PlGATA1 and PlGATA6 Antagonistically Regulate ABA homeostasis to modulate Bud Dormancy Induction in Tree Peony

PlGATA1 and PlGATA6 Antagonistically Regulate ABA homeostasis to modulate Bud Dormancy Induction in Tree Peony | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL Plant, Cell & Environment This is a preprint and has not been peer reviewed. Data may be preliminary. 18 July 2025 V1 Latest version Share on PlGATA1 and PlGATA6 Antagonistically Regulate ABA homeostasis to modulate Bud Dormancy Induction in Tree Peony Authors : Ziwen Geng , Chunyan He , Fangting Qi , Jianing Han , Lei Zhang , and Fangyun Cheng 0000-0002-3928-5731 [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.175282409.95767052/v1 292 views 159 downloads Contents Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Bud dormancy critically affects tree peony growth and flowering, particularly for off-season forcing cultivation. GATA transcription factors are known regulators of ABA-mediated seed dormancy in plants. However, their potential role in bud dormancy remains unexplored. In this study, we systematic identified GATA family in tree peony genome and revealed key regulators PlGATA1 and PlGATA6 . Comparing the gene expressions in dormant and non-dormant buds in Paeonia × lemoinei ’High Noon’, we observed starkly contrasting expression patterns for PlGATA1 and PlGATA6 . Exogenous ABA treatment induced PlGATA1 expression while suppressing PlGATA6 , indicating the functional divergence in their regulatory pathways. Overexpression in Arabidopsis demonstrated that PlGATA1 induced seed dormancy, while PlGATA6 inhibits its. In tree peony, overexpression of PlGATA1 or silencing of PlGATA6 induced bud dormancy and modulated ABA metabolism-related gene expression. Critically, in vivo and in vitro binding assays confirmed that PlGATA1 activates the PlABI5 promoter, while PlGATA6 activates PlCYP707A1 promoter. Collectively, our findings demonstrate that PlGATA1 and PlGATA6 serve as a ’molecular switch’ governing bud dormancy induction in tree peony through antagonistically regulating ABA homeostasis. These results provide novel insights into the transcriptional regulatory mechanisms of bud dormancy and identify potential target genes to improve off-season cultivation in woody plants. PlGATA1 and PlGATA6 Antagonistically Regulate ABA homeostasis to modulate Bud Dormancy Induction in Tree Peony Ziwen Geng 1 , Chunyan He 1 , Fangting Qi 1 , Jianing Han 1 , Lei Zhang 1* and Fangyun Cheng 1* 1 State Key Laboratory of Efficient Production of Forest Resources, National Engineering Research Center for Floriculture, Peony International Institute, School of Landscape Architecture, Beijing Forestry University, Beijing 10083, China. * Corresponding Author: Fangyun Cheng and Lei Zhang Email address: [email protected] ; [email protected] Abstract Bud dormancy critically affects tree peony growth and flowering, particularly for off-season forcing cultivation. GATA transcription factors are known regulators of ABA-mediated seed dormancy in plants. However, their potential role in bud dormancy remains unexplored. In this study, we systematic identified GATA family in tree peony genome and revealed key regulators PlGATA1 and PlGATA6 . Comparing the gene expressions in dormant and non-dormant buds in Paeonia × lemoinei ’High Noon’, we observed starkly contrasting expression patterns for PlGATA1 and PlGATA6 . Exogenous ABA treatment induced PlGATA1 expression while suppressing PlGATA6 , indicating the functional divergence in their regulatory pathways. Overexpression in Arabidopsis demonstrated that PlGATA1 induced seed dormancy, while PlGATA6 inhibits its. In tree peony, overexpression of PlGATA1 or silencing of PlGATA6 induced bud dormancy and modulated ABA metabolism-related gene expression. Critically, in vivo and in vitro binding assays confirmed that PlGATA1 activates the PlABI5 promoter, while PlGATA6 activates PlCYP707A1 promoter. Collectively, our findings demonstrate that PlGATA1 and PlGATA6 serve as a ’molecular switch’ governing bud dormancy induction in tree peony through antagonistically regulating ABA homeostasis. These results provide novel insights into the transcriptional regulatory mechanisms of bud dormancy and identify potential target genes to improve off-season cultivation in woody plants. Keywords: tree peony; PlGATA1 ; PlGATA6 ; antagonistic regulation; bud dormancy induction; ABA homeostasis Introduction Bud dormancy represents a crucial physiological adaptation for most temperate perennial plants (Yang et al., 2021). Based on the key regulatory factors governing dormancy induction and release, bud dormancy is classified into three distinct types: endodormancy, paradormancy, and ecodormancy (Gillespie and Volaire, 2017). Among these, endodormancy is particularly significant as it imposes an essential chilling requirement that strictly governs key phenological events including bud break, vegetative growth, and floral transition (Lundell et al., 2019; Yamane et al., 2023). Despite its evolutionarily advantageous, this chilling requirement posed major challenges for modern horticulture, particularly in off-season production. As a model temperate perennial woody species, tree peony ( Paeonia suffruticosa ) exhibits endodormancy, requiring extended chilling to fulfill dormancy prior to bud break (Huang et al., 2008). Winter forcing cultivation is essential for off-season flower production in tree peony. However, tree peony’s bud dormancy induction poses a critical bottleneck to industrial-scale implementation (Bonhomme et al., 2000; Gai et al., 2013; Yuan et al., 2024). Interestingly, the cultivar Paeonia × lemoinei ’High Noon’ presents a unique natural system for studying dormancy regulation, as it simultaneously produces both dormant buds (requiring chilling-mediated dormancy release) and non-dormant buds (developing without chilling) (Chang et al., 2022). This intrinsic comparison within a single genotype offers a powerful system for investigating the regulatory mechanisms of bud dormancy induction, which is scientifically critical and directly addressing the forcing cultivation bottleneck in tree peony. The regulation of bud dormancy in tree peony involves a complex molecular network that maintains dormancy through multi-layered control mechanisms. These involves ABA-dominated hormone signaling, Ca²⁺ signaling, and cell cycle-related gene expression (Gai et al., 2013; Gai et al., 2024; Zhang et al., 2024a). Within this regulatory network, abscisic acid (ABA) serves as a pivotal phytohormone, with studies establishing a significant correlation between dynamic ABA levels and dormancy progression: ABA accumulates markedly during dormancy induction and declines progressively during release. This hormonal shift exhibits antagonistic interplay with gibberellin (GA) (Gao et al., 2023; Mornya and Cheng, 2018; Singh et al., 2018; Yuan et al., 2024). Recent advances have elucidated key components of ABA-mediated dormancy regulation in tree peony. Notably, the R2R3-MYB transcription factor PsMYB306 upregulates the ABA biosynthetic gene PsNCED3 , thereby negatively regulating ABA-dependent dormancy release (Yuan et al., 2024). Although the ABA-mediated regulatory network has been partially characterized, current insights derive predominantly from dormancy release. However, the transcriptional mechanisms governing bud dormancy induction remain largely unexplored. GATA transcription factors (TFs) are regulatory proteins defined by a conserved type IV zinc finger domain featuring the CX₂CX₁₇₋₂₀CX₂C motif (Bi et al., 2005). This domain enables specific recognition and binding of the WGATAR ((A/T) GATA(A/G)) cis-element in target gene promoters (Lowry and