Seedling raising method for breaking dormancy of tephrosia vogelii

Abstract

This invention discloses a method for breaking seed dormancy in *Palmeria nivea* seeds. This invention rapidly breaks seed dormancy through minimally invasive seed coat incision, combined with a fungicide-treated seedling tray device. This allows for seed germination control within 25 days from seed collection, with a rooting rate of 92.4% and a seedling survival rate of 83.69% after 45 days. After 45 days of cultivation in the fungicide-treated seedling tray, without the need for water or fertilizer, the average plant height is 8.46 cm, branch nodes reach 3 nodes, and root system length reaches 5.89 cm. This invention achieves rapid breaking of seed dormancy in *Palmeria nivea*, efficiently promotes seed germination and seedling growth, and reduces seedling management time. It achieves the goals of rapid seed germination and efficient seedling growth, providing a strategy for the protection and population expansion of the endangered plant *Palmeria nivea*.

Description

Technical Field

[0001] This invention belongs to the field of endangered plant conservation and seedling propagation technology, specifically relating to a seedling cultivation method for breaking the dormancy of Dalbergia odorifera seeds. Background Technology

[0002] The rare and endangered Ormosia henryi Prain, a Class II protected plant in my country, is endemic to the country. It is naturally distributed in subtropical provinces such as Anhui, Fujian, Jiangxi, Hubei, Hunan, Guangdong, and Hainan, primarily in mountainous areas, streamsides, and valley forests at altitudes of 600-1200 meters. Ormosia henryi Prain has demanding ecological requirements, resulting in scarce and unevenly distributed wild resources. Its wood is of excellent quality, making it a valuable timber for high-end furniture, craft carvings, and special decorative items. It is also an important medicinal herb. Furthermore, its beautiful tree shape, evergreen foliage, abundant blossoms, and red pods make it an excellent landscaping tree. Due to its superior wood quality, large trees are frequently felled, leading to poor natural regeneration. Additionally, its narrow distribution area and demanding ecological requirements contribute to the dwindling wild population. The seed coat of *Palmaria dentata* is hard and dense, with poor water and air permeability. Substances inhibiting seed germination are present in both the seed coat and embryo, resulting in a long dormancy period and difficulty in germination, leading to weak natural reproductive capacity. In conventional asexual reproduction methods, stem cuttings are difficult to root, and other methods besides root cuttings are extremely ineffective. However, the availability of root cutting materials is affected by root growth and development; only a small number of suitable roots can be obtained from older trees, making it difficult to propagate from young trees. The seed yield of *Palmaria dentata* pods is low, with an empty pod rate of 51.3%. Of the seeds that develop normally within the pods, 45.3% are susceptible to insect damage. The hard and dense seed coat of *Palmaria dentata* seeds, due to mechanical barriers and inhibitory substances, results in a low germination rate of only 16% under natural conditions. The reproductive capacity of *Palmaria dentata* is low (1.6), only 1.8% of the ideal reproductive capacity. Under natural conditions, the germination rate of rosewood seeds after burying them for one year is only 16% (Reference: Wei Xiaoli, Meng Xianshuai, Deng Zhao. Relationship between the reproductive ecology of rare tree species rosewood seeds and their endangerment[J]. Seeds, 2014, 33(1):82-86.). Low reproductive capacity is one of the important reasons for the endangerment of rosewood.

[0003] Currently, physicochemical methods are mostly used to treat *Palmeria nivea* seeds to promote germination. One method involves soaking the seeds in wood ash for 1 day, then in warm water for 3 days, followed by stratification in moist sand for 3-4 days. Additionally, a 500 mg / L solution has been found to be effective. －1Soaking *Palmeria nivea* seeds in gibberellin for 12 hours followed by 45 days of low-temperature, sand-mixed, moist storage at 4℃ broke seed dormancy, increasing germination rate and germination potential by 36.6% and 32.7%, respectively, compared to the control. Furthermore, 2 hours of concentrated sulfuric acid etching effectively improved seed coat permeability, enhancing water absorption without damaging the seeds. Overall, chemical treatment and variable-temperature methods effectively improved seed coat permeability and increased water absorption in *Palmeria nivea*. Mixed sand and wet storage treatment can improve the activity of related enzymes, which plays a key role in breaking the dormancy of Dalbergia odorifera seeds (References: Deng Zhao, Wei Xiaoli, Meng Xianshuai, et al. Preliminary study on dormancy and germination of Dalbergia odorifera seeds [J]. Guizhou Agricultural Sciences, 2011, 39(5):69~72. Jiang Changzhi. Dalbergia odorifera seedling technology [J]. Practical Forestry Technology, 2004, (9):26. Shen Shaonan, Liu Shanggui, Cai Huanliu. High-yield cultivation technology of precious tree species Dalbergia odorifera [J]. Modern Agricultural Science and Technology, 2009, (1):81~84.). In the existing rapid propagation system of Dalbergia odorifera, Dalbergia odorifera tissue culture uses MS medium as the basic medium. When using Dalbergia odorifera seedling stem segments as explants, 2.0 mg·L⁻¹ is added to the MS medium. －1 6-BA, 0.5 mg·L －1 NAA, 8g·L －1 Agar, 30 g·L －1 Sucrose, adjusted to pH 5.8–6.0, germinated after 45 days of cultivation at approximately 26°C; germination was achieved using 1 / 2 WPM + IBA 1.0 mg·L⁻¹. －1 +NAA 2.0 mg·L －1 +8g / L agar －1 +10g / L sucrose －1 Rooting culture was carried out in the rooting medium at pH 6.0–6.2 and 23℃ for about 15 days (Reference: Qiao Dong. Research on tissue culture technology of Dalbergia odorifera [D]. Guizhou University, 2016).

[0004] Currently, among the existing conventional asexual propagation methods, only root cutting has been reported with good results; other asexual propagation techniques have not been reported. For *Palmeria nivea* seed roots, collection should be done from mid-March to early April on healthy *Palmeria nivea* trees that are over 10 years old and have a diameter at breast height (DBH) of 8-28 cm. The small end of the root should have a diameter of at least 0.5 cm, and the root segment should be 8-15 cm long. Root cuttings place extremely high demands on the mother tree and are difficult to propagate rapidly.

