Seeds

Populus alba clone ‘Villafranca’ (white poplar), highly suitable for biomass production and ecosystem restoration, is a model system for molecular and physiological studies, but no reports are available concerning seed quality. Although clonal propagation is the preferred approach for commercial purposes, attention should be given to face genetic variability losses in the existing germplasm. To address this challenge, new populations should be developed starting from seeds, overcoming the issues of low germinability and viability during storage. This study proposes to develop tailored treatments to improve the germination of long-term stored white poplar seeds. Priming and soaking protocols, based on the use of water or spermidine (Spd, 50 and 100 μM), were tested. Treatment efficacy was assessed based on germination parameters, reactive oxygen species (ROS) profiles, and the expression patterns of genes with key roles in early seed germination. Soaking with 100 μM Spd for 4 h significantly enhanced germination percentage and speed. Low ROS levels were evidenced in the Spd-treated seeds, compared to water-soaked seeds. High expression of genes involved in desiccation tolerance acquisition, polyamine biosynthesis, and antioxidant defense was observed only in dry seeds. The results are discussed in view of the potential protective role of Spd. Full article

Seed science is the comprehensive study of seeds. It encompasses their biology, production, technology, genetics, physiology, ecology, and applications in agriculture and conservation. Seed science has undergone transformative advancements through the integration of microbial technologies, with beneficial microorganisms emerging as critical tools for enhancing germination, seedling vigor, and crop resilience. Research demonstrates that microbial treatments improve nutrient uptake, hormonal regulation, and stress tolerance while establishing early symbiotic relationships with plants. This review synthesizes recent advances in understanding the roles of beneficial microbes in seed science, focusing on their impact on seed germination, seedling growth, and plant health. We explore the composition and transmission of seed microbiomes, highlighting the vertical transfer of microbes from parent plants to seeds and the influence of environmental factors on microbial community structure. The review also discusses innovative approaches to seed microbiome engineering. Particular attention is given to seed biopriming with plant growth-promoting bacteria (PGPB), which has shown significant potential in improving germination rates, seedling vigor, and crop productivity. Specific microbial strains, such as Trichoderma species and Pseudomonas fluorescens, are discussed with emphasis on their mechanisms of action in enhancing plant performance. The review also addresses the impact of breeding on seed microbiomes and explores emerging research directions, including the development of tailored microbial inoculants and the investigation of intracellular seed bacteria. By synthesizing these findings, this review aims to provide a comprehensive summary of the current state of seed microbiome research and its implications in seed science for sustainable agriculture. Full article

Seed propagation is the primary means of reproducing many native and endemic species, including garberia [Garberia heterophylla (W. Bartram) Merrill & F. Harper]. This attractive pollinator plant, native to Florida, is scarcely found in nursery production and largely unknown to the gardening community. To better understand the seed biology of garberia, a series of experiments were conducted to evaluate the effects of population on seed viability and germination response to four seasonal temperatures, as well as the effects of time on seed storability. Initial seed viability was 49% and 60% for Central and North Florida populations, respectively. Seeds germinated readily, indicating non-dormancy, with significant effects of population and temperature observed. Overall, on day 28, a greater germination proportion was observed from seeds collected from North Florida than Central Florida across temperatures, except for winter (11/22 °C), where responses were similar. The greatest germination proportion for seeds collected from North Florida was observed at 15/27 °C (fall) and 19/29 °C (spring), whereas the greatest germination from Central Florida was observed at 11/22 °C (winter), with the steepest decline observed at summer temperatures (24/33 °C). Further, it was observed that garberia seeds are intolerant of long-term storage, losing viability as early as 3 months under conventional cold or room temperature storage and decreasing substantially more after 6 months. These findings contribute to the overall understanding of the seed biology of underutilized species such as garberia, key to the development of efficient and reliable propagation systems for our nursery industry. Full article

