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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.

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.

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 recognized as an important oilseed tree peony species with potential as a raw material source for cosmetic and health care products, strong seed setting capacity, high seed oil yield, and abundant omega-3 polyunsaturated fatty acids. P. ostii, commonly called Fengdan, is widely cultivated in China. The cultivation method difference has an important influence on the oil-use feature, which is a key index for evaluating the quality of oilseed crops. This study aimed to select an optimal cultivation method to provide the first reference data for high-yield and high-quality seed oil harvesting and to facilitate the understanding of the quality difference in the formation mechanism of seed oil in cultivated P. ostii. This study selected five representative cultivation methods, open field cultivation, sunny slope cultivation, shady slope cultivation, understory intercropping cultivation, and high-altitude cultivation, and investigated the influence of cultivation method differences on the oil-use feature based on the three aspects of seed yield traits, oil yield, and fatty acid compositions. Six seed yield traits (fruit pod length, fruit pod width, fruit pod thickness, number of fruit pods per plant, 1000-grain weight, and seed yield per plant) and oil yield reached the maximum values of 7.75 cm, 6.99 cm, 1.57 cm, 11.33, 290.45 g, 85.8 g, and 30.41%, respectively, using the understory intercropping cultivation method. Fatty acid compositions were mainly identified as α-linolenic acid, linoleic acid, oleic acid, stearic acid, and palmitic acid, with significant content differences among different cultivation methods (P < 0.05). Functional component α-linolenic acid and total amounts of unsaturated fatty acids reached the maximum values of 46.85% and 65.23%, respectively, with high-altitude cultivation and understory intercropping cultivation. The seed yield traits, oil yield, and total amounts of unsaturated fatty acids were optimal with the understory intercropping cultivation method, whereas high-altitude cultivation was conducive to the accumulation of α-linolenic acid. Understory intercropping cultivation with appropriate altitude increases is recommended as a preponderant cultivation method for high-yield and high-quality seed oil harvesting for this crop.

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 s...

Paeonia ostii is a woody oil crop with potential value as an edible oil. With the aim of acquiring systematic knowledge of the development of P. ostii seeds, the oil content, biomass, and water content of the seeds were determined. Changes in the distribution of hydrogen protons in P. ostii seeds during follicle development were traced using magnetic resonance imaging (MRI). The formation of oil bodies in the endosperm and embryo was observed using transmission electron microscopy (TEM). Dynamic changes in oleic acid, linoleic acid, and α-linolenic acid contents were assessed by gas chromatography-mass spectrometry (GC-MS). The magnetic resonance images showed that, during early follicle development [45–85 days after flowering (DAF)], a greater quantity of liquid mucus was present within the seeds, and seeds in the same follicle developed at different rates. At 95 to 115 DAF, proton density was distributed evenly in all areas of the seed. A small dark area appeared in the center of the seed, and mucus in the follicles and water in the pericarp disappeared gradually. TEM observations showed that at 45 DAF, a few oil bodies were scattered at the cell periphery in the endosperm, and oil bodies were more numerous in the embryo. With the progression of seed development, the number and size of oil bodies in the embryo and endosperm continued to increase. The fresh and dry mass of P. ostii seeds increased from 45 to 105 DAF, then decreased after 105 DAF. The moisture content decreased, whereas the oil content increased and attained 33.1% at seed maturity. The three predominant unsaturated fatty acids accumulated simultaneously and showed stages of initial accumulation (45–65 DAF) and rapid accumulation (65–105 DAF). The results suggest that 65 to 105 DAF is a critical period for unsaturated fatty acid accumulation in P. ostii seeds.

Graphene oxide (GO) is considered to be an emerging environmental pollutant with its inevitable release into the environment. Thus, its potential environmental risks and biosafety are receiving increased attention. In this study, Paeonia ostii was exposed to GO under drought stress. The results demonstrated that GO prevented soil water from evaporating due to its hydrophilic oxygen-containing functional groups and did not change the soil pH. Moreover, GO treatment resulted in lower increases in reactive oxygen species, relative electrical conductivity and free proline content, and greater increases in the antioxidant enzyme activities of P. ostii under drought stress compared with those in the control. And under drought stress, higher photosynthesis, more intact mesophyll cells and organelles and open stomata were found in P. ostii under GO treatment. Furthermore, GO treatment induced greater changes in the expression patterns of genes required for lignin biosynthesis, photosynthesis-antenna proteins, carbon fixation in photosynthetic organisms, and glyoxylate and dicarboxylate metabolism. Additionally, GO did not accumulate in P. ostii due to the soil environment and the electrostatic repulsion between GO and the roots. GO did not have toxic effects on P. ostii and was an effective soil water retention agent; therefore, it could be economically beneficial for the production of plants under drought stress.

