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(1) Tree peony (Paeonia suffruticosa Andrews) is a woody ornamental plant originating from China, and beloved by people worldwide. Non-structural carbohydrates (NSCs) play a crucial role in regulating the flowering quality of tree peonies in both field and potted conditions. However, the effects of NSCs accumulation and allocation in various organs during the vegetative growth stage on the flowering quality of tree peony under forcing culture remains unclear. (2) Two-year-old grafted seedlings of tree peony cv. 'Luoyanghong' were subjected to orthogonal treatments to investigate the role of NSCs accumulation in plants' developmental process. We measured leaf photosynthetic capacity, NSCs accumulation in the organs of seedlings, observed key ornamental characteristics of flowering quality under forcing culture conditions, and evaluated the qualities of seedlings and flowers using the seedling index (SI) and flowering index (FI), respectively. (3) There was a significant positive correlation between leaf photosynthetic capacity and NSCs accumulation in both the whole plant and roots of potted tree peony. Roots were identified as the primary organs for NSCs accumulation in potted tree peonies. Sufficient NSCs accumulation in the plant, particularly in the roots during the defoliation period, was essential not only for enhancing the seedling quality of potted tree peonies but also for improving the flowering quality under forcing culture conditions. Both the seedling index (SI) and flowering index (FI) exhibited a significant dose-response with increasing root NSCs accumulation at defoliation. The T3 group, which involved slight root pruning (by 25%), combined with a high-concentration rooting agent (750 mg·L−1) and Metarhizium anisopliae (20 million U·mL−1), resulted in the highest photosynthetic capacity, SI, FI and NSCs accumulation status (NSCAR), making it the optimal treatment combination. (4) This finding indicates that increasing NSCs accumulation in the roots of potted tree peonies is a crucial biological foundation for producing high-quality potted flowers under forcing culture conditions, which provide new insights into the importance of NSCs in tree peony flowering and may improve the production technology for high-quality potted tree peony flowers under forcing culture conditions. [ABSTRACT FROM AUTHOR]

The tree peony (Paeonia ostii), a newly recognized woody oil plant endemic to China, is noteworthy for its high content of unsaturated fatty acids (UFA), particularly alpha-linolenic acid (ALA). Fatty acid desaturases (FADs) are integral to plant development and defense mechanisms. Nonetheless, there is limited understanding of (i) the molecular mechanism underlying FA biosynthesis in various varieties during seed maturation and (ii) a genome-wide analysis of FAD family genes within the tree peony. We selected three distinct cultivars of tree peony for transcriptome sequencing and performed an extensive analysis of PoFAD genes. In total, 67,542 unigenes were acquired and annotated with six protein databases available to the public. Forty-one differentially expressed genes (DEGs) pertinent to FA biosynthesis and lipid metabolism were identified in this study. Notably, genes such as PoFAD2, PoFAD6, and PoSAD were found to be significantly upregulated, contributing to a differential linolenic acid and linoleic acid content across the three cultivars. Herein, 24 PoFADs from the P. ostii genome were recognized and categorized into four distinct clusters according to their conserved structural features. The distribution of PoFADs was found to be random and uneven across five chromosomes, indicating a complex genomic architecture. Six colinear gene pairs were found between P. ostii and V. vinifera, indicating a potential link due to their close relationship. Together, these findings significantly enhance our knowledge of the molecular processes governing fatty acid synthesis, elucidate the functional roles of the FAD gene family, and lay the groundwork for using genetic manipulation to boost lipid levels. [ABSTRACT FROM AUTHOR]

Peony is a flower species with great ornamental value. Germplasm innovation is urgently needed due to the rapid development of peony industrialization around the world. However, the occurrence of seed abortion restricts the hybrid breeding programs of peony, significantly impeding the development of new cultivars. Hence, gaining a comprehensive understanding of the molecular mechanism of seed abortion in peony will provide opportunities to overcome this challenge of seed abortion in hybrid breeding programs for peony. In this study, we conducted physiological and transcriptomic analyses on peony seeds at three distinct time points (14d, 18d, and 28d) after pollination, aiming to investigate the mechanism underlying seed abortion. The results indicated that the regulation of seed abortion in peony involves multiple metabolic pathways, which are regulated by a plenty of differentially expressed genes. Specifically, we successfully identified 25 differentially expressed genes involved in sucrose catabolism as well as starch synthesis and catabolism, indicating sucrose and starch metabolic pathways play significant roles in the occurrence of seed abortion during peony hybridization. Six genes associated with sucrose transport and catabolism consistently exhibited the same expression pattern across different time points. This suggested that the modifications in enzyme activities contribute to the change of starch and sucrose contents during seed development. Additionally, the activities of peroxidase and superoxide dismutase changed during seed abortion. Certain proteins encoded by genes associated with starch and sucrose metabolism may potentially interact with SOD-related proteins. However, the precise mechanisms of these interactions require further validation. Finally, we identified several transcription factor families that are also implicated in seed abortion, highlighting the intricate gene regulatory network associated with this process. These results provide a theoretical foundation for determining the optimal period of seed embryo salvage and present a novel approach to enhance the seed set rate of 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]

Background: Black spot disease in tree peony caused by the fungal necrotroph A. alternata, is a primary limiting factor in the production of the tree peony. The intricate molecular mechanisms underlying the tree peony resistance to A. alternata have not been thoroughly investigated. Results: The present study utilized high-throughput RNA sequencing (RNA-seq) technology to conduct global expression profiling, revealing an intricate network of genes implicated in the interaction between tree peony and A. alternata. RNA-Seq libraries were constructed from leaf samples and high-throughput sequenced using the BGISEQ-500 sequencing platform. Six distinct libraries were characterized. M1, M2 and M3 were derived from leaves that had undergone mock inoculation, while I1, I2 and I3 originated from leaves that had been inoculated with the pathogen. A range of 10.22–11.80 gigabases (Gb) of clean bases were generated, comprising 68,131,232 − 78,633,602 clean bases and 56,677 − 68,996 Unigenes. A grand total of 99,721 Unigenes were acquired, boasting a mean length of 1,266 base pairs. All these 99,721 Unigenes were annotated in various databases, including NR (Non-Redundant, 61.99%), NT (Nucleotide, 45.50%), SwissProt (46.32%), KEGG (Kyoto Encyclopedia of Genes and Genomes, 49.33%), KOG (clusters of euKaryotic Orthologous Groups, 50.18%), Pfam (Protein family, 47.16%), and GO (Gene Ontology, 34.86%). In total, 66,641 (66.83%) Unigenes had matches in at least one database. By conducting a comparative transcriptome analysis of the mock- and A. alternata-infected sample libraries, we found differentially expressed genes (DEGs) that are related to phytohormone signalling, pathogen recognition, active oxygen generation, and circadian rhythm regulation. Furthermore, multiple different kinds of transcription factors were identified. The expression levels of 10 selected genes were validated employing qRT-PCR (quantitative real-time PCR) to confirm RNA-Seq data. Conclusions: A multitude of transcriptome sequences have been generated, thus offering a valuable genetic repository for further scholarly exploration on the immune mechanisms underlying the tree peony infected by A. alternata. While the expression of most DEGs increased, a few DEGs showed decreased expression. [ABSTRACT FROM AUTHOR]

An important family of transcription factors (TFs) in plants known as NAC (NAM, ATAF1/2, and CUC2) is crucial for the responses of plants to environmental stressors. In this study, we mined the NAC TF family members of tree peony (Paeonia suffruticosa Andrews) from genome-wide data and analyzed their response to heat and waterlogging stresses in conjunction with transcriptome data. Based on tree peony's genomic information, a total of 48 PsNAC genes were discovered. Based on how similar their protein sequences were, these PsNAC genes were divided into 14 branches. While the gene structures and conserved protein motifs of the PsNAC genes within each branch were largely the same, the cis-acting elements in the promoter region varied significantly. Transcriptome data revealed the presence of five PsNAC genes (PsNAC06, PsNAC23, PsNAC38, PsNAC41, PsNAC47) and one PsNAC gene (PsNAC37) in response to heat and waterlogging stresses, respectively. qRT-PCR analysis reconfirmed the response of these five PsNAC genes to heat stress and one PsNAC gene to waterlogging stress. This study lays a foundation for the study of the functions and regulatory mechanisms of NAC TFs in tree peony. Meanwhile, the NAC TFs of tree peony in response to heat and waterlogging stress were excavated, which is of great significance for the selection and breeding of new tree peony varieties with strong heat and waterlogging tolerance. [ABSTRACT FROM AUTHOR]