Atchley, 2000). In plants, the GATA family is classified into four subfamilies (Clades I–IV), with members within each clade exhibiting high structural conservation across phylogenetically diverse species (Zhang et al., 2024b). Critically, members of Clade I and II GATAs have been directly implicated in regulating seed dormancy through the ABA signaling pathway in Arabidopsis , wheat, and rice (Cheng et al., 2021; Liu et al., 2005). Given the highly conserved regulatory pathways between seed and bud dormancy (Ahmad et al., 2024; Gao et al., 2025; Graeber et al., 2012; Ma et al., 2024; Nv et al., 2019; Sano and Marion-Poll, 2021), Clade I/II subfamilies of GATA TFs likely occupy a central position in the bud dormancy network, potentially acting as upstream initiators of ABA-mediated induction. However, their role in regulating bud dormancy induction in tree peony remains a critical knowledge gap. In this study we characterized the evolutionary features of GATA gene family in tree peony. We established that the paralogous genes PlGATA1 and PlGATA6 , derived from a segmental duplication event, participate in ABA-mediated signaling pathways during bud dormancy induction. We demonstrated that this paralog exhibits differential responses to exogenous ABA treatment ( PlGATA1 is induced while PlGATA6 is suppressed) and functionally associate with the core dormancy signaling network. Evolutionary analysis further indicated that functional divergence post-duplication has enabled this TF pair to act as an antagonistic molecular switch regulating dormancy induction. Collectively, our findings not only elucidate a novel mechanism of antagonistic GATA regulation in bud dormancy induction, advancing fundamental understanding of dormancy physiology in perennial plants, but also identify PlGATA1 and PlGATA6 as promising targets for overcoming technical barriers in off-season production of tree peony. Materials & Methods Plant materials and treatment Tree peony cultivar P. × lemoinei ‘High Noon’ was obtained from Guose Peony Nursery (Yanqing district, Beijing, China, 40◦462 N, 116◦073 E). The dormant and non-dormant buds at flower bud differentiation were used to regarding the expression pattern analysis of the PlGATA genes of Clade II. Three replicates were set up, and each replicate included three plants. Fresh non-dormant buds were collected for transient transformation. Buds are collected according to morphological description of bud in ‘High Noon’ (Zhou et al., 2013). For expression analysis of genes in response to exogenous hormones, the non-dormant buds were treated with 0.02% (v/v) Triton X+300 ABA µM (RYON, Shanghai, China), and 0.02% Triton X-100 as a control (mock). The apical buds were collected after ABA treatment for 0, 3, 6, 12 h, and stored at -80℃ for further analysis. Identification of PlGATA gene family in tree peony The GATA domain hidden Markov model (HMM) profile (PF00320) was retrieved from the Pfam database (https://www.ebi.ac.uk/interpro/) (Mistry et al., 2020). This profile was subsequently used to screen Tibetan wild tree peony ( Paeonia ludlowii ) reference genome (Xiao et al., 2023) using HMMER 3.0 (Eddy, 1998) with a stringent E-value cutoff of < 1e−5. The presence of the GATA domain in all putative proteins was then manually confirmed using the SMART (http://smart.embl-heidelberg.de) (Letunic et al., 2020) and conserved Domain Databases (CDD) (https://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi) (Lu et al., 2020). A range of GATA protein properties, including molecular weight, isoelectric points, instability index, aliphatic index and GRAVY, were determined using the ExPASy ProtParam tool (http://web.expasy.org/protparam/) (Artimo et al., 2012), and protein subcellular localizations were predicted using WoLF PSORT (https://wolfpsort.hgc.jp) (Horton et al., 2007). Phylogeny, chromosomal localization and synteny analysis of PlGATA genes A phylogenetic tree was constructed using the Neighbor-Joining approach method (NJ) with default settings (neighbor‐joining; parameters: Bootstrap 1000) in MEGA version 11 (Tamura et al., 2021). The GATA protein sequences from Arabidopsis thaliana ( AtGATA ) (Reyes et al., 2004), grapevine ( Vitis vinifera L.) ( VviGATA ) (Zhang et al., 2023) and wheat ( Triticum aestivum L.) ( TaGATA ) (Cheng et al., 2021) were downloaded from the genome databases corresponding to each species. The constructed phylogenetic tree was visualized and refined using iTOL version 6 (Letunic and Bork, 2021). The PlGATA genes were mapped onto chromosomes using P. ludlowii genome (Xiao et al., 2023). The chromosomal location of each PlGATA gene was identified using the physical location information from P. ludlowii genome (Xiao et al., 2023). The synteny blocks of tree peony GATA genes, as well as between tree peony and grapevine, A. thaliana, wheat genes were analyzed using MCScanX (Wang et al., 2012), and globe plot diagrams were made using Circos-0.69-6 (http://circos.ca) (Krzywinski et al., 2009) and TBtools (Chen et al., 2020). Exon–intron structure, conserved domains, conserved motif and cis‑acting regulatory element analysis Exon and intron structures of the confirmed GATA genes were determined based on CDS and each full-length sequence in P. ludlowii genome (Xiao et al., 2023). Conserved domains of GATA were predicted using the CD-search website (https://www. ncbi.nlm.nih.gov/Structure/BWRPSB/BWRPSB.Cgi). Multiple sequence alignments of the conserved GATA domain were performed using DNAMAN (v7.0.2, Lynnon Biosoft) (Crooks et al., 2004). Motif analysis of the PlGATA amino acid sequences was performed using MEME (http://alternate.meme-suite.org/tools/meme) (Bailey et al., 2015). The upstream 2000 nucleotide sequences of the PlGATA genes were extracted using TBtools. To investigate the potential regulatory mechanism of PlGATA gene on environmental stress, the PlantCARE database (http://bioinformatics.psb.ugent.be/webtools/plantcare/html/) was used to identify the cis ‐acting regulatory elements of the PlGATA gene (Lescot et al., 2002). These results were visualized using TBtools (v2.069) (Chen et al., 2020). Subcellular localization and construction of transgenic A. thaliana Using the designed primers (Additional file 1: Table S1), the full‐length coding regions of PlGATA1 and PlGATA6 were cloned by PCR and ligated to pTOPO‐T vector for transformation into competent Escherichia coli cells. NcoI restricted enzymes was used for cleaveged pCAMBIA1302‐GFP. The PlGATA1 and PlGATA6 gene fragment was ligated into pCAMBIA1302‐GFP and transformed into E. coli to obtain the recombinant plasmid pCAMBIA1302‐ PlGATA1/6 ‐GFP. The pCAMBIA1302‐GFP and pCAMBIA1302‐ PlGATA1/6 ‐GFP plasmids were transferred into Agrobacterium GV1301 and suspended in infection buffer. They were incubated at 28℃ for 3 h, and the infection buffer was injected into tobacco leaves using a syringe. After 3 days of dark treatment, the localization results were observed under confocal microscopy. Transgenic Arabidopsis was obtained through the floral dip method (Clough and Bent, 2010). Agrobacterium containing the full‐length PlGATA1 and PlGATA6 construct were inoculated in 100 mL of LB liquid medium and cultured for 16 h at 200 rpm and 28℃. The culture was then centrifuged at 5000 rpm for 10 min, and the collected cells were resuspended in a 5% sucrose solution to an OD600 of 0.6-0.8, with 0.01% Silwet‐77 surfactant added. After the stratification of Arabidopsis seeds at 4°C for 3 days, they were sown in potting soil and grown in a growth chamber at 22℃ with a 16‐h light/8‐h dark photoperiod. Flower buds at the early flowering stage were immersed in the transformation solution for 2 min, with dipping repeated three times over a