A seedling raising method suitable for oryza sativa and application thereof

Abstract

The application relates to the technical field of seed seedling raising, and provides a seedling raising method suitable for seeds of the genus Ormosia and application thereof, the seedling raising method comprises the following steps: (1) in early March, the seeds of the genus Ormosia are put into hot water with a temperature of 60-80 DEG C, the water is naturally cooled, the seeds are taken out after being absorbed, and the seeds are sprayed with water to keep moist; (2) step (1) is repeated once a day, and the process is continued for 5-7 days, so that the seeds are obtained after preliminary treatment; (3) the seeds after preliminary treatment are soaked in a suspension seed coating agent for 10-18 hours, so that the seeds after soaking are obtained; (4) the seeds after soaking are inoculated into a substrate to accelerate germination for 30-50 days. The seedling raising method can improve the germination rate and germination uniformity of the seeds of the genus Ormosia, and simultaneously improve the quality of seedlings, increase the height and the number of lateral roots of the seedlings. The seedling raising method is low in cost, wide in applicability, easy to popularize, and can provide reliable technical support for seedling raising of the seeds of the genus Ormosia of the legume family.

Description

Technical Field

[0001] This invention relates to the field of seed and seedling technology, and in particular to a seedling method applicable to seeds of the genus *Ormosia* and its application. Background Technology

[0002] The Pterocarpus henryi Prainensis, Ormosia hosiei Hemsl. & E.H. Wilson, and Ormosia semicastrata Hance are all trees belonging to the genus Ormosia in the legume family. These three species produce high-quality timber, making them valuable for timber production, and they also possess significant medicinal and economic value. Their seeds are bright red and glossy, giving them considerable ornamental value. However, due to scarce wild resources and limited artificial cultivation, these three species are facing extinction.

[0003] The seeds of *Palmaria*, *Ormosia henryi*, and *Ormosia rubescens* are all hard seeds. Previous studies have primarily used methods such as hot water soaking, physical damage, or fungal soaking to promote seed germination. However, these methods are often only applicable to a single tree species, have a narrow scope of application, and are not suitable for large-scale seedling production. Therefore, there is an urgent need in this field to develop an artificial propagation and large-scale seedling production method applicable to various *Ormosia* species, including *Palmaria*, *Ormosia henryi*, and *Ormosia rubescens*. Summary of the Invention

[0004] The purpose of this invention is to provide a seedling cultivation method and its application for seeds of the genus *Ormosia*. This method can effectively improve the germination rate, uniformity of germination, and seedling quality of *Ormosia* seeds, providing technical support for their artificial propagation, thereby promoting the protection and cultivation of rare and endangered tree species. Furthermore, this seedling cultivation method has the advantages of low cost, wide applicability, and ease of promotion, and will provide reliable technical guidance for the seedling cultivation of rare *Ormosia* tree species, assisting in their protection and propagation.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0006] This invention provides a method for raising seedlings of the genus *Ormosia*, comprising the following steps:

[0007] (1) In early March, put the seeds of the genus *Ormosia* into hot water at 60-80°C and let the water cool naturally. After the seeds have absorbed water and swelled, take them out and spray them with water to keep them moist.

[0008] (2) Repeat step (1) once a day for 5 to 7 days to obtain pre-treated seeds;

[0009] (3) Soak the pre-treated seeds in a suspension seed coating agent for 10-18 hours to obtain soaked seeds;

[0010] (4) Soak the seeds and inoculate them into the substrate to germinate for 30 to 50 days.

[0011] Preferably, the water level of the hot water in step (1) is 5-7 cm higher than the seeds of the genus *Gynostemma*.

[0012] Preferably, in step (3), the liquid level of the suspension seed coating agent during soaking is 3-5 cm higher than the seeds. The active ingredients of the suspension seed coating agent are metalaxyl and fludioxonil. The concentration of metalaxyl in the suspension seed coating agent is 18.5-74 mg / L, and the concentration of fludioxonil is 12.5-50 mg / L.