• The field cultivation of oil tree peony experienced a "medium" pollination deficit. • Excluding pollinator pollination reduced the yield of oil tree peony by 44.24%, and oil tree peony showed a high dependence on bumblebee pollination. • Supplemental bumblebee pollination increased the seed yield of oil tree peony by 82.87% and the oil yield by 10.21%, which reduced the degree of pollination defects to a lower level. • Pollination by bumblebees ensures the stability of yield and quality of oil tree peony over time. Oil tree peony is a unique woody oil crop found exclusively in China. Its seed oil is a highly nutritious edible vegetable oil. However, its seed production varies greatly and is often low. Oil tree peony is a plant that relies on insects for pollination, but inadequate pollination is a common occurrence. In this study, we focused on the Paeonia ostii 'Fengdan' species and conducted a five-year study (from 2017 to 2022) in Luoyang, China. Our objective was to assess the extent of pollination deficiency and dependence, as well as quantify the contribution of supplemented bumblebees (Bombus terrestris) to the yield of oil tree peony. We also examined the effect of supplemented bumblebees on the temporal variation in yield through various pollinator survey designs, including supplemented bumblebees (BP), open pollination (OP), pollinator exclusion (EP), self-pollination treatments (SP), and artificial cross-pollination (CP). The results revealed a medium pollination deficit (D = 0.50) at the study sites for three consecutive years from 2020 to 2022, indicating limited pollination for oil tree peony under natural conditions. Importantly, BP significantly compensated for this deficiency (D = 0.22). Over the course of five years, BP increased the yield of oil tree peony by 82.87% on average compared to EP. Furthermore, the yield variation coefficient of EP (CVt=0.20) increased by 100% compared to BP (CVt=0.10). And we also observed significant variability in other yield parameters. Additionally, oil tree peony production was reduced by 44.24% in the absence of flower visitors, and the dependence on BP reached a "high" level. The fatty acid content in the seed oil did not show significant differences across treatments, indicating its stability. Overall, our pollination practices demonstrate that supplementing bee pollination can provide sufficient and stable pollination services, resulting in a higher and more stable yield of oil tree peony seeds. [ABSTRACT FROM AUTHOR]

Copyright of Bulletin of Botanical Research is the property of Bulletin of Botanical Research Editorial Department 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.)

Abstract Background Black spot disease in tree peony caused by the fungal necrotroph A. alternata, is a primary limiting factor in the production of the tree peony. The intricate molecular mechanisms underlying the tree peony resistance to A. alternata have not been thoroughly investigated. Results The present study utilized high-throughput RNA sequencing (RNA-seq) technology to conduct global expression profiling, revealing an intricate network of genes implicated in the interaction between tree peony and A. alternata. RNA-Seq libraries were constructed from leaf samples and high-throughput sequenced using the BGISEQ-500 sequencing platform. Six distinct libraries were characterized. M1, M2 and M3 were derived from leaves that had undergone mock inoculation, while I1, I2 and I3 originated from leaves that had been inoculated with the pathogen. A range of 10.22–11.80 gigabases (Gb) of clean bases were generated, comprising 68,131,232 − 78,633,602 clean bases and 56,677 − 68,996 Unigenes. A grand total of 99,721 Unigenes were acquired, boasting a mean length of 1,266 base pairs. All these 99,721 Unigenes were annotated in various databases, including NR (Non-Redundant, 61.99%), NT (Nucleotide, 45.50%), SwissProt (46.32%), KEGG (Kyoto Encyclopedia of Genes and Genomes, 49.33%), KOG (clusters of euKaryotic Orthologous Groups, 50.18%), Pfam (Protein family, 47.16%), and GO (Gene Ontology, 34.86%). In total, 66,641 (66.83%) Unigenes had matches in at least one database. By conducting a comparative transcriptome analysis of the mock- and A. alternata-infected sample libraries, we found differentially expressed genes (DEGs) that are related to phytohormone signalling, pathogen recognition, active oxygen generation, and circadian rhythm regulation. Furthermore, multiple different kinds of transcription factors were identified. The expression levels of 10 selected genes were validated employing qRT-PCR (quantitative real-time PCR) to confirm RNA-Seq data. Conclusions A multitude of transcriptome sequences have been generated, thus offering a valuable genetic repository for further scholarly exploration on the immune mechanisms underlying the tree peony infected by A. alternata. While the expression of most DEGs increased, a few DEGs showed decreased expression.

The effectiveness of oil-in-water (O/W) microemulsion (ME) in enhancing the oxidation and the storage stability of peony seed oil (PSO), its fatty acid mixtures (FAM-1, FAM-2), and α-linolenic acid (ALA) were investigated in this work. Storage stability was assessed by measuring iodine value, acid value, and fatty acid composition under varying temperature conditions devoid of oxygen. Additionally, antioxidant stability of both the oil phase and ME was evaluated by measuring the peroxide value, with oxidation kinetics modeled using the Arrhenius equation to predict shelf life. The results demonstrated that the change of iodine values for PSO, FAM-1, FAM-2 and ALA at 4, and -20 °C for 90 days was not obvious. At 25°C, the ALA content in PSO, FAM-1, FAM-2 and ALA samples decreased by 4.81%, 3.94%, 2.86%, and 0.12% after 90 days. Therefore, low temperature without oxygen environment is more suitable for the storage of oil and fatty acid. Embedding PSO in ME can effectively control the generation rate of peroxides. At 4°C, the shelf life of PSO, FAM-1 and FAM-2 encapsulated in MEs was prolonged by 28.36%, 9.89% and 4.24%, respectively. Thes results confirm that O/W ME can improve the oxidation stability and prolong the storage period of fat-soluble compounds. These findings provide valuable insights into enhancing the oxidative stability of PSO and its fatty acids, thereby facilitating the development of ME products loaded with fatty acids in the future. [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]

Summary: Tree peony (Paeonia suffruticosa) undergoes bud endodormancy, and gibberellin (GA) pathway plays a crucial role in dormancy regulation. Recently, a key DELLA protein PsRGL1 has been identified as a negative regulator of bud dormancy release. However, the mechanism of GA signal to break bud dormancy remains unknown.In this study, yeast two‐hybrid screened PsSOC1 interacting with PsRGL1 through its MADS domain, and interaction was identified using pull‐down and luciferase complementation imaging assays Transformation in tree peony and hybrid poplar confirmed that PsSOC1 facilitated bud dormancy release. Transcriptome analysis of PsSOC1‐overexpressed buds indicated PsCYCD3.3 and PsEBB3 were its potential downstream targets combining with promoter survey, and they also accelerated bud dormancy release verified by genetic analysis.Yeast one‐hybrid, electrophoretic mobility shifts assays, chromatin immunoprecipitation quantitative PCR, and dual luciferase assays confirmed that PsSOC1 could directly bind to the CArG motif of PsCYCD3.3 and PsEBB3 promoters via its MADS domain. PsRGL1–PsSOC1 interaction inhibited the DNA‐binding activity of PsSOC1. Additionally, PsCYCD3.3 promoted bud dormancy release by rebooting cell proliferation.These findings elucidated a novel GA pathway, GA‐PsRGL1‐PsSOC1‐PsCYCDs, which expanded our understanding of the GA pathway in bud dormancy release. [ABSTRACT FROM AUTHOR]

Copyright of Science & Technology of Food Industry is the property of Science & Technology of Food Industry Editorial Office 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.)