week to produce T1 generation plants. These plants were screened, and if T2 generation plants showed 100% hygromycin resistance in the T3 generation, they were considered homozygous transformants. RNA extraction and quantitative real‐time PCR analysis Total RNA was extracted from tree peony floral bud using the Trizol method. The RNA was reverse‐transcribed into cDNA using the M‐MLV reverse transcriptase kit (Takara Bio, Kyoto, Japan). RT‐PCR was performed using the 2×Phanta Flash Master Mix (Takara Bio, Kyoto, Japan). Quantitative real‐time PCR (qRT‐PCR) was carried out using the CFX96 Real‐Time PCR System (Applied Biosystems). The melting curve was determined at the end of the reaction. The 2‐∆CT method was used for statistical analysis of the results (Livak and Schmittgen 2001), with Ubiquitin ( UBQ ) serving as the reference gene. Each sample was run in triplicate, and the CT values were calculated. Phenotypic analysis of Arabidopsis transformed with the PlGATA1 and PlGATA6 genes Germination assays were done on Murashige and Skoog (MS) medium (Caisson Labs) containing 1% sucrose and 0.5% GelRite (Plant media). All assays were done with freshly harvested (FH) seeds and after-ripened (AR) seeds (4-week storage collected at the same time from mature vector controls (VC) and transgenic plants grown together in the same tray under controlled conditions (at 23℃ and 75% RH under long-day conditions). Seeds were dried for 5 days over silica beads before using them in the assays. After-ripening of seeds was done by drying the seeds over silica beads for 4 weeks at room temperature. For cold stratification, the plates were covered with aluminum foil and kept at 4℃ for 3 days, then transferred to a tissue culture room set at 24℃ under a 16 h light/8 h dark cycle. Percentage germination was scored based on the emerging of radical and greening of cotyledon was considered seed germination from at least 16 or 25 seeds per replicate and three independent experiments. Construction of TRV2-PlGATA6 silencing vector and transformation Based on the results of alignment of PlGATA6 with other DELLA proteins, the specific fragment of PlGATA6 was inserted into TRV2 vector to obtain the TRV2- PlGATA6 recombinant vector as described by Zhang et al. (Zhang et al., 2019). TRV2 and TRV2- PlGATA6 were mixed with TRV1 at a volume ratio of 1:1 and placed for 4-6 h in a dark room. The bud scales were removed, and they were sterilized and submerged in infiltration buffer for 3-4 min in a vacuum dryer at 0.3 MPa. The buds were then transferred to 1/2 MS medium containing 200 μM acetosyringone. After 4 d of dark treatment (8℃, 3 d followed by 22℃, 1 d), the buds were transferred into MS medium (200 mg. L −1 ticarcillin, and 0.5 M MES) and cultured in an incubator (22℃, 16 h light/8 h dark). After being infected for 7 d, total RNA was extracted, qRT-PCR was performed to analyze the silencing efficiency, and the expression of genes associated with dormancy, including PlCYP707A1 , PlCYP707A2-1 , PlCYP707A2-2 , PlPP2C , PlPYL2 , PlSARK1 , PlSARK2 , PlSDR2A and PlABI5 were analyzed. Morphological changes, including the relative increase in bud width and height, were measured after 15 days. A total of 15 buds were used per transformation, of which five were used for detection of expression and 10 for morphological observation. Construction of PlGATA1 overexpression vector and transformation Super1300- PlGATA1 recombinant plasmid was constructed and transformed into Agrobacterium EHA105 (empty Super1300 plasmid as a control). Under sterile conditions, tree peony buds were infiltrated as described by Zhang et al. (Zhang et al., 2019). The buds were immersed in the bacterial solution and infiltrated at a vacuum of 0.7 atm for 5 min, then slowly deflated to allow the bacterial solution to enter the buds. The infiltration step was repeated two times. After releasing the vacuum, the buds were washed for 4-5 times with sterile water before inoculating into MS medium, cultured in the dark for 3 d at 8℃, and then cultured in an incubator (22℃, 16 h light/8 h dark) as described above. After infiltration for 7 d, the relative expression levels of PlGATA1 were analyzed by qRT-PCR, and those higher than 1.5 times were used to analyze the expression of dormancy marker genes. The flower bud phenotype (relative growth in bud length and width) was investigated after 15 days. Yeast 1-hybrid (Y1H) and dual-luciferase reporter assay For the dual-luciferase reporter assay, the PlGATA1 , PlGATA6 , PlCYP707A1 , PlCYP707A2-1 , PlPP2C , PlPYL2 , PlSARK1 , PlSARK2 , and PlABI5 promoter fragment was cloned into the pGreenII 0800-LUC vector and the recombinant plasmid was transformed into Agrobacterium GV3101 and cultured overnight to an OD600 of 0.8; it was then mixed with pGreenII62-SK- PlGATA1 and pGreenII62-SK- PlGATA6 Agrobacterium solution at a ratio of 1:5 and 1:8, respectively. The pGreenII 0800-LUC empty vector was used as a control and then injected into N. benthamiana leaves (Yao et al., 2020). After 2 to 3 d of darkness, the leaves were dipped into D-luciferin sodium salt (Vazyme Biotech, Nanjing, China) and observed using an SH-Compact 523 chemiluminescence imaging system (Shenhua, Hangzhou, China). Then the LUC and REN activities were measured using a luminometer (Männedorf, Switzerland) and shown as the LUC/REN ratio. The Y1H assay was performed as described by (Yu et al., 2021). The whole promoter sequence (full) of PlABI5 and PlCYP707A2-1 was divided into 3 regions (R1-3), and were introduced into the HindⅢ-SaiⅠ sites of the pAbAi vector, which were referred to as the bait constructs and co-transformed into yeast Y1HGold. The complete coding sequence of PlGATA1 and PlGATA6 was introduced into the NdeI-BamHI sites of the pGADT7-Rec vector harboring the GAL4 activation domain to generate the pray construct and co-transformed into yeast Y1HGold. The transformed candidates were grown on SD/-Ura medium in the presence of 0-, 100-, 180- or 200-ng mL −1 AbA for 3 d, and pGADT7+pAbA-bait was used as a control. Statistical analysis IBM SPSS v25.0 (SPSS, Illinois, USA) was used to conduct statistical analyses. One-way analyses of variance (ANOVAs) with Duncan’s multiple range tests ( P < 0.05) or independent-samples t-tests were conducted in at least 3 independent experiments. Characterization of PlGATAs in tree peony To investigate the expansion patterns and functional conservation of the GATA family in tree peony, we conducted a comprehensive analysis, encompassing phylogeny, chromosomal localization, and comparative genomic. Phylogenetic reconstruction using full-length GATA protein sequences from multiple plant species resolved 19 PlGATA genes (Additional file 1: Table S2) into four well-supported clades (I-IV; Figure 1A). Clade I was the largest group, containing nine members, followed by Clade II (six members), Clade III (three members), and Clade IV (one members) (Figure 1B). Notably, several PlGATA proteins, particularly within Clade II, clustered closely with orthologs from A. thaliana , T. aestivum and V. vinifera (Figure 1A, Additional file 1: Table S2), suggesting function conservation across these species. Conserved motif analysis identified 10 significant motifs (E-value<0.05) with distinct clade distributions (Figure 1C; Additional file 2: Table S3). Domain analysis confirmed the presence of the conserved GATA domain in all PlGATA proteins (Figure 1D). Zinc finger motif analysis showed that the canonical C-X 2 -CX 18 -C-X 2 -C type predominated in Clades I, II, and IV. However, within Clade II, the gene PL-1G249490 exhibited a divergent C-X 2 -C-X 18 -R-X 2 -S motif. Clade III members uniquely possessed extended zinc fingers (C-X 2 -C-X 20 -C-X 2 -C) (Additional