthase (TPS) as a key enzyme in trehalose metabolism plays important roles in metabolic regulation and abiotic stress tolerance in many species. In our previous study, 10 TPS family members in Paeonia ostii have been identified, and among them PoTPS1 and PoTPS5 were regarded as critical genes in regulating growth and development of P. ostii. In this study, the full-length of cDNAs with 1698 nucleotides encoding PoTPS1 and 2571 nucleotides encoding PoTPS5 from P. ostii were cloned. The sequence analysis revealed that PoTPS1 protein belongs to the Class I group and PoTPS5 was a Class II TPS protein, and they possess highly conserved residues. The expression levels of PoTPS1 and PoTPS5 were induced by sugar and abiotic stress, especially under glucose and high temperature treatments. Then, PoTPS1 and PoTPS5 protein were expressed at high level in Escherichia coli and purified by affinity chromatography. The optimum temperature of PoTPS1 and PoTPS5 were 50 ℃ and 60 ℃, and the optimum pH for both PoTPS1 and PoTPS5 was 6.0. Metal cations such as Mg2+ and Zn2+ stimulated PoTPS1 activity significantly, and the Mg2+, Cu2+ and Zn2+ motivated PoTPS5 activity tremendously. The addition of chondroitin sulfate was shown to stimulate enzyme activity.Key message: We purified the tehalose-6-phosphate synthase PoTPS1 and PoTPS5 from Paeonia ostii to study their enzymatic characteristics and analyzed their expression levels under abiotic stress. [ABSTRACT FROM AUTHOR]

Paeonia ostii is an important economic oil and medicinal crop. Its anthers are often used to make tea in China with beneficial effects on human health. However, the metabolite profiles, as well as potential biological activities of P. ostii anthers and the pollen within anthers have not been systematically analyzed, which hinders the improvement of P. ostii utilization. With comprehensive untargeted metabolomic analysis using UPLC-QTOF-MS, we identified a total of 105 metabolites in anthers and pollen, mainly including phenylpropanoids, polyketides, organic acids, benzenoids, lipids, and organic oxygen compounds. Multivariate statistical analysis revealed the metabolite differences between anthers and pollen, with higher carbohydrates and flavonoids content in pollen and higher phenolic content in anthers. Meanwhile, both anthers and pollen extracts exhibited antioxidant activity, antibacterial activity, α-glucosidase and α-amylase inhibitory activity. In general, the anther stage of S4 showed the highest biological activity among all samples. This study illuminated the metabolites and biological activities of anthers and pollen of P. ostii, which supports the further utilization of them. [ABSTRACT FROM AUTHOR]

While there is long‐standing interest in the role of inter‐ and transgenerational plasticity via the maternal line, it rarely has been studied via the paternal line. Thus, consequences of the paternal environment for parental fitness and for performance of offspring in the environments experienced by either fathers or mothers are not known. We studied the intergenerational plasticity (IGP) of the plant species Paeonia ostii (Paeoniaceae) and tested the hypothesis that exposure of fathers to environmental stress (i) reduces parental fitness and performance of offspring grown under non‐stressful conditions, but (ii) mitigates the negative effects of environmental stress on fitness of parents and performance of offspring.Crosses were made in a greenhouse within six families of P. ostii between parents grown under drought and in a mesic environment, and the offspring of each cross were grown under both dry and mesic conditions. Production and germination of seeds and morphological and physiological traits of offspring were measured as indicators of parental fitness and offspring performance, respectively.Paternal drought decreased seed number per fruit, except when maternal plants also were grown in drought conditions. Offspring drought decreased seedling performance. However, when fathers experienced drought this negative effect on the offspring was partly mitigated, in particular when mothers also had experienced drought. In contrast, offspring grown in mesic conditions had improved seedling performance, especially when either parent (or both) also were grown in mesic conditions. Such statistical differences remained when seed mass was included as a covariate.Overall, paternal pollen of P. ostii mediated IGP to drought almost as well as it did for maternal ovules. IGP was adaptive if environments remained constantly dry across generations but maladaptive if environments changed. Hence, under future climate changes, paternal IGP might be both a blessing and a curse, with the blessing occurring when the focal habitat becomes drier and pollen comes from already‐dry places, while the curse may dominate in predictably moist habitats surrounded by drier habitats. Read the free Plain Language Summary for this article on the Journal blog. [ABSTRACT FROM AUTHOR]

The effects of selenite (0, 15, 30, 45 mg L–1) on physiological characteristics and medicinal components of Paeonia ostii were analyzed. The results showed that selenite application promoted the activity of superoxide dismutase and the contents of soluble sugar, proline, carotenoids, total flavonoids, and total polyphenols, and decreased the contents of reactive oxygen species, relative electrical conductivity, and malondialdehyde. In addition, selenite also increased chlorophyll content, improved electron transfer ability, PSI and PSII performance, and the coordination between PSI and PSII, which significantly improved photosynthetic capacity. Moreover, selenite treatment also greatly increased the contents of gallic acid, catechin, albiflorin, paeoniflorin, benzoic acid, and paeonol in Moutan cortex radicis (MCR). These results showed that selenite effectively protected the photosynthetic apparatus from photooxidative damage by enhancing antioxidant capacity, improving photosynthetic capacity, and increasing the content of the medicinal compounds in MCR. [ABSTRACT FROM AUTHOR]

Paeonia ostii, a plant of substantial economic significance, continues to face constraints in achieving large-scale propagation. In vitro propagation offers a promising avenue for the production of disease-free plants and the genetic transformation of peonies to instill novel traits. However, significant challenges persist in tissue culture, particularly with regards to the reproduction coefficient of shoots and the rooting process. This study reports an efficacious protocol for P. ostii micropropagation, focusing on in vitro root development facilitated through the application of phloroglucinol (PG). Furthermore, the study unveils the molecular signature of P. ostii during in vitro root development. The results indicate that the modified Y3 medium (Y3M), supplemented with 1 mg/L 6-benzyladenine (BA) and 0.1 mg/L α-naphthaleneacetic acid (NAA), is optimal for adventitious bud induction, achieving a 96.67% induction rate and an average of 16.03 adventitious shoots per sample. The highest elongation percentage (92.15%) and the longest average shoot length (3.87 cm) were obtained with Y3M containing 0.3 mg/L BA and 0.03 mg/L NAA. Additionally, the optimal medium for inducing root formation in P. ostii was identified as WPM supplemented with 3 mg/L indole-3-butyric acid (IBA) and 100 mg/L phloroglucinol (PG). Lignin content detection, microscope inspection, and molecular signature results demonstrated that PG enhanced lignin biosynthesis, thereby promoting in vitro rooting of P. ostii.