[0005] The hard, dense seed coat of *Palmeria nivea* (Chinese palm) results in poor water and air permeability, and due to dormancy, germination is slow, with a long germination period and low germination rate, posing major challenges to artificial cultivation. Under most germination-inducing conditions, the germination rate of *Palmeria nivea* seeds is not ideal, and germination is uneven. *Palmeria nivea* seedlings grow very slowly, have poor drought and frost resistance, and are prone to death, significantly impacting the expansion of *Palmeria nivea* populations through seed propagation. The hairy surface of *Palmeria nivea* leads to severe endophytic contamination when using explants from naturally germinated plants or mature plants for tissue culture. Therefore, it is essential to select suitable and effective explants and employ appropriate sterilization techniques to establish a rapid tissue culture propagation system for *Palmeria nivea*. However, tissue culture technology has limitations in terms of propagation materials, and most tissue-cultured seedlings rooted in test tubes are weak or otherwise unsuitable, resulting in poor growth after transplanting to nurseries. Currently, *Palmeria nivea* cutting and grafting techniques are not mature, making asexual reproduction difficult to achieve. Breaking the dormancy of rosewood seeds caused by inhibitory substances, and specifically initiating dormancy to shorten the dormancy period, is a critical bottleneck in the propagation of rosewood seeds.

[0006] Low reproductive capacity is one of the important reasons for the endangerment of the Dalbergia odorifera species. In the future, the protection of Dalbergia odorifera should strengthen theoretical and technical research on seed abortion mechanism, pest and disease control during flowering and fruiting period, breaking seed dormancy, and improving seed germination rate and seedling survival rate. Summary of the Invention

[0007] As an endangered and valuable timber species, *Dalbergia odorifera* has great development potential. Currently, its natural forest resources are nearing depletion, and it is listed as a national second-class protected plant. To solve the problem of seed dormancy in *Dalbergia odorifera* and achieve rapid seed germination and efficient seedling growth, this invention provides a highly efficient seedling cultivation method that rapidly breaks seed dormancy and promotes seed germination and seedling growth. The method of this invention is simple to operate, utilizing a seedling tray with bacterial filtration to provide an excellent growth environment for seed germination and development, and the seedling management cost is extremely low.

[0008] The seedling cultivation method for breaking the dormancy of *Palmeria nivea* seeds of the present invention includes the following steps:

[0009] S1. After the pods of the rosewood mature to a dark black color, pick them from the tree, remove the seeds, select healthy and plump rosewood seeds with a thousand-seed weight of not less than 200g, and dry them at room temperature within 2-5 days; wash off any adhering substances with detergent, rinse thoroughly with running water, and then wash 3-5 times with sterile water, and blot dry the seed surface with sterile filter paper; S2. Remove a small area of ​​the seed coat, about 2*2mm in size, from the hilum of the seed, being careful not to damage the cotyledons, hypocotyl, plumule, and radicle at the hilum; do not remove the rest of the seed coat; then soak the seeds in an appropriate amount of sterile water in the dark for 24 hours, changing the sterile water every 6 hours during this period;

[0010] S3. Sow the seeds obtained in step S2 into the nutrient substrate of the seedling box at a depth of 1.5-2cm, and then cover the outer cup tightly with the outer cup lid; place the seedling box with seeds in a 15-18℃ environment with a light / dark duration of 12 / 12 hours for cultivation.

[0011] S4. After the seed cotyledons emerge from the soil, adjust the cultivation conditions to a temperature of 23-26℃ and a light / dark duration of 16 / 8 hours. When the seedling branches reach 2-3 nodes, remove the seedlings from the seedling box and transplant them.

[0012] Preferably, the seedling box includes an outer cup, an outer cup lid, and an inner cup. The bottom of the outer cup is filled with sterile water to a certain height. The inner cup is fixed in the lower middle part of the outer cup, and the bottom of the inner cup is higher than the sterile water level. A thread hole is provided at the bottom of the inner cup, through which the absorbent thread passes and can be tightly sealed. The absorbent thread on the outside of the inner cup hangs down naturally to the bottom of the outer cup, and the absorbent thread inside the inner cup extends along the inner cup wall to the outside of the inner cup opening. The inner cup is filled with nutrient substrate. The outer cup lid is provided with bacterial filtration and ventilation holes, and the outer cup lid is used to tightly fit over the outer cup opening.

[0013] Preferably, all parts of the seedling box have undergone sterilization treatment.

[0014] Preferably, the outer cup and outer cup lid are made of transparent material, and the inner cup is a paper cup with a polyethylene coating on the inner wall.

[0015] Preferably, the outer cup is made of PS (polystyrene) material with a thickness of 0.2cm, and the outer cup mouth is a threaded mouth; the outer cup lid is made of PP (polypropylene) material, and is a threaded lid that fits onto the outer cup mouth.

[0016] Preferably, there are two or more absorbent threads, and the absorbent threads are cotton threads with a thickness of 0.4 mm.

[0017] Preferably, the nutrient substrate is peat moss with fiber length of 5-20 mm, which is fully moistened with sterile water and filled to the point that it does not drip water when squeezed into a ball by hand.

[0018] Preferably, the inner cup is fixed in the lower middle part of the outer cup, that is, the mouth of the inner cup is abutted against the inner cup and the abutting position is located in the middle of the outer cup.

[0019] Preferably, the room temperature drying in step S1 involves air-drying the freshly extracted seeds from the pods or placing them in a drying box for thorough drying.

[0020] Preferably, the sowing of the seeds obtained in step S2 into the nutrient substrate of the seedling tray is carried out at a rate of one seed per seedling tray.

[0021] In the seedling box described above, after the inner cup containing sterilized nutrient substrate is filled into the outer cup, the mouth of the inner cup automatically locks into a fixed position on the inside of the outer cup according to the design dimensions. The two cotton threads at the bottom of the inner cup naturally hang down into the sterile water at the bottom of the outer cup. The water is drawn into the inner cup through the natural siphon principle of the cotton threads, keeping the nutrient substrate in the inner cup moist.

[0022] All operational steps, as well as the filling of substrate and water, are sterilized to minimize contamination from external bacteria and pests, and to prevent problems such as fly larvae eating the cotyledons or cotyledon contamination and rotting during the germination process of the rosewood.

[0023] Using the seedling box of this invention for cultivation, there is no need to open the seedling box for management during the entire cultivation period, saving time and effort. It can be continuously cultivated for about 45 to 60 days. When the seedling leaves have grown to the third node, in order to meet the nutrient and space requirements of the rosewood seedlings, the seedlings can be transplanted into larger pots for further growth.