file 2: Figure S1) and additional TIFY and CCT domains (Figure 1D). These domains, also found in CONSTANS (CO), CO-like (COL), and TIMING OF CAB 1 (TOC1), are predicted to mediate protein-protein interaction and nuclear localization (Robson et al., 2002). Gene structure analysis revealed that PlGATAs in Clade I and II generally have simpler organizations (2-4 exons) compared to those in Clades III and IV (Figure 1E). These structural variations indicate significant functional diversification within the PlGATAs , particularly in DNA-binding specificity and developmental regulation. Chromosomal localization analysis demonstrated that the 18 PlGATA genes are unevenly distributed across five chromosomes ( PL-UG217660 is on Scaffold2552), with chromosome 3 harboring the highest number of genes (six genes; Figure 1F). Notably, on chromosome 3, two closely related Clade II members, PlGATA1 ( PL-3G231960 ) and PlGATA6 ( PL-3G268170 ), exhibit high sequence similarity and conserved synteny, strongly suggesting their origin from a local duplication event (Figure 1F). Furthermore, no tandem duplication events were detected within the PlGATA family. Comparative genomic analysis with A. thaliana , V. vinifera , and T. aestivum revealed varying degrees of synthetic conservation (Figure 1G). The strongest collinearity was observed with V. vinifera (20 syntenic gene pairs), followed by A. thaliana (14 pairs), while only one syntenic pair was detected with T. aestivum . This pattern reflects the closer evolutionary relationship between tree peony and other dicot species, particularly the woody perennial grapevine. Figure 1. Comprehensive genomic and evolutionary characterization of the PlGATA family in tree peony. A Phylogenetic analysis of GATA proteins from P. ludlowii , V. vinifera , A. thaliana and T. aestivum L. The phylogenetic tree was constructed based on the full-length amino acid sequences (Additional file 1: Table S2) using MEGA 11 with the Neighbor-Joining method and 1,000 bootstrap replicates. B Phylogenetic relationship among the identified PlGATA proteins. C Conserved motif analysis of PlGATA proteins. Ten predicted motifs are represented by different colored boxes, with detailed motif information in Additional file 2: Table S3. D Conserved protein domain analysis of PlGATAs. E Exon-intron structure analysis of PlGATAs . Green boxes and black lines represent exons and introns, respectively. Each pattern’s length is shown in proportion. F Genomic distribution and collinearity analysis of PlGATA genes in tree peony. The sliding window size set is 100 kb, with red to blue indicating gene density from high to low. G Collinearity analysis of GATA genes between tree peony and Arabidopsis, wheat and grapevine species. Gray lines represent all collinear gene pairs; red lines represent the collinear of PlGATA gene pairs. Expression analysis of PlGATAs genes To elucidate the potential transcriptional regulation mechanisms of PlGATAs in Clade II , we performed a comprehensive analysis of cis -acting regulatory elements within their promoter regions (Additional file 1: Table S4). The identified elements were classified into four major functional categories: phytohormone, development, light responsiveness, and biotic/abiotic stress (Figure 2A). Notably, phytohormone-related cis -elements constituted the predominant category (25-54% of total elements) (Figure 2B), which involved in ABA response (AAGAA-motif, ABRE, G-Box, ABRE3a, ABRE4, AP-1), salicylic acid response (TCA-element), MeJA-response (CGTCA-motif, TGACG-motif), gibberellin-response (TATC-box, P-box, GARE-motif), auxin response (AuxRR-core, TGA-element), cytokinin response (CARE, CCGTCC motif) and ethylene (ERE) (Figure 2A). Significantly, ABA-responsive elements were ubiquitously present in all promoters of all Clade II PlGATA members. Further examination of Clade II promoters identified conserved ABA-responsive elements alongside other motifs potentially relevant to bud dormancy, such as light-responsive motifs and stress-responsive elements. The universal presence of ABA-responsive elements across PlGATA family, including Clade II members, combined with the identification of other dormancy-associated elements (light-responsive), specifically enriched in Clade II promoters, supports the potential involvement of these genes in ABA-mediated processes critical for bud dormancy regulation in tree peony. Expression of Clade II PlGATAs genes was profiled in two contrasting bud types at the floral differentiation stage of P. × lemoinei ‘High Noon’: dormant and non-dormant buds (Figure 2C). Comparative analysis revealed distinct expression patterns. PlGATA6 and PlGATA11 showed relatively high expression in non-dormant bud, whereas PlGATA1 exhibited lower levels. Most Clade II PlGATAs genes displayed differential expression patterns between these two bud types, suggesting functional involvement in bud dormancy regulation in tree peony. Notably, qRT-PCR analysis showed significant expression differences between the segmental duplicate gene pair PlGATA1 and PlGATA6 , which are orthologous to TaGATA1 involved in ABA-mediated dormancy regulation (Wei et al., 2023) (Figure 2C). Given these striking expression differences and their potential functional significance, PlGATA1 and PlGATA6 were selected for subsequent functional characterization. Figure 2. Expression analysis of PlGATAs genes. Analysis of cis -acting regulatory elements in PlGATAs promoters was showed in A and B. A Functional category (phytohormones, abiotic and biotic stresses, light responsive elements, plant growth and development) and the numbers of cis -acting regulatory elements in different PlGATA family members. B Total number of cis -acting regulatory elements in each category. The red font represents ABA response elements, and the yellow font represents light response elements. C qRT-qPCR analysis of Clade II PlGATAs of P. × lemoinei ‘High Noon’ buds. Antagonistic regulation of bud dormancy by ABA-responsive PlGATA1 and PlGATA6 We successfully cloned PlGATA1 and PlGATA6 from P. × lemoinei ‘High Noon’. Full-length proteins consist of 159 and 138 amino acid residues for PlGATA1 and PlGATA6, respectively (Additional file 2: Figure S2 and Figure S3). Comparative analysis of orthologs ( A. thaliana AtGATA15/16 , V. vinifera VviGATA16a , and T. aestivum TaGATA1 ) confirmed both possess the canonical C-X₂-C-X₁₈-C-X₂-C zinc finger motif and fully conserved LCNACGI signature sequence, characteristic of Clade II plant GATA factors (Reyes et al., 2004). Despite zinc finger conservation, PlGATA6 exhibits a unique lysine (K) substitution adjacent to the third cysteine residue (Figure 3B). Domain architecture analysis further showed divergent regulatory regions: PlGATA1 contains an acidic N-terminus enriched in aspartate/glutamate (D/E), whereas PlGATA6 features a basic region rich in lysine/arginine (K/R) (Figure 3B). To determine the subcellular localization of PlGATA1 and PlGATA6, we performed transient expression assays in Nicotiana benthamiana leaves using GFP-tagged fusion proteins. Co-expression with the nuclear marker H2B-mCherry revealed complete colocalization of both GFP signals (green) with the nuclear mCherry fluorescence (red) (Figure 3A), demonstrating exclusive nuclear localization for PlGATA1 and PlGATA6. This supports their role as transcriptional regulators in the nucleus. Given ABA’s established role in bud dodormancy, we further investigated the temporal expression of PlGATA1 and PlGATA6 in floral buds following exogenous ABA treatment (0, 3, 6, and 12 h). qRT-PCR analysis revealed antagonistic responses: PlGATA1 significantly upregulated, while PlGATA6 rapidly downregulated (Figure 3C). These results