The brief longevity of tree peony blossoms constrains its ornamental value and economic worth. Gibberellins (GAs) are crucial in the modulation of flower senescence, and GA 20-oxidase (GA20ox), GA 3-oxidase (GA3ox), and GA 2-oxidase (GA2ox) catalyze the synthesis and deactivation of bioactive GAs. In Paeonia ostii, a total of three PoGA20ox, ten PoGA3ox, and twelve PoGA2ox proteins were identified and comprehensively analyzed. The analysis of the gene structures, conserved domains, and motifs revealed structural similarities and variances among the GA20ox, GA3ox, GA2ox-A, and GA2ox-B subfamilies. The synteny analysis indicated a scarcity of collinear blocks within the P. ostii genome, with no tandem or whole-genome duplication/segmental duplications found in PoGAoxs. The investigation into the binding of transcription factors to PoGAox promoters and the assessments of the expression levels suggest that PoGA2ox1 and PoGA2ox8.1 are promising candidate genes implicated in the regulation of floral senescence. Further, Pos.gene61099 (BPC6) and Pos.gene61094 (CIL2) appear to modulate PoGA2ox1 transcription in a positive and negative manner, respectively, while Pos.gene38359 (DDF1) and Pos.gene17639 (DREB1C) likely enhance PoGA2ox8.1's expression. This study lays a foundation for an in-depth understanding of PoGAox functions and the development of strategies to delay flower senescence in tree peony. [ABSTRACT FROM AUTHOR]

Paeonia ostii 'Feng Dan' is widely cultivated in China for its ornamental, medicinal, and edible properties. The whole plant of tree peony is rich in bioactive substances, while the comprehensive understanding of metabolites in the leaves is limited. In this study, an untargeted metabolomics strategy based on UPLC-ESI-TOF-MS was conducted to analyze the dynamic variations of bioactive metabolites in P. ostii 'Feng Dan' leaves during development. A total of 321 metabolites were rapidly annotated based on the GNPS platform, in-house database, and publications. To accurately quantify the selected metabolites, a targeted method of HPLC-ESI-QQQ-MS was used. Albiflorin, paeoniflorin, pentagalloylglucose, luteolin 7-glucoside, and benzoylpaeoniflorin were recognized as the dominant bioactive compounds with significant content variations during leaf development. Metabolite variations during the development of P. ostii 'Feng Dan' leaves are greatly attributed to the variations in antioxidant activities. Among all tested bacteria, the leaf extract exhibited exceptional inhibitory effects against Streptococcus hemolytis-β. This research firstly provides new insights into tree peony leaves during development. The stages of S1–S2 may be the most promising harvesting time for potential use in food or pharmaceutical purposes. [ABSTRACT FROM AUTHOR]

Tree peony is an emerging oil crop that has received widespread attention in recent years. Studying the nutritional composition and loss patterns of tree peony seeds is not only a fundamental requirement for the tree peony planting and processing industry, but also of great significance for the development of related industries such as tree peony medicine and food. Traditionally, the number of related studies is relatively small, and the available research data are even rarer. This article mainly takes mature seeds of characteristic peonies from the Tongling area and the main production area Bozhou as samples. Firstly, the authenticity and viability of the seeds are identified using microscopy and TTC staining, and then the seeds are powdered for future use. Secondly, the soxhlet extraction method was used to measure the crude fat oil content in peony seeds, which ranges from 22.67% to 29.66%. The protein content in peony seeds was measured to be around 11.76%-21.17% using the second method of food protein determination standard method - ultraviolet spectrophotometry. After methyl esterification of fresh oil from tree peony seed oil, it was separated and detected by GC-MS (GC7890B-MSD5977A gas chromatography-mass spectrometry). Further retrieval was conducted using the NISTsearch2.0 standard spectral library, and artificial spectral analysis was conducted in conjunction with relevant literature to determine the 31 fatty acid compositions of tree peony seed oil. Finally,? the relative content of each component was calculated using peak area normalization method. After acid hydrolysis of fresh oil from tree peony seeds, 16 amino acids and their contents were measured using Hitachi L-8900 high-speed amino acid analyzer. The dataset for determining the nutritional composition of tree peony seeds provides preliminary information on the nutritional composition of tree peony seeds under different production and storage conditions, which has direct academic reference value for tree peony seed germination, seed processing, and reasonable storage. [ABSTRACT FROM AUTHOR]

SERK is a marker gene for early somatic embryogenesis. We screened and functionally verified a SERK-interacting protein to gain insights into tree-peony somatic embryogenesis. Using PoSERK as bait, we identified PorbcL (i.e., the large subunit of Rubisco) as a SERK-interacting protein from a yeast two-hybrid (Y2H) library of cDNA from developing tree-peony somatic embryos. The interaction between PorbcL and PoSERK was verified by Y2H and bimolecular fluorescence complementation analyses. PorbcL encodes a 586-amino-acid acidic non-secreted hydrophobic non-transmembrane protein that is mainly localized in the chloroplast and plasma membrane. PorbcL was highly expressed in tree-peony roots and flowers and was up-regulated during zygotic embryo development. PorbcL overexpression caused the up-regulation of PoSERK (encoding somatic embryogenesis receptor-like kinase), PoAGL15 (encoding agamous-like 15), and PoGPT1 (encoding glucose-6-phosphate translocator), while it caused the down-regulation of PoLEC1 (encoding leafy cotyledon 1) in tree-peony callus. PorbcL overexpression led to increased indole-3-acetic acid (IAA) content but decreasing contents of abscisic acid (ABA) and 6-benzyladenosine (BAPR). The changes in gene expression, high IAA levels, and increased ratio of IAA to ABA, BAPR, 1-Aminocyclopropanecarboxylic acid (ACC), 5-Deoxystrigol (5DS), and brassinolide (BL) promoted embryogenesis. These results provide a foundation for establishing a tree-peony embryogenic callus system.