Heavy metal composite pollution, such as Cd–Pb, occurs commonly in agriculture soils around the world, potentially harming the ecological environment and human health via food chain/network migration. There are numerous strategies for treating and rehabilitating polluted farmland today, including the use of attractive plants for phytoremediation. However, it is unclear how to use attractive plants to properly treat and fully utilize Cd–Pb composite damaged farms. The combined Cd–Pb pollution of agriculture in Xieping village, Huixian county, Gansu province is increasingly prevalent. Therefore, based on the results of literature research and the actual situation of the study area, nine kinds of ornamental plants with Cd–Pb composite pollution remediation potential and comprehensive utilization value were first selected in this paper, and the greenhouse test was carried out in the Cd–Pb composite pollution farmland in the study area. Second, the bioconcentration factor (BCF), transfer coefficient (TF), and economic feasibility principle were utilized to select the best treatment plants, and a novel model of effective remediation and comprehensive exploitation of Cd–Pb composite damaged farmland was developed. The results showed that the nine tested plants grew well under the Cd–Pb combined pollution stress, and all of them showed certain tolerance. However, the biomass, Cd–Pb accumulation, enrichment and transport capacity of the different plants were significantly different. Among them, Paeonia suffruticosa Andr., Rumex acetosa L., Calendula officinalis L., Rose chinensis Jacq., Tagetes erecta L. and Impatiens balsamina L. show excellent restorative abilities. Combining the ornamental value, safety and economy of flowering plants, P. suffruticosa, R. chinensis, C. officinalis, T. erecta and I. balsamina were finally selected as the plants for the effective remediation and comprehensive utilisation of Cd–Pb composite pollution in farmland. In conclusion, this study proposes a new model for the effective remediation and comprehensive utilisation of heavy metal composite pollution in farmland by flowering plants with economic feasibility. [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]

Tissue culture is preferred for solving the shortcomings of low efficiency of conventional propagation in tree peony (Paeonia sect. Moutan), an economically important woody plant in China with various uses. Paeonia ostii ‘Feng Dan’ is one of the varieties with strong adaptability and high oil productivity. However, in vitro regeneration via organogenesis is hard to obtain. For instance, the indirect organogenesis from callus is a lengthy process, and the direct organogenesis is lacked. This study presents a protocol of direct organogenesis using cotyledon explant for in vitro regeneration of P. ostii ‘Feng Dan’, and includes two pathways. The explants need pretreated in the optimal dedifferentiation induction medium (DIM) [Murashige and Skoog (MS) medium with 2.27 µM thidiazuron (TDZ) + 5.37 µM α-naphthylacetic acid (NAA)] for 10 days, and then the cotyledons without callus induced were transferred to differentiation medium (DM) with 6 subcultures, 90 days in total. The best DM for the frequency of leaf clusters induction (66.67%) was Woody plant medium (WPM) containing 4.04 µM N-(2-chloro-4-pyridyl)-N-phenylurea (CPPU) + 4.54 µM TDZ, and for the frequency of MNs induction (41.67%) was WPM containing 2.02 µM CPPU + 2.27 µM TDZ. For two pathways, mean number of regenerated shoots (≈ 4 shoots per explant), the frequency of root induction (≈ 50%) and the frequency of survival (≈ 40%) were similar. Histological studies confirmed the process of direct organogenesis, and revealed that shoots and MNs were originated from the increased division of meristematic cells under cortical tissue, as well as from actively dividing meristematic cells around the vascular center. Moreover, shoots regenerated through MNs differentiation were originated from the epidermal and subepidermal cells. This study is supplements in the field of in vitro regeneration in tree peony, and is an improvement to current protocols of clonal micropropagation, at the same time facilitating fundamental studies of developmental biology and genetic transformation.Key message: Plant regeneration through two pathways of direct organogenesis using cotyledon explant was first presented in Paeonia ostii ‘Feng Dan’. [ABSTRACT FROM AUTHOR]

(1) Background: Tree peonies display extensive cultivar diversity due to widespread hybridization, resulting in a complex genetic architecture. This complexity complicates the selection of universal reference genes across different cultivars for qRT-PCR analyses. Paeonia suffruticosa 'Doulv', notable for its unique green blooms in China, exhibits chlorosis post-flowering and features petaloid stamens and pistils. (2) Methods: Based on published literature and RNA-seq data from 'Doulv', nine candidate reference genes—ACT (Actin), TUB (β-Tubulin), UBC (Ubiquitin Conjugating Enzyme), UBQ (Ubiquitin), UPL (Ubiquitin Protein Ligase), PP2A (Protein Phosphatase 2A), PP2C (Protein Phosphatase 2C), MBF1A (Multiprotein Bridging Factor 1A), and GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase)—were selected. Their expression stability was assessed across various tissues and developmental stages of 'Doulv' flowers using qRT-PCR, with evaluations conducted via GeNorm_v3.5, NormFinder_v20, and BestKeeper_v1.0. Gene cloning and expression analyses of PsCUC3, including its subcellular localization, were performed. (3) Results: GAPDH and ACT were identified as the most stable reference genes in petaloid stamens across various developmental stages of 'Doulv', whereas UBC and MBF1A were optimal across different tissues. Notably, specific conserved amino acids in PsCUC3 from 'Doulv' diverged from those in NAM/CUC3 proteins of other species, impacting its protein structure. PsCUC3 expression analysis revealed no correlation with chlorophyll content in petaloid stamens but an association with petaloid organ development. Furthermore, PsCUC3 was predominantly localized in the nucleus. (4) Conclusions: This study comprehensively evaluated suitable reference genes using GeNorm_v3.5, NormFinder_v20, and BestKeeper_v1.0 software, establishing a robust qRT-PCR detection system for 'Doulv' peony. These results provide a solid experimental foundation for further research on 'Doulv' peony. Building on this experimental foundation, the functional analysis of the PsCUC3 gene was conducted. The findings suggest a potential association between the PsCUC3 gene and floral morphology alterations in 'Doulv', identifying PsCUC3 as crucial for understanding the molecular mechanisms influencing floral structure in tree peonies. [ABSTRACT FROM AUTHOR]

Bud dormancy is crucial for woody perennial plants to resist low-temperature stress in winter. However, the molecular regulatory mechanisms underlying bud dormancy release are largely unclear. Here, a tree peony (Paeonia suffruticosa) transcript ARABIDOPSIS TOXICOS EN LEVADURA 33 (PsATL33), encoding a RING-H2 finger protein, was selected from previously generated RNA sequencing data of chilling-treated buds. The objective of this study is to investigate the role of PsATL33 in the regulation of cold-induced bud dormancy release. Subcellular localization assay revealed that PsATL33 was localized to the nucleus and plasma membrane. Reverse transcription-quantitative PCR analysis showed that PsATL33 was dramatically upregulated during cold-triggered bud dormancy release. Exogenous treatments with gibberellin (GA3) increased, but abscisic acid (ABA) inhibited the transcription of PsATL33. Ectopic transformation assay indicated that overexpression of PsATL33 in petunia promoted seed germination, plant growth, and axillary bud break. Silencing of PsATL33 in tree peony through virus-induced gene silencing assay delayed bud dormancy release. tobacco rattle virus (TRV)-PsATL33-infected buds exhibited reduced expression levels of dormancy break-related genes EARLY BUD-BREAK 1 (PsEBB1) and CARBOXYLESTERASE 15 (PsCXE15). Silencing of PsATL33 decreased the accumulation of bioactive GAs, GA1 and GA3, rather than ABA. Transcript levels of several genes involved in GA biosynthesis and signaling, including GA20-OXIDASE 1 (PsGA20ox1), GA3-OXIDASE 1 (PsGA3ox1), PsGA3ox3, GA2-OXIDASE 1 (PsGA2ox1), and GAINSENSITIVE 1A (PsGAI1A), were changed by PsATL33 silencing. Taken together, our data suggest that PsATL33 functions as a positive regulator of cold-induced bud dormancy release by modulating GA production. [ABSTRACT FROM AUTHOR]