demonstrate that the segmentally duplicated PlGATA1 and PlGATA6 underwent evolutionary neofunctionalization, retaining conserved DNA-binding domains but acquiring divergent regulatory architectures and antagonistic ABA-responsive expression profiles. This functional divergence establishes them as a pair of antagonistic transcriptional regulators modulating bud dormancy in tree peony. Figure 3. Sequence structure and expression pattern analysis of PlGATA genes. A Subcellular localization of PlGATA1-GFP and PlGATA6-GFP in N. benthamiana leaves. H2B-mCherry markS nuclei. Scale bars: 25μm (35S: GFP/PlGATA6-GFP), 50μm (PlGATA1/6-GFP). B Multiple sequence alignment of PlGATA1 and PlGATA6 with orthologs from A. thaliana (AtGATA15/16), V. vinifera (VviGATA16a) and T. aestivum (TaGATA1). C qRT-qPCR analysis of PlGATA1 and PlGATA6 in non-dormant buds treated with 100 μM ABA. Asterisks or letters indicate statistical significance as calculated by Student’s t test (*P < 0.05, **P < 0.01). Overexpression of PlGATA1 induces seed and bud dormancy To reveal the role of PlGATA1 in bud dormancy regulation, we employed a heterologous expression system in A. thaliana , leveraging conserved ABA signaling pathways between bud and seed dormancy (Xu et al., 2025). Three independent transgenic lines ( PlGATA1 -OE1/2/3) were selected for phenotypic analysis (Additional file 2: Figure S4). Germination assays were performed under three conditions: FH seeds and AR seeds (4-week storage), and cold-stratified seeds (4℃, 3 days). Notably, FH seeds of PlGATA1 -OE lines exhibited significantly delayed germination compared to vector controls (VC) (Figure 4A and B). Although stratification partially alleviated this dormancy phenotype, PlGATA1 -OE germination rate remained significantly lower than VC (Figure 4A and C), demonstrating PlGATA1 ’s conserved role in dormancy maintenance. For functional validation in tree peony, we generated PlGATA1- overexpressing transgenic plants via Agrobacterium -mediated transformation of non-dormant buds of P. × lemoinei ’High Noon’. qRT-PCR confirmed significant PlGATA1 upregulation in three independent overexpression lines ( PlGATA1 -OE) after 7 days (Figure 4F). These lines were subsequently employed to assess transcriptional regulation of paralog PlGATA6 and key dormancy-related markers ( PlCYP707A1 , PlCYP707A2-1 , PlCYP707A2-2 , PlPP2C , PlPYL2 , PlSARK1 , PlSARK2 , PlSDR2A and PlABI5 ). PlGATA1 -OE lines showed concurrent downregulation of PlGATA6 and ABA catabolism gene PlCYP707A1 , alongside upregulation of ABA signaling components ( PlSARK1 , PlSARK2 , PlPYL2 , and PlABI5 ) (Figure 4G). After 15 days, phenotypic analysis revealed that PlGATA1 -OE buds exhibited reduced longitudinal and lateral dimensions compared to controls (Figure 4D and E). These results indicated that PlGATA1 may modulate bud dormancy through ABA-dependent regulatory pathways. Figure 4. PlGATA1 Overexpression induces seed and bud dormancy. A Germination phenotype of FH and AR seeds from VC and PlGATA1 -OE A. thaliana on MS medium. B-C Germination rates of FH and AR seeds of VC and PlGATA1 -OE lines at different time points. D Phenotype of sprouting buds 15 days after infiltrated with super1300 empty vector or super1300- PlGATA1 . Isolated buds shown for comparison. Scale bar=5.0 mm. E Relative growth rate (height and width) of buds 15 days post-infiltration. F qRT-PCR analysis of PlGATA1 expression in buds 7 days post-infiltration. G qRT-qPCR analysis of paralog PlGATA6 and ABA metabolism genes 7 days post-infiltration. Transcript abundances were normalized to PsUBQ . Error bars represent SE of the mean from 3 biological replicates. Statistical significance was verified using Student’s t test and denoted by asterisks (*P < 0.05, **P < 0.01). Red pentagrams represent genes with significant differential expression. PlGATA6 inhibits seed and bud dormancy To examine functional divergence between segmentally duplicated paralogs PlGATA1 and PlGATA6 in dormancy regulation, we compared germination phenotyping using two distinct seed physiological states: FH and AR seeds. Three independent PlGATA6 -overexpressing transgenic lines (PlGATA6-OE1/2/3) and VC were analyzed to ensure biological reproducibility (Additional file 2: Figure S4). Germination assays revealed that FH PlGATA6 seeds exhibited significantly accelerated germination kinetics compared to VC under non-stratified conditions (Figure 5A and B). This phenotypic distinction was abolished following cold stratification (4℃ for 3 days), with both genotypes achieving comparable germination rates (Figure 5A and C), suggesting PlGATA6’s specific regulates primary dormancy rather than stratification-dependent secondary dormancy. To further investigate PlGATA6 role in bud dormancy, we employed a TRV-based VIGS system for knock-down in tree peony buds (Additional file 2: Figure S3). Compared to empty vector controls, TRV- PlGATA6 -infected buds showed delayed sprouting and inhibited growth at 15 days post-inoculation (DPI) (Figure 5D). Morphometric analysis confirmed that PlGATA6 -silenced buds had a significantly reduced width (Figure 5E). Efficient PlGATA6 knockdown was validated by qRT-PCR at 5 DPI (Figure 5F). Molecular analysis revealed that silencing of PlGATA6 led to downregulation of ABA catabolic gene ( PlCYP707A2-1 ) and signaling repressor ( PlPP2C1 ), while upregulation of ABA biosynthetic genes ( PlSARK1 and PlSARK2 ) and ABA-responsive TF ( PlABI5 ) (Figure 5F). Additionally, paralog PlGATA1 was significantly upregulated in the gene-silenced buds. These findings suggest that PlGATA6 modulates bud dormancy through ABA homeostasis and signaling, potentially through transcriptional regulation of ABA metabolic and response genes. Figure 5. PlGATA6 overexpression inhibits seed and bud dormancy. A Germination phenotype of FH and AR seeds from VC and PlGATA6 -OE Arabidopsis plants on MS medium and MS+100 μ M ABA medium. B-C Germination rates of FH and AR seeds of VC and PlGATA6 -OE lines on MS medium at different time points. D Phenotypes of P. × lemoinei ‘High Noon’ buds 15 days post Agrobacterium infiltration with TRV empty vector or TRV- PlGATA6 . Isolated buds showed for comparison. Scale bar=5.0 mm. E Growth widths measurements at 15 days. F qRT-PCR analysis of PlGATA6, paralog PlGATA1 and ABA metabolism genes expression in the buds inoculated at 5 days. Transcript abundances were normalized to PsUBQ. Error bars represent SE of the mean from 3 biological replicates. Statistical significance was verified using Student’s t test and denoted by asterisks (*P < 0.05, **P < 0.01). Red pentagrams represent genes with significant differential expression PlGATA1 directly binds and upregulates PlABI5 to modulate ABA-mediated bud dormancy Based on previous reports that Arabidopsis GATA15 binds WGATAR motifs (Scazzocchio, 2000), we cloned promoter regions of differentially expressed genes from overexpressing buds containing predicted GATA-binding sites to identify PlGATA1 targets. Using a dual-luciferase reporter system, we tested paralog PlGATA6 and five ABA-related genes ( PlCYP707A1 , PlSARK1 , PlSARK2 , PlPYL2 , and PlABI5 ) (Figure 6A), PlGATA1 specifically transactivated the PlABI5 promoter, increasing firefly luciferase activity 5.22-fold (p<0.01) with enhanced luminescence compared to controls (Figure 6B and C). To validate this activation, another reporter system involving a Y1H assay was employed. Three fragments in PlABI5 promoter region containing varying GATAs motifs (P1-3) were used for further binding analysis (Additional file 2: Figure S6 and 6D). Our results showed that PlGATA1 could bind to all fragment of P1, P2 and P3 (Figure 6E). These observations indicate that PlGATA1 directly