Acacia species are important trees in arid ecosystems due to their diverse ecological roles, such as providing vegetation cover, community structures, food resources for animals, soil stabilization, and erosion prevention. However, in the Arabian Peninsula, Acacia species are declining due to climate change, overgrazing, and fuelwood harvesting. This study evaluates the effectiveness of various pre-sowing treatments—sulfuric acid soaking and tap and hot water soaking—on breaking seed dormancy to enhance germination in nine Acacia species native to the Al-Baha region of Saudi Arabia. The key germination indicators assessed were the mean germination time (MGT), germination percentage (GP), and germination index (GI). Sulfuric acid treatments for 10–15 min reduced the MGT and increased the GP for A. etbaica, A. hamoulosa, and A. tortilis, while A. origena responded best to 1 min of hot water soaking. Conversely, A. asak, A. ehrenbergiana, and A. johnwoodii showed little to no germination improvement with treatment and A. oerfota and A. gerrardii showed no germination improvement, indicating the need for alternative methods. These findings indicate that the seed germination requirements vary within Acacia spp. from the same geographic region and similar climatic conditions. Further work is required for five of the species tested to develop better seed germination techniques, given the potential utility of Acacia spp., in ecological restoration and sustainable land management in arid regions. Full article

Crops are continually subjected to frequent and extreme changes in climate, such as high temperatures and soil water deficits. Many studies have shown the individual effects of these factors on plants, but their combined effects on reproductive growth and subsequent seed germinability have received little attention. In this study, we used canola (Brassica napus) plants and grew them through their lifecycle under two temperature regimes (20/10 °C and 24/14 °C, 16 h light/8 h dark) in controlled-environment growth chambers. Half of the plants were watered to field capacity (well-watered) and the other half at wilting point (water-stressed). During the reproductive stage, the flower, silique, and seed traits were measured. Higher temperatures decreased the petal width by 1.17 times but increased petal anthocyanins by 1.03 times. The water deficit decreased the silique length and total seed number by 1.21 and 1.32 times, respectively, but increased nectar sugar concentration by 1.28 times. The total volume of nectar was affected by the interaction of temperature and water. The nectar volume was lowest in the water-stressed plants under higher temperatures (2.66 ± 0.29 µL per flower) but highest in the well-watered plants under the same temperature regime (5.73 ± 0.37 µL per flower). In conclusion, the combined effects of temperature and water were less pronounced than the individual effects of these factors on canola reproductive yield. Full article

This paper presents a pilot project conducted by ISPRA and ARPA Campania to develop a monitoring protocol to detect the presence of genetically modified (GM) oilseed rape (Brassica napus) plants resulting from accidental seed dispersal during transportation from entry points to storage and processing facilities; the project has been implemented in Italy’s Campania region. The unintentional dispersal of GM oilseed rape seeds and the potential establishment of feral populations have been identified as environmental concerns in various countries, even when GM oilseed rape is imported solely for processing and not for cultivation. The project activities were designed, taking into account the characteristics of the Italian environment and infrastructures. Multiple sampling campaigns were conducted in autumn 2018, spring 2019, and autumn 2019 to validate the selected transects and assess the presence of Brassicaceae species, with a particular focus on oilseed rape. These efforts involved direct monitoring and sample collection along transport routes from the port of Salerno to seed companies in the provinces of Benevento and Caserta. Field observations and import data revealed a decrease in oilseed rape movement at the port of Salerno in the years preceding the survey, while seed companies near Benevento remained active sites for white mustard (Sinapis alba). The presence of S. alba and the simultaneous occurrence of oilseed rape and Raphanus raphanistrum—a species with high hybridization potential—support the hypothesis that seed companies may act as hotspots for accidental seed dispersal and that potential interspecific gene flow can occur. The study also validated the adopted sampling and molecular analysis methods, including DNA extraction and PCR, for the detection of the Cruciferin A (CruA) gene in all Brassica species collected. These findings highlight the need to strengthen post-marketing monitoring plans, even when GM rapeseed is imported solely for processing, to mitigate the potential risks associated with unintended gene flow. Full article

The taxonomic complexity of Phoenix palms in the Canary Islands, where multiple morphotypes representing at least four taxa currently exist, presents significant challenges for archaeobotanical identification. We developed a Bayesian probabilistic framework to identify archaeological Phoenix seeds within the context of genus-wide morphological diversity. Our analysis incorporated thousands of specimens including modern reference collections, archaeological materials from pre-Hispanic sites in Gran Canaria and La Gomera (3–16th centuries CE), and fossil remains. We recorded quantitative measurements and qualitative characteristics for each specimen. To understand taphonomic effects, we conducted experimental carbonization of modern P. canariensis seeds and documented the resulting morphological alterations. We performed a hierarchical cluster analysis using Ward’s minimum variance method and calculated taxonomic assignment probabilities for archaeological specimens using Bayesian inference, where likelihood was derived from taxon proportions within assigned clusters. The results indicated a high probability (0.69–1.00) that the archaeological specimens belong to P. canariensis var. canariensis, with no evidence for P. dactylifera presence. These findings provide critical insights into pre-Hispanic exploitation of Phoenix palms, particularly the endemic P. canariensis, which served as a vital resource, providing food, fiber, and construction materials. Our methodological approach offers a robust framework for addressing taxonomic uncertainty in archaeobotanical research while enhancing understanding of historical palm biogeography and resource use patterns in the Canary Islands. Full article