The tree peony, a traditional flower in China, has a short and concentrated flowering period, restricting the development of the tree peony industry. To explore the molecular mechanism of tree peony flowering-stage regulation, PoEP1, which regulated the flowering period, was identified and cloned based on the transcriptome and degradome data of the early-flowering mutant Paeonia ostii 'Fengdan' (MU) and Paeonia ostii 'Fengdan' (FD). Through bioinformatics analysis, expression pattern analysis, and transgene function verification, the role of PoEP1 in the regulation of tree peony flowering was explored. The open-reading frame of PoEP1 is 1161 bp, encoding 386 amino acids, containing two conserved domains. PoEP1 was homologous to the EP1 of other species. Subcellular localization results showed that the protein was localized in the cell wall and that PoEP1 expression was highest in the initial decay stage of the tree peony. The overexpression of PoEP1 in transgenic plants advanced and shortened the flowering time, indicating that PoEP1 overexpression promotes flowering and senescence and shorten the flowering time of plants. The results of this study provide a theoretical basis for exploring the role of PoEP1 in the regulation of tree peony flowering. [ABSTRACT FROM AUTHOR]

The low survival rate of transplanted plantlets, which has limited the utility of tissue-culture-based methods for the rapid propagation of tree peonies, is due to plantlet dormancy after rooting. We previously determined that the auxin response factor PsARF may be a key regulator of tree peony dormancy. To clarify the mechanism mediating tree peony plantlet dormancy, PsARF genes were systematically identified and analyzed. Additionally, PsARF16a was transiently expressed in the leaves of tree peony plantlets to examine its regulatory effects on a downstream gene network. Nineteen PsARF genes were identified and divided into four classes. All PsARF genes encoded proteins with conserved B3 and ARF domains. The number of motifs, exons, and introns varied between PsARF genes in different classes. The overexpression of PsARF16a altered the expression of NCED, ZEP, PYL, GA2ox1, GID1, and other key genes in abscisic acid (ABA) and gibberellin (GA) signal transduction pathways, thereby promoting ABA synthesis and decreasing GA synthesis. Significant changes to the expression of some key genes contributing to starch and sugar metabolism (e.g., AMY2A, BAM3, BGLU, STP, and SUS2) may be associated with the gradual conversion of sugar into starch. This study provides important insights into PsARF functions in tree peonies. [ABSTRACT FROM AUTHOR]

With the changes of people's lifestyle, hyperlipidemia and hyperglycemia which were induced from a diet high in both fat and sugar have become serious health concerns. Tree peony seed oil (PSO) is a novel kind of edible oil that shows great potential in the food industry because of its high constituent of unsaturated fatty acids. Based 16S rRNA and gut untargeted metabolomics, this study elucidated that the mechanism of PSO regulating blood glucose (Glu) and lipids. The impact of PSO on gut microbiota balance and gut metabolites of mice with a high‐fat diet (HFD) was evaluated. The findings indicated that PSO decreased HFD mice's body weight and fat accumulation, ameliorating the levels of blood lipid, reduced liver fat vacuole levels. What's more PSO modulated the proportion of gut microbiota in HFD mice and enhanced the abundance of probiotics. Furthermore, untargeted metabolomic analysis revealed that PSO not only impacted the generation of short‐chain fatty acids (SCFAs) by gut microorganism and altered metabolic pathway but exerted influence on secondary bile acids (BA), amino acid metabolism, and various other metabolites. These results suggested that PSO has the potential function for mitigating HFD‐induced hyperlipidemia and hyperglycemia by regulating gut microbiota and host metabolism. [ABSTRACT FROM AUTHOR]

PIN-FORMED (PIN) proteins, which function as efflux transporters, play many crucial roles in the polar transportation of auxin within plants. In this study, the exogenous applications of auxin IAA and TIBA were found to significantly prolong and shorten the florescence of tree peony (Paeonia suffruticosa Andr.) flowers. This finding suggests that auxin has some regulatory influence in petal senescence and abscission. Further analysis revealed a total of 8 PsPINs distributed across three chromosomes, which could be categorized into two classes based on phylogenetic and structural analysis. PsPIN1, PsPIN2a-b, and PsPIN4 were separated into the "long" PIN category, while PsPIN5, PsPIN6a-b, and PsPIN8 belonged to the "short" one. Additionally, the cis-regulatory elements of PsPIN promoters were associated with plant development, phytohormones, and environmental stress. These genes displayed tissuespecific expression, and phosphorylation sites were abundant throughout the protein family. Notably, PsPIN4 displayed distinct and elevated expression levels in roots, leaves, and flower organs. Expression patterns among the abscission zone (AZ) and adjacent areas during various flowering stages and IAA treatment indicate that PsPIN4 likely influences the initiation of peony petal abscission. The PsPIN4 protein was observed to be co-localized on both the plasma membrane and the cell nucleus. The ectopic expression of PsPIN4 reversed the premature flower organs abscission in the Atpin4 and significantly protracted florescence when introduced to Col Arabidopsis. Our findings established a strong basis for further investigation of PIN gene biological functions, particularly concerning intrinsic relationship between PIN-mediated auxin polar. [ABSTRACT FROM AUTHOR]

The present study focused on water-soluble essential oil recovered from the hydrolate of ten Paeonia × suffruticosa cultivars. Thirty-seven components, mostly oxygenated compounds (94.6–99.6%), were detected by gas chromatography–mass spectrometry (GC-MS) and GC–flame ionisation detector (GC-FID). The geranic acid chemotype was discovered (in cultivar 'Lan BaoShi'). Eight key oxygenated components were analysed in silico with antidepressant targets sodium-dependent serotonin transporter (SERT), 5-hydroxytryptamine receptor 1 A (5-HT1A), and monoamine oxidase type A (MAO-A). Geraniol, nerol, citronellol, and geranic acid presented superior docking properties. Phenylethyl alcohol and 1,3,5-trimethoxybenzene were also well docked. These molecules were bound to the active sites successfully (with partial occupancy in SERT). They might increase serotonin level or mimic its effect in central nervous system. (Z)-3-Hexen-1-ol and 1-hexanol showed weak binding. The in silico analysis revealed for the first time that the key water-soluble essential oil components of P. × suffruticosa potentially targeted antidepressant targets. [ABSTRACT FROM AUTHOR]

The tree peony, a novel woody oil crop extensively cultivated in China, necessitates further investigation into artificial pollination technology to enhance seed yield. In this study, we conducted artificial pollination experiments with 6-year-old Paeonia ostii 'Feng Dan' seedings for suitable pollen sources, pollen concentration, pollination timing, and pollination frequency. By evaluating seed yields, active ingredients, and oil quality, we derived the following significant conclusions. Firstly, compared to natural pollination, artificial pollination could significantly increase the fruit diameter by 13.94–27.58%, seed yields by 35.17–58.99%, and oil content by 6.45–7.52% in tree peonies. In active ingredients, seeds produced by pollen from Hantai County significantly enhanced starch content (by 48.64%), total phenols (by 41.18%) and antioxidant capacity (by 54.39%). In oil quality, seeds produced by pollen from Heyang County exhibited the highest α-linolenic acid and total fatty acid content with enhancements of 1.68%, 7.41%, and 8.48%. Secondly, hand pollination with pure pollen significantly increased seed yield by 58.99%, total phenol content by 40.97%, antioxidant capacity by 54.39%, and oil content by 1.53% compared to natural pollination. Thirdly, pollination at 2/3 bloom range significantly increased seed number by 63.08% and yield by 45.61% compared to natural pollination. Finally, the effect of one, two, and three pollination events had no difference in seed yield. So, to summarize, applying a 100% concentration of allochthonous pollen once is recommended when the bloom range is more than two thirds. [ABSTRACT FROM AUTHOR]