[0024] This invention rapidly breaks the dormancy of *Palmeria nivea* seeds through minimally invasive seed coat incision. Combined with a fungicide-treated seedling tray, it enables the control of *Palmeria nivea* seed germination from seed harvesting to emergence within 25 days, with a rooting rate of 92.4% and a germination rate of 83.69% after 45 days. Cultivated in the fungicide-treated seedling tray for 45 days without water or fertilizer supplementation, the average plant height is 8.46 cm, with branch nodes reaching 3 nodes and root length reaching 5.89 cm. This invention achieves rapid breaking of *Palmeria nivea* seed dormancy, efficiently promotes seed germination and seedling growth, and reduces seedling management time, achieving the goals of rapid seed germination and efficient seedling growth. It provides a strategy for the protection and population expansion of the endangered plant *Palmeria nivea*. Attached Figure Description

[0025] Figure 1 This is a structural diagram of the seedling box in Example 1; where the attached diagram is labeled as follows: 1: outer cup lid, 2: filter and vent hole, 3: outer cup, 4: nutrient substrate, 5: inner cup, 6: sterile water, 7: sowing or seedling growth position, 8: water absorption line, 9: threading hole.

[0026] Figure 2 This is a physical image of the outer cup lid with filter and ventilation holes of the seedling box in Example 1.

[0027] Figure 3 This is a physical diagram of the seedling box in Example 1, showing the outer cup and outer cup lid in use.

[0028] Figure 4 This is a physical image of the inner cup of the seedling box in Example 1.

[0029] Figure 5 These are the size parameters of each component in the overall structure of the seedling box in Example 1.

[0030] Figure 6 These are images of Dalbergia odorifera seed treatment. Figures 1-6 represent: 1: Front view of the hilum of a dried Dalbergia odorifera seed; 2: Front view of a dried Dalbergia odorifera seed after the seed coat at the hilum has been removed; 3: Inside view of the seed coat at the hilum of a dried Dalbergia odorifera seed; 4: Side view of a dried Dalbergia odorifera seed; 5: Side view of a dried Dalbergia odorifera seed after the seed coat has been removed; 6: Image of a Dalbergia odorifera seed after 24 hours of imbibition and removal of the seed coat.

[0031] Figure 7 These are comparative images of the cotyledons and seed coats of dried and imbibition seeds of *Dalbergia odorifera*. The left side of the image shows the cotyledons and seed coat of dried *Dalbergia odorifera* seeds after the seed coat has been removed. The right side of the image shows the cotyledons and seed coat of *Dalbergia odorifera* seeds after the seed coat has been removed 24 hours after imbibition.

[0032] Figure 8 These are comparative images of the areas where the seed coat of dried rosewood seeds has been minimally invasively removed; Figure A shows a schematic diagram of a dried rosewood seed with the hilum removed; Figure B shows a schematic diagram of a dried rosewood seed with the seed coat removed from the top opposite the hilum.

[0033] Figure 9 These are images showing the germination and growth of *Palmaria rubra* seedlings using a seedling tray; the left side shows the growth after 30 days of cultivation, and the right side shows the growth after 45 days. Detailed Implementation

[0034] The following embodiments are further illustrations of the present invention, but not limitations thereof.

[0035] The method for breaking the dormancy of rosewood seeds in this invention is achieved by combining the seedling box of Example 1.

[0036] Example 1

[0037] Please refer to Figure 1 The seedling box in this embodiment includes an outer cup 3, an outer cup lid 1, and an inner cup 5. The bottom of the outer cup 3 is filled with a certain height of sterile water 6. The inner cup 5 is fixed in the lower middle part of the outer cup 3, and the bottom of the inner cup 5 is higher than the surface of the sterile water 6. The bottom of the inner cup 5 is provided with a thread hole 9. Two (or more) absorbent threads 8 pass through the thread hole 9 and can just fill the gap of the thread hole 9. The absorbent threads 8 on the outside of the inner cup 5 naturally hang down to the bottom of the outer cup 3. The absorbent threads inside the inner cup 5 extend along the cup wall of the inner cup 5 to the outside of the cup opening of the inner cup 5. The inner cup 5 is filled with nutrient substrate 4. The outer cup lid 1 is provided with a filter and ventilation hole 2. Figure 2 The outer cup lid 1 is used to tightly fit over the mouth of the outer cup 3. Figure 3 ).

[0038] The outer cup 3 is made of PS (polystyrene) with a threaded opening. The threaded opening is designed to work with the outer cup lid 1, effectively preventing the intrusion of miscellaneous bacteria. The outer cup 3 is made of transparent, thickened PS material (0.2cm thick), which facilitates multi-directional and three-dimensional light exposure for the seedlings after they sprout, and also allows growers to easily observe the seedling growth. The thickened PS material also provides rigidity, effectively protecting the seedling box from external impacts and ensuring a good growth space for the seedlings inside the inner cup during transport. The transparent and smooth inner wall of the outer cup allows condensed water droplets to quickly slide off into the nutrient substrate inside the inner cup, achieving water recycling. The outer cup lid 1 is made of PP (polypropylene) and is a transparent, threaded cap. The outer cup lid 1 has filter and ventilation holes 2 (made of tetrafluoroethylene, 0.22μm permeability) to maintain air circulation between the seedling box and the outside environment. Combined with the threaded opening design of the lid, this effectively prevents miscellaneous bacteria from entering the seedling box. The transparent material also facilitates light exposure for the seedlings after they sprout, promoting seedling growth.

[0039] Inner cup 5 is a thickened paper cup with an inner wall coated with PE (polyethylene). Figure 4 This design effectively prevents moisture from seeping from the filling substrate to the outer wall, keeping the nutrient substrate moist and preventing the cup from deforming during long-term cultivation. It's worth noting that the inner cup is made of paper instead of plastic because when the outside temperature is high, a lot of moisture accumulates inside the seedling box. The paper inner cup can absorb some of the moisture condensed on the outer cup wall, regulating the humidity inside the seedling box. The thickened paper cup provides a dark environment for germination, ensuring successful sprouting. Additionally, the waterproof inner membrane keeps the nutrient substrate in the inner cup sufficiently moist. The rigidity of the thickened paper cup ensures that the inner cup will not deform throughout the seedling period and remains firmly secured in the appropriate position within the outer cup.