With persistent elevation in global temperature, water scarcity becomes a major threat to plant growth and development, yield security, agricultural sustainability, and food production. Proline, as a key osmolyte and antioxidant, plays a critical role in regulating drought tolerance in plants, especially its key biosynthetic enzyme, delta-1-pyrroline-5-carboxylate synthase (P5CS), which always positively responds to drought stress. As an important woody oil crop, the expansion of Paeonia ostii cultivation needs to address the issue of plant drought tolerance. Here, we isolated a PoP5CS gene from P. ostii, with an open reading frame of 1842 bp encoding 613 amino acids. PoP5CS expression progressively increased in response to increasing drought stress, and it was localized in the cytoplasm. Silencing of PoP5CS in P. ostii reduced drought tolerance, accompanied by decreased proline content, elevated reactive oxygen species (ROS) accumulation, and increased relative electrical conductivity (REC) and malondialdehyde (MDA) levels. Conversely, overexpression of PoP5CS in Nicotiana tabacum plants enhanced drought resistance, manifested by increased proline levels, reduced ROS accumulation, and lower REC and MDA contents. This study isolates PoP5CS from P. ostii and validates its role in regulating drought tolerance, providing valuable genetic resources and theoretical insights for the development of drought-resistant P. ostii cultivars.

Introduction: Paeonia ostii T. Hong & J.X. Zhang (s.s.) (Chinese name, Fengdan) is a widely cultivated food-medicine plant in China, in which root bark, seed kernels, and flowers are utilized for their medicinal and edible values. However, other parts of the plant are not used efficiently, in part due to a poor understanding of their chemical composition and potential biological activity. Methods: Untargeted ultra-performance liquid chromatography-quadrupole time of flight-mass spectrometry (UPLC-Q-TOF-MS) metabolomics was applied to characterize the metabolic profiles of 10 different parts of P. ostii. Results and discussion: A total of 160 metabolites were alternatively identified definitely or tentatively, which were significantly different in various plant parts by multivariate statistical analysis. Quantitative analysis showed that underutilized plant parts also contain many active ingredients. Compared with the medicinal part of root bark, the root core part still contains a higher content of paeoniflorin (17.60 ± 0.06 mg/g) and PGG (15.50 ± 2.00 mg/g). Petals, as an edible part, contain high levels of quercitrin, and stamens have higher methyl gallate and PGG. Unexpectedly, the ovary has the highest content of methyl gallate and rather high levels of PGG (38.14 ± 1.27 mg/g), and it also contains surprisingly high concentrations of floralalbiflorin I. Paeoniflorin (38.68 ± 0.76 mg/g) is the most abundant in leaves, and the content is even higher than in the root bark. Branches are also rich in a variety of catechin derivatives and active ingredients such as hydrolyzable tannins. Seed kernels also contain fairly high levels of paeoniflorin and albiflorin. Fruit shells still contain a variety of components, although not at high levels. Seed coats, as by-products removed from peony seeds before oil extraction, have high contents of stilbenes, such as trans-gnetin H and suffruticosol B, showing significant potential for exploitation. Except for the seed kernels, extracts obtained from other parts exhibited good antioxidant activity in DPPH, ABTS, and ferric ion reducing antioxidant power (FRAP) assays (0.09-1.52 mmol TE/g). Five compounds (gallic acid, PGG, trans-resveratrol, kaempferol, and quercitrin) were important ingredients that contributed to their antioxidant activities. Furthermore, P. ostii seed cakes were first reported to possess agonistic activity toward CB1/CB2 receptors. This study provides a scientific basis for the further development and utilization of P. ostii plant resources. [ABSTRACT FROM AUTHOR]

Paeonia ostii is a worldwide ornamental flower and an emerging oil crop. Zyotic embryogenesis is a critical process during seed development, and it can provide a basis for improving the efficiency of somatic embryogenesis (SE). In this study, transcriptome sequencing of embryo development was performed to investigate gene expression profiling in P. ostii and identified Differentially expressed genes (DEGs) related to transcription factors, plant hormones, and antioxidant enzymes. The results indicated that IAA (Indole-3-acetic acid), GA (Gibberellin), BR (Brassinosteroid) and ETH (Ethylene) were beneficial to early embryonic morphogenesis, while CTK (Cytokinin) and ABA (Abscisic Acid) promoted embryo morphogenesis and maturation. The antioxidant enzymes' activity was the highest in early embryos and an important participant in embryo formation. The high expression of the genes encoding fatty acid desaturase was beneficial to fast oil accumulation. Representative DEGs were selected and validated using qRT-PCR. Protein-protein interaction network (PPI) was predicted, and six central node proteins, including AUX1, PIN1, ARF6, LAX3, ABCB19, PIF3, and PIF4, were screened. Our results provided new insights into the formation of embryo development and even somatic embryo development in tree peonies. [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]