Calcium plays a crucial role in plant growth and development, yet little is known about its function in endodormancy regulation. Tree peony (Paeonia suffruticosa), characterized by compound buds and large flowers, is well-known for its ornamental and medicinal value. To break bud dormancy release is a prerequisite of flowering and forcing culture, particularly during the Spring Festival. In this study, the Ca2+ chelator EGTA and Ca2+ channel blocker LaCl3 were applied, resulting in a significant delay in budburst during both chilling- and gibberellin (GA)- induced dormancy release in a dosage-dependent manner. As expected, the retardation of bud break was recovered by the supplementation of 30 mM CaCl2, indicating a facilitating role of calcium in dormancy release. Accordingly, several calcium-sensor-encoding genes including Calmodulin (CaM) and Ca2+-dependent protein kinases (CDPKs) were significantly upregulated by prolonged chilling and exogenous GAs. Ultrastructure observations revealed a decline in starch grains and the reopening of transport corridors following prolonged chilling. Calcium deposits were abundant in the cell walls and intercellular spaces at the early dormant stage but were enriched in the cytosol and nucleus before dormancy release. Additionally, several genes associated with dormancy release, including EBB1, EBB3, SVP, GA20ox, RGL1, BG6, and BG9, were differentially expressed after calcium blocking and recovery treatments, indicating that calcium might partially modulate dormancy release through GA and ABA pathways. Our findings provide novel insights into the mechanism of dormancy release and offer potential benefits for improving and perfecting forcing culture technology in tree peonies. [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. [ABSTRACT FROM AUTHOR]

The RNA-Seq and gene expression data of mature leaves under high temperature stress of Paeonia suffruticosa 'Hu Hong' were used to explore the key genes of heat tolerance of peony. The weighted gene co-expression network analysis (WGCNA) method was used to construct the network, and the main modules and core genes of co-expression were screened according to the results of gene expression and module function enrichment analysis. According to the correlation of gene expression, the network was divided into 19 modules. By analyzing the expression patterns of each module gene, Blue, Salmon and Yellow were identified as the key modules of peony heat response related functions. GO and KEGG functional enrichment analysis was performed on the genes in the three modules and a network diagram was constructed. Based on this, two key genes PsWRKY53 (TRINITY_DN60998_c1_g2, TRINITY_DN71537_c0_g1) and PsHsfB2b (TRINITY_DN56794_c0_g1) were excavated, which may play a key role in the heat shock response of peony. The three co-expression modules and two key genes were helpful to further elucidate the heat resistance mechanism of P. suffruticosa 'Hu Hong'. [ABSTRACT FROM AUTHOR]

Tree peony (Paeonia suffruticosa), a traditional Chinese cultivated flowering shrub and a prominent landscaping plant, is subject to root rot. The key symptoms of root rot include black roots and the yellowing of leaves. Biological control is an effective and eco-friendly strategy for managing this disease. This study focused on the isolation, identification, and biological control of the pathogens causing peony root rot. Utilizing tissue isolation, pathogenicity determination, and rDNA-ITS sequence analysis, Fusarium solani and Fusarium oxysporum were determined to be the primary causal agents of peony root rot. Illumina MiSeq high-throughput sequencing of 16 s-rDNA was used to investigate and characterize bacterial community structure and diversity in healthy and diseased peony roots, rhizosphere, and bulk soil. Results indicated that the presence of pathogenic fungi influences the structure of the rhizosphere bacterial community and that peony roots exhibit a strong selective effect on root bacterial colonization. Variations in the composition of the endophytic microbial community in healthy and diseased roots exceeded the variation in the rhizosphere. Proteobacteria and Actinobacteria are the dominant taxa in the rhizosphere and among root endophytes, comprising 90–96 % of the bacterial microbiota. The rhizosphere of healthy plants exhibits a significant enrichment in Proteobacteria (79.6 %) and Actinobacteria (14.9 %), while root endophytes in healthy plats exhibit enrichment in Proteobacteria (83.5 %). In contrast, the rhizosphere and root endophytes in diseased plants are abundant in Proteobacteria (69 % and 66.1 %, respectively). Notably, nine strains of biocontrol bacteria were isolated. Three synthetic bacterial consortia were then constructed based on inhibitory assays and the dissolving rates of phosphorus and potassium exhibited by the candidate bacteria. The synthetic bacterial consortia were evaluated for biocontrol and growth promotion properties. The constructed synthetic consortium with the best performance reduced the average morbidity and mortality of treated plants by 27.59 % and 55.56 %, respectively, compared to the untreated control. In summary, synthetic bacterial consortium I (SCI) exhibited the best disease control and growth-promoting effects on tree peony. Using a synthetic bacterial consortium represents a new and novel approach to the biological control of peony root rot.

Tree peony (Paeonia suffruticosa Andrews) is a well-known ornamental plant with high economic value, but the short fluorescence is a key obstacle to its ornamental value and industry development. High temperature accelerates flower senescence and abscission, but the associated mechanisms are poorly understood. In this study, the tandem mass tag (TMT) proteome and label-free quantitative ubiquitome from tree peony cut flowers treated with 20 °C for 0 h (RT0), 20 °C or 28 °C for 60 h (RT60 or HT60) were examined based on morphological observation, respectively. Totally, 6970 proteins and 1545 lysine ubiquitinated (Kub) sites in 844 proteins were identified. Hydrophilic residues (such as glutamate and aspartate) neighboring the Kub sites were in preference, and 36.01% of the Kub sites were located on the protein surface. The differentially expressed proteins (DEPs) and Kub-DEPs in HT60 vs RT60 were mainly enriched in ribosomal protein, protein biosynthesis, secondary metabolites biosynthesis, flavonoid metabolism, carbohydrate catabolism, and auxin biosynthesis and signaling revealed by GO and KEGG analysis, accompanying the increase of endogenous abscisic acid (ABA) accumulation and decrease of endogenous indoleacetic acid (IAA) level. Additionally, the expression patterns of six enzymes (SAMS, ACO, YUC, CHS, ANS and PFK) putatively with Kub modifications were analyzed by proteome and real-time quantitative RT-PCR. The cell-free degradation assays showed PsSAMS and PsACO proteins could be degraded via the 26 S proteasome system in tree peony flowers. Finally, a working model was proposed for the acceleration of flower senescence and abscission by high temperature. In summary, all results contributed to understanding the mechanism of flower senescence induced by high temperature and prolonging fluorescence in tree peony.

The tree peony (Paeonia ostii), a newly recognized woody oil plant endemic to China, is noteworthy for its high content of unsaturated fatty acids (UFA), particularly alpha-linolenic acid (ALA). Fatty acid desaturases (FADs) are integral to plant development and defense mechanisms. Nonetheless, there is limited understanding of (i) the molecular mechanism underlying FA biosynthesis in various varieties during seed maturation and (ii) a genome-wide analysis of FAD family genes within the tree peony. We selected three distinct cultivars of tree peony for transcriptome sequencing and performed an extensive analysis of PoFAD genes. In total, 67,542 unigenes were acquired and annotated with six protein databases available to the public. Forty-one differentially expressed genes (DEGs) pertinent to FA biosynthesis and lipid metabolism were identified in this study. Notably, genes such as PoFAD2, PoFAD6, and PoSAD were found to be significantly upregulated, contributing to a differential linolenic acid and linoleic acid content across the three cultivars. Herein, 24 PoFADs from the P. ostii genome were recognized and categorized into four distinct clusters according to their conserved structural features. The distribution of PoFADs was found to be random and uneven across five chromosomes, indicating a complex genomic architecture. Six colinear gene pairs were found between P. ostii and V. vinifera, indicating a potential link due to their close relationship. Together, these findings significantly enhance our knowledge of the molecular processes governing fatty acid synthesis, elucidate the functional roles of the FAD gene family, and lay the groundwork for using genetic manipulation to boost lipid levels.

