(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.

(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.

Selenium (Se) is an essential micronutrient for human health, yet its deficiency remains prevalent worldwide. Biofortification through foliar Se application is an effective strategy to enhance Se levels in crops. Paeonia ostii ‘Fengdan’ is a multifunctional woody plant with potential for Se enrichment, though its Se uptake and transformation mechanisms remain unclear. This study systematically investigated the effects of foliar-applied Na₂SeO₃ (0–200 mg L⁻¹) on Se uptake, accumulation, speciation, and nutritional quality in tree peony. Results showed that Se uptake increased with higher Na₂SeO₃ concentrations, displaying a clear dose-dependent pattern across all organs. Se accumulation significantly enhanced, with a pronounced shift in distribution towards above-ground organs under experimental conditions. Notably, tree peony exhibited strong biotransformation capacity, converting over 73% of Se in leaves and over 81% in seeds into organic forms, primarily SeCys₂ and SeMet, with minor MeSeCys. Comprehensive evaluation indicated that 100 mg L⁻¹ Na₂SeO₃ yielded optimal results, significantly enhancing leaf and seed biomass, increasing seed nutrient contents (soluble proteins, sugars, phenolics), and improving the unsaturated fatty acid profile of seed oil. These findings highlight tree peony’s potential as an efficient bioreactor for organic Se and provide a theoretical foundation for developing Se-enriched products from tree peony.

Potted cultivation serves as a vital strategy for industrialized production of standardized tree peonies, engineering seedlings capable of year-round and off-site transplantation. However, the limited root zone in potted conditions restricts root development, resulting in suboptimal seedling quality and hindering commercial-scale production. This study aimed to investigate the relationship between the accumulation characteristics of non-structural carbohydrates (NSCs) and growth performance in potted tree peonies, while also optimizing the transplantation technologies for potted cultivation. Using two-year-old grafted seedlings of ‘Luoyanghong’ as experimental material, the effects of root pruning, rooting agent, and Metarhizium anisopliae application on morphological development and NSCs accumulation in potted tree peony seedlings were investigated. The results showed that old roots serve as the primary storage organs for NSCs in the potted tree peony. Slight root pruning (25%) was beneficial for fibrous root growth, whereas excessive root pruning (50%) resulted in reduced biomass and NSCs accumulation. The application of a high concentration of rooting agents effectively promoted root growth and mitigated the adverse effects of root pruning. Furthermore, Metarhizium anisopliae significantly increased the stem number in potted tree peonies. The optimal protocol identified through range analysis involved 25% root pruning, followed by irrigation with a solution containing 750 mg·L⁻¹ rooting agent and 20 million spores·mL⁻¹ of Metarhizium anisopliae. The rational distribution of NSCs and coordinated growth across different organs enhanced NSCs accumulation in potted tree peonies. These results demonstrate that combining root pruning with the application of rooting agent and Metarhizium anisopliae can effectively increase NSCs accumulation, optimize plant morphology, and ultimately improve the quality of potted tree peony seedlings.

Corynebacterium glutamicum is an important industrial production strain that is widely used in amino acid fermentation, biopharmaceuticals, and other fields. It is particularly important to develop efficient genome editing methods for the targeted modification of C. glutamicum production strains. Currently, the gene editing system of C. glutamicum is inefficient and time-consuming. In this paper, we reported on a Francisella novicida (Fn) CRISPR-Cpf1-based system for genome editing. The system used linear DNA detached from the plasmid, and, with the assistance of the recombinase RecET, gene deletion was achieved by simultaneous electrotransformation of linear DNA with a plasmid carrying the FnCpf1 and crRNA expression cassette for double-strand breaks of the genome. Compared with previous all-in-one plasmids, this system reduced the time for one round of constructing recombinant plasmids and shortened the editing cycle by about 24 h. Finally, we successfully constructed an engineered strain (X−2) with high L-valine production by using the linear DNA-mediated gene deletion system. This method is of great importance for accelerating the process of metabolic engineering modification of C. glutamicum and its further application in high value-added products.

Tree peony (Paeonia sect. Moutan) is one of the most important traditional ornamental woody flowers in China. However, its cultivation is often challenged by intense light and high temperatures during summer, leading to severe photo-oxidative damage and leaf senescence. In this study, we investigated the protective effects of polyolefin (PO) film on potted tree peony leaves under summer intense light and high temperature stress. Using tree peony ‘Luoyanghong’ as experimental material, we systematically compared the effects of two controls under natural light (CK1 and CK2, housed in separate greenhouses), single-layer PO film (PO1), and double-layer PO film (PO2) treatments. Microenvironment monitoring showed that single-layer and double-layer PO films reduced light intensity by 21% and 49%, respectively, while having limited effects on temperature. Morphological and physiological analyses indicated that PO film treatments effectively alleviated leaf yellowing and withering, maintained normal physiological morphology, and increased chlorophyll (Chl) and carotenoid (Car) content. The net photosynthetic rates of PO1 and PO2 plants were 18–36% higher than those of the control groups (CK1 and CK2). Evans blue and NBT staining revealed that PO film reduced cellular damage and reactive oxygen species (ROS) accumulation, while decreasing malondialdehyde (MDA) levels and increasing catalase (CAT) activity. Furthermore, qRT-PCR results showed that stress-responsive genes (Hsp70, Hsp90, and ATG5) and photosynthetic genes (RbcS and RbcL) were upregulated under PO film treatment. Principal component analysis (PCA) indicated that high light intensity, rather than temperature, was the primary factor causing leaf damage in potted tree peonies. The results show that PO film is an effective and low-cost agronomic measure, which can alleviate the intense light stress under high temperature conditions, relieve photo-oxidative damage, maintain photosynthetic performance, and increase the survival rate of potted tree peonies in summer.