[0013] Preferably, the moisture content of the substrate in step (4) is 50-60%, the temperature of the substrate is 25-30°C, and the substrate is composed of raw materials in the following volume ratio: peat:vermiculite:perlite = 0.8-1.2:0.8-1.2:0.8-1.2, and the volume ratio of the seeds to the substrate is 0.8-1.2:2-4.

[0014] Preferably, the substrate in step (4) includes a disinfection step before use: first disinfecting the substrate with potassium permanganate solution, and then disinfecting it a second time with carbendazim solution.

[0015] Preferably, the concentration of potassium permanganate in the potassium permanganate solution is 0.05% to 1 wt%, and the concentration of carbendazim in the carbendazim solution is 0.2% to 0.5 wt%.

[0016] Preferably, the germination method described in step (4) is stratification germination.

[0017] The present invention also provides the application of the described seedling raising method in seed raising of the genus *Ormosia*.

[0018] Preferably, the seeds of the genus *Ormosia* are one or more of the following: *Palmaria* seeds, *Ormosia* seeds, and *Ormosia henryi* seeds.

[0019] The beneficial effects of this invention are as follows:

[0020] 1. This invention significantly improves the seed germination rates of *Palmaria dentata*, *Ormosia henryi*, and *Ormosia rubescens*, reaching 89%, 94.33%, and 74.33%, respectively. Compared to traditional single seedling cultivation methods, the germination rate of *Palmaria dentata* is increased by 9% (Deng Zhao, Preliminary Study on Seed Dormancy and Germination of *Palmaria dentata*, 2011), the germination rate of *Ormosia henryi* is increased by 21% (Kong Ting, Research on Seed Germination Experiment and Seedling Technology of *Ormosia henryi*, 2022), and the germination rate of *Ormosia rubescens* is increased by 4.33% (Huang Honglan, Research on Seed Characteristics and Seedling Growth Characteristics of *Ormosia rubescens*, 2025). Furthermore, the seedling cultivation method of this application also improves the uniformity of seed germination, with germination uniformity of 80%, 81.2%, and 71.8% for *Palmaria dentata*, *Ormosia henryi*, and *Ormosia rubescens*, respectively.

[0021] 2. This invention improves the quality of *Palmaria lobata*, *Ormosia henryi*, and *Ormosia rubescens* seedlings. Compared with pure vermiculite as the substrate, *Palmaria lobata* seedlings show a 20% increase in height and a 110% increase in the number of lateral roots; *Ormosia henryi* seedlings show a 42% increase in height and a 110% increase in the number of lateral roots; and *Ormosia rubescens* seedlings show a 21% increase in height and a 115% increase in the number of lateral roots.

[0022] 3. This invention is applicable to the cultivation of seedlings from hard seeds of three tree species: Dalbergia odorifera, Ormosia henryi, and Ormosia rubescens. It breaks through the limitation of existing technologies that are only applicable to a single tree species and provides a unified technical solution for the protection and propagation of rare tree species of the Ormosia genus.

[0023] 4. The method of this invention is simple and easy to implement, and can be mastered by ordinary forestry workers. It is conducive to its widespread application in forestry production and is of great significance for the protection of rare tree species and the improvement of the ecological environment. Attached Figure Description

[0024] Figure 1 This is a comparison chart of the germination rates of *Palmaria lobata*, *Ormosia henryi*, and *Ormosia rubescens* seeds on different substrates in Example 3.

[0025] Figure 2 The images show the germination status of *Palmaria dentata* seeds on different substrates in Example 3.

[0026] Figure 3 The diagram shows the germination status of red bean seeds on different substrates in Example 3;

[0027] Figure 4 The images show the germination status of soft-podded red bean seeds on different substrates in Example 3.

[0028] Figure 5 The image shows the growth of seedlings of *Palmaria dentata*, *Lysimachia foenum-graecum*, and *Lysimachia foenum-graecum* on different substrates in Example 3. Detailed Implementation

[0029] This invention provides a method for raising seedlings of the genus *Ormosia*, comprising the following steps:

[0030] (1) In early March, put the seeds of the genus *Ormosia* into hot water at 60-80°C and let the water cool naturally. After the seeds have absorbed water and swelled, take them out and spray them with water to keep them moist.

