Shui-Yan Yu

Paeoniflorin is a pharmacologically important cage-like monoterpene glycoside characteristic of Paeonia plants, yet its biosynthetic pathway has remained largely unresolved, hindering sustainable production. Here, we confirmed that paeoniflorin biosynthesis originates from α-pinene and identified 3 novel cytochrome P450 enzymes that catalyze pinene oxidation. CYP71AN126 catalyzes the hydroxylation of α-pinene at positions C4 and C10, followed by further oxidation of the alcohol to a ketone at C4, whereas CYP76A225/226 exclusively catalyze C10 hydroxylation. Virus-induced gene silencing assays demonstrated that silencing CYP71AN126, but not CYP76A225 and CYP76A226, significantly reduced the paeoniflorin content, indicating that C4 hydroxylation plays an important role in paeoniflorin biosynthesis, whereas C10 hydroxylation is not. Through the analysis of natural sequence and activity divergence among CYP71AN126 and CYP76A225/226, combined with protein structure prediction and site-directed mutagenesis, we identified L493 as a critical residue involved in regulating catalytic site specificity and substrate specificity of CYP71AN126. Mutation of L493 reduced or eliminated the formation of undesired C10 hydroxylation side product and enhanced substrate specificity. These findings establish C4 oxidation of α-pinene as the critical committed step in paeoniflorin biosynthesis. Our study lays a foundation for elucidating the complete biosynthetic pathway of paeoniflorin in Paeonia and provides a target for enzyme engineering of CYP71AN126 aimed at the efficient production of paeoniflorin via synthetic biology approaches.

Triacylglycerol (TAG), the primary storage lipid in plants, determines oil quality through its fatty acid composition. This review focuses on the biosynthesis of TAG, systematically analyzing the mechanistic similarities and differences between the acyl-CoA-dependent Kennedy pathway (catalyzed by the rate-limiting enzyme DGAT) and the acyl-CoA-independent pathway (regulated by the rate-limiting enzyme PDAT). By integrating functional studies, evolutionary analyses, and lipidomic data, we reveal the distinct substrate preferences of DGAT and PDAT, their differential contributions to TAG synthesis, and their synergistic mechanisms in shaping triacylglycerol diversity. This work establishes a theoretical framework for the targeted engineering of plant oils with enhanced nutritional and industrial value through optimized fatty acid profiles.

Root zone restriction (RZR) technology optimizes plant growth and quality. However, the fleshy root system of Paeonia ostii exhibits sensitivity to spatial constraints, and research on the plasticity of its root architecture and adaptation mechanisms remains inadequate. This study provides a functional analysis of biomass allocation and root architectural responses to the root-zone restriction (RZR) in P. ostii, comparing three container volumes (8.5, 17, and 34 L). While the total biomass increased with root zone volume (e.g., shoot biomass rose from 9.30 g to 59.94 g), RZR induced a 44.8% increase in root-to-shoot ratio, indicating carbon reallocation to enhance belowground resource acquisition. The principal component analysis identified root biomass, volume, and surface area as key plasticity drivers. Optimal root efficiency occurred at 26.09–28.23 L, where root length and tip/fork numbers peaked. Mechanistically, RZR elevated superoxide dismutase (SOD) activity by 49.74% but reduced catalase (CAT) by 74.24%, disrupting H₂O₂ homeostasis. Concurrently, auxin transporter genes (PIN1, AUX1) were upregulated, promoting root elongation and lateral branching through auxin redistribution. We hypothesize that ROS–auxin crosstalk mediates architectural reconfiguration to mitigate spatial stress, with thickened roots enhancing structural stability in restricted environments. The study underscores the need to optimize root zone volume in woody species cultivation, providing thresholds (e.g., >28 L for mature plants) to balance biomass yield and physiological costs in horticultural management.

The tree peony holds significant historical, cultural, and practical value. P. ostii is extensively cultivated in China, where it represents the primary oil-producing variety of tree peonies. However, the current nutrient supplementation system for P. ostii lacks an empirical basis, resulting in frequent wastage of nutrients during daily production. In this study, varying ratios and quantities of nitrogen, phosphorus, and potassium were administered to P. ostii at distinct phases of its annual growth cycle, specifically during the bud sprouting, post-flowering, and dormancy periods. The results showed that during the bud sprouting period, the plants treated with a high nitrogen and potassium ratio (a high N&K ratio, N–P–K = 35%–20%–35%) had better flowering traits than those treated with a high phosphorus ratio (a high P ratio, N–P–K = 20%–35%–20%). Under the standard application amount, plants treated with a high N&K ratio outperformed those treated with a high P ratio in terms of flowering duration, net photosynthetic rate, and flowering biomass, with increases of 20.9%, 10.7%, and 32.9%, respectively. During the post-flowering period, all plants died when treated with 1.5 × standard amounts of the above ratios. At this period, the all-high ratio with N–P–K = 45%–45%–45% resulted in a 70.4% increase in fruit set, a 43.8% increase in seed number, and a 153.8% increase in biomass compared with the high N&K ratio. During the dormancy period, nutrient supplementation with the standard amount of P led to higher increases in ground diameter and biomass. Therefore, in subsequent tree peony production, particular attention should be paid to nutrient supplementation during the post-flowering period to prevent excessive fertilizer application and safeguard the plants’ normal growth and development.