[0040] For more specific details, please refer to Figure 5The outer cup 3 is 0.2cm thick, and the thickened (0.05mm thick) paper inner cup 5 has a cup mouth diameter of 11cm and a height of 8cm, so that the cup mouth of the inner cup 5 fits snugly against the inner cup 3 at a vertical height of about 11cm. There is a height space of about 10cm between the cup mouth of the inner cup 5 and the cup mouth of the outer cup 3, and a height space of 2-2.5cm between the bottom of the inner cup 5 and the bottom of the inner cup of the outer cup 3. The bottom of the outer cup 3 is filled with 2cm of sterilized distilled water to maintain the water supply throughout the growth period of the seedling box, which can achieve up to 45 days without the need for additional watering. The absorbent thread 8 is a cotton thread with a thickness of 0.4mm. Two absorbent threads 8 pass through the thread hole 9 and can fit tightly into the gap of the thread hole 9 to prevent peat soil from falling out. They also use the siphon principle to supply water to the nutrient substrate. The length of a single cotton thread is 18-20cm. The absorbent threads 8 are evenly placed on both sides of the inner cup 5 to ensure the balance of moisture in the nutrient substrate. The nutrient substrate 4 is peat moss with fiber lengths of 5-20 mm. It is thoroughly moistened with sterile water until it can be formed into a ball without dripping, and then filled up to the inner cup 5, which already has absorbent lines 8. After inserting the inner cup 5, filled with the nutrient substrate, into the outer cup 3, the outer edge of the inner cup 5 should fit snugly inside the outer cup according to the designed dimensions. The two absorbent lines 8 at the bottom of the inner cup 5 naturally descend into the sterile distilled water at the bottom of the outer cup 3, drawing water to the inner cup through the natural siphon effect of the cotton threads, maintaining the moisture of the nutrient substrate in the inner cup. The pre-treated, budding *Palmaria* seeds are sown at the sowing or seedling growth position 7 on the nutrient substrate (planting depth 1.5-2 cm). Finally, the outer cup 5 is tightened with the outer cup cap 1, and the entire seedling tray is placed under the following conditions for cultivation:

[0041] The seed germination period was set at 15–18℃ (approximately 10 days). Lower temperatures can prevent seeds from becoming moldy, but have no significant effect on the development of the hypocotyl and plumule.

[0042] After the seeds germinate, they enter the growth period. Adjusting the temperature to 23-26℃ can rapidly promote leaf development and growth. No need to open the seedling tray for management throughout the entire cultivation period, saving time and effort. The seedlings can be continuously cultivated for about 45 days. When the seedlings' leaf branches have grown to the third node, to meet the nutrient and space requirements of the rosewood seedlings, they can be transplanted into larger pots for further growth.

[0043] Example 2

[0044] 1. Materials and Methods

[0045] 1.1 Seed Screening and Treatment Experiment of *Palmaria rubra* var. *marginata* var. *marginata*, approximately 75 years old, collected on November 15, 2021, from naturally matured pods with dark black / blackish-brown pericarps. Plump, round, and uniformly colored seeds were immediately collected and dried in a drying box for 2 days. Seeds with plump appearance, uniform color, and a thousand-seed weight of not less than 200g were selected for subsequent germination. The seeds were quickly and thoroughly washed for 3-5 minutes with detergent (1mL of Libai dishwashing liquid diluted in 500mL of tap water) to remove impurities and adhering substances. They were then rinsed thoroughly with running water and repeatedly washed 3-5 times with sterile water. The seeds were then blotted dry with sterile filter paper. After uniform washing, the seeds were treated to promote seed imbibition.

[0046] Non-seedling minimally invasive excision treatment group:

[0047] (1)T1: After cleaning, the seeds of the rosewood were directly spread in a sterile petri dish, and an appropriate amount of sterile distilled water was added, with the water level at least covering the seeds. The seeds were soaked in the dark for 24 hours, and the sterile distilled water was changed every 6 hours. Then, the subsequent comparative experiment was carried out.

[0048] (2)T2: After cleaning, the seeds of the rosewood were directly spread in a sterile petri dish, and an appropriate amount of sterile distilled water was added. The dish was placed in a 60°C incubator for constant temperature soaking (i.e., warm water soaking method). The water surface should at least cover the seeds. The dish was soaked in the dark for 24 hours, and the sterile distilled water was changed every 6 hours. Then, the subsequent comparative experiment was carried out.

[0049] Minimally invasive excision treatment group for seed coat:

[0050] (3) T3: Use a blade to reference the cleaned rosewood seeds. Figure 8 Standard B involves removing the top seed coat (approximately 2*2 mm) opposite the hilum, taking care not to damage the cotyledons, hypocotyl, plumule, or radicle at the hilum. For *Palmaria* seeds with only the top seed coat removed from the hilum, lay them flat in a sterile petri dish, add an appropriate volume of sterile distilled water (enough to completely cover the seeds), and soak in the dark for 24 hours, changing the sterile distilled water every 6 hours. Subsequent comparative experiments are then conducted.

[0051] (4) T4: Use a blade to reference the cleaned rosewood seeds. Figure 8For standard B, the apical seed coat opposite the hilum (approximately 2*2 mm) is removed. The area of ​​seed coat removal is similar to that of treatment T3, but care must be taken not to damage the cotyledons, hypocotyl, plumule, or radicle. For *Dalbergia odorifera* seeds with only the apical seed coat opposite the hilum removed, they are directly spread flat in a sterile petri dish, an appropriate volume of sterile distilled water is added, and the dish is placed in a 60℃ incubator for constant temperature soaking, ensuring the water level at least covers the seeds. The solution is soaked in the dark for 24 hours, with the sterile distilled water changed every 6 hours. Subsequent comparative experiments are then conducted.

[0052] (5) T5: Use a blade to reference the cleaned rosewood seeds. Figure 8 According to standard A, the seed coat (approximately 2*2mm) at the hilum of the seed is removed, but care should be taken not to damage the cotyledons, hypocotyl, plumule, and radicle at the hilum. Figure 6 For *Palmaria rubra* seeds with only the seed coat removed from the hilum, they were directly spread in a sterile petri dish, and an appropriate amount of sterile distilled water was added, ensuring that the water level at least covered the seeds. The seeds were then soaked in the dark for 24 hours, with the sterile distilled water being changed every 6 hours. Subsequent comparative experiments were then conducted.