[0031] (2) Repeat step (1) once a day for 5 to 7 days to obtain pre-treated seeds;

[0032] (3) Soak the pre-treated seeds in a suspension seed coating agent for 10-18 hours to obtain soaked seeds;

[0033] (4) Soak the seeds and inoculate them into the substrate to germinate for 30 to 50 days.

[0034] In this invention, the water level of the hot water in step (1) is preferably 5-7 cm above the seeds of the red bean genus, and more preferably 6 cm above the seeds.

[0035] In this invention, during the soaking process in step (3), the liquid level of the suspension seed coating agent is preferably 3-5 cm above the seeds, more preferably 4 cm above the seeds. The active ingredients of the suspension seed coating agent are preferably metalaxyl and fludioxonil. The final concentration of metalaxyl in the suspension seed coating agent is preferably 18.5-74 mg / L, more preferably 37 mg / L, and the final concentration of fludioxonil is preferably 12.5-50 mg / L, more preferably 25 mg / L.

[0036] In this invention, the moisture content of the substrate in step (4) is preferably 50-60%, more preferably 55%, the temperature of the substrate is preferably 25-30°C, more preferably 27°C, and the substrate is preferably composed of raw materials in the following volume ratio: peat:vermiculite:perlite = 0.8-1.2:0.8-1.2:0.8-1.2, more preferably composed of raw materials in the following volume ratio: peat:vermiculite:perlite = 1:1:1, and even more preferably composed of raw materials in the following volume ratio: peat:vermiculite:perlite = 0.8:1:1.2; the volume ratio of the seeds to the substrate is preferably 0.8-1.2:2-4, more preferably 1:3, and even more preferably 0.8:2.5.

[0037] In this invention, the substrate in step (4) preferably includes a disinfection step before use: first disinfecting the substrate with potassium permanganate solution, and then disinfecting it a second time with carbendazim solution.

[0038] In this invention, the concentration of potassium permanganate in the potassium permanganate solution is preferably 0.05% to 1 wt%, more preferably 0.08 wt%, and the concentration of carbendazim in the carbendazim solution is preferably 0.2% to 0.5 wt%, more preferably 0.3% to 0.4 wt%.

[0039] In this invention, the germination method in step (4) is preferably stratification germination. The preferred step of stratification germination is: placing the seeds and substrate in a seedling tray and watering once every 2 to 4 days. The seedling tray is a seedling tray with a neat grid at the bottom and a size of 40 to 45 cm × 40 to 45 cm × 5 to 10 cm. The number of seed layers is 1 to 3. More preferably, the seedling tray is a seedling tray with a neat grid at the bottom and a size of 43 cm × 43 cm × 7 cm. The number of seed layers is 2.

[0040] This invention also provides the application of the described seedling raising method in seed raising of the genus *Ormosia*.

[0041] In this invention, the seeds of the genus *Ormosia* are preferably one or more of the following: seeds of *Palmaria*, seeds of *Ormosia* tree, and seeds of *Ormosia henryi*.

[0042] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0043] Example 1 Seed Collection and Processing

[0044] (1) Seeds were collected between October and November 2024. The seeds of *Palmaria dentata* were collected from Daozhen, Guizhou; the seeds of *Ormosia henryi* were collected from Chengdu, Sichuan; and the seeds of *Ormosia rubescens* were collected from Qianxinan, Guizhou. After cutting the pods with pruning shears, the seeds were extracted, placed in breathable cloth bags, and then brought back to the laboratory for air-drying. The air-drying time was 60 days. Afterward, the seeds were placed in breathable cloth bags and stored in a dry place for later use.

[0045] (2) On March 7, 2025, 900 seeds each of the stored Dalbergia odorifera, Ormosia henryi and Ormosia lentigines were placed in hot water at 70°C and allowed to cool naturally. After the seeds swelled, they were taken out and sprayed with water to keep them moist. The water level of the hot water was 6 cm above the seeds.