activates PlABI5 transcription through binding GATA cis -elements, thereby mechanistically linking GATA-mediated regulation to ABA signaling in bud dormancy. Figure 6. PlGATA1 directly transactivates PlABI5 promoter. A Schematic diagram of effector-reporter constructs for dual-luciferase reporter system. REN, Renilla luciferase; LUC, firefly luciferase. B-C Dual-luciferase assay of candidate promoters co-infiltrated with pGreenⅡ-62-SK-PlGATA1 in tobacco leaves. A colour bar indicates luciferase intensity from weak (blue) to strong (red). LUC empty vector was used as the control. Data present mean ± SD of three independent biological replicates. Error bars represent SE of the mean from 5 biological replicates. Statistical significance was verified using Student’s t test (*P < 0.05, **P < 0.01) and denoted by asterisks. D Schematic diagrams of bait-pray constructs for YIH assay. E The R1, R2 and R3 regions are shown in the complete (full) promoter of PlABI5 . Yeast cells were diluted with distilled water (10 −1 to 10 −3 ) and grown on SD/-Leu medium with 100 or 180 mg mL −1 Aureobasidin A (AbA). PlGATA6 directly binds and upregulates PlCYP707A2-1 to modulate ABA-mediated bud dormancy To systematically identify downstream targets of PlGATA6, we cloned promoter regions of differentially expressed genes (DEGs) from our TRV-VIGS screen that contained predicted GATA cis -elements. Using a dual-luciferase reporter system, we examined the potential interaction between PlGATA6 and full-length promoters of paralog PlGATA1 , PlCYP707A2-1 , PlPP2C , PlSARK1 , PlSARK2 , and PlABI5 (Figure 7A). PlGATA6 specifically activate the PlCYP707A2-1 promoter, evidenced by enhanced luciferase signal intensity (Figure 7B) and a 9.62-fold increase in the activities of firefly luciferase (LUC; Figure 7C). This activation was validated by Y1H assay. Four fragments in the 2k bp PlCYP707A2-1 promoter region (P1-3) were analyzed for binding (Figure 7D). PlGATA6 bound to the fragment of P1, P2 and P3 (Figure 7E; Additional file 2: Figure S7). These results establish PlGATA6 as a direct transcriptional activator of PlCYP707A2-1 , which encodes a key ABA catabolic enzyme, ultimately modulating bud dormancy. Figure 7. PlGATA6 directly transactivates PlCYP707A2-1 promoter. A Schematic diagram of effector-reporter constructs for dual-luciferase reporter system. REN, Renilla luciferase; LUC, firefly luciferase. B-C Dual-luciferase assay of candidate promoters co-infiltrated with pGreenⅡ-62-SK-PlGATA6 in tobacco leaves. A colour bar indicates luciferase intensity from weak (blue) to strong (red). LUC empty vector was used as the control. Data present mean ± SD of three independent biological replicates. Error bars represent SE of the mean from 5 biological replicates. Statistical significance was verified using Student’s t test (*P < 0.05, **P < 0.01) and denoted by asterisks. D Schematic diagrams of bait-pray constructs for Y1H assay. The R1, R2 and R3 regions are shown in the complete (full) promoter of PlCYP707A2-1 . E Yeast cells were diluted with distilled water (10 −1 to 10 −3 ) and grown on SD/-Leu medium with 200 mg mL −1 Aureobasidin A (AbA). Discussion Bud dormancy imposes a critical constraint on off-season production in forcing cultivation of tree peony. Although recent advances have elucidated dormancy release mechanisms (Niu et al., 2025; Yuan et al., 2024; Zhang et al., 2024b; Zhang et al., 2024c), the transcriptional regulation governing dormancy induction remains elusive. Given the central role of ABA signaling, identifying its upstream regulators during dormancy initiation is essential. Building on phylogenetic and functional evidence implicating Clade II GATA TFs in ABA-mediated dormancy across plants, we investigated these subfamilies in tree peony. Our results reveal a novel antagonistic regulatory module comprising segmentally duplicated paralogs PlGATA1 and PlGATA6 , which coordinately fine-tune ABA homeostasis to control bud dormancy induction in tree peony (Fig. 8): PlGATA1 promotes dormancy by directly binding and activating the promoter of PlABI5 (a core ABA signaling effector)) (Figure 8), thereby enhancing ABA responsiveness. This functional conservation aligns with its wheat ortholog TaGATA1 (Wei et al., 2023). Conversely, PlGATA6 suppresses dormancy by directly activating the ABA catabolic gene PlCYP707A2-1 , reducing ABA accumulation (Figure 8). Critically, these paralogs exhibit reciprocal transcriptional antagonism. PlGATA1 overexpression suppresses PlGATA6 expression, while PlGATA6 silencing upregulates PlGATA1 (Figure 4 and 5). Though neither directly transactivates the other’s promoter (Figure 6 and 7), this cross-regulation suggests indirect reinforcement of their opposing functions. Notably, this module further exhibits ABA-responsive reinforcement: ABA induces PlGATA1 while repressing PlGATA6 (Figure 3D and E), creating a self-amplifying loop that enhances ABA signaling. This mechanism mirrors Arabidopsis seed dormancy, where ABI4-mediated repression of CYP707A boosts ABA accumulation (Shu et al., 2013). This functional divergence reflects neofunctionalization following segmental duplication (Panchy et al., 2016). PlGATA1 and PlGATA6 retain high sequence conservation in core DNA-binding GATA domains, including the Clade II-specific LCNACGI motif (Reyes et al., 2004). However, PlGATA1 and PlGATA6 exhibit significant divergence in regulatory regions (Figure L3). PlGATA1 possesses an acidic N-terminus typical of transcriptional activation domains (Boija et al., 2018), while PlGATA6 features a basic motif potentially enhancing DNA affinity or binding cis -elements like light-responsive sequences (Waters et al., 2009). This structural divergence underpins functional specialization: PlGATA1 acts as a transcriptional activator promoting dormancy, whereas PlGATA6 activates genes suppressing dormancy. Such antagonism within duplicated GATA pairs is evolutionarily conserved, as observed in Arabidopsis GNC/GNL (Richter et al., 2013). Moreover, the principle of antagonistic TF pairs regulating dormancy through ABA signaling extends beyond GATA. For example, antagonistic bHLH pairs similarly balance ABA/GA crosstalk during dormancy cycling (Xu, F et al., 2022), underscoring a recurrent evolutionary strategy for environmental adaptation. The PlGATA1 and PlGATA6 module could function as a central hub integrating endogenous (ABA feedback) and likely environmental cues to orchestrate bud dormancy induction in tree peony. While we demonstrate that PlGATA1 and PlGATA6 primarily fine-tune ABA homeostasis, coordination between ABA and other phytohormones (GA), is also critical for regulating tree peony bud dormancy (Yuan et al., 2024). More importantly, whether PlGATA1 and PlGATA 6 similarly modulate other hormones or signaling pathways in bud dormancy induction still remains to be investigated. In summary, this study establishes PlGATA1 and PlGATA6 as evolutionarily diverged paralogs forming an antagonistic switch regulating ABA-mediated bud dormancy induction. By activating opposing ABA network ( PlGATA1 reinforcing signaling via PlABI5 , and PlGATA6 promoting catabolism via PlCYP707A2-1 ) while reciprocally repressing each other under ABA-responsive feedback, they create a robust regulatory system governing dormancy transition. Our work bridges a key knowledge gap in bud dormancy regulation in tree peony and provides both conceptual frameworks and actionable targets for manipulating dormancy in horticulture. Figure 8. Proposed model for PlGATA1 and PlGATA6 antagonistically regulating bud dormancy induction in tree peony. PlGATA1 enhances ABA biosynthesis by directly activating PlABI5 transcription. PlGATA6 promotes ABA catabolism through