[0053] (6)T6: Use a blade to reference the cleaned rosewood seeds. Figure 8 According to standard A, the seed coat (approximately 2*2mm) at the hilum of the seed is removed, but care should be taken not to damage the cotyledons, hypocotyl, plumule, and radicle at the hilum. Figure 6 For *Palmaria rubra* seeds with only the seed coat removed from the hilum, they were directly spread flat in a sterile petri dish, an appropriate amount of sterile distilled water was added, and the dish was placed in a 60°C incubator for constant temperature soaking, with the water level at least covering the seeds. The solution was soaked in the dark for 24 hours, with the sterile distilled water being changed every 6 hours. Subsequent comparative experiments were then conducted.

[0054] Seed coat completely removed treatment group:

[0055] For seeds treated with T1-T6, the seed coat was completely removed from each seed. Figure 7 Subsequent experiments were conducted after the seed coat was removed, and the treatment groups were numbered Tb1-Tb6 accordingly.

[0056] 1.2 Design of planting methods

[0057] (1) Seedling cultivation using the seedling tray in Example 1

[0058] method:

[0059] Seeds treated with T1-T6 were cultured using this method, with 15 seeds per treatment and 3 biological replicates per treatment. Before assembling the seedling trays, the outer cups were thoroughly washed with detergent, rinsed 2-3 times with sterile water, and then air-dried. The cup lids were sterilized at 121℃ under high pressure. Both the outer cups and lids were reusable. The inner cups were for single use only, filled with fully moistened peat moss that had been sterilized under high pressure. The peat moss should be moist enough to clump together without dripping water. Sterile distilled water was added to the bottom of the outer cup to a depth of 2cm, ensuring it did not submerge the bottom of the inner cup. The absorbent cotton thread leaking from the bottom of the inner cup should be fully submerged in the water. After assembling the seedling trays, one swollen *Paederia scandens* seed was planted in the nutrient substrate of the inner cup of each seedling tray, at a planting depth of 1.5-2cm. Finally, tighten the outer cup lid and place the seedling box in a greenhouse at 15-18℃ with a light / dark cycle of 12 / 12 hours. The sterilization requirements at each stage are designed to minimize the impact of external pests, diseases, and contaminating bacteria on the germination process of *Palmeria nivea* seeds, creating a low-microbial environment for seed germination. Setting a lower growth temperature during seed germination effectively prevents the rapid growth of contaminating bacteria within the seed coat. The continued 12-hour light cycle also inhibits the growth of contaminating bacteria on the substrate surface. The darkness provided by the inner cup is sufficient for the dark culture required for seed germination. Once the cotyledons emerge from the soil after germination, the temperature can be adjusted to 23-26℃, with a light / dark cycle of 16 / 8 hours to promote photosynthesis and accelerate the growth of *Palmeria nivea* seedlings.

[0060] (2) Planting method for tissue culture seedlings:

[0061] Seeds treated with T1-T6 were cultured using this method, with 15 seeds per treatment and 3 biological replicates per treatment. The method for cultivating tissue culture seedlings is referenced in: Wu Gaoyin. Study on the induction and mechanism of somatic embryogenesis in *Dalbergia odorifera* [D]. Guizhou University, 2021. DOI:10.27047 / d.cnki.ggudu.2021.000028. Seeds were treated with concentrated H2SO4 for 1 hour, then rinsed off the concentrated H2SO4 under tap water. They were then treated with 75% ethanol for 1 minute, 2% NaClO for 8 minutes, rinsed 5 times with sterile water, and soaked in sterile water for 24 hours to allow for imbibition. Mature embryos and cotyledons could then be extracted. The imbibition seeds were cultured in MS medium containing BA (1.0 mg / L) and NAA (0.1 mg / L).

[0062] (3) Conventional tray seedling raising method:

[0063] Seeds treated with T1-T6 were used for seedling cultivation according to this method, with 15 seeds per treatment and 3 biological replicates for each treatment. Reference for tray seedling cultivation method: Duan Ruyan, Wei Xiaoli, Zhang Yi, Wang Yinglin, Liang Tangyan. Substrate selection for container seedling cultivation of *Dalbergia odorifera* [J]. Forestry Science and Technology Development, 2015, 29(04):27-31. DOI:10.13360 / j.issn.1000-8101.2015.04.006. The optimal substrate formula (peat:perlite:vermiculite = 2:1:1) was used for preparation, with 2% superphosphate and 20g·m³ of carbendazim added to the substrate. -3 The seedling container is a 12cm×12cm×15cm black non-woven bag. Seedlings are cultured under the same conditions as those grown in seedling boxes.

[0064] 2 Results and Analysis

[0065] (1) Analysis of imbibition effect of different treatments (T1-T6) on Pterocarya spp. seeds

[0066] After treating Pterocarya stenoptera seeds with treatments T1-T6 for 24 hours, the imbibition effect was compared. The results are shown in Table 1. The comparison revealed that minimally invasive seed coat treatment improved the imbibition rate of Pterocarya stenoptera seeds. Seeds without invasive treatment had dense seed coats with poor permeability, making water absorption difficult within the same treatment time. Even with warm water soaking, the imbibition rate after 24 hours was low, though slightly better than room temperature soaking. In the treatments with minimally invasive seed coat treatment, all seeds achieved good imbibition after 24 hours. Among the two minimally invasive treatments—removing the seed coat opposite the hilum (T3, T4) and removing the seed coat at the hilum (T5, T6)—the imbibition effect was particularly rapid with removal of the seed coat at the hilum (T5, T6), showing a noticeable imbibition effect within 30 minutes of soaking. Figure 8 The seeds can achieve complete imbibition within 6 hours, and reach full plumpness after 24 hours, at which point their volume stops increasing. A comparison of the imbibition effects of the T5 and T6 treatments in water at different temperatures showed no significant difference in appearance.

[0067] Table 1 Comparison of the effects of different seed treatments on promoting water absorption

[0068] Seed pretreatment 30min 6h 24h T1 No change No change 8.32% seed imbibition T2 No change No change 13.21% seed imbibition T3 No change No change Seeds fully absorb water T4 No change Seed coat slightly swollen Seeds fully absorb water T5 Seed coat swelling Seed imbibition Seeds fully absorb water T6 Seed coat swelling Seed imbibition Seeds fully absorb water

[0069] (2) Comparison of radicles and germination of Dalbergia odorifera seeds after T1-T6 treatment.