Annual grass weeds can provide significant competition to an establishing sown tropical perennial grass pasture. At least two years of grass weed control prior to sowing is required to reduce the weed seed bank. Pre-emergent herbicides used in summer cereals, such as atrazine or s-metolachlor with metcamifen seed safener, may reduce this preparation time. Two controlled-environment experiments were conducted to assess the potential for these pre-emergent herbicides to be used with several tropical perennial grasses. Experiment 1 tested the effect of metcamifen (400 g L⁻¹ a.i. at 0–2× label rate) on the emergence and vigor of Chloris gayana, Dichanthium aristatum, Digitaria eriantha and Panicum coloratum, with Sorghum bicolor as the control. Experiment 2 tested the effect of s-metolachlor (960 g ha⁻¹ a.i.) with metcamifen-treated or untreated seed, and atrazine (1800 g ha⁻¹ a.i.) on the emergence and early growth of the grasses. Metcamifen did not inhibit emergence or vigor of the grasses. Without metcamifen seed treatment, s-metolachlor reduced the growth of the tropical perennial grasses by 47–100%, while it had no such effect on S. bicolor. In contrast, there was no effect of atrazine on shoot yields of the grasses, nor of s-metolachlor when D. aristatum, D. eriantha and P. coloratum seed had been treated with metcamifen. The collective results indicate that the herbicide safener metcamifen does not reduce the viability of tropical perennial grass seed and provides some protection against s-metolachlor, albeit not complete protection at the rates used in our study. Atrazine did not affect emergence or early growth of the grasses. Full article

Bioactive compounds in plants, such as carrots, have been widely used for their benefits. In agriculture, their potential as biostimulants still needs to be investigated, especially for their possible antioxidant action in plants subjected to abiotic stresses, such as salinity. This work aimed to evaluate the elicitor potential of carrot extract in alleviating salt stress in rice plants. This study aimed to evaluate the elicitor potential of carrot extract in alleviating saline stress in the rice cultivars BRS Querência and BRS 358. Aqueous extracts of carrot roots at concentrations of 0% (water), 25%, 50%, and 100% were used to soak rice seeds for 48 h, which were then subjected to different concentrations of NaCl (0, 25, 75, and 150 mM). To determine the effect of carrot extract as an elicitor under saline stress conditions, the following tests were conducted: germination, seedling length, dry mass, and oxidative stress through the activity of antioxidant enzymes, superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), peroxide content, and lipid peroxidation (hydrogen peroxide H₂O₂ and malonaldehyde MDA). Carrot extract increased the germination rate and maintained germination even under increased salinity rates in both cultivars. The application of 25 mM NaCl also boosted germination rates, followed by a significant decrease due to increased salinity rates. Shoot and root lengths and dry mass parameters showed a linear decrease in response to increasing NaCl concentrations. The activities of superoxide dismutase (SOD), ascorbate peroxidase (APx), and catalase (CAT) enzymes tended to decrease as the concentration of carrot extract increased, whereas the opposite was observed with NaCl application. Based on the combined analysis of the evaluated parameters, carrot extract application under the tested conditions was efficient in mitigating oxidative stress caused by high salinity conditions. Full article

Kentucky bluegrass (KBG) has poor seed establishment in the fall when used as a perennial groundcover in corn production. This study was conducted to investigate the effect of various seed treatments and soil amendments on the establishment of KBG under drought and non-drought conditions, simulated in a growth chamber. The effect of seed treatments, soil amendments, and irrigation frequency on KBG germination and shoot dry weight were measured over 21 days in a controlled environment at 21 °C, 50% RH, and exposure to a constant red light. The treatments were the Hydroloc seed treatment, a lime soil amendment, the Pivot Bio seed treatment, an ammonium nitrate soil amendment, a gibberellic acid seed treatment, osmotic seed priming, and an untreated control. The layout was a randomized complete block design, with two irrigation frequencies (restricted and full irrigation) and four replications (blocks). The irrigation treatments were applied to whole plots and the seed treatments were applied to subplots. The entire experiment was repeated four times. Irrigation affected the germination of all the seed treatments, but the size of the effect depended on the seed treatment applied. The control and Hydroloc treatments did not have significantly different dry shoot weights, while all the other treatments had significantly different dry shoot weights when comparing the irrigation regimes. The Hydroloc treatment significantly outperformed all the other treatments in regard to the restricted and full irrigation regime. These results indicate that the Hydroloc seed treatment improves KBG germination and shoot dry weight in drought and non-drought conditions, promoting KBG establishment in a wide range of soil moisture conditions. Full article