direct transcriptional regulation of PlCYP707A2-1 . ABA positively regulates PlGATA1 expression while suppressing PlGATA6 , establishing a self-reinforcing loop. Solid arrows indicate activation; T-bars denote repression. Acknowledgements This work was supported by the National Natural Science Foundation of China (31972446), the National Key Research and Development Program of China (2023YFD1200105) and the 5·5 Engineering Research & Innovation Team Project of Beijing Forestry University (No: BLRC2023A06). Conflicts of Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Data Availability Statement The data that support the findings of this study are available on request from the corresponding author. The data are not publicly available due to privacy or ethical restrictions. References Ahmad, S., Lu, C., Gao, J., et al. (2024). Integrated Proteomic, Transcriptomic, and Metabolomic Profiling Reveals that the Gibberellin–Abscisic Acid Hub Runs Flower Development in the Chinese Orchid Cymbidium sinense . Horticulture Research, 11 (5). https://doi.org/10.1093/hr/uhae073 Artimo, P., Jonnalagedda, M., Arnold, K., et al. (2012). ExPASy: SIB Bioinformatics Resource Portal. Nucleic Acids Research, 40 (W1), W597. https://doi.org/10.1093/nar/gks400 Bailey, T. L., Johnson, J., Grant, C. E., et al. (2015). The MEME Suite. Nucleic Acids Research, 43 (W1), W39-W49. https://doi.org/10.1093/nar/gkv416 Bi, Y. M., Zhang, Y., Signorelli, T., et al. (2005). Genetic Analysis of Arabidopsis GATA Transcription Factor Gene Family Reveals a Nitrate-Inducible Member Important for Chlorophyll Synthesis and Glucose Sensitivity. The Plant Journal, 44 (4), 680-692. https://doi.org/10.1111/j.1365-313X.2005.02568.x Boija, A., Klein, I. A., Sabari, B. R., et al. (2018). Transcription Factors Activate Genes through the Phase-Separation Capacity of Their Activation Domains. Cell, 175 (7), 1842-1855.e1816. https://doi.org/10.1016/j.cell.2018.10.042 Bonhomme, M., Rageau, R., Gendraud, M. (2000). Influence of Temperature on the Dynamics of Atp, Adp and Non-Adenylic Triphosphate Nucleotides in Vegetative and Floral Peach Buds During Dormancy. Tree Physiology, 20 (9), 615-621. https://doi.org/10.1093/treephys/20.9.615 Chang, Y., Zhang, W., Ma, Y., et al. (2022). Transcriptome Analysis of Floral Bud Development and Function Analysis of a Novel Co Gene in Paeonia × lemoinei ‘High Noon’. Scientific Reports, 12 (1). https://doi.org/10.1038/s41598-022-22195-z Chen, C., Chen, H., Zhang, Y., et al. (2020). TBtools: An Integrative Toolkit Developed for Interactive Analyses of Big Biological Data. Molecular Plant, 13 (8), 1194-1202. https://doi.org/10.1016/j.molp.2020.06.009 Cheng, X., Tian, B., Gao, C., et al. (2021). Identification and Expression Analysis of Candidate Genes Related to Seed Dormancy and Germination in the Wheat GATA Family. Plant Physiology and Biochemistry, 169 , 343-359. https://doi.org/10.1016/j.plaphy.2021.11.012 Clough, S. J., and Bent, A. F. (2010). Floral Dip: A Simplified Method for Agrobacterium -Mediated Transformation of Arabidopsis thaliana . Plant Journal, 16 (6), 735-743. https://doi.org/10.1046/j.1365-313x.1998.00343.x Crooks, G. E., Hon, G., Chandonia, J.-M., et al. (2004). WebLogo: A Sequence Logo Generator: Figure 1. Genome Research, 14 (6), 1188-1190. https://doi.org/10.1101/gr.849004 Eddy, S. (1998). Profle hidden Markov models. Bioinformatics, 14 (9), 755–763. https://doi.org/10.1093/bioinformatics/14.9.755 Gai, S., Zhang, Y., Liu, C., et al. (2013). Transcript Profiling of Paoenia ostii during Artificial Chilling Induced Dormancy Release Identifies Activation of GA Pathway and Carbohydrate Metabolism. PLoS ONE, 8 (2). https://doi.org/10.1371/journal.pone.0055297 Gai, W., Liu, C., Yang, M., et al. (2024). Calcium Signaling Facilitates Chilling- and Ga- Induced Dormancy Release in Tree Peony. Frontiers in Plant Science, 15 . https://doi.org/10.3389/fpls.2024.1362804 Gao, H., Li, J., Yu, J., et al. (2025). Germination Fitness Differs Between Penoxsulam Resistant and Susceptible Echinochloa Phyllopogon Biotypes. Agriculture Communications, 3 (2). https://doi.org/10.1016/j.agrcom.2025.100088 Gao, L., Niu, D., Chi, T., et al. (2023). PsRGL1 Negatively Regulates Chilling- and Gibberellin-Induced Dormancy Release by PsF-box1-Mediated Targeting for Proteolytic Degradation in Tree Peony. Horticulture Research, 10 (5). https://doi.org/10.1093/hr/uhad044 Gillespie, L. M., and Volaire, F. A. (2017). Are Winter and Summer Dormancy Symmetrical Seasonal Adaptive Strategies? The Case of Temperate Herbaceous Perennials. Annals of Botany, 119 (3), 311-323. https://doi.org/10.1093/aob/mcw264 Graeber, K. A. I., Nakabayashi, K., Miatton, E., et al. (2012). Molecular Mechanisms of Seed Dormancy. Plant, Cell & Environment, 35 (10), 1769-1786. https://doi.org/10.1111/j.1365-3040.2012.02542.x Horton, P., Park, K.-J., Obayashi, T., et al. (2007). WoLF PSORT: Protein Localization Predictor. Nucleic Acids Research, 35 (Web Server issue), 585-587. https://doi.org/10.1093/nar/gkm259 Huang, X., Xue, T., Dai, S., et al. (2008). Genes Associated With the Release of Dormant Buds in Tree Peonies ( Paeonia suffruticosa ). Acta Physiologiae Plantarum, 30 (6), 797-806. https://doi.org/10.1007/s11738-008-0184-0 Krzywinski, M., Schein, J., Birol, İ., et al. (2009). Circos: An Information Aesthetic for Comparative Genomics. Genome Research, 19 (9), 1639-1645. https://doi.org/10.1101/gr.092759.109 Lescot, M., Déhais, P., Thijs, G., et al. (2002). Plantcare, a Database of Plant Cis -Acting Regulatory Elements and A Portal to Tools for In Silico Analysis of Promoter Sequences. Nucleic Acids Research, 30 (1), 325-327. https://doi.org/10.1093/nar/30.1.325 Letunic, I., and Bork, P. (2021). Interactive Tree Of Life (iTOL) v5: An Online Tool for Phylogenetic Tree Display and Annotation. Nucleic Acids Research, 49 (W1), W293-W296. https://doi.org/10.1093/nar/gkab301 Letunic, I., Khedkar, S., Bork, P. (2020). SMART: Recent Updates, New Developments and Status in 2020. Nucleic Acids Research, 49 (D1), D458–D460. https://doi.org/10.1093/nar/gkaa937 Liu, P. P., Koizuka, N., Martin, R. C., et al. (2005). The BME3 ( Blue Micropylar End 3 ) GATA Zinc Finger Transcription Factor is a Positive Regulator of Arabidopsis Seed Germination. The Plant Journal, 44 (6), 960-971. https://doi.org/10.1111/j.1365-313X.2005.02588.x Livak, K. J., & Schmittgen, T. D. (2001). Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT Method. Methods, 25(4), 402-408. https://doi.org/10.1006/meth.2001.1262 Lowry, J. A., and Atchley, W. R. (2000). Molecular Evolution of the GATA Family of Transcription Factors: Conservation Within the DNA-Binding Domain. Journal of Molecular Evolution, 50 (2), 103-115. https://doi.org/10.1007/s002399910012 Lu, S., Wang, J., Chitsaz, F., et al. (2020). CDD/SPARCLE: the Conserved Domain Database in 2020. Nucleic Acids Research, 48 (D1), D265-D268. https://doi.org/10.1093/nar/gkz991 Lundell, R., Hänninen, H., Saarinen, T., et al. (2019). Beyond Rest and Quiescence (Endodormancy and Ecodormancy): a Novel Model for Quantifying Plant–Environment Interaction in Bud Dormancy Release. Plant, Cell & Environment, 43 (1), 40-54. https://doi.org/10.1111/pce.13650 Ma, M.-M., Zhang, H.