Tree peony (Paeonia suffruticosa Andr.) is a traditional Chinese flower with significant ornamental and medicinal value. Its growth and development process is regulated by some internal and external factors, and the related regulatory mechanism is largely unknown. Myelocytomatosis transcription factors (MYCs) play significant roles in various processes such as plant growth and development, the phytohormone response, and the stress response. As the identification and understanding of the MYC family in tree peony remains limited, this study aimed to address this gap by identifying a total of 15 PsMYCs in tree peony and categorizing them into six subgroups based on bioinformatics methods. Furthermore, the gene structure, conservative domains, cis-elements, and expression patterns of the PsMYCs were thoroughly analyzed to provide a comprehensive overview of their characteristics. An analysis in terms of gene structure and conserved motif composition suggested that each subtribe had similarities in function. An analysis of the promoter sequence revealed the presence of numerous cis-elements associated with plant growth and development, the hormone response, and the stress response. qRT-PCR results and the protein interaction network further demonstrated the potential functions of PsMYCs in the growth and development process. While in comparison to the control, only PsMYC2 exhibited a statistically significant variation in expression levels in response to exogenous hormone treatments and abiotic stress. A promoter activity analysis of PsMYC2 revealed its sensitivity to Flu and high temperatures, but exhibited no discernible difference under exogenous GA treatment. These findings help establish a basis for comprehending the molecular mechanism by which PsMYCs regulate the growth and development of tree peony. [ABSTRACT FROM AUTHOR]

Tree peony (Paeonia suffruticosa) is a significant medicinal plant. However, the low rooting number is a bottleneck problem in the micropropagation protocols of P. ostii 'Fengdan'. The activity of superoxide dismutase (SOD) is closely related to root development. But research on the SOD gene's impact on rooting is still lacking. In this study, RNA sequencing (RNA-seq) was used to analyze the four crucial stages of root development in P. ostii 'Fengdan' seedlings, including the early root primordium formation stage (Gmfq), root primordium formation stage (Gmf), root protrusion stage (Gtq), and root outgrowth stage (Gzc). A total of 141.77 GB of data were obtained; 71,718, 29,804, and 24,712 differentially expressed genes (DEGs) were identified in the comparison groups of Gmfq vs. Gmf, Gmf vs. Gtq, and Gtq vs. Gzc, respectively. Among the 20 most highly expressed DEGs in the three comparison groups, only the CuZnSOD gene (SUB13202229, PoSOD) was found to be significantly expressed in Gtq vs. Gzc. The overexpression of PoSOD increased the number of adventitious roots and promoted the activities of peroxidase (POD) and SOD in P. ostii 'Fengdan'. The gene ADVENTITIOUS ROOTING RELATED OXYGENASE1 (PoARRO-1), which is closely associated with the development of adventitious roots, was also significantly upregulated in overexpressing PoSOD plants. Furthermore, PoSOD interacted with PoARRO-1 in yeast two-hybrid (Y2H) and biomolecular luminescence complementation (BiFC) assays. In conclusion, PoSOD could interact with PoARRO-1 and enhance the root development of tube plantlets in P. ostii 'Fengdan'. This study will help us to preliminarily understand the molecular mechanism of adventitious root formation and improve the root quality of tree peony and other medicinal plants. [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]

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BABY BOOM (BBM) is one of the members of the plant-specific APETALA2/ethylene-responsive factor (AP2/ERF) transcription factor superfamily. It acts as a key regulator of plant cell pluripotency, playing a significant role in promoting somatic embryogenesis. In this study, a BBM gene named PoBBM was screened, cloned, and identified from the third-generation full-length transcriptome data of Paeonia ostii. Its open reading frame was 2136 bp, encoding 711 amino acids. Sequence feature analysis revealed that it possessed two AP2 conserved domains and eight motifs, including bbm-1. The phylogenetic tree indicated that PoBBM clusters with AtBBM in the euANT group of the Arabidopsis AP2 family, which is most closely related to grape VvBBM and may have the same ancestry as grape. Subcellular localization demonstrated that the PoBBM protein was localized in the nucleus. Semi-quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to assess the PoBBM transcript levels during ten developmental stages of somatic embryos and in five tissue types of peonies. The results indicate that PoBBM was highly expressed in the early stages of peony somatic embryo development. The expression on 0–15 d was the highest and decreased gradually with somatic embryogenesis. The gene is almost not expressed after 40 d since somatic embryo formation. PoBBM was expressed in roots, stems, leaves, seeds, and calli, with the highest levels in seeds, followed by leaves and calli. The PoBBM protein displayed transcriptional self-activation activity, which may facilitate further research on its relationships with other proteins. The above results provide a key gene PoBBM for somatic embryogenesis in peonies, which is significant for advancing the establishment of a stable and efficient regeneration and genetic transformation system for peonies. [ABSTRACT FROM AUTHOR]

Aleuroglyphus ovatus (Acari: Acaridae) is a major pest mite of stored grains that is distributed worldwide. Paeonol, a phenolic component of the essential oil extracted from the Chinese herb Paeonia moutan, possesses a range of biological activities, including antiviral, antifungal and acaricidal activity. This study investigated the bioactivity of paeonol against A. ovatus and its effect on the activity of detoxification enzymes. The bioactivity of paeonol against A. ovatus was determined by contact, fumigation and repellency bioassays, and the mechanism was preliminarily explored via morphological observation of the color changes of mite epidermis and determination of the changing trend of some important enzymes associated with acaricidal efficacy in the mites. The results showed that the median lethal concentration (LC50) in the contact and fumigation bioassays was 9.832 μg/cm2 and 14.827 μg/cm3, respectively, and the acaricidal activity of paeonol was higher under direct contact than under fumigation. Dynamic symptomatology studies registered typical neurotoxicity symptoms including excitation, convulsion and paralysis in A. ovatus treated with paeonol. The enzyme activity of catalase (CAT), nitric oxide synthase (NOS) and glutathione-S-transferase (GST) was higher, whereas the activity of superoxide dismutase (SOD) and acetylcholinesterase (AChE) was lower, compared to the control group. CAT, NOS and GST were activated, whereas SOD and AChE activities were inhibited after paeonol intervention. Our findings suggest paeonol has potent acaricidal activity against A. ovatus and thus may be used as an agent to control the stored-product mite A. ovatus. [ABSTRACT FROM AUTHOR]

Abstract Background Tree peony (Paeonia sect. Moutan DC.) is a famous flower native to China with high ornamental, medicinal, and oil value. However, the low regeneration rate of callus is one of the main constraints for the establishment of a genetic transformation system in tree peony. By histomorphological observation, transcriptomic analysis and metabolite determination, we investigated the molecular mechanism of somatic embryogenesis after the establishment of a culture system and the induction of somatic embryo(SE) formation. Results We found that SE formation was successfully induced when cotyledons were used as explants. A total of 3185 differentially expressed genes were screened by comparative transcriptomic analysis of embryogenic callus (EC), SE, and non-embryogenic callus (NEC). Compared to NEC, the auxin synthesis-related genes GH3.6 and PCO2 were up-regulated, whereas cytokinin dehydrogenase (CKX6) and CYP450 family genes were down-regulated in somatic embryogenesis. In SE, the auxin content was significantly higher than the cytokinin content. The methyltransferase-related gene S-adenosylmethionine synthase (SAMS) and the flavonoid biosynthesis-related gene (ANS and F3’5’H) were down-regulated in somatic embryogenesis. The determination of flavonoids showed that rhoifolin and hyperoside had the highest content in SE. The results of transcriptome analysis were consistent with the relative expression of 8 candidate genes by quantitative polymerase chain reaction analysis. Conclusion The results revealed that auxin and cytokinin may play a key role in ‘Fengdan’ somatic embryogenesis. The genes related to somatic embryogenesis were revealed, which has partly elucidated the molecular mechanism of somatic embryogenesis in ‘Fengdan’.

The tree peony, a traditional flower in China, has a short and concentrated flowering period, restricting the development of the tree peony industry. To explore the molecular mechanism of tree peony flowering-stage regulation, PoEP1, which regulated the flowering period, was identified and cloned based on the transcriptome and degradome data of the early-flowering mutant Paeonia ostii ‘Fengdan’ (MU) and Paeonia ostii ‘Fengdan’ (FD). Through bioinformatics analysis, expression pattern analysis, and transgene function verification, the role of PoEP1 in the regulation of tree peony flowering was explored. The open-reading frame of PoEP1 is 1161 bp, encoding 386 amino acids, containing two conserved domains. PoEP1 was homologous to the EP1 of other species. Subcellular localization results showed that the protein was localized in the cell wall and that PoEP1 expression was highest in the initial decay stage of the tree peony. The overexpression of PoEP1 in transgenic plants advanced and shortened the flowering time, indicating that PoEP1 overexpression promotes flowering and senescence and shorten the flowering time of plants. The results of this study provide a theoretical basis for exploring the role of PoEP1 in the regulation of tree peony flowering.