Paeonia ostii is an important woody oil crop mainly cross-pollinated. However, the low yield has become an important factor restricting the industrial development of P. ostii. Cross-pollination has become one of the important measures to increase the seed yield. Therefore, conservation of pollen with high vitality is crucial to ensure successful pollination of P. ostii. In this study, we found an effective methodological system to assess the viability, ability to germinate, and optimal storage conditions of P. ostii pollen grains. The optimal medium in vitro was 50 g/L sucrose, 100 mg/L boric acid, 50 g/L PEG6000, 100 mg/L potassium nitrate, 300 mg/L calcium nitrate, and 200 mg/L magnesium sulfate at pH 5.4. Optimal germination condition in vitro was achieved at 25 °C for 120 min, allowing easy observation of the germination percentage and length of the pollen tubes. In addition, the viability of pollen grains was assessed by comparing nine staining methods. Among them, MTT, TTC, benzidine-H2O2, and FDA were effective to distinguish between viable and non-viable pollen, and the results of the FDA staining method were similar to the pollen germination percentage in vitro. After evaluation of pollen storage, thawing and rehydration experiments showed that thawing at 4 °C for 30 min and rehydration at 25 °C for 30 min increased the germination percentage of pollen grains stored at low temperatures. The low-temperature storage experiments showed that 4 °C was suitable for short-term storage of P. ostii pollen grains, while −80 °C was suitable for long-term storage. This is the first report on the in vitro germination, viability tests, and storage of P. ostii pollen grains, which will provide useful information for P. ostii germplasm conservation and artificial pollination. [ABSTRACT FROM AUTHOR]

Non-structural carbohydrates (NSC) as resource reserves of plants play important roles in energy supply for normal growth and reproduction under environmental stress. The yield of perennial crops is mainly determined by the carbohydrate production and allocation in the current growth season, as well as the re-allocation of NSC reserves. However, the contribution of NSC to crop yield has not been fully determined. Fengdan (Paeonia ostii) is a variety of oil Peony that is newly developed in China. The effects of tree age and NSC on yield were examined by investigated the variations of biomass, soluble sugars, starch, and NSC in the organ and whole tree levels in the dormant and ripening stages of Fengdan populations with 4-, 6-, and 8-year-old in 2020 and 5-, 7-, and 9-year old in 2021. Results showed that the biomass, yield (seed biomass), soluble sugars, starch, and NSC reserve of Fengdan at the whole tree level increased with the increase in age. Although consistent correlations were observed between soluble sugars, starch and NSC storage, and yield among the plants with different ages, Fengdan showed allometric growth relationships between the accumulation of soluble sugars, starch, and NSC and yield and biomass (standardized major axis analyses slope b ≠ 1). Tree age significantly affected biomass and its allocation and NSC levels, especially the yield of Fengdan plants. The results of the investigation of the variations in the relationships between the yield and seasonal fluctuations of NSC and biomass indicate that roots is the key storage structure, whereas stems both serve as sink and/or source functions for the adult (7-9a) plants. NSC level, particularly the concentration of soluble sugars, in stems mainly influences Fengdan yield. These findings together provide new insights into the mechanisms underlying the yield formation of Fengdan and have implications for manipulating sink-source relationship to achieve high yield. [ABSTRACT FROM AUTHOR]

The brief longevity of tree peony blossoms constrains its ornamental value and economic worth. Gibberellins (GAs) are crucial in the modulation of flower senescence, and GA 20-oxidase (GA20ox), GA 3-oxidase (GA3ox), and GA 2-oxidase (GA2ox) catalyze the synthesis and deactivation of bioactive GAs. In Paeonia ostii, a total of three PoGA20ox, ten PoGA3ox, and twelve PoGA2ox proteins were identified and comprehensively analyzed. The analysis of the gene structures, conserved domains, and motifs revealed structural similarities and variances among the GA20ox, GA3ox, GA2ox-A, and GA2ox-B subfamilies. The synteny analysis indicated a scarcity of collinear blocks within the P. ostii genome, with no tandem or whole-genome duplication/segmental duplications found in PoGAoxs. The investigation into the binding of transcription factors to PoGAox promoters and the assessments of the expression levels suggest that PoGA2ox1 and PoGA2ox8.1 are promising candidate genes implicated in the regulation of floral senescence. Further, Pos.gene61099 (BPC6) and Pos.gene61094 (CIL2) appear to modulate PoGA2ox1 transcription in a positive and negative manner, respectively, while Pos.gene38359 (DDF1) and Pos.gene17639 (DREB1C) likely enhance PoGA2ox8.1’s expression. This study lays a foundation for an in-depth understanding of PoGAox functions and the development of strategies to delay flower senescence in tree peony.

Two new glycosides 2-hydroxy-4-methoxy-acetophenone-3-O-[ß-D-apiofuranosyl (1→6)-ß-Dglucoside (1) and 4-hydroxy-2-O-ß-rutinosyl acetophenone (2), along with two known compounds, paeonolide (3), involcranoside B (4) were isolated from the roots of Paeonia ostii. In addition, compound 4 was isolated from this genus for the first time. Their structures were established on the basis of spectral and chemical evidence. All the compounds showed inactive nitric oxide (NO) inhibitory effects. [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]

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]

Paeonia ostii is an emerging woody oil crop, but the high temperature in summer is extremely unfavorable for its growth and development. Understory intercropping cultivation would provide shaded environments which could effectively reduce the ambient temperature. In order to explore whether understory intercropping cultivation would reduce the effects of high-temperature stress on P. ostii, the changes of leaf physiological indicators and leaf microstructures of sole-cropping and intercropping P. ostii were investigated. P. ostii that intercropping cultivated under Illinois pecans (Carya illinoinensis (Wangenh.) K. Koch) with 4 m × 4 m and 4 m × 8 m rowing spaces were used as samples in this study. The results showed that with continuous high temperature, the high-temperature damage index of P. ostii kept increasing, whereas the leaf relative water content continued to decline. Compared to sole-cropping, the high-temperature damage index, relative electrical conductivity, proline content, antioxidant enzymes superoxide dismutase (SOD), and ascorbate peroxidase (APX) activities of intercropping P. ostii under C. illinoinensis were significantly decreased, whereas the leaf relative water content was higher. Moreover, compared to sole-cropping, intercropping P. ostii under C. illinoinensis increased SPAD and chlorophyll contents, made mesophyll cell ultrastructures more intact, and made the chloroplasts rounder and more filled with starch granules and lipid globules, leading to enhanced photosynthesis (Pn) and transpiration rates (Tr). Notably, the reduction of high-temperature damage on intercropping P. ostii under C. illinoinensis with 4 m × 4 m rowing spaces was more significant than that under C. illinoinensis with 4 m × 8 m rowing spaces. This research provides some reference values for efficient plantation of P. ostii in the middle and lower reaches of the Yangtze River in China. [ABSTRACT FROM AUTHOR]