Oil peony (Paeonia spp.) is a new type of woody oil crop in China with a large cultivation area. Inadequate pollination is one of the main reasons for low seed yield. A pollination net room was built at an oil tree peony base, the numbers of honeybees (Apis mellifera L.) and ground bumblebees (Bombus terrestris L.) were artificially increased and the foraging behaviors and daily activities of the two bees on the plants were observed. Four different pollination methods (honeybee pollination, ground bumblebee pollination, natural field pollination and pollination without insects) were applied. The visit interval, single-flower visit time, number of single-flower visits, number of flowers visited per minute and number of stigma contacts were compared. Meanwhile, the effects of honeybee and bumblebee pollination on the oil yield and quality of peony seeds were compared. There were noticeable differences in daily activity between honeybees and ground bumblebees. Significant differences in the single-flower visit time, visit interval and visit frequency were also observed; honeybee and ground bumblebee pollination increased the seed yield of oil tree peony by 78.74% and 31.88%, respectively. Therefore, both honeybees and ground bumblebees are effective pollinators of oil tree peony. These results provide a theoretical basis for further utilization of bee resources for oil tree peony pollination.

Paeonia ostii T. Hong & J.X. Zhang, an emerging oil crop, was intercropped with 3-year-old plantations of pawpaw [Chaenomeles sinensis (Thouin.) Koehne], Chinese toon (Toona sinensis Roem.), and walnut (Juglans regia L.). In order to achieve a better production system, we studied the effects of intercropping on the photosynthesis, growth, and yield of P. ostii. The results showed that different pattern of agroforestry systems changed microclimatic and growing site conditions in comparison to the control treatment. The correlation analysis demonstrated that both similarities and differences were found in relationship between net photosynthetic rate (PN) and main ecophysiological factors at different treatments. Agroforestry systems significantly reduced transpiration rate and increased water-use efficiency (WUE), maximal quantum yield of PSII photochemistry, chlorophyll (Chl) a and total Chl contents, whereas there were no differences between PN, intercellular CO2 concentration, photochemical efficiency of PSII in the light, and plant height in these systems. The obviously exponential relationship between PN and PAR during a day were observed in Chinese toon and walnut treatments. The highest effective quantum yield of PSII photochemistry, electron transport rate, and photochemical quenching coefficient were observed in walnut treatment. The higher WUE and SPAD value, the thinnest stem, the biggest crown, the lowest stomatal conductance, and Chl a/b ratio as well as the fewest pods and harvest seed yield were observed in pawpaw treatment. In addition, there was a significant correlation between SPAD value and Chl (a+b) of P. ostii in agroforestry system. This study could be an important contribution for the science of land management in oil peony and other understory crops.

Paeonia ostii is an industrial crop with broad prospects, and folks have habit of drinking its stamen tea in China, but its beneficial healthy effects remain largely unclear. Here, we identified its main active components, evaluated its antioxidant activity, and examined its safety property and its beneficial effects in the model animal Caenorhabditis elegans. The results showed that P. ostii stamen (POS) contained many active components with high antioxidant activity. Moreover, POS tea did not cause lethality, influence growth, locomotion behavior and reproduction, and induce intestinal autofluorescence in C. elegans. Furthermore, 1.2 mg·mL-1 of POS tea treatment significantly extended the lifespan and improved growth, locomotion behavior and intestinal autofluorescence of C. elegans, while its reproduction and Escherichia coli OP50 growth were not affected. In addition, POS tea treatment significantly induced stress resistance to extend the lifespan of C. elegans under heat stress and oxidative stress conditions. All these suggested that POS tea was safe and had beneficial healthy effects, which could provide a theoretical basis for its production and popularization.