In vitro pollen germination is considered the most efficient method to assess pollen viability. The pollen germination frequency and pollen tube length, which are key indicators of pollen viability, should be accurately measured during in vitro culture. In this study, a Mask R-CNN model trained using microscopic images of tree peony (Paeonia suffruticosa) pollen has been proposed to rapidly detect the pollen germination rate and pollen tube length. To reduce the workload during image acquisition, images of synthesized crossed pollen tubes were added to the training dataset, significantly improving the model accuracy in recognizing crossed pollen tubes. At an Intersection over Union threshold of 50%, a mean average precision of 0.949 was achieved. The performance of the model was verified using 120 testing images. The R 2 value of the linear regression model using detected pollen germination frequency against the ground truth was 0.909 and that using average pollen tube length was 0.958. Further, the model was successfully applied to two other plant species, indicating a good generalizability and potential to be applied widely. [ABSTRACT FROM AUTHOR]

Background: Tree peony (Paeonia sect. Moutan DC.) is a famous flower native to China with high ornamental, medicinal, and oil value. However, the low regeneration rate of callus is one of the main constraints for the establishment of a genetic transformation system in tree peony. By histomorphological observation, transcriptomic analysis and metabolite determination, we investigated the molecular mechanism of somatic embryogenesis after the establishment of a culture system and the induction of somatic embryo(SE) formation. Results: We found that SE formation was successfully induced when cotyledons were used as explants. A total of 3185 differentially expressed genes were screened by comparative transcriptomic analysis of embryogenic callus (EC), SE, and non-embryogenic callus (NEC). Compared to NEC, the auxin synthesis-related genes GH3.6 and PCO2 were up-regulated, whereas cytokinin dehydrogenase (CKX6) and CYP450 family genes were down-regulated in somatic embryogenesis. In SE, the auxin content was significantly higher than the cytokinin content. The methyltransferase-related gene S-adenosylmethionine synthase (SAMS) and the flavonoid biosynthesis-related gene (ANS and F3'5'H) were down-regulated in somatic embryogenesis. The determination of flavonoids showed that rhoifolin and hyperoside had the highest content in SE. The results of transcriptome analysis were consistent with the relative expression of 8 candidate genes by quantitative polymerase chain reaction analysis. Conclusion: The results revealed that auxin and cytokinin may play a key role in 'Fengdan' somatic embryogenesis. The genes related to somatic embryogenesis were revealed, which has partly elucidated the molecular mechanism of somatic embryogenesis in 'Fengdan'. [ABSTRACT FROM AUTHOR]

Tree peony (Paeonia suffruticosa Andr.) is a traditional Chinese flower, which prefers cool weather. However, high temperature in summer in the middle and lower reaches of the Yangtze River restricts its growth and development. In this study, osmotic regulation, antioxidant enzyme activities, and photosynthetic characteristics of tree peony in response to high-temperature stress were investigated. The results showed that high-temperature stress had destroyed the cell membrane, manifested as the increased relative electrical conductivity and malondialdehyde content. Moreover, high-temperature stress led to excessive accumulation of reactive oxygen species, thereby, activating antioxidant enzyme activities. Also, photosynthetic parameters and chlorophyll fluorescence parameters directly reflected the damage to the photosystem II reflection center under high-temperature stress. In addition, high-temperature stress led to stomatal closure and chloroplast damage. This study revealed the physiological responses of tree peony to high-temperature stress, laying a foundation for the promotion of tree peony in high-temperature areas and the improvement of high-temperature resistance. [ABSTRACT FROM AUTHOR]

Two carboxymethylated polysaccharides (CPSP30, CPSP60) were obtained from the petals of Paeonia x suffruticosa Andrews (PSP30, PSP60, respectively) with different molecular weight. Chemical characterization, morphological properties and antioxidant activities of PSP and CPSP were determined. PSP were mainly composed of galactose, glucose, fructose and galacturonic with different molar percentages. CPSP were mainly composed of arabinose, galactose, glucose with different molar percentages. The signals of carboxymethyl groups were found in Fourier transform infrared spectroscopy (FT-IR) and 13C nuclear magnetic resonance (NMR) spectra. Scanning electron microscopy (SEM) indicated that the original polysaccharides had a smooth surface, while the modified polysaccharides had an irregular lamellar structure. Thermal analysis indicated that the modified polysaccharides (CPSP30, CPSP60) presented higher thermal stability than the original polysaccharides. All polysaccharides presented typical structures of non-crystalline compounds. Carboxymethylation did not destroy the triple-helical structures of the polysaccharides. With its lower molecular weight, CPSP30 had higher scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH), OH, and Fe2+ chelating ability than PSP30. The scavenging DPPH and OH abilities of both derivatives were higher than that of original polysaccharides. This study demonstrates that carboxymethylation improved the solubilities and antioxidant abilities of the original polysaccharides, thus enhancing their potential for application in food and pharmaceutical industry. [Display omitted] • Two polysaccharides with different molecular weights from Paeonia x suffruticosa petals were obtained. • Their carboxymethylated derivatives were successfully performed. • Their carboxymethylated derivatives were more thermally stable. • The carboxymethylated derivatives had higher scavenging DPPH and OH abilities. [ABSTRACT FROM AUTHOR]

Background: Skin pigmentation is modulated by various processes, with melanogenesis playing a key role. Melanin is synthesized by the catalysis of melanogenesis‐related enzymes, such as tyrosinase and tyrosine‐related proteins TRP‐1 and TRP‐2. Paeoniflorin is the main bioactive component of Paeonia suffruticosa Andr., Paeonia lactiflora., or Paeonia veitchii Lynch and has been used for centuries for its anti‐inflammatory, anti‐oxidant, and anti‐carcinogenic properties. Aims & methods: In this study, melanin biosynthesis in mouse melanoma (B16F10) cells was induced using α‐melanocyte‐stimulating hormone (α‐MSH), and then cells were co‐treated with paeoniflorin to evaluate its potential anti‐melanogenic effect. Results: α‐MSH stimulation increased melanin content, tyrosinase activity, and melanogenesis‐related markers in a dose‐dependent manner. However, treatment with paeoniflorin reversed α‐MSH‐induced upregulation of melanin content and tyrosinase activity. Furthermore, paeoniflorin inhibited cAMP response element‐binding protein activation and TRP‐1, TRP‐2, and microphthalmia‐associated transcription factor protein expression in α‐MSH‐stimulated B16F10 cells. Conclusion: Overall, these findings show the potential of paeoniflorin as a depigmenting agent for cosmetic products. [ABSTRACT FROM AUTHOR]

Tree peony (Paeonia section Moutan DC.) is a traditional ornamental flower of China, which has thousands of varieties with different flower colors and types after a long history of natural selection and artificial breeding. However, tree peony is a perennial woody plant with a long breeding, and there are still significant challenges to accelerate the process of genetic improvement of important ornamental traits. In this study, a total of sixteen primer pairs with high polymorphism and good universality were selected from the initial pool of 115 SSR markers. The SSR marker set was derived from published papers on the genetic linkage map and association analysis of tree peony. Furthermore, we conducted a genetic diversity and population structure analysis on 322 tree peony cultivars using molecular markers with functional. These SSRs amplified a total of 391 alleles, the average number of different alleles was 5.113 alleles across all loci. The average Shannon's information index, gene diversity and polymorphism information content were 0.842, 0.532, and 0.503 over all loci, respectively. Population genetic diversity analysis indicated that the average expected heterozygosity of the total population was larger than the observed heterozygosity, showing the presence of a certain degree of heterozygous deletion phenomenon. The Japan varieties had the richest diversity with the highest H (0.508) and PIC (0.479) values. The Zhongyuan varieties showed the greatest variation may be related to its longstanding cultivation history. Moreover, the STRUCTURE and principal coordinate analyses indicated that 322 tree peony individuals from five populations were grouped into two clusters. An analysis of molecular variance demonstrated significant genetic diversity among different populations. This research may contribute to the sustainable management, conservation, and utilization of tree peony resources. [ABSTRACT FROM AUTHOR]