-F., Tian, Q., et al. (2024). MIKC Type Mads-Box Transcription Factor LcSVP2 is Involved in Dormancy Regulation of The Terminal Buds in Evergreen Perennial Litchi ( Litchi chinensis Sonn. ). Horticulture Research, 11 (7). https://doi.org/10.1093/hr/uhae150 Mistry, J., Chuguransky, S., Williams, L., et al. (2020). Pfam: The protein families database in 2021. Nucleic Acids Research, 49 (D1), D412–D419. https://doi.org/10.1093/nar/gkaa913 Mornya, P. M. P., and Cheng, F. (2018). Effect of Combined Chilling and GA 3 Treatment on Bud Abortion in Forced ‘Luoyanghong’ Tree Peony (Paeonia suffruticosa Andr.). Horticultural Plant Journal, 4 (6), 250-256. https://doi.org/10.1016/j.hpj.2018.09.003 Niu, D., Liu, F., Gao, L., et al. (2025). MADS-Domain Transcription Factor AGAMOUS LIKE-9 Participates in the Gibberellin Pathway to Promote Bud Dormancy Release of Tree Peony. Horticulture Research, 12 (5), uhaf043. https://doi.org/10.1093/hr/uhaf043 Nv, O., Obroucheva, N., Lityagina, S., et al. (2019). Distinct Pathways of Regulation Seed Dormancy and Germination by Abscisic Acid. International Journal of Cell Science & Molecular Biology, 5 (4). https://doi:10.19080/ijcsmb.2019.05.555667 Panchy, N., Lehti-Shiu, M., Shiu, S.-H. (2016). Evolution of Gene Duplication in Plants. Plant Physiology, 171 (4), 2294-2316. https://doi:10.1104/pp.16.00523 Reyes, J. C., Muro-Pastor, M. I., Florencio, F. J. (2004). The GATA Family of Transcription Factors in Arabidopsis and Rice. Plant Physiology, 134 (4), 1718-1732. https://doi.org/10.1104/pp.103.037788 Richter, R., Behringer, C., Zourelidou, M., et al. (2013). Convergence of Auxin and Gibberellin Signaling on the Regulation of the GATA Transcription Factors Gnc and Gnl in Arabidopsis thaliana . Pnas, 110 (32), 13192-13197. https://doi.org/10.1073/pnas.1304250110 Robson, F., Costa, M. M. R., Hepworth, S. R., et al. (2002). Functional Importance of Conserved Domains in the Flowering‐Time Gene Constans Demonstrated by Analysis of Mutant Alleles and Transgenic Plants. The Plant Journal, 28 (6), 619-631. https://doi.org/10.1046/j.1365-313x.2001.01163.x Sano, N., and Marion-Poll, A. (2021). ABA Metabolism and Homeostasis in Seed Dormancy and Germination. International Journal of Molecular Sciences, 22 (10). https://doi.org/10.3390/ijms22105069 Scazzocchio, C. (2000). The Fungal GATA Factors. Curr. Opin. Microbiol., 3 (2), 126-131. https://doi.org/10.1016/S1369-5274(00)00063-1 Shu, K., Zhang, H., Wang, S., et al. (2013). ABI4 Regulates Primary Seed Dormancy by Regulating the Biogenesis of Abscisic Acid and Gibberellins in Arabidopsis. PLOS Genetics, 9 (6). https://doi.org/10.1371/journal.pgen.1003577 Singh, R. K., Maurya, J. P., Azeez, A., et al. (2018). A Genetic Network Mediating the Control of Bud Break in Hybrid Aspen. Nature Communications, 9 (1). https://doi.10.1038/s41467-018-06696-y Tamura, K., Stecher, G., Kumar, S., et al. (2021). MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Molecular Biology and Evolution, 38 (7), 3022-3027. https://doi.org/10.1093/molbev/msab120 Wang, Y., Tang, H., DeBarry, J. D., et al. (2012). MCScanX: A Toolkit for Detection and Evolutionary Analysis of Gene Synteny and Collinearity. Nucleic Acids Research, 40 (7), e49-e49. https://doi.org/10.1093/nar/gkr1293 Waters, M. T., Wang, P., Korkaric, M., et al. (2009). GLK Transcription Factors Coordinate Expression of the Photosynthetic Apparatus in Arabidopsis . The Plant Cell, 21 (4), 1109-1128. https://doi.org/10.1105/tpc.108.065250 Wei, X., Li, Y., Zhu, X., et al. (2023). The GATA Transcription Factor TaGATA1 Recruits Demethylase TaELF6‐A1 and Enhances Seed Dormancy in Wheat by Directly Regulating TaABI5 . Journal of Integrative Plant Biology, 65 (5), 1262-1276. https://doi.org/10.1111/jipb.13437 Xiao, P.-X., Li, Y., Lu, J., et al. (2023). High-quality Assembly and Methylome of a Tibetan Wild Tree Peony Genome ( Paeonia ludlowii ) Reveal the Evolution of Giant Genome Architecture. Horticulture Research, 10 (12). https://doi.org/10.1093/hr/uhad241 Xu, F., Tang, J., Wang, S., et al. (2022). Antagonistic Control of Seed Dormancy in Rice by Two BHLH Transcription Factors. Nature Genetics, 54 (12), 1972-1982. https://doi.org/10.1038/s41588-022-01240-7 Xu, S., He, Y., Zhou, Z., et al. (2025). Transcriptome Analysis Reveals the Key Roles of TaSMP1 and ABA Signaling Pathway in Wheat Seed Dormancy and Germination. Planta, 261 (4), 1-20. https://doi.org/10.1007/s00425-025-04667-4 Yamane, H., Andrés, F., Bai, S., et al. (2023). Editorial: Environmental and Molecular Control of Bud Dormancy and Bud Break in Woody Perennials: An Integrative Approach. Frontiers in Plant Science, 14 . https://doi.org/10.3389/fpls.2023.1104108 Yang, Q., Gao, Y., Wu, X., et al. (2021). Bud Endodormancy in Deciduous Fruit Trees: Advances and Prospects. Horticulture Research, 8 . https://doi.org/10.1038/s41438-021-00575-2 Yao, J., Shen, Z., Zhang, Y., et al. (2020). Populus euphratica WRKY1 Binds the Promoter of H+-ATPase Gene to Enhance Gene Expression and Salt Tolerance. Journal of Experimental Botany, 71 (4), 1527-1539. https://doi.org/10.1093/jxb/erz493 Yu, M., Liu, J., Du, B., et al. (2021). NAC Transcription Factor PwNAC11 Activates ERD1 by Interaction with ABF3 and DREB2A to Enhance Drought Tolerance in Transgenic Arabidopsis. International Journal of Molecular Sciences, 22 (13). https://doi.org/10.3390/ijms22136952 Yuan, Y., Zeng, L., Kong, D., et al. (2024). Abscisic Acid–Induced Transcription Factor PsMYB306 Negatively Regulates Tree Peony Bud Dormancy Release. Plant Physiology, 194 (4), 2449-2471. https://doi.org/10.1093/plphys/kiae014 Zhang, S., Feng, M., Chen, W., et al. (2019). In Rose, Transcription Factor PTM Balances Growth and Drought Survival via PIP2;1 Aquaporin. Nature Plants, 5 (3), 290-299. https://doi.org/10.1038/s41477-019-0376-1 Zhang, T., Wang, X., Yuan, Y., et al. (2024a). PsmiR159b- PsMYB65 Module Functions in the Resumption of Bud Growth after Endodormancy by Affecting the Cell Cycle in Tree Peony. Horticulture Research, 11 (4). https://doi.org/10.1093/hr/uhae052 Zhang, X., Fan, R., Yu, Z., et al. (2024b). Genome-Wide Identification of Gata Transcription Factors in Tetraploid Potato and Expression Analysis in Differently Colored Potato Flesh. Frontiers in Plant Science, 15 . https://doi.org/10.3389/fpls.2024.1330559 Zhang, X., Ma, J., Yang, S., et al. (2023). Analysis of GATA Transcription Factors and Their Expression Patterns under Abiotic Stress in Grapevine ( Vitis vinifera L.). BMC Plant Biology, 23 (1). https://doi.org/10.1186/s12870-023-04604-1 Zhang, Y., Niu, D., Yuan, Y., et al. (2024c). PsSOC1 is Involved in the Gibberellin Pathway to Trigger Cell Proliferation and Budburst During Endodormancy Release in Tree Peony. New Phytologist, 243 (3), 17. https://doi.org/10.1111/nph.19893 Zhou, H., Cheng, F.-Y., Wang, R., et al. (2013). Transcriptome Comparison Reveals Key Candidate Genes Responsible for the Unusual Reblooming Trait in Tree Peonies. PLoS ONE, 8 (11). https://doi.org/10.1371/journal.pone.0079996 Supplementary Material File (image1.gif) Download 3.97 MB Information & Authors Information Version history V1 Version 1 18 July 2025 Copyright This work is licensed under a Non Exclusive No Reuse License. Collection Plant, Cell & Environment Keywords aba homeostasis antagonistic regulation bud dormancy induction hormones signaling tree peony Authors Affiliations Ziwen Geng Beijing Forestry University School of Landscape Architecture View all articles by this author Chunyan He Beijing Forestry University School of Landscape Architecture View all articles by this author Fangting Qi Beijing Forestry University School of Landscape Architecture View all articles by this author Jianing Han Beijing Forestry University School of Landscape Architecture View all articles by this author Lei Zhang Beijing Forestry University School of Landscape Architecture View all articles by this author Fangyun Cheng 0000-0002-3928-5731 [email protected] Beijing Forestry University School of Landscape Architecture View all articles by this author Metrics & Citations Metrics Article Usage 292 views 159 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Ziwen Geng, Chunyan He, Fangting Qi, et al. 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