Tree peony is a famous flower plant in China, but the short and concentrated flowering period limits its ornamental value and economic value. Brassinolide (BR) plays an important role in plant growth and development including flowering. There have been a large number of reports on the molecular aspects of the flowering process, but the genetic mechanism that was responsible for miRNA-guided regulation of tree peony is almost unclear. In this study, the leaves of tree peony cultivar, ‘Feng Dan’, were sprayed with different concentrations of BR, and the obvious bloom delay was found at the treatment with BR 50 μg/L. The small RNA sequencing and transcriptome sequencing were performed on the petals of tree peony under an untreated control (CK) and the treatment with BR 50 μg/L during four consecutive flowering development stages. A total of 22 known miRNAs belonging to 12 families were identified and 84 novel miRNAs were predicted. Combined with transcriptome data, a total of 376 target genes were predicted for the 18 differentially expressed known miRNAs and 177 target genes were predicted for the 23 differentially expressed novel miRNAs. Additionally, the potential miRNAs and their target genes were identified, including miR156b targeting SPL, miR172a_4 targeting AP2 and four novel miRNAs targeting SPA1, and revealed that they might affect the flowering time in tree peony. Collectively, these results would provide a theoretical basis for further analysis of miRNA-guided regulation on flowering period in tree peony.

Tree peony (Paeonia ostii) is an important ornamental plant with high oil and medical values and is widely distributed in China. In order to explore the effects of sucrose treatment on the growth of the P. ostii seeds and identify key genes involved in sucrose metabolism and transport during the early development of the P. ostii seeds, the comparative transcriptome sequencing of the P. ostii seed coats and endosperms at 50 days after flowering (DAF) with and without sucrose supply were conducted using Illumina HiSeq platform. A total of 97,976 unigenes were obtained and annotated with seven public function protein databases. There were 22,779, 22,632, 16,700, and 36,233 differentially expressed genes (DEGs) identified in the four comparisons, respectively. A total of 77 DEGs associated with the sucrose metabolism and transporter were identified in this study. Among them, sucrose transporter protein (SUT) and sweet sugar transporter (SWEET) were predicted to participate in sucrose transport from seed coat to embryo, and invertase (INV) was identified to be involved in sucrose degradation. In addition, sucrose synthase (SUS) and sucrose phosphate synthase (SPS) related to sucrose synthesis were highly expressed in seeds with sucrose supply. Finally, nine differentially expressed genes were selected randomly to analyze their expression levels using qRT-PCR, which were consistent with the transcriptome sequencing results. Above all, these results and analyses provide valuable gene resources and directions for further studying the effects of sucrose supply on the molecular mechanism of the sucrose metabolism and transporter during the development of P. ostii seeds. [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.

Paeonia ostii is an important woody oil crop mainly cross-pollinated. However, the low yield has become an important factor restricting the industrial development of P. ostii. Cross-pollination has become one of the important measures to increase the seed yield. Therefore, conservation of pollen with high vitality is crucial to ensure successful pollination of P. ostii. In this study, we found an effective methodological system to assess the viability, ability to germinate, and optimal storage conditions of P. ostii pollen grains. The optimal medium in vitro was 50 g/L sucrose, 100 mg/L boric acid, 50 g/L PEG6000, 100 mg/L potassium nitrate, 300 mg/L calcium nitrate, and 200 mg/L magnesium sulfate at pH 5.4. Optimal germination condition in vitro was achieved at 25 °C for 120 min, allowing easy observation of the germination percentage and length of the pollen tubes. In addition, the viability of pollen grains was assessed by comparing nine staining methods. Among them, MTT, TTC, benzidine-H2O2, and FDA were effective to distinguish between viable and non-viable pollen, and the results of the FDA staining method were similar to the pollen germination percentage in vitro. After evaluation of pollen storage, thawing and rehydration experiments showed that thawing at 4 °C for 30 min and rehydration at 25 °C for 30 min increased the germination percentage of pollen grains stored at low temperatures. The low-temperature storage experiments showed that 4 °C was suitable for short-term storage of P. ostii pollen grains, while −80 °C was suitable for long-term storage. This is the first report on the in vitro germination, viability tests, and storage of P. ostii pollen grains, which will provide useful information for P. ostii germplasm conservation and artificial pollination.

Tree peony (sect. Moutan) is a kind of Traditional Chinese Medicine and ornamental plant, which has been widely cultivated and utilized for thousands of years. To further study the active components of Paeonia ostii (Moutan , Fengdan), six fractions (soluble free (F), soluble esterification, soluble glycosylation, insoluble bound, insoluble esterification and insoluble glycosylation) were extracted from the leaves and roots by alkaline and acid treatment for the first time. Twenty-one typical compounds were identified and quantified by HPLC-MS. The results showed that total phenolic content (TPC) in peony roots (PR) and peony leaves (PL) were as high as 125.48 and 280.38 mg GAE·g−1 dw, which maximizes the extraction efficiency of phenolic compounds, especially leaves, compared with the conventional method. PR-F and PL-F had the highest TPC, antioxidant and anti-tyrosinase activities. Paeoniflorin was the main compounds in PL and PR. It and pentagalloylglucose (PGG) almost reached the anti-tyrosinase level of kojic acid, but they showed different inhibitory mechanisms by molecular docking. On the whole, PR-F, PL-F, PGG and paeoniflorin might be potential for skin whitening products. [Display omitted] • Paeonia ostii leaves contained high polyphenol may be underestimated for a long time. • PGG and paeoniflorin almost reached the tyrosinase inhibitory strength of kojic acid. • PR-F and PL-F had the highest TPC, antioxidant and anti-tyrosinase activities. • PR-F, PL-F, PGG and paeoniflorin might be potential materials for whitening products. • PGG, paeoniflorin and kojic acid show diverse anti-tyrosinase mechanisms by docking. [ABSTRACT FROM AUTHOR]