Tree peony is a unique oil plant resource in China, and tree peony seed oil is one of the healthy edible oils with a very promising future. However, the main oil tree peony cultivars promoted in China are Paeonia ostii 'Fengdan' and Paeonia rockii. In order to explore new oil tree peony cultivars, 68 tree peony cultivars were investigated and cultivars with oil potential were selected by cluster analysis and grey relational analysis (GRA) in this study. The results demonstrated that the 68 cultivars varied significantly in terms of agronomic characteristics (p < 0.05), with the coefficient of variation in seed yield per plant reaching a high of 75.36%. The oil content of 46 cultivars was higher than 'Fengdan' (20.87 ± 0.26%) and 'Zibanbai' (21.24 ± 1.01%), while the alpha-linolenic acids and total unsaturated fatty acid contents of 26 cultivars were higher than 'Fengdan' (39.79 ± 1.13% and 88.99 ± 0.47%) and 'Zibanbai' (40.51 ± 0.09% and 93.59 ± 0.09%). Finally, three cultivars with better integrated traits were selected by cluster analysis and grey relational analysis (GRA), comprising of 'Changshoule', 'Xianchizhenghui', and 'Yupantuojin'. The contents of alpha-linolenic acids and total unsaturated fatty acids in 'Changshoule' (47.98 ± 0.17% and 93.60 ± 0.08%), 'Xianchizhenghui' (49.44 ± 0.63% and 93.80 ± 0.06%), and 'Yupantuojin' (40.46 ± 0.26% and 93.58 ± 0.06%) were higher than that of 'Fengdan' (39.79 ± 1.13% and 88.99 ± 0.47%). In general, these cultivars can be used as hybrid parental materials for breeding new excellent oil tree peony cultivars. [ABSTRACT FROM AUTHOR]

Tree peonies are an economically important crop with flowers of high ornamental value. Most tree peony cultivars in gardens are once-flowering, and the continuous flowering (CF) trait has been revealed only in a few tree peony cultivars, such as 'High Noon' ('HN'). However, the molecular mechanism underlying its CF remains unclear. Here, we demonstrated that PsTFL1 functions as a floral inhibitor via the ectopic expression of PsTFL1 in transgenic Arabidopsis thaliana plants. Our findings suggest that PsFT and PsTFL1 interact with PsFD, and the detected interactions may occur in the nucleus. Compared with the non-CF variety, the gene expression patterns of PsFT, PsTFL1, and PsFD during the flower development indicate that these three genes may be related to the CF habit in tree peony 'HN'. These findings will aid future investigations of CF behavior and promote the breeding of tree peonies and other perennial woody plants. [ABSTRACT FROM AUTHOR]

This study aimed at chemical variability of flower essential oils of 10 well-known cultivars of Paeonia × suffruticosa Andr. The essential oil yields ranged as 0.19–0.67% (w/w). The chemical compositions varied with cultivars. 'Xueying Taohua' essential oil had predominance of citronellol (56.4%). 'Guifei Chacui' essential oil was rich in citronellol (35.9%) and phloroglucinol trimethyl ether (25.3%). 'Sheng Gejin' and 'Yuhou Fengguang' essential oils were rich in phloroglucinol trimethyl ether (36.5%, 26.8%). 'Yinhong Qiaodui' essential oil contained geraniol + nerol (23.9%). Those from the other five cultivars did not present typical chemotypes. Nevertheless several compounds were noted, mainly germacrene D, heptadeca-6,9-diene, (Z)-hex-3-en-1-ol and linalool oxide (furan). 'Luhe Hong' essential oil contained the most unique components, including α-cadinol, τ-muurolol and δ-cadinene. Lack of phenylethyl alcohol was noticeable in some essential oil samples as compared with hydrolate extracts investigated previously. Multivariate analyses revealed four distinct clusters from all the essential oil samples. [ABSTRACT FROM AUTHOR]

SUMMARY: Seed oil not only provides energy for seed postgermination development but also provides essential nutrients and raw materials for human products. However, the transcriptional regulatory mechanism controlling seed oil accumulation remains largely unknown. Tree peony (Paeonia rockii) is an emerging woody oilseed crop in China that is known for its high‐quality seed oil. Here, we revealed that a tree peony nuclear factor Y transcription factor, PrNF‐YC2, is expressed predominantly in developing seeds and functions as an essential positive regulator of seed oil accumulation. PrNF‐YC2 promoted oil accumulation in both transient ectopic overexpression Nicotiana benthamiana leaves and stable transgenic Arabidopsis thaliana seeds, globally upregulating the expression of genes involved in oil accumulation. In contrast, PrNF‐YC2‐silenced tree peony leaves using a virus‐induced gene silencing system showed reduced oil content and expression of oil synthesis‐related genes, including four master positive regulators contributing to oil accumulation, namely, LEAFY COTYLEDON1 (LEC1), ABSCISIC ACID INSENSITIVE3 (ABI3), FUSCA3 (FUS3), and WRINKLED1 (WRI1). We demonstrated that PrNF‐YC2 directly activates PrLEC1 and PrABI3 alone and indirectly activates PrFUS3 and PrWRI1 by interacting with PrLEC1. Moreover, interaction with PrLEC1 also enhances the activation capacity of PrNF‐YC2. The activation of these four master positive regulators by PrNF‐YC2 triggered the upregulation of numerous oil synthesis‐related genes, thus promoting oil accumulation. These findings provide new insights into the regulatory mechanism of seed oil accumulation and manipulation of PrNF‐YC2 may be beneficial for enhancing oil yield in tree peony and other oilseed crops. Significance Statement: We show that the tree peony NF‐Y transcriptional factor PrNF‐YC2 directly activates PrLEC1 and PrABI3 alone and indirectly activates PrFUS3 and PrWRI1 by recruiting PrLEC1, thus plays an essential role in seed oil accumulation. To our knowledge, this study demonstrates for the first time that NF‐YC transcription factor is a major gene that regulates seed oil accumulation via dual modes. This study also enriches our knowledge of the transcriptional regulation of LEC1. [ABSTRACT FROM AUTHOR]

miRNA plays an important role in plant growth and development and in response to various stresses. Quantitative real-time PCR (qRT-PCR) technology is often used to detect the expression level of miRNAs and genes by comparing with reference genes. In order to screen out the optimal reference miRNAs in different tree peony varieties, the petals of 42 different early- and late-flowering tree peony varieties were used as experimental materials, and geNorm, NormFinder, Bestkeeper, and RefFinder software were used to evaluate the stability of 16 candidate reference miRNAs. The results showed that the average Ct values of all candidate reference miRNAs were between 15.34 ± 0.29 and 32.64 ± 0.38. The optimal number of reference miRNAs was four, which were PsPC-5p-19095, PsPC-3p-51259, PsmiR159a, and PsPC-3p-6660 in geNorm. The stability of PsPC-3p-6660 was the highest in the analysis results of NormFinder software. Among the analysis results of Bestkeeper software, PsMIR319-p5 has the highest stability. Among the results of comprehensive evaluation and analysis of several software using RefFinder, the candidate reference miRNA with the highest stability was PsPC-3p-6660. When PsPC-3p-6660 was used as the reference miRNA, the expression of PomiR171 and PomiR414 in response to different flowering times of tree peony was relatively stable in 42 tree peony varieties, indicating that PsPC-3p-6660 was stable and reliable. The results of this study provide a reference miRNA for studying the expression changes of miRNA in different tree peony varieties and further exploring the regulatory mechanism of miRNA in different peony varieties. [ABSTRACT FROM AUTHOR]