Paeonia ostii is the representative of oil-utilized peony, its roots (PR) and leaves (PL) are discarded as by-products, resulting in a waste of resources. The exploration of extraction process of active ingredients from PR/PL is beneficial to the comprehensive utilization. In this work, the optimum process of Ultrasound-Assisted Extraction (UAEo) of total flavonoids content (TFC) from PR and PL was determined by single factor and response surface methodology. The results showed that UAEo was 80 min, 1:5 g/mL, 250 W, 33.83 mg Rutin/g dw for PR and 60 min, 1:10 g/mL, 250 W, 25.85 mg Rutin/g dw for PL. Then, Homogenization-Assisted Extraction (HAE), Homogenization-Ultrasound-Assisted Extraction (HUAEo) and Ultrasound-Homogenization-Assisted Extraction (UHAEo) were further analyzed. The highest PR(PL)-TFC by HUAEo at homogenization 5 min was 49.58 ± 0.25 mg/g with an increase of 46.6% (33.02 ± 0.04 mg/g with an increase of 27.7%). The highest PR/PL-total phenolic content by the HUAEo reached 77.84 ± 0.52 mg/g dw and 146.62 ± 2.77 mg/g dw for homogenization 3 min. However, there was no significant difference between HUAEo and UHAEo. In conclusion, the TFC increased with the extension of HAE time, and the combined extraction was higher yield than the single extraction.

Multi-walled carbon nanotubes (MWCNTs) are considered important nanomaterials with rapidly growing applications. They are inevitably released into the environment, which has attracted considerable attention for their potential threats to ecosystems. In this study, Paeonia ostii T. Hong et J. X. Zhang was exposed to MWCNTs at different concentrations under high temperature. The results showed that high temperature-induced P. ostii damage was prevented by MWCNTs, and 200 mg/L was the most effective concentration. First, MWCNTs prevented increases in reactive oxygen species, relative electrical conductivity and free proline content, and reduced decreases in SPAD, chlorophyll and carotenoid contents. Moreover, the ascorbate-glutathione (ASA-GSH) cycle was activated in response to the MWCNTs treatments, whereas the superoxide dismutase and catalase activities were inhibited. And the MWCNTs treatments also resulted in higher photosynthesis and more intact anatomical structures. Furthermore, the metabolome also confirmed that the ASA-GSH cycle played a critical role in P. ostii high-temperature tolerance, and other biological processes also responded to the MWCNTs treatments. Additionally, the genes involved in the P. ostii ASA-GSH cycle were highly expressed in response to the MWCNTs treatments. These results elucidated the beneficial role of MWCNTs in P. ostii growth under high temperature.

As emerging woody oil crops, the tree peony seeds recently have been attracting great attention for their metabolites and bioactivities. In this research, the phytochemical profiles of the seed coats of tree peonies from different production regions were investigated systematically. Twelve phytochemicals were separated and prepared, mainly belonging to stilbenes. A great variation in stilbene content was detected in the three Paeonia plants, and Paeonia ostii seed coats (POSC) had significantly higher contents of the stilbene compounds than other species. There were nineteen significant correlations between ecogeographical factors and the predominant compounds. A clear discrimination among the species was observed in their HPLC fingerprint and chemometric analysis. Furthermore, POSC extracts could significantly reduce the starch mediated PBG (postprandial blood glucose) levels in normal/diabetic mice. Meanwhile, in vitro enzyme tests revealed that the predominant compounds, suffruticosol B and ampelopsin D, could effectively and competitively inhibit α-glucosidase, indicating that POSC could be a natural source of hypoglycemics in the food and drug fields.

Background Selenium is an essential micronutrient in human metabolism. However, the Se level in foods is far below the standards due to the fact that it is deficient in two thirds of arable soils. Therefore, consuming Se-enriched plant products which convert inorganic Se into organic Se might be a safe and effective way to supplement Se for the human body. Paeonia ostii is a unique medicinal and edible plant in China, and also a new special woody oil crop. It serves as a perfect mediator for Se biofortification. In this report the effects of nano-Se foliar application on photosynthetic parameters, physiological characteristics and Se-enrichment effect in various plant organs of P. ostii cultivar Feng Dan were investigated in randomized complete block design experiments with different doses of spray in Se-deficient soils.Results By contrast to the control, nano-Se sprays enhanced net photosynthetic rate, transpiration rate, stomatal conductance, intercellular CO2 concentration, and chlorophyll content of Feng Dan. The content of soluble sugar, soluble protein, proline and activities of superoxide dismutase and peroxidase were increased while malondialdehyde content was significantly reduced. The stimulatory effect at concentrations of 6.00 and 8.25 g·hm-2 was the best in this study. nano-Se application significantly increased the Se content in all organs of Fengdan except the root compared to the control, and Se contents in various organs were increased paralleled with the increase of concentrations applied, and ordered as follows under suitable concentration, Testa > leaf > kernel > stem > root.ConclusionsThe above results show that nano-Se sprays at concentrations of 6.00-8.25 g·hm-2 stimulated the growth of Feng Dan and achieved the best Se-enrichment effect in this study. It also proved that the Se biofortification effect depends on its dosage applied and types of plant organs. These results provide guidance on Se enrichment of horticultural crops.