Paeoniflorin biosynthesis, O-benzoyltransferase, monoterpene, Paeonia ostii, BAHD acyltransferase Keywords: O-benzoyltransferase; paeoniflorin biosynthesis; monoterpene; Paeonia ostii; BAHD acyltransferase EN O-benzoyltransferase paeoniflorin biosynthesis monoterpene Paeonia ostii BAHD acyltransferase 14 16 3 01/11/23 20230101 NES 230101 PoDPBT, an O-benzoyltransferase belonging to the BAHD family, can catalyze the benzoylation of 8-debenzoylpaeoniflorin to paeoniflorin. [Extracted from the article]

The Paeonia ostii , also known as "Feng Dan" have a crucial role in folk medicine to treat lumbar muscles strain, knee osteoarthritis and cervical spondylosis. In this study, four new phenolic compounds, specifically Paeoniaostiph A-E ( 1 - 4 ) phenolic compounds were characterised through spectroscopic techniques, including 1D and 2D NMR, HRESIMS, UV, IR, and electronic circular dichroism computations to explore their structures. Cytotoxicity and NO production inhibition of the new phenolic compounds were also studied. The results of the cytotoxicity experiment showed that compound 1 is cytotoxic to two human cancer cell lines with IC 50 values ranging from 13.3 to 13.5 μM. Compounds 1 and 2 showed certain inhibitory activity on NO production. This is the first report on isolating the components from natural sources.

Paeonia ostii is an emerging woody oil crop, but the high temperature in summer is extremely unfavorable for its growth and development. Understory intercropping cultivation would provide shaded environments which could effectively reduce the ambient temperature. In order to explore whether understory intercropping cultivation would reduce the effects of high-temperature stress on P. ostii, the changes of leaf physiological indicators and leaf microstructures of sole-cropping and intercropping P. ostii were investigated. P. ostii that intercropping cultivated under Illinois pecans (Carya illinoinensis (Wangenh.) K. Koch) with 4 m × 4 m and 4 m × 8 m rowing spaces were used as samples in this study. The results showed that with continuous high temperature, the high-temperature damage index of P. ostii kept increasing, whereas the leaf relative water content continued to decline. Compared to sole-cropping, the high-temperature damage index, relative electrical conductivity, proline content, antioxidant enzymes superoxide dismutase (SOD), and ascorbate peroxidase (APX) activities of intercropping P. ostii under C. illinoinensis were significantly decreased, whereas the leaf relative water content was higher. Moreover, compared to sole-cropping, intercropping P. ostii under C. illinoinensis increased SPAD and chlorophyll contents, made mesophyll cell ultrastructures more intact, and made the chloroplasts rounder and more filled with starch granules and lipid globules, leading to enhanced photosynthesis (Pn) and transpiration rates (Tr). Notably, the reduction of high-temperature damage on intercropping P. ostii under C. illinoinensis with 4 m × 4 m rowing spaces was more significant than that under C. illinoinensis with 4 m × 8 m rowing spaces. This research provides some reference values for efficient plantation of P. ostii in the middle and lower reaches of the Yangtze River in China.

Tree peony (Paeonia ostii) is an ornamental flowering plant that is generally recalcitrant to establishment of a mature somatic embryo regeneration system in vitro. Glucose-6-phosphate translocator (GPT) plays an important regulatory role in embryogenesis of plants. In this study, PoGPT1 was cloned, and a bioinformatic analysis and functional verification of the gene were performed. The results showed that PoGPT1 encoded a polypeptide of 392 amino acids, which was a basic non-secreted hydrophobic transmembrane protein, and was mainly localized in the plastids. PoGPT1 was highly expressed in tree peony leaves, and its transcript abundance increased with the progression of zygotic embryo development. Overexpression of PoGPT1 caused up-regulation of leafy cotyledon 1 (PoLEC1), somatic embryogenesis receptor-like kinase (PoSERK), and agamous-like15 (PoAGL15) in tree peony callus. In addition, PoGPT1 overexpression promoted the increase in indole-3-acetic acid (IAA), 5-deoxystrigol (5DS), and brassinolide (BL) contents, especially of IAA, but reduced the contents of abscisic acid (ABA), 6-benzyladenosine (BARP), and 1-aminocyclopropanecarboxylic acid (ACC). The present research showed that PoGPT1 synergistically regulated the contents of endogenous hormones and expression levels of embryogenesis-related genes to promote the embryonic development of tree peony. The results provide theoretical and technical support for the establishment of a tree peony embryogenic callus formation and subsequent research on somatic embryogenesis.

Background The tree peony (Paeonia section Moutan DC), one of the traditional famous flowers with both ornamental and medicinal value, was widely used in China. Surprisingly little is known about the full-length transcriptome sequencing in tree peony, limiting the research on its gene function and molecular mechanism. The trehalose phosphate phosphatase (TPS) family genes has been found to affect plant growth and development and the function of TPS genes in Paeonia ostii is unknown. Methods In our study, we performed single molecule, full-length transcript sequencing in P. ostii. 10 TPS family members were identified from PacBio sequencing for bioinformatics analysis and transcriptional expression analysis. Results A total of 230,736 reads of insert (ROI) sequences and 114,215 full-Length non-chimeric reads (FLNC) were obtained for further ORFs and transcription factors prediction, SSR analysis and lncRNA identification. NR, Swissprot, GO, COG, KOG, Pfam and KEGG databases were used to obtain annotation information of transcripts. 10 TPS family members were identified with molecular weights between 48.0 to 108.5 kD and isoelectric point between 5.61 to 6.37. Furthermore, we found that TPS family members contain conserved TPP or TPS domain. Based on phylogenetic tree analysis, PoTPS1 protein was highly similar to AtTPS1 protein in Arabidopsis. Finally, we analyzed the expression levels of all TPS genes in P. ostii and found PoTPS5 expressed at the highest level. In conclusion, this study combined the results of the transcriptome to systematically analyze the 10 TPS family members, and sets a framework for further research of this important gene family in development of tree peony.