[0070] Rosewood seeds treated with T1-T6 were treated under each treatment condition for 24 hours, and their germination effects were compared. The germination process of rosewood seeds begins with the elongation of the radicle, followed by germination. The germination results are shown in Table 2. Comparative analysis revealed that during the 24-hour soaking period, the radicles of all treatments did not show significant elongation; only the overall seed imbibition effect was observed. On day 7, in tissue culture, some seeds treated with T1, T2, and T3 showed slight elongation of the radicle in some seeds. Seeds treated with T5 and T6 showed slight radicle elongation in seedling trays and plug seedlings. On day 14, in tissue culture, some seeds treated with T1, T2, T3, and T4 showed slight radicle elongation in some seeds; seeds treated with T5 and T6 showed more slight radicle elongation in seedling trays and plug seedlings. On day 25, all seed treatments, except for T5 and T6 which showed no change in the seeds during tissue culture, exhibited radicle growth and elongation. Furthermore, seeds treated with T5 and T6 began to germinate in seedling trays and plug seedlings. On day 45, all seed treatments, except for T5 and T6 which showed no change in the seeds during tissue culture, exhibited radicle growth and seedling germination.

[0071] Table 2 shows that treatments T5 and T6 exhibited the earliest radicle growth and development in the seedling tray method. T5 showed the highest germination rate among all treatments at 25 days, and the best germination effect was observed after 45 days. However, treatments T5 and T6 are not suitable for tissue culture seedlings. Damaged seed hilum areas, during subsequent tissue culture sterilization and bottle placement, resulted in the radicle and plumule being affected by the sterilizing agent, almost completely losing their germination viability. T3 and T4 also used minimally invasive seed coat treatment, but the location of the minimally invasive treatment differed. While some germination was achieved in tissue culture, radicle growth and germination rate were still affected compared to seeds treated with uninvasive seed coats. The sterilization process caused chemical damage to the exposed cotyledons, which affected seed viability in subsequent tissue culture. T1 and T2 methods involved uninvasive seed coat treatment, achieving better germination results in tissue culture seedlings. Tissue culture requires several sterilization steps; retaining the intact seed coat maximizes the protection of cotyledons, radicles, and plumules, ensuring seed germination viability.

[0072] Regarding room temperature soaking and hot water soaking treatments, there was no significant difference in the final germination effect among the non-invasive seed coat treatment methods (T1, T2). Among the minimally invasive seed coat treatment methods (T3-T6), there was no significant difference in the final germination effect between treatments T3 and T4. Treatments T5 and T6 showed a significant difference in the final germination effect, with T6 exhibiting a poorer germination effect. This is because treatments T5 and T6 exposed the radicle and plumule, and the continuous temperature during the hot water soaking process negatively impacted seed viability.

[0073] From the perspective of whether or not the seed coat undergoes minimally invasive treatment, in substrate-based seedling methods (seedling boxes and plug trays), minimally invasive seed coat treatment can indeed achieve better germination results, promote seed imbibition, and facilitate rapid germination. Among these methods, minimally invasive treatment at the seed hilum is particularly effective. The exposed radicle and plumule can rapidly absorb water, and the cotyledons also achieve rapid imbibition. This significantly promotes the rapid breaking of seed dormancy, resulting in radicle growth rate and efficiency superior to minimally invasive treatment at other parts of the seed, and even better than exposing the radicle tip. The mechanical resistance from the seed coat during germination is minimal, and germination efficiency is significantly improved.

[0074] Table 2 Comparison of germination effects of different seed treatments

[0075]

[0076]

[0077] (3) Comparison of seed germination effects after all seed coats have been removed

[0078] For seeds treated with T1-T6, the seed coats were completely removed before subsequent planting experiments were conducted. The treatment groups after seed coat removal were numbered Tb1-Tb6, and the statistical data are shown in Table 3.

[0079] Seeds treated with T1-T6 underwent complete seed coat removal and were designated as Tb1-Tb6. For Tb1 and Tb2, due to incomplete seed imbibition after 24 hours of soaking, some cotyledons were damaged during seed coat removal. Tb3 and Tb4 also experienced partial cotyledon damage during seed coat removal, but their overall seed coat imbibition performance was better than Tb1 and Tb2. The seed coats of Tb5 and Tb6 were extremely easy to remove, and the seeds fully imbibed.

[0080] In terms of cultivation methods, since Tb1-Tb6 were treated after all seed coats were removed, no development and germination effects were achieved in tissue culture. The sterilization process of tissue culture affected both the imbibition of seeds after complete removal of the seed coat (Tb5, Tb6) and the imbibition of seeds without imbibition (Tb1-Tb4), which may have resulted in the complete loss of seed viability.

[0081] Comparison of seeds with completely removed seed coats revealed that the initial minimally invasive treatment promoted seed imbibition, and even after complete removal of the seed coat, optimal radicle growth and germination were still observed (Tb5, Tb6), with radicle growth beginning on day 7. In contrast, for seeds without imbibition (Tb1-Tb4), even after complete removal of the seed coat, the radicle growth rate and germination were significantly worse than the combination treated with the initial minimally invasive seed coat, with radicle growth only beginning to appear on day 14.

[0082] In terms of planting methods, since the tissue culture method did not achieve seed germination, the seedling box method still showed better seedling effect when comparing the seedling tray method and the plug tray method, with the "Tb5-seedling box" showing the best effect.

[0083] Table 3 Comparison of germination effects of different seed coat treatments

[0084]

[0085]

[0086] After combining the above-mentioned different treatment states (T1-T6) of *Dalbergia odorifera* seeds with different planting methods, the seed mold contamination was statistically analyzed after 14 days (Table 4). Both minimally invasive and non-invasive seed coat treatments (T1-T6) achieved good bacterial isolation in the seedling tray method. In the tissue culture method, the original non-invasive seed coat treatment (T1-T2) achieved the lowest contamination rate after the tissue culture disinfection process. However, the minimally invasive seed coat treatments (T3-T6) in the early stages of the tissue culture process showed high contamination rates, especially the treatment method after complete seed imbibition (T5-T6), which showed the highest contamination rate in the later stages of tissue culture. Early seed coat breakage and imbibition may lead to the invasion of exogenous pathogens or active growth of endophytes, which, despite the tissue culture disinfection process, still increased the contamination rate. In plug tray seedling cultivation, a similar contamination trend as in tissue culture seedling cultivation has emerged, but the contamination rate has increased significantly. This is because the substrate in plug tray seedling cultivation not only contains residual pathogens but also some insect eggs. As a result, some seeds are eaten by insect larvae during cultivation, which accelerates the rate of pathogen contamination and leads to a higher contamination rate. In particular, the T5-T6 treatment, which involves exposed embryos and radicles, is extremely susceptible to pathogen contamination and insect larvae, resulting in the highest contamination rate.