This is the first report concerning the sequence of morpho-histological and cytological events occurring during organogenesis from cotyledon-derived meristematic nodules (MNs) in Paeonia ostii ‘Feng Dan’. Sections were made and studies were carried out with dissecting microscope, light microscope, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) observation. Histological studies revealed a complex developmental process of morphogenesis that including five stages: (1) callus originated from cell division in both cambial and cortical regions, and type I - yellow compact callus with densely arranged clumps were identified as embryogenic callus. (2) pre-nodular structure consisted of organization center (a central area of vascularization surrounded by meristematic cell layers) and an epidermis-like layer; (3) independent MNs comprised of organization center, a cortical-like area of parenchymatous cells and an epidermal-like area; (4) nodular clusters displayed vigorously internal meristematic cell division and generated a relative movement towards the nodules periphery, establishing vascular connection with primordia; (5) successive new elongated shoots with complete vascular system and axillary bud primordia were developed. SEM observations showed three types of extracellular matrix (ECM), a smooth membranous layer, fibrillar structures and granular mucilage-like secretions on embryogenic callus, and recorded its dynamic morphological changes. Ultrastructural analysis revealed striking changes of chloroplast morphology and starch content during MNs morphogenesis. This study allows a better understanding of in vitro regeneration via MN culture and provides references for protocol optimization and genetic transformation.

Paeonia ostii ‘Feng dan’ is recommended as a new promising oil crop because of the nutrient elements in its seed; however, the development of this new oil crop is now limited due to its low seed setting rate. This study identified ways to improve the cross pollination of Feng dan as the maternal parent, which has been proved to be a suitable oil crop, so as to improve the seed setting rate of ‘Feng dan’, and to predict the functional relationship between pollen vigor and seed setting rate. improving the efficiency of cross pollination. Three major steps were conducted, first, to screen the appropriate method of measuring pollen vigor of the paternal parent, which could have a strong relationship with seed setting rate. Second, to obtain the corresponding seed setting rate by cross pollination. Third, the functional relationship between two indexes was founded which could be used in cross pollination practice. Results indicated that the best paternal parents of ‘Feng dan’ were ‘Feng dan’ cultivars from other different cultivation populations; The most suitable medium concentration for pollen germination was 100~150 g/L sucrose, 0.06~0.08 g/L boric acid, and 50~200 g/L PEG-6000. Both the seed setting rate and the number of seed grains were positively correlated with the pollen activity, and the seed setting rate was negatively correlated with the local precipitation.

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.

Paeoniaceae is an abundant germplasm resource with significant medicinal values in China, the principal medicinal components of which include paeoniflorin and paeonol. These compounds are typically obtained from air-dried root samples, which the use of freeze-drying as an alternative method has not been tested. Additionally, the presence of these two compounds in various wild Paeoniaceae species has not been previously explored, nor have the differences between various plant organs been fully evaluated. Here, freeze-drying and air-drying methods were compared to assess the changes in paeoniflorin and paeonol in root samples using ultra-performance liquid chromatography-mass spectrometer. The contents of these compounds in the roots, leaves, stems, and petals were then tested in freeze-dried materials. We also quantitatively detected the paeoniflorin and paeonol contents in the roots of 14 species collected from 20 natural habitats. Results indicated that the paeoniflorin content decreased under air-drying in comparison to freeze-drying, while the opposite trend was observed for paeonol. Our findings also demonstrated that the root xylem of species in Section Moutan, particularly Paeonia ostii, contains considerable paeonol and paeoniflorin and should thus be fully utilized as a medicinal resource. Furthermore, paeonol was mainly detected in the roots, while paeoniflorin was widely distributed in different organs; the highest content was in the leaf at the budding stage, suggesting that the leaves should be developed as a new paeoniflorin resource. Paeoniflorin contents were also found to be higher at earlier development stages. Based on the standards of the Chinese Pharmacopoeia, five species of Section Moutan and six species of Section Paeonia could be used as potential traditional Chinese medicinal resources. These findings of this study enhance our understanding of these two medicinal compounds and provide a foundation for the further development and utilization of Paeoniaceae as medicinal plant resources.

The cortex root of Paeonia ostii, is used as a traditional Chinese medicine for treating female diseases. Phytochemical investigation of the water-soluble fraction of the plant led to the isolation of two new acetoisovanillone glycosides: acetoisovanillone-3-O-β-D-glucopyranoside (1) and 2-hydroxy-acetoisovanillone-3-O-β-D-glucopyranoside (2). Their structures were elucidated by extensive spectroscopic methods.