[0046] (3) From March 8, 2025 to March 12, 2025, repeat step (2) once a day for the seeds of the genus *Aegilops* treated the previous day: put the seeds of the genus *Aegilops* treated the previous day into hot water at 70°C, let the water cool naturally, and take them out after they have absorbed the water and swelled. Spray water on the seeds to keep them moist, and finally obtain the seeds that have undergone preliminary treatment.

[0047] (4) Soak the pre-treated seeds in the suspension seed coating agent for 12 hours to obtain the soaked seeds for later use.

[0048] During soaking, the liquid level of the suspension seed coating agent needs to be 4 cm higher than the seeds. The active ingredients of the suspension seed coating agent are metalaxyl and fludioxonil. The final concentration of metalaxyl in the suspension seed coating agent is 37 mg / L, and the final concentration of fludioxonil in the mixed suspension seed coating agent is 25 mg / L. The suspension seed coating agent is prepared by diluting a mixed solution of metalaxyl at a concentration of 37 mg / L and fludioxonil at a concentration of 25 mg / L by 1000 times.

[0049] Experiment Example 2: Seed Quality Index Determination

[0050] According to the seed moisture content and weight determination methods in the "Forest Tree Seed Inspection Procedures (GB / T 2772-1999)," the moisture content (%) and weight (g) of seeds of *Dalbergia odorifera*, *Ormosia henryi*, and *Ormosia rubescens* were determined. Three replicates were set up for each tree species, with 20 seeds per replicate. Twenty seeds were randomly selected from each tree species, and the transverse and longitudinal diameters were measured using calipers. The transverse-to-longitudinal diameter ratio was calculated (Xie Shiyang, Study on the Effects of Different Treatments on Seedling Cultivation of *Ormosia rubescens*, 2024), to understand the characteristics of hard-seeded seeds in the three tree species.

[0051] Table 1 shows that there were significant differences in the 100-seed weight, moisture content, transverse diameter, longitudinal diameter, and aspect ratio of the three tree species (p<0.05). Among them, the red bean tree seeds had the highest 100-seed weight (83.857g) and moisture content (13.468%), while the soft-pod red bean seeds had the lowest 100-seed weight (33.269g) and the highest aspect ratio (1.1).

[0052] Table 1. Seed quality of three tree species

[0053] species Rosewood Red Bean Tree Soft-pod red beans Weight of 100 grains (g) 40.967±0.687b 83.857±2.74a 33.269±0.77b Moisture content (%) 12.992±0.94b 13.468±0.11a 12.464±0.15b transverse diameter (mm) 10.33±0.76b 12.91±1.02a 9.02±0.65c Longitudinal diameter (mm) 8.79±0.52b 13.04±1.86a 9.96±0.55b Aspect Ratio 0.85±0.05b 1.01±0.17a 1.1±0.07a

[0054] Example 3: Seedling cultivation experiment with different substrates

[0055] (1) This invention designs three different matrix ratio schemes:

[0056] ① Pure vermiculite;

[0057] ② Vermiculite and perlite are mixed in a 1:1 volume ratio;

[0058] ③ Mix peat, vermiculite and perlite in a volume ratio of 1:1:1.

[0059] In each substrate formulation, 100 seeds each of the soaked and swollen *Palmaria rubra*, *Lysimachia foenum-graecum*, and *Lysimachia foenum-graecum* were placed, as described in Example 1, and three replicates were set up, with each replicate cultured for 50 days.

[0060] All of the above-mentioned substrates include a disinfection step before use: first, the substrate is disinfected with potassium permanganate solution, and then disinfected a second time with carbendazim solution. The concentration of potassium permanganate in the potassium permanganate solution is 0.08 wt%, and the concentration of carbendazim in the carbendazim solution is 0.3 wt%.

[0061] Stratification for germination: Select a seedling tray with a neat grid at the bottom and a size of 43cm×43cm×7cm; mix seeds and sterilized substrate at a volume ratio of 1:3 for germination, and place the seeds in 2 layers.