[0087] Table 4. Mold contamination rates under different seed treatment and planting methods (T1-T6)

[0088] Seed pretreatment Seedling tray (contamination rate) Tissue culture (contamination rate) Pit contamination rate T1 0 3.23％ 15.89％ T2 0 2.29％ 14.13％ T3 1.32％ 8.67％ 18.52％ T4 0 7.44％ 19.44％ T5 0 13.82％ 31.40％ T6 1.06％ 12.67％ 35.56％

[0089] After combining different treatment states (Tb1-Tb6) of *Palmeria nivea* seeds with different planting methods, the seed mold contamination was statistically analyzed after 14 days (Table 5). In the control group where the seed coat of *Palmeria nivea* was completely removed, all three planting methods showed varying degrees of contamination rates, indicating that retaining the seed coat of *Palmeria nivea* played a certain protective role in isolating or preventing pathogens and pests from invading the seeds during germination. Regarding the method of completely removing the seed coat to break dormancy, Tb5 and Tb6 showed higher contamination rates. The early imbibition process of the seeds accelerated the reproduction rate of internal pathogens and the attachment of exogenous bacteria, leading to earlier mold growth in subsequent planting methods.

[0090] Table 5. Mold contamination rates under different seed treatment and planting methods (Tb1-Tb6)

[0091] Seed pretreatment Seedling tray (contamination rate) Tissue culture (contamination rate) Pit contamination rate Tb1 2.14％ 3.23％ 27.64％ Tb2 1.30％ 2.29％ 23.52％ Tb3 1.89％ 5.45％ 25.79％ Tb4 2.08％ 2.98％ 26.97％ Tb5 5.37％ 10.16％ 44.12％ Tb6 4.79％ 9.46％ 47.32％

[0092] Comparing the results in Tables 4 and 5, it can be seen that when the seed coat of *Dalbergia odorifera* seeds is treated, the method of completely removing the seed coat without any damage achieves better germination results in tissue culture seedling cultivation; the method of removing the seed coat at the hilum with minimal damage achieves better germination results in the "T5-seedling box method"; and the method of completely removing the seed coat achieves the best germination results when combined with the seedling box method.

[0093] (4) Screening of the most optimal and efficient method for quickly breaking dormancy in Pterocarya stenoptera seeds

[0094] The optimal choices for different combinations of seed treatment and planting methods for Dalbergia odorifera were obtained through the analysis of Table 2.

[0095] Among the non-invasive seed treatment methods, the "T1-group culture seedling method" showed the best results; among the minimally invasive seed coat treatment methods, the "T5-seedling box method" showed the best results. Comparing whether or not the seed coat underwent minimally invasive treatment, in substrate-based seedling methods, minimally invasive seed treatment achieved better rooting and germination results than non-invasive treatment.

[0096] Among the planting methods, the "T1-tissue culture seedling method" showed the best results in tissue culture seedling cultivation; the "T5-seedling box method" showed the best results in seedling tray methods; and the "T5-plug tray method" showed the best results in plug tray seedling cultivation. Therefore, the "T5-plug tray method" showed the best results in the selection of substrate-based planting methods.

[0097] Regarding the comparison of soaking temperatures, there were no significant differences between the non-invasive seed treatment methods T1 and T2; there were no significant differences between the minimally invasive seed coat treatments T3 and T4; however, there were significant differences between the minimally invasive seed hilum treatments T5 and T6. This indicates that warm water soaking is more suitable for treatment methods T1-T4, and not suitable for seed coat treatment methods that directly expose the radicle and plumule, as this would reduce seed viability.

[0098] After 45 days of planting, the overall germination efficiency of the seeds using the "T5-seedling box method" was significantly better than that of other seed treatments, different temperature soaking methods, and planting methods.

[0099] The analysis in Table 3 shows that the "Tb5-seedling box method" achieved better germination results. However, the final germination rate was much lower than that of the "T5-seedling box method".

[0100] Based on the analysis of Tables 2-5 and the final statistical germination efficiency, the "T5-seedling box method" achieved the highest germination rate among all seed treatment and planting methods. After 45 days of planting, the rooting rate reached 92.4%, and the germination and seedling rate reached 83.69%. After 45 days of cultivation in the self-made filter-treated seedling box without the need for water or fertilizer, the average plant height was 8.46 cm. Figure 9 The branch nodes have grown to 3 nodes, and the root system is up to 5.89 cm long.

[0101] Dalbergia odorifera is a legume, and its seeds are rich in nutrients. However, during open-field sowing, it is highly susceptible to attack by larvae of flies and other insects in the soil. Pathogenic bacteria also easily attach to and grow on the seed surface. Furthermore, due to its dense seed coat and extremely long dormancy period, seeds often rot before germination even begin, often due to pathogen infestation. Tables 1, 2, and 3 show that the T1 seed treatment method achieved better results in tissue culture seedling cultivation, consistent with previously reported literature. The T5 seed treatment method achieved the best results in seedling tray cultivation. This indicates that the minimally invasive seed coat treatment (T5) not only quickly breaks seed dormancy and promotes rapid cotyledon imbibition in a moist environment, allowing the embryo to acquire external moisture and nutrients in a short time, but also provides good protection for the cotyledons during seed germination, preventing mold and rot caused by exogenous pathogens and damage from pests and diseases remaining in the culture medium. In particular, the use of seedling trays provides low-bacteria, naturally suitable germination conditions for *Palmeria nivea* seeds. The seedling trays maintain the seeds' need for air and moisture during germination, while reducing management costs and providing a stable microenvironment. Previous literature reports that tissue culture is also effective for *Palmeria nivea* seed germination, avoiding the invasion of exogenous bacteria. However, experimental comparisons in this invention revealed that due to the high requirements for a sterile environment, *Palmeria nivea* seeds still exhibit significant endophytic contamination after sterilization and placement in the culture bottle. Furthermore, the endophytic fungi within the seeds themselves cannot be completely eliminated, making contamination during the tissue culture process extremely easy. The application of T5 seed coat micro-invasive treatment to the tissue culture process did not achieve optimal results. After micro-invasive treatment and subsequent sterilization, the exposed embryo portion was easily affected by the disinfectant, leading to loss of activity and germination failure. Although the T1 treatment can achieve better germination results of Pterocarya stenoptera seeds in tissue culture, the results of the above comparative experiments show that the germination efficiency, germination time period, and seedling growth of Pterocarya stenoptera seeds under tissue culture conditions are generally worse than those under seedling tray conditions.