Mineral malnutrition negatively impacts almost 30% of the world population and could result in anemia, blindness, and stunted growth. Seed traits involve nutritional, physical, and physiological characteristics that play a key role in achieving seed quality. Chickpea (Cicer arietinum), the second most important food legume crop worldwide, is characterized by its rich nutrient profile, fiber, and antioxidant content. However, seed quality traits composition and daily nutritional value in chickpea seeds remain largely unknown. The objectives of the current study were to determine the variability in concentrations of macro and micronutrients, oil, protein, their correlations, and daily nutritional values (% DV) in the seeds of 61 chickpea genotypes. The results demonstrate substantial variation among chickpea genotypes in most nutrients, protein, and oil content. Moreover, this study identifies 12 promising chickpea genotypes that indicate large nutrient content and potential usage in chickpea biofortification and breeding programs. Our results further show that consumption of 100 g chickpea seeds could provide 122% DV of manganese (Mn), 77% DV of copper (Cu), 43% DV of magnesium (Mg), 32% DV of phosphorus (P), 27% DV of zinc (Zn), 26% DV of iron (Fe), and 5% DV of calcium (Ca). Overall, these findings improve our understanding of chickpea seed nutrient content as well as global food security by combatting mineral malnutrition. Full article

Seed priming is a simple, inexpensive, and effective pre-sowing technique that enables plants to better tolerate abiotic stresses, including high soil salinity, which is a major limiting factor in the establishment of halophytes for saline agriculture, as germinating seeds and early seedlings of many halophytes are sensitive compared to the mature vegetative stage. This article attempts to provide an overview of the research on the seed priming effects on halophyte seeds and subsequent seedlings/plants. Different physio-chemical and molecular processes, including the induction of priming/stress memory, which enhance salinity tolerance following seed priming, have also been discussed. This review also covers the aspects of reactive oxygen species (ROS), and nitric oxide (NO) signaling(s) that are activated as a result of seed priming. Finally, the limitations and prospects of seed priming to enhance the agronomic potential of halophytes for saline agriculture have been discussed. Full article

The cost of producing organic crops is increasing. Agricultural wastes can be used as biostimulants to increase plant growth and productivity and reduce the dependence on chemical fertilizers. A pouch assay and a potted greenhouse experiment were conducted to investigate the effect of pyroligneous acid (PA) and sea lettuce (SL) on kale (Brassica oleracea subsp. acephala (DC.) Metzg) seed germination and growth. Although previous studies have demonstrated that these two biostimulants could promote plant germination and growth, there is little research to compare their effects on seed germination and plant growth. The pouch assay showed that PA liquid affected the seed germination rate under different concentrations; the seed germination rate decreased as the concentration of PA liquid increased. However, the effect of seed germination was less pronounced in SL liquids. Kale seeds treated with 0.01% PA showed the best elongation and seedling growth performance. Moreover, the greenhouse experiment indicates that SL liquids significantly (p < 0.05) affected kale growth production, while PA liquid had less difference on kale growth under various concentrations. The 0.25% PA and 1% SL increased the aboveground fresh weight by ca. 26% and 29%, respectively. Also, the phytochemical contents of kale leaves, including phenolics, flavonoids, ascorbate, and protein, were significantly increased with 0.25% PA and 1% SL application. These results suggest that low concentrations of PA are more suitable for seedling root growth in kale and 1% SL had the most significant growth-promoting effect on kale. Hydrothermal carbonization sea lettuce liquid can be used as a good biostimulant for agricultural production to improve kale germination and growth. Full article