[0062] Routine Management: Place the seedling trays flat inside the greenhouse, construct an arched shed 1m wide and 0.6m high, and cover it with white plastic arched shed film and a single layer of shade netting. Water every 3 days to maintain a relative humidity of 55±2% for the substrate moisture content, and a temperature of 27±2℃ for the substrate and inside the greenhouse.

[0063] (2) Germination index

[0064] Seed germination was determined by the emergence of the radicle ("white root") (Li Zhiping, Effects of NaCl and Na2CO3 stress on seed germination and seedling growth of *Quercus variabilis*, 2015). Germination rate was calculated after 30 days. The formula for calculating germination rate is:

[0065] Germination rate % = (Number of normally germinated seeds / Total number of seeds tested) × 100%; Formula 1

[0066] (3) Germination uniformity

[0067] Based on the current seedling status, the germinated seeds were divided into 6 categories. After 30 days, the percentage of different types of seeds in the total number of germinated seeds was counted, and the total percentage of categories 3 to 6 was also counted to observe the uniformity of germination.

[0068] Category 1: Radicle ≤ 1cm;

[0069] Category 2: 1cm < radicle ≤ 3cm;

[0070] Category 3: 3cm ≤ radicle, or 3cm ≤ radicle and plumule protruding;

[0071] Category 4: Both radicle and plumule are present, and the plumule is ≤1.5cm;

[0072] Category 5: Both radicle and plumule are present, and the plumule is 1.5cm < plumule ≤ 3cm;

[0073] Category 6: Both radicle and plumule are present, with plumule >3cm, and young leaves are exposed or unfolded.

[0074] (4) Seedling growth indicators

[0075] Fifty days after seedling cultivation, 10 seedlings were randomly selected from each treatment group. The seedling height (cm) and root length (cm) of the *Dalbergia odorifera*, *Ormosia henryi*, and *Ormosia rubescens* seedlings were measured using a steel tape measure. Their fresh weight (g) was measured using a benchtop balance with an accuracy of 0.01%, and the number of lateral roots was counted.

[0076] Depend on Figure 1 It was found that the germination rates of *Palmeria nivea*, *Ormosia henryi*, and *Ormosia rubescens* differed significantly under different substrates (p<0.05). The highest germination rates were observed in a substrate composed of a 1:1:1 mixture of peat, vermiculite, and perlite, with 89% for *Palmeria nivea*, 94.33% for *Ormosia henryi*, and 74.33% for *Ormosia rubescens*.

[0077] Table 2 shows that there were significant differences in seed types among *Dalbergia odorifera*, *Ormosia henryi*, and *Ormosia rubescens* grown on different substrates (p<0.05). Among these, *Dalbergia odorifera*, *Ormosia henryi*, and *Ormosia rubescens* had the lowest proportion of germination in categories 1 and 2 on a peat:vermiculite:perlite = 1:1:1 substrate, while the combined proportion of germination in categories 3 to 6 was the highest, indicating that the germination uniformity of the three tree species was highest on this substrate.

[0078] from Figure 2 , Figure 3 , Figure 4 It can be seen that the germination states of *Dalbergia odorifera*, *Ormosia henryi*, and *Ormosia rubescens* seeds differed on different substrates. *Dalbergia odorifera*, on both the vermiculite:perlite = 1:1 and peat:vermiculite:perlite = 1:1:1 substrates, showed true leaves in all germination stages (Category 6). *Ormosia henryi*, on the vermiculite:perlite = 1:1:1 substrate, showed true leaves in both germination stages (Category 5 and Category 6). *Ormosia rubescens*, on the other hand, showed true leaves in its Category 6 germination stage on the vermiculite:perlite = 1:1:1 substrate. This indicates that the vermiculite:perlite = 1:1:1 substrate formula is beneficial for the formation of true leaves in seedlings.

[0079] Table 2. Statistics on germination status of three tree species on different substrates.

[0080]

[0081] Note: The one-way ANOVA test showed a significant difference at the p<0.05 level.