Tree peony (Paeonia section Moutan DC.) is an excellent ornamental plant, of which Paeonia ostii (P. ostii) has a high oil value. It is widely cultivated in China, but severe drought affects its growth. In this study, the effects of exogenous calcium on drought-induced damage of P. ostii were studied. The results showed that under drought stress, leaf water content showed a downward trend, while reactive oxygen species (ROS), relative electrical conductivity (REC), proline (Pro) content, and related antioxidant enzymes increased significantly. Spraying CaCl2 could effectively slow leaf wilting and water loss, induced an increase in enzyme activity of the antioxidant enzyme system, and reduced the accumulation of ROS caused by drought stress. Simultaneously, REC and Pro content could be alleviated, and the degree of cell membrane damage could be reduced. In addition, CaCl2 improved photosynthetic characteristics and chlorophyll fluorescence parameters. These results indicated that CaCl2 reduced the harmful effects of drought stress on the growth of P. ostii by regulating infiltration, activating photosynthesis, and enhancing the antioxidant system. These findings suggested that CaCl2 can be used to manage drought stress in P. ostii cultivation.

Paeonia ostii seeds have recently been identified as a new source of α-linolenic acid in China. Studying the gene expression patterns of unsaturated fatty acid-related genes would be helpful for understanding the mechanism of α-linolenic acid accumulation. Quantitative real-time polymerase chain reaction (qRT-PCR) is a useful method for reliably evaluating gene expression, and it is necessary to select reliable reference genes for data normalization in qRT-PCR analysis. In this study, we evaluated the expression stability of 12 candidate reference genes using four mathematical algorithms (∆Ct, BestKeeper, NormFinder, and geNorm). The web-based tool RefFinder was used to integrate the results and to provide a comprehensive ranking order. The expression stability ranking orders of reference genes were different caculated by these four algorithms, and the ranking order analyzed by the RefFinder was UBQ > Tip41 > UCE > EF-1α > α-TUB > PP2A > ACT > GAPDH > SAM > CYP > β-TUB > 18S at the different seed development stages, and UBQ > Tip41 > EF-1α > α-TUB > PP2A > UCE > GAPDH > SAM > ACT > CYP > 18S > β-TUB in P. ostii tissues. UBQ and Tip41 are the two most stable whereas 18S and β-TUB are the two least stable reference genes for gene expression in various tissues and seeds at different developmental stages in P. ostii.

Two tree peony species, Paeonia ostii and Paeonia rockii, from eight different provenances in China (four provenances for each species) were collected for their photosynthetic characteristics. Their diurnal variations of photosynthesis, the light- and CO2-response curves and chlorophyll content were determined. Among all eight provenances, Ziwuling P. rockii had a significantly higher average daily net photosynthetic rate, showing a superior photosynthetic capacity, followed by Fengxiang P. ostii and Xunyang P. ostii. In addition, Ziwuling P. rockii had the highest maximum net photosynthetic rates, thus, presented a strong photosynthetic potential. Furthermore, Xunyang P. ostii, Fengxiang P. ostii, and Heze P. ostii also exhibited a great photosynthetic capacity and light adaptability, which was consistent with their high chlorophyll a/b values. Taken together, Ziwuling P. rockii had the strongest photosynthetic capacity among all tested sources, followed by Fengxiang P. ostii, and Xunyang P. ostii, which are recommended to be used as seed sources for oil peony cultivation and cross-breeding.

Paeonia ostii is one of the important woody oleaginous plants, and is cultivated on a large scale in China. The main purpose of this study is to evaluate the characteristic difference of P. ostii seed and oil sourced different geographical regions in China. The thousand seed weight (TSW), oil content and kernel content of the P. ostii seed, the composition and content of the main fatty acids, and the physicochemical characteristics of the seed oil of 62 P. ostii seed samples from different cultivation regions were monitored and evaluated in this study. The results showed that there were extremely significant difference among the TSW and the contents of the main fatty acids in P. ostii seed and oil. The geographical regions were related to the variability in TSW, the contents of the main fatty acids (FAs) and the total content of the main fatty acids (TCMFA), perhaps due to the influence of environmental factors, and the water, fertilizer application, crop management, or other factors. P. ostii would become a kind of woody oleaginous plant and an industrial crop with broad prospects for its high content of alpha-linolenic acid (ALA) and the high oil content of the seeds.