[0102] In summary, this invention utilizes a minimally invasive physical method involving the seed coat, combined with the seedling box of Example 1, to achieve rapid dormancy breaking and germination of *Palmeria nivea* seeds and efficient seedling growth. The physical treatment involves minimally invasive removal of part of the seed coat at the hilum, naturally exposing the embryo and rapidly breaking seed dormancy. The remaining seed coat provides maximum protection for the cotyledons without affecting water and nutrient absorption. Combined with the seedling box device, it provides a stable, low-bacterial microenvironment with optimal humidity during seed germination, closely mimicking the external environmental conditions required for natural seed germination, thus promoting rapid seed germination and efficient seedling growth in *Palmeria nivea*. The use of seedling trays greatly reduces the invasion of exogenous bacteria and pests during seed germination. Compared with tissue culture, it is lower in cost and simpler in operation. Moreover, the entire process does not involve direct application of disinfectants to the rosewood seeds, thus maximizing seed viability and shortening the time for rosewood seeds to break dormancy in the seedling tray. This achieves a highly efficient process from seed collection to germination within 15-20 days, with a rooting rate as high as 92.4%, which is significantly better than the experimental results of rosewood seedling cultivation reported to date.

[0103] In this invention, *Palmeria nivea* seeds were cultured for 45 days in a self-made bacterial culture tray without water replenishment. The germination and seedling emergence rate reached 83.69%, with an average plant height of 8.46 cm, branching nodes reaching 2-3 nodes, and root length up to 5.89 cm. The experimental results of this invention are significantly superior to existing methods in both seed germination efficiency and seedling growth, and the entire germination and seedling cultivation cycle is significantly shortened, reducing costs and increasing efficiency. *Palmeria nivea* is an endangered protected plant, making seed collection relatively difficult and seeds precious. Efficient seed propagation can maximize the rapid propagation of different seed sources. From the perspective of seed and seedling germination and propagation alone, the seedling tray combined with minimally invasive seed coat propagation yields more efficient results than tissue culture and traditional substrate propagation. Using the method of this invention for *Palmeria nivea* seed propagation can also provide a richer source of plant stem segments for tissue culture propagation, improving the conservation effect of the endangered plant *Palmeria nivea*.

Claims

1. A seedling raising method for breaking dormancy of a Terminalia ferdinandiana seed, characterised in that, The method comprises the following steps: S1. After the fruiting of the fruit of the palm tree is matured to dark black, the seeds are peeled off from the tree, and healthy and full palm seeds with a thousand seed weight of not less than 200 g are selected and dried at room temperature within 2-5 days; the surface attachments are washed with a detergent, washed clean with flowing water, then washed 3-5 times with sterile water, and the surface moisture of the seeds is absorbed with sterile filter paper; S2. The seed coat of a 2*2 mm size micro region at the seed navel is cut off without damaging the cotyledon, embryo axis, germ and radicle at the seed navel, and the rest of the seed coat is not cut off; then the seed is soaked in a proper amount of sterile water for 24 hours in the dark, and the sterile water is replaced every 6 hours during the soaking; S3. The seed obtained in step S2 is sown in the nutrient medium of the seedling box, and the sowing depth is 1.5-2 cm, then the outer cup is tightly covered with the outer cup cover; the sowed seedling box is placed in a culture condition of 15-18℃, light / dark length of 12 / 12 hours; S4. After the seed cotyledon breaks the soil, the culture condition is adjusted to a temperature of 23-26℃, light / dark length of 16 / 8 hours; when the branch node of the seedling is 2-3 nodes long, the seedling is taken out from the seedling box and transplanted; The seedling box comprises an outer cup, an outer cup cover and an inner cup, the outer cup bottom is filled with sterile water of a certain height, the inner cup is fixed at a middle-lower position in the outer cup, the outer bottom of the inner cup is higher than the liquid level of the sterile water, the inner cup bottom is provided with a threading hole, the water absorption line passes through the threading hole and can tightly plug the threading hole gap, the water absorption line outside the inner cup naturally falls to the outer cup bottom, the water absorption line inside the inner cup extends along the inner cup wall to the outside of the inner cup mouth, and the inner cup is filled with nutrient medium; the outer cup cover is provided with a bacteria filtering and air permeable hole, and the outer cup cover is used for tightly covering the outer cup mouth; The water absorption line is two or more.

2. The seedling raising method for breaking seed dormancy of Vismia sp. according to claim 1, characterized in that, Each part of the seedling box is sterilized.

3. The seedling raising method for breaking seed dormancy of Vismia sp. according to claim 1, characterized in that, The outer cup and the outer cup cover are transparent materials, and the inner cup is a paper cup with a polyethylene film on the inner wall.

4. The seedling raising method for breaking seed dormancy of Vismia sp. according to claim 3, characterized in that, The outer cup is made of PS material with a thickness of 0.2 cm, and the outer cup mouth is a threaded cup mouth; the outer cup cover is made of PP material, which is a threaded cover matched with the outer cup mouth.

5. The seed germination method according to claim 1, wherein The water absorption line is a cotton line with a thickness of 0.4 mm.

6. The seedling raising method for breaking seed dormancy of Vismia sp. according to claim 1, characterized in that, The nutrient medium is peat soil with a fiber length of 5-20 mm, which is fully moistened with sterile water and filled to a degree that the hand-formed group does not drip water.

7. The seedling raising method for breaking seed dormancy of Vismia sp. according to claim 1, characterized in that, The inner cup is fixed at a middle-lower position in the outer cup, that is, the inner cup mouth is clamped in the outer cup and the clamping position height is located in the middle of the outer cup.

8. The seed germination method according to claim 1, wherein The room temperature drying in step S1 is natural air drying or full drying in a drying box.

9. The seedling raising method for breaking seed dormancy of Vismia sp. according to claim 1, characterized in that, The sowing of the seed obtained in step S2 in the nutrient medium of the seedling box is performed according to one seed per seedling box.

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