To select tree peony varieties with high ornamental value suitable for the hot and humid environment of the south Yangtze River area, 74 tree peony varieties from the Chenshan Botanical Garden in Shanghai were selected as experimental materials. The experiment involved initial selection for leaf humidity and heat tolerance, assessing the ornamental value during the flowering period of the following year and measuring a series of indicators such as the relative chlorophyll content of the leaves. Methods including principal component analysis, cluster analysis, and comprehensive membership function were employed. The results indicated that among the varieties assessed for humidity and heat tolerance, 43 obtained a composite score of 3.75 or above. Upon further assessment of the flowering period for these 43 varieties in the subsequent year, 14 varieties clustered in the top tier based on their scoring for ornamental value. Then, during the summer season, physiological indicators were measured for the previously selected 14 tree peony varieties. Combining these data with the comprehensive membership function, the humidity and heat tolerance of each variety were quantified. The synthesis of scores, combined with cluster analysis, revealed that varieties including 'Yinhong Qiaodui' , 'Shengge Jin' , 'Linghua Zhanlu' , 'Baixue Gongzhu' , 'Di Guan' , and 'Cunsong Ying' exhibit both high ornamental value and tolerance to humidity and heat, making them suitable for cultivation in the south Yangtze River area.

Tree peony (Paeonia suffruticosa Andr.), which is a traditional flower cultivated in China, is rapidly becoming an important species in the cut flower industry. Thus, extending the vase life of tree peony cut flowers is a major goal in the cut flower industry. Melatonin, which is a new type of antioxidant, plays an important regulatory role in the preservation of cut flowers. Therefore, this study employed the cut flower of tree peony “Diguan” as the test material to investigate the preservative effects of the antioxidant melatonin on the cut flower of tree peony “Diguan”. We examined tree peony cut flowers in terms of their morphology, lifespan, relative fresh weight, relative diameter, and water balance value after treatments with different melatonin concentrations (0.2, 0.3, 0.4, and 0.5 mg·L⁻¹) to select the optimal treatment concentration. Considered together, these analyses clarified the effects of melatonin on the preservation of “Diguan” tree peony cut flowers. Specifically, the exogenous application of melatonin positively affected the preservation of tree peony cut flowers by improving the water balance value and increasing the soluble protein content and antioxidant enzyme activities, thereby prolonging the ornamental period of tree peony cut flowers. The fresh weight of flower branches is significantly positively correlated with soluble protein, and cut flower lifespan increases with the values of soluble protein and the fresh weight of flower branches, with a large correlation coefficient. It can be used as an important indicator to measure cut flower lifespan in subsequent research. The 0.4 mg L⁻¹ melatonin treatment was optimal for preserving tree peony cut flowers because of its positive effects on the duration of the ornamental period and ornamental quality.

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

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.

Tree peony (Paeonia suffruticosa Andr.), a unique traditional flower in China, is famous for its ornamental value, medical use, and edible oil production. Traditional propagation methods, such as sowing, dividing, and grafting do not allow the large-scale production of selected peony varieties. Therefore, the objective of our study is to evaluate an air-layering technique on the rooting success of three tree peony cultivars (‘Baoqing Hong’, ‘Quehao’, and ‘Xishi’). The experiments were established through consideration of the influence of the time of the year the rooting was performed (mid-May, mid-June, or mid-July) and the growth regulators (1-naphthaleneacetic acid-NAA and indole-3-butyric acid-IBA) applied at different concentrations (1000 mg/L, 1500 mg/L, 2000 mg/L). The results showed that the rooting rate was the highest when the air-layering time occurred in mid-June, and the rooting rate of ‘Quehao’ was found to be the most significant, reaching 100%. The rooting percentages of 82.86% and 77.14% were obtained for ‘Baoqing Hong’ and ‘Xishi’, respectively. The growth regulators affected the rooting performance of the three cultivars differently. The rooting parameters of ‘Quehao’ were negatively correlated with the concentration of NAA but positively correlated with IBA, whereas the ‘Baoqing Hong’ and ‘Xishi’ cultivars showed no dose dependence for the supplied growth regulators. Root number, root tip number, and maximum root length in ‘Quehao’ were higher than those of the other two cultivars. The conclusion of our study is that the air-layering technique is a suitable method for achieving satisfactory propagation of selected tree peony cultivars.

Lipid components in the developing kernel of Paeonia ostii were determined, and the fatty acid (FA) distributions in triacylglycerol and phospholipids were characterized. The lipids in the kernel were mainly phospholipids (43%), neutral glycerides (24%), fatty acyls (26%), and sphingolipids (4.5%). The dominant neutral glycerides were TAG and diacylglycerol. The PL components included phosphatidic acid, phosphatidyl glycerol, phosphatidyl choline, phosphatidyl serine, phosphatidyl inositol, and phosphatidyl ethanolamine. As the kernel developed, the profiles of the molecular species comprising TAG and PL changed, especially during the earlier phases of oil accumulation. During rapid oil accumulation, the abundances of sphingosine-1-phosphate, pyruvic acid, stearic acid, and alpha-linolenic acid changed significantly; the sphingolipid metabolism and unsaturated FAs biosynthesis pathways were significantly enriched in these differentially abundant metabolites. Our results improve our understanding of lipid accumulation in tree peony seeds, and provide a framework for the analysis of lipid metabolisms in other oil crops.