The herbaceous peony (Paeonia lactiflora Pall.) plant is world-renowned for its ornamental, medicinal, edible, and oil values. As global warming intensifies, its growth and development are often affected by high-temperature stress, especially in low-latitude regions. Superoxide dismutase (SOD) is an important enzyme in the plant antioxidant systems and plays vital roles in stress response by maintaining the dynamic balance of reactive oxygen species (ROS) concentrations. To reveal the members of then SOD gene family and their potential roles under high-temperature stress, we performed a comprehensive identification of the SOD gene family in the low-latitude cultivar ‘Hang Baishao’ and analyzed the expression patterns of SOD family genes (PlSODs) in response to high-temperature stress and exogenous hormones. The present study identified ten potential PlSOD genes, encoding 145–261 amino acids, and their molecular weights varied from 15.319 to 29.973 kDa. Phylogenetic analysis indicated that PlSOD genes were categorized into three sub-families, and members within each sub-family exhibited similar conserved motifs. Gene expression analysis suggested that SOD genes were highly expressed in leaves, stems, and dormancy buds. Moreover, RNA-seq data revealed that PlCSD1-1, PlCSD3, and PlFSD1 may be related to high-temperature stress response. Finally, based on the Quantitative Real-time PCR (qRT-PCR) results, seven SOD genes were significantly upregulated in response to high-temperature stress, and exogenous EBR and ABA treatments can enhance high-temperature tolerance in P. lactiflora. Overall, these discoveries lay the foundation for elucidating the function of PlSOD genes for the thermotolerance of herbaceous peony and facilitating the genetic breeding of herbaceous peony cultivars with strong high-temperature resistance.

Gene function verification is a crucial step in studying the molecular mechanisms regulating various plant life activities. However, a stable and efficient homologous genetic transgenic system for herbaceous peonies has not been established. In this study, using virus-induced gene silencing technology (VIGS), a highly efficient homologous transient verification system with distinctive advantages was proposed, which not only achieves true “intact-plant” infiltration but also minimizes the operation. One-year-old roots of the representative species, Paeonia lactiflora Pall., were used as the materials; prechilling (4 °C) treatment for 3–5 weeks was applied as a critical precondition for P. lactiflora to acquire a certain chilling accumulation. A dormancy-related gene named HOMEOBOX PROTEIN 31 (PlHB31), believed to negatively regulate bud endodormancy release (BER), was chosen as the target gene in this study. GFP fluorescence was detected in directly infiltrated and newly developed roots and buds; the transgenic plantlets exhibited remarkably earlier budbreak, and PlHB31 was significantly downregulated in silenced plantlets. This study established a homologous transient silencing system featuring intact-plant infiltration and minimized manipulation for gene function research, and also offers technical support and serves as a theoretical basis for gene function discovery in numerous other geophytes.

The MIKC-type MADS-box (MIKC) gene family is essential for controlling various plant developmental processes, including flowering time and dormancy transitions. Although the MIKC gene family has been widely studied across different plants, its characterization and functional study in herbaceous peony remain limited. In this study, 19 Paeonia lactiflora Pall. MIKC-type (PlMIKC) genes were identified from the transcriptome of a low-chilling requirement Paeonia lactiflora Pall. cultivar ‘Hang Baishao’. These MIKC genes were categorized into seven clades: six were classified as MIKC^C-type, including FUL/AP1, DAM, PI, AGL18, AGL12, AG, and SOC1, and one, AGL30, was classified as MIKC*-type. Notably, the FLC clade genes were absent in Paeonia lactiflora Pall. The PlMIKC genes were predominantly localized to the nucleus, and their sequences contained highly conserved MADS and K-domains. Phylogenetic analysis demonstrated that PlMIKC genes share a strong evolutionary affinity with the MIKC genes from grapevine (Vitis vinifera) and poplar (Populus trichocarpa). A low-temperature-induced bud dormancy transition (BDT) experiment revealed that PlMIKC genes, such as PlFUL and PlDAM, were highly expressed during dormancy maintenance, while PlSOC1, PlAGL12, and PlAGL30 were upregulated during BDT. Additionally, the transient overexpression of PlSOC1 in ‘Hang Baishao’ significantly accelerated BDT and promoted bud break, suggesting that SOC1, traditionally linked to flowering regulation, also plays a key role in dormancy transition. Since limited literature on the MIKC gene family is currently available in herbaceous peony, this study expands the knowledge of the MIKC genes in Paeonia lactiflora Pall. and offers valuable insights into the molecular regulation of bud dormancy in response to low temperatures.