Background: Paeonia ostii seeds were identified as novel sources of edible plant oil with a high proportion of α-linolenic acid, a type of n-3 fatty acid with many health benefits. Due to the unreliability of seed oil content and quality, it is necessary to discover the mechanism underlying lipid biosynthesis in Paeonia ostii seeds. Objectives: This study aimed to identify the key genes involved in lipid biosynthesis in Paeonia ostii seeds by analyzing the relationship among the seed characteristics and the expression patterns of lipid genes in Paeonia ostii during seed development. Methods: Preliminary research on Paeonia ostii seed development was carried out from 10 days after pollination until maturity, focusing on phenology, oil content and lipid profiles. In addition, we investigated the spatiotemporal expression of 36 lipid biosynthetic genes in Paeonia ostii by using quantitative real-time PCR. Results: The results suggested that the development of Paeonia ostii seeds from pollination to maturity could be divided into three periods. The 36 lipid genes showed various spatiotemporal expression patterns and five gene groups with distinct temporal patterns during seed development were identified by clustering analysis of expression data. Furthermore, the relationships between gene expression and lipid/fatty acid accumulation and some candidate key lipid genes were discussed. Conclusions: This study provided the global patterns of fatty acid and lipid biosynthesis-related gene expression, which are critical to understanding the molecular basis of lipid biosynthesis and identifying the lipid accumulation rate-limiting genes during seed development. [ABSTRACT FROM AUTHOR]

• A drought-related bZIP transcription factor PobZIP4 was identified in paeonia ostii. • PobZIP4 positively responded to drought stress with enhanced expression. • PobZIP4 enhanced drought tolerance in plants by scavenging ROS. Drought, as a natural disaster of force majeure, significantly disrupts the physiological balance of plants and crop yields. The basic leucine zipper (bZIP) transcription factors assume the pivotal role of paramount regulators in orchestrating drought response. In this study, a bZIP transcription factor PobZIP4 was isolated on the base of Paeonia ostii transcriptome, and its function in response to drought stress was verified. PobZIP4 had a 579 bp open reading frame, encoded 192 amino acids, and existed in a conserved structural domain typical of bZIP, and was highly expressed when P. ostii was stressed by drought. Moreover, the subcellular localization of PobZIP4 was observed with fluorescent protein labeling method and it was found that PobZIP4 localized in the Nicotiana benthamiana epidermal cell nucleus. Then, we silenced PobZIP4 in P. ostii using virus-induced gene silencing technology, and found that PobZIP4 -silenced plants showed significantly less tolerant to drought stress than the negative control. The accumulation of reactive oxygen species was found to be heightened, while the photosynthetic capacity, proline content, and protective enzyme activities exhibited a significant decline upon silencing PobZIP4 , which unequivocally demonstrated that PobZIP4 amplifies the drought tolerance of P. ostii. This study highlighted the positive role of PobZIP4 in mitigating drought stress in P. ostii , thereby establishing a molecular basis for enhancing the drought tolerance of P. ostii. [ABSTRACT FROM AUTHOR]

Tree peony seeds oil (TPSO, Paeonia ostii) extracted at different roasting temperatures were studied for color, acid values, total phenols, fatty acid composition, antioxidant capacity, volatiles, and benzoic acid. Roasting led to increased antioxidant capacity and acid values. Headspace analysis of raw peony seed oil showed only six volatile compounds, while after roasting volatiles such as pyrazines, aldehydes and furans were observed and contributed to nutty aromas. Unsaturated fatty acids constituted more than 90% of the total fatty acids with α-linolenic acid as the predominant fatty acid of TPSO and was not significantly changed with roasting temperature. Benzoic acid concentration increased significantly as roasting temperature increased. Although the occurrence of benzoic acid in virgin oil might be a concern, it can be removed through refining for commercial production.

Tree peony (Paeonia ostii) is an important ornamental plant with high oil and medical values and is widely distributed in China. In order to explore the effects of sucrose treatment on the growth of the P. ostii seeds and identify key genes involved in sucrose metabolism and transport during the early development of the P. ostii seeds, the comparative transcriptome sequencing of the P. ostii seed coats and endosperms at 50 days after flowering (DAF) with and without sucrose supply were conducted using Illumina HiSeq platform. A total of 97,976 unigenes were obtained and annotated with seven public function protein databases. There were 22,779, 22,632, 16,700, and 36,233 differentially expressed genes (DEGs) identified in the four comparisons, respectively. A total of 77 DEGs associated with the sucrose metabolism and transporter were identified in this study. Among them, sucrose transporter protein (SUT) and sweet sugar transporter (SWEET) were predicted to participate in sucrose transport from seed coat to embryo, and invertase (INV) was identified to be involved in sucrose degradation. In addition, sucrose synthase (SUS) and sucrose phosphate synthase (SPS) related to sucrose synthesis were highly expressed in seeds with sucrose supply. Finally, nine differentially expressed genes were selected randomly to analyze their expression levels using qRT-PCR, which were consistent with the transcriptome sequencing results. Above all, these results and analyses provide valuable gene resources and directions for further studying the effects of sucrose supply on the molecular mechanism of the sucrose metabolism and transporter during the development of P. ostii seeds.