[0082] Depend on Figure 5 Table 3 shows that the seedling height, root length, and number of lateral roots of *Dalbergia odorifera* and *Ormosia henryi* differed significantly on the three different substrates (p<0.05); the seedling height, root length, number of lateral roots, and fresh weight of *Ormosia henryi* seedlings also differed significantly on the three different substrates (p<0.05). All three species grew best on a peat:vermiculite:perlite = 1:1:1 substrate, especially in terms of seedling height and number of lateral roots.

[0083] Table 3. Seedling growth indicators of three tree species on different substrates

[0084] Note: The one-way ANOVA test showed a significant difference at the p<0.05 level.

[0085] As can be seen from the above embodiments, the present invention provides a seedling cultivation method for seeds of the genus *Ormosia* and its application. The seedling cultivation method includes the following steps: (1) In early March, *Ormosia* seeds are placed in hot water at 60-80°C, and the water is allowed to cool naturally. After the seeds have swelled, they are taken out and sprayed with water to keep them moist; (2) Step (1) is repeated once a day for 5-7 days to obtain pre-treated seeds; (3) The pre-treated seeds are soaked in a suspension seed coating agent for 10-18 hours to obtain soaked seeds; (4) The soaked seeds are inoculated into a substrate to germinate for 30-50 days. The seedling cultivation method of the present invention can improve the germination rate and uniformity of *Ormosia* seeds, while improving seedling quality, increasing seedling height and the number of lateral roots. It can be seen that the seedling cultivation method of this application is low in cost, widely applicable, and easy to promote, and can provide reliable technical support for the seedling cultivation of rare tree species of the genus *Ormosia* in the legume family.

[0086] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for raising seedlings of the genus *Ormosia*, characterized in that, Includes the following steps: (1) In early March, put the seeds of the genus *Ormosia* into hot water at 60-80°C and let the water cool naturally. After the seeds have absorbed water and swelled, take them out and spray them with water to keep them moist. (2) Repeat step (1) once a day for 5 to 7 days to obtain preliminarily treated seeds; (3) Soak the pre-treated seeds in a suspension seed coating agent for 10-18 hours to obtain soaked seeds; (4) Soak the seeds and inoculate them into the substrate to germinate for 30-50 days; The active ingredients of the suspension seed coating agent in step (3) are metalaxyl and fludioxonil. The concentration of metalaxyl in the suspension seed coating agent is 18.5~74 mg / L, and the concentration of fludioxonil is 12.5~50 mg / L. The substrate in step (4) is composed of raw materials in the following volume ratio: peat:vermiculite:perlite = 0.8~1.2:0.8~1.2:0.8~1.2, and the volume ratio of the seeds to the substrate is 0.8~1.2:2~4.

2. The seedling raising method according to claim 1, characterized in that, In step (1), the water level of the hot water should be 5-7 cm above the seeds of the red bean genus.

3. The seedling raising method according to claim 2, characterized in that, In step (3), the liquid level of the seed coating agent during soaking should be 3-5 cm higher than the seeds.

4. The seedling raising method according to claim 3, characterized in that, The water content of the substrate in step (4) is 50-60%, and the temperature of the substrate is 25-30℃.

5. The seedling raising method according to claim 4, characterized in that, The substrate described in step (4) also includes a disinfection step before use: first disinfect the substrate with potassium permanganate solution, and then disinfect it a second time with carbendazim solution.

6. The seedling raising method according to claim 5, characterized in that, The concentration of potassium permanganate in the potassium permanganate solution is 0.05%~1wt%, and the concentration of carbendazim in the carbendazim solution is 0.2%~0.5wt%.

7. The seedling raising method according to claim 6, characterized in that, The germination method described in step (4) is stratification germination.

8. The application of the seedling raising method according to any one of claims 1 to 7 in the seed raising of the genus *Ormosia*.

9. The application according to claim 8, characterized in that, The seeds of the genus *Ormosia* are one or more of the following: seeds of *Palmaria*, seeds of *Ormosia* tree, and seeds of *Ormosia henryi*.

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