Insect-based (silkworm cocoons) and plant-based (cotton wool pads and gauzes) fiber substrates were used to support and ameliorate seed germination originating from trifoliate orange (Poncirus trifoliata) and pomegranate (Punica granatum) trees. Three different commercial formulations of beneficial microorganisms (Bacillus spp.-Azotobacter spp., Saccharomyces boulardii, and Saccharomyces cerevisiae) were administered to seeds in order to evaluate their contribution to germination and growth. The silkworm cocoons provided better germination rates for P. trifoliata seeds (83.33%) among the tested media without any microbial supplementation. These rates increased towards the absolute maximum (100%) when Bacillus spp.-Azotobacter spp., S. boulardii and S. cerevisiae were applied. Furthermore, inoculums of Bacillus spp.-Azotobacter spp. 2 mL and S. cerevisiae 3 g raised the pomegranate seed germination ability by 30–33.33% and 50–67.7%, respectively, on silkworm cocoon substrates when compared to plant-derived, cellulosic fiber substrates under the same biotic exposure. On increasing the size of applied microbial inoculums, seed germination moved from optimum to suboptimum for all germination media. Examination of multipartite pH compatibility (between seeds, microorganisms, and germination media) was beneficial and of functional value. In conclusion, the germination rates of both tree species can be raised using bacterial and yeast supplementation, including medical-grade S. boulardii, on environmentally friendly materials such as insect- and plant-based fiber substrates. Full article

Azadirachta indica (Neem tree) has become widely naturalised and invasive across many countries and regions including northern Australia. To aid management of A. indica where it has become a weed, a series of studies were undertaken to determine its potential soil seed bank persistence. In a field trial, packets of seeds were buried, retrieved periodically over two years and the seed viability assessed. Viability declined rapidly, with a single viable seed retrieved after 12 months burial and none thereafter. Burial depth, soil type, and pasture cover (present and excluded) significantly influenced viability (%) at 3- and 6-month retrievals. Similar data were obtained from repeated runs of a controlled ageing laboratory experiment, which categorized seeds as forming a ‘transient’ seed bank. In a third trial, fresh fruits were placed on the soil surface in replicated field enclosures over two consecutive years and seedling emergence monitored fortnightly. In both years there was no emergence from pasture excluded soil plots and emergence ceased after 2.3 and 8.4 months in plots with pasture present. A fourth (glasshouse) trial found most seeds will emerge from the soil when buried from 1 to 4 cm. However, more fatal germination than successful emergence was recorded for seeds buried at 8 cm. Seed desiccation and fatal germination are factors in A. indica developing a transient soil seed bank, and infestations require shorter-term control programs where seed input is prevented. Full article

The foxtail millet exhibits a diverse range of hull colors, which are crucial indicators for assessing its nutritional and economic value. However, the molecular regulatory mechanisms that govern the hull color of foxtail millet are largely unknown at present. This gap in knowledge significantly impedes efforts to enhance the quality traits of foxtail millet. This study utilized a population of 250 F₆ recombinant inbred lines (RILs) generated from a cross between two foxtail millet varieties: Yugu18 (with light yellow seeds) and Hongjiugu19 (with red seeds). Four methods, the visual grouping method (I), the visual colorimetric method (II), the Lab determination method (III), and the RGB determination method (IV), were employed to determine the hull color of each line across four environments and QTL identification were conducted subsequently. It showed that there were 10, 12, 69 and 56 QTLs were detected for hull color through four methods, and these QTLs were integrated into 4, 6, 27 and 25 unique QTLs, respectively. There were three, four, four and four major QTLs. Of which, three major QTLs (qHC1.1, qHC1.2 and qHC9.3) on chromosomes 1 and 9 could be detected by all 4 methods. qHC9.1 was detected by all four methods except for method I. There were also one, one, seven and four minor identity QTLs identified across the 4 methods. Four minor QTLs (qHC3.1, qHC3.3, qHC4.1 and qHC5.1) can be stably detected only in method III, and two minor QTLs (qHC8.2 and qHC9.2) can be stably detected only in method IV. Generally, method I is fast, efficient and cost-effective, which is suitable for the rapid detection of hull color. Method II is also low-cost; however, it can detect more QTL for hull color, making it suitable for identifying major QTL loci in large populations. Methods III and IV can map more minor QTL and are more accurate in hull color characterization. This study identified four important hull color QTL for foxtail millet, which largely align with those reported in previous research. These findings establish a foundation for characterizing hull color indices and further advancing QTL mapping for grain color. Full article