Chalcone synthase (CHS) is the key enzyme in the flavonoid biosynthetic pathway and has been studied in many plants, but the function of the CHS gene has not been well characterized in Paeonia ostii. In this study, we obtained a CHS homolog gene from P. ostii, which possessed the putative conserved amino acids of chalcone synthase by multiple alignment analysis and demonstrated the highest expression in developing seeds. In vitro assays of the recombinant PoCHS protein confirmed enzymatic activity using malonyl‐CoA and 4‐coumaroyl‐CoA as substrates, and the optimal pH and reaction temperature were 7.5 and 40 °C, respectively. Furthermore, ectopic over‐expression of PoCHS in Arabidopsis up‐regulated the expression levels of genes involved in seed development (ABI), glycolysis (PKp2, PDH‐E1a, and SUS2/3), and especially fatty acid biosynthesis (BCCP2, CAC2, CDS2, FatA, and FAD3). This resulted in an increased unsaturated fatty acid content, especially α‐linolenic acid, in transgenic Arabidopsis seeds. In this study, we examined the functions of CHS homolog of P. ostii and demonstrated its new function in seed fatty acid biosynthesis. [ABSTRACT FROM AUTHOR]

MicroRNA (miRNA) belongs to a class of non-coding small RNAs that post-transcriptionally regulate gene expression. In this study, we employed transcriptome, small RNA, and degradome sequencing to systematically investigate the miRNAs in Paeonia ostii. In total, 113 novel miRNA hairpin precursors were predicted, and 123 novel potential miRNAs were identified. Among them, five novel miRNAs and 23 known miRNAs were verified. Expression analyses further reveal that the levels of Pos-miR121, Pos-miR156, and Pos-miR 393 were negatively correlated with the age of Paeonia ostii in leaves, whereas those of Pos-miR159, Pos-miR165, Pos-miR166, Pos-miR172, and Pos-miR319 were gradually increased with age, implying that these miRNAs may play important roles in the acquisition of the floral competence in Paeonia ostii. Taken together, our results provide an important resource for further studies of the molecular basis of floral transition in Paeonia ostii. [ABSTRACT FROM AUTHOR]

Many factors may affect reproduction of animal‐pollinated species. In this study, the effects of pollen limitation, attractive traits (flower number, plant height and flower width) and flowering phenological traits (flowering onset, duration and synchrony) on female reproduction, as well as the patterns of variation in fruit and seed production within plants, were investigated in Paeonia ostii "Feng Dan" over two flowering seasons (2018 and 2019). Fruit set was very high (90%), and pollen supplementation did not increase fruit and seed production in either year, indicating no pollen limitation. Fruit set, ovule number per fruit and mean individual seed weight per fruit were not affected by any of the six attractive and phenological traits in either year, whereas seed number per fruit was related to the three attractive traits in one or both years. Seed number per plant was positively affected by the three attractive traits and best explained by flower number in both years, but the effect of each of the three phenological traits on seed number per plant differed between years. Within plants, the fruit set, ovule number, seed set and seed number per fruit declined from early‐ to late‐opening flowers, presumably because of resource preemption, but the mean individual seed weight did not vary across the flowering sequence. Our study shows that attractive traits of Paeonia ostii "Feng Dan" are more important than flowering phenological traits in the prediction of total seed production per plant. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Henan Agricultural Sciences is the property of Editorial Board of Journal of Henan Agricultural Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Tree peony seeds are rich in α-linolenic acid (ALA), and the peony seed oil is now being produced in China. Paeonia ostii is the most widely used tree peony species for oil extraction, which is commercially called Fengdan and treated as a single cultivar. Here, 50 P. ostii individuals from the same population in northern China were randomly selected for fatty acids (FAs) analysis. Thirteen FAs were isolated, and the most abundant five were palmitic acid (5.31–6.99%), stearic acid (1.22–2.76%), oleic acid (18.78–28.15%), linoleic acid (11.86–26.10%), and ALA (41.11–57.51%). There were significant individual differences of plants in FA quality and quantity and the linoleic acid content in Plant No. 48 even exceeded the scope of 1–99%. Further statistical analysis indicated that most of the individual FAs, saturated FAs, unsaturated FAs, and total FAs levels showed significant positive correlations to each other, whereas the seed yield per plant was independent and not correlated to the factors mentioned above. Ward's hierarchical clustering results grouped the 50 plants into four clusters based on FA contents and seed yield, and the seven plants in Cluster IV were identified as good candidates for oil production. Our results confirmed that the individual differences did occur in P. ostii and Fengdan cannot be simply treated as one uniform cultivar. Also, these results may help simplify the selection of plants for oil peony breeding and accelerate the development of the oil peony industry.

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Tree peony is a perennial deciduous shrub with great ornamental and medicinal value. A limitation of its current functional genomic research is the lack of effective molecular genetic tools. Here, the first application of a Tobacco rattle virus (TRV)-based virus-induced gene silencing (VIGS) in the tree peony species Paeonia ostii is presented. Two different approaches, leaf syringe-infiltration and seedling vacuum-infiltration, were utilized for Agrobacterium-mediated inoculation. The vacuum-infiltration was shown to result in a more complete Agrobacterium penetration than syringe-infiltration, and thereby determined as an appropriate inoculation method. The silencing of reporter gene PoPDS encoding phytoene desaturase was achieved in TRV-PoPDS-infected triennial tree peony plantlets, with a typical photobleaching phenotype shown in uppermost newly-sprouted leaves. The endogenous PoPDS transcripts were remarkably down-regulated in VIGS photobleached leaves. Moreover, the green fluorescent protein (GFP) fluorescence was detected in leaves and roots of plants inoculated with TRV-GFP, suggesting the capability of TRV to silence genes in various tissues. Taken together, the data demonstrated that the TRV-based VIGS technique could be adapted for high-throughput functional characterization of genes in tree peony.

Two new glycosides, ethyl- O-β-D -furanosyl-(1→6)- O-β-D- glucopyranoside ( 1 ) and (5-'')-galloyl-ethyl- O-β-D -furanosyl-(1→6) -O-β-D -glucopyranoside ( 2 ), together with eight known compounds ( 3 - 10 ) were obtained from the n -BuOH extraction of Paeonia ostii . Their structures were identified via the extensive spectroscopic analysis. Compounds 1 , 3-10 exhibited the anti-inflammation activities, which inhibited the production of NO, TNF- α and IL-1 β in LPS-induced RAW264.7 cells with IC 50 values ranging from 6.00 to 86.78 µ Μ.