The chemical quality of fenugreek seeds is a critical factor influencing their nutritional value, medicinal properties, and suitability for agricultural and industrial applications, making it essential to evaluate their biochemical composition and functional potential. This study evaluated the effects of salinity and different fertilization types on seed quality, early growth parameters, and key biochemical characteristics during fenugreek (Trigonella foenum-graecum L.) cultivation. A field experiment was established at the Agricultural University of Athens during the cropping period 2019–2020 (Year A) and 2020–2021 (Year B) in a split-plot design with the five main fertilization treatments Biocyclic-Vegan Humus Soil (BVH), Farmyard Manure (FM), Organic Compost (OC), Inorganic Fertilizer (IF; 11-14-14), and No Treatment Control (NTC) and two treatments: Elevated Salinity (ES) and Normal Salinity (NS). Fertilization significantly influenced various agronomic traits and seed compositions across both experimental years. The evaluation showed that organic fertilization with BVH yielded the best results among the treatments studied. The BVH × NS treatment consistently produced the highest plant height, seed protein content, seed yield, and mineral content, including nitrogen, calcium, and potassium, compared to the other treatments. ES impacted the concentrations of several elements, namely N (%), Ca (g/100 g), K (g/100 g), Fe (ppm), P (%), and Mg (g/100 g). Salinity also affected certain traits, such as Seed Total Ash and Dry Matter content, with significant interactions observed. These findings highlight the critical role of fertilization, especially organic fertilization, in improving both agronomic performance and seed nutritional quality in the studied crops, as well as in mitigating the adverse effects of salinity. Full article

Downy mildew caused by the oomycete Pseudoperonospora cubensis is a devastating disease of cucurbits. Cucumis species are attacked by pathotype 3 (clade 2) of the pathogen, while Cucurbita species are attacked by pathotype 6 (clade 1). The Sikkim-type cucumbers PI 197088 and PI 330628 express high levels of resistance against both pathotypes (clades) of the pathogen but no green-fruit cucumber cultivars resistant to the disease are available on the market. Here we report on several non-Sikkim accessions of cucumber that show resistance against downy mildew in four consecutive seasons. Mean % foliage attacked with downy mildew in the susceptible controls Ilan and SMR-18 was 93% and 71%, respectively, as against 0.2% and 1.8% in the Sikkim-type resistant controls PI 197088 and PI 330628, respectively. Twenty-four green fruit accessions were significantly more resistant than the susceptible cucumber controls. Five accessions showed less than 10% infected leaf area with downy mildew as follows: PI 432870—5%, PI 390266—7.5%, PI 418964—8.5%, PI 390258—8.8%, and G12—10%. PI 390258 and PI 390266 were susceptible to race 1 of powdery mildew but resistant to race 2, whereas PI 418964 was resistant to both races. These accessions may be used in breeding programs to accelerate the production of green-fruit, disease-resistant cucumbers. Full article

Herbaceous peonies, specifically the Balkan–Anatolian Paeonia peregrina Miller, are species with various uses such as ornamental and garden purposes, or they can be cut as flowers or potted, or they can be eaten or used for medicinal purposes due to the rich nutritional content of their seeds. However, conventional propagation methods, including rhizome division, grafting, and layering, are slow, while seed propagation is challenging due to double morpho-physiological dormancy. This study therefore evaluated the in vitro germination potential of P. peregrina seeds in darkness under different culture conditions, including different temperature regimes (constant at 15 °C or alternating from 22 °C to 15 °C); incubation periods [120 days: 22 °C (14 d) to 15 °C (0–105 d); 120 days: 22 °C (33 d) to 15 °C (0–87 d); 90 or 140 days at 15 °C]; seed cold storage period (none; 30 days; 3 months; or 2, 5, and 8 years); and gibberellic acid (GA₃) concentrations (0, 250, 500, 750, and 1000 mg L⁻¹), as dormancy release methods. The results indicated that 60-day-stored seeds (30 d at 15 °C and 30 d cold-storaged at 4–5 °C) exhibited 100% germination within an 80-day culture under 250 mg L⁻¹ GA₃ at 15 °C. A lower and constant temperature of 15 °C, a shorter 30-day seed cold storage period, and the lowest GA₃ concentration of 250 mg L⁻¹ comprised the most effective combination treatment for dormancy release and germination acceleration. An understanding of the underlying mechanisms of seed dormancy removal is imperative for successful germination, growth rate and seedling establishment, shortened breeding cycles, and germplasm conservation, leading to the field cultivation and economic production of these peony plants. Full article