Method for accelerating germination and improving seedling emergence rate of phoebe sheareri

Abstract

The application belongs to the technical field of plant seedling, and more particularly relates to a method for rapidly germinating and improving the germination rate of Phellodendron amurense seeds. The method comprises the following steps: disinfecting the Phellodendron amurense seeds with alcohol and sodium hypochlorite solution, and then naturally storing the seeds in sand at low temperature for 87 days. The germination rate and the germination speed of the Phellodendron amurense seeds are significantly improved by optimizing the storage germination conditions and the ratio of the seedling substrate, and the germination rate can reach 73.67 %. The method is simple in operation, low in cost, and environmentally friendly, and is suitable for large-scale seedling and afforestation of Phellodendron amurense.

Description

Technical Field

[0001] This invention belongs to the field of plant seedling technology, and more specifically, relates to a method for rapidly germinating and improving the germination rate of *Pterocarya stenoptera* seeds. Background Technology

[0002] The tree, taxonomically named Prunusbuergeriana *Prunus mume*, a deciduous tree belonging to the genus *Prunus* in the family Rosaceae, is widely distributed in mountainous areas throughout my country. It is a tall tree with good timber quality, and its round drupes turn blackish-red when ripe, attracting some birds. *Prunus mume* is not only an excellent native timber species but can also be used as a shade tree or ornamental tree to create distinctive rural landscapes, making it a multi-purpose native tree species with development potential. Currently, apart from a small-scale application in furniture making, *Prunus mume* has not been formally cultivated or utilized, and research on it is extremely scarce; only one previous study on *Prunus mume* seeds exists. Studies by Chen et al. (Chen SY, Li WY, Han MC, et al. Association of abscisic acid and gibberellins with dormancy and germination in seeds of Prunus buergeriana miq[J]. Taiwan Journal of Forest Science, 2005, 20(3): 227-237.) have shown that fresh seeds of Prunus buergeriana exhibit dormancy due to high concentrations of abscisic acid (ABA). Dormancy could be broken by cold stratification, treatment at 4℃ for 12 weeks, or treatment with 500 ppm of exogenous gibberellin GA3. However, cold stratification showed higher germination efficiency, with an average germination time 40 days shorter than GA3 treatment. While warm stratification reduced ABA content to near germination levels (approximately 24 ng / g), it lacked significant accumulation of endogenous gibberellins GA1, GA3, GA4, and their precursor GA20, resulting in seed failure to germinate. Cold stratification, on the other hand, not only reduced ABA to similarly low levels but also significantly activated the GA biosynthesis pathway. The study confirmed that the germination of *Pterocarya stenoptera* seeds requires dual regulation by ABA inhibition and GA promotion. This research revealed the dormancy characteristics of *Pterocarya stenoptera* seeds, providing a theoretical basis for their sexual reproduction. However, to date, no reports have been found regarding storage-induced germination and propagation techniques for *Pterocarya stenoptera* seeds. Summary of the Invention

[0003] The purpose of this invention is to provide a method for rapidly germinating and increasing the germination rate of *Pterocarya stenoptera* seeds.

[0004] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0005] This invention provides a method for rapidly germinating and improving the germination rate of *Pterocarya stenoptera* seeds, comprising the following steps: soaking *Pterocarya stenoptera* seeds in water for disinfection, mixing them with moist sand and placing them in a seed bag, burying the seed bag in the moist sand for natural low-temperature storage until the germination rate of the *Pterocarya stenoptera* seeds reaches 73.67±5.31%, and obtaining germinated seeds; then sowing the germinated seeds in a seedling substrate for seedling cultivation. The natural low-temperature conditions are as follows during storage: daytime temperature not exceeding 21.01℃ and not lower than -2.1℃, nighttime temperature not exceeding 17.15℃ and not lower than -2.11℃.

[0006] In the method provided by this invention, the germination rate of *Pterocarya stenoptera* seeds that have undergone natural low-temperature sand stratification is the highest, and the seedling emergence rate can reach as high as 71.11±10.70% 20 days after sowing. Furthermore, when *Pterocarya stenoptera* seeds that have undergone natural low-temperature sand stratification are sown in composite nutrient soil or fine sand, it is beneficial for the seeds to germinate quickly, and the seedling emergence rate is significantly increased.

[0007] Furthermore, the soaking time is 24 hours.

[0008] Furthermore, the disinfection process includes: soaking in 75% alcohol for 10 seconds, then soaking in 10% sodium hypochlorite solution for 10 minutes, and finally rinsing five times with sterile water.

[0009] Furthermore, the seed bag is buried at a depth of 15cm.

[0010] Furthermore, the moisture content of the wet sand is 30%.

[0011] Furthermore, the volume ratio of the tilapia seeds to the wet sand in the seed bag is 1:5.

[0012] Furthermore, the seedling substrate is fine sand or a mixed substrate, wherein the mixed substrate is prepared by mixing nutrient soil and vermiculite in a volume ratio of 1:1.

[0013] Furthermore, the vermiculite particle size is 3mm~6mm.

[0014] The present invention has the following beneficial effects:

[0015] The method provided by this invention clarifies the appropriate storage and germination methods for *Pteris vittata* seeds, as well as suitable sowing substrates, filling the gap in *Pteris vittata* seed propagation technology and laying a technical foundation for the large-scale seedling cultivation and development of the native plant *Pteris vittata*. Attached Figure Description

[0016] Figure 1 This is a statistical graph showing the effects of different sowing substrates on the leaf growth of *Pterocarya stenoptera* seedlings. Different letters represent significant differences between treatment groups at the 0.05 level. Detailed Implementation

[0017] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments, but this should not be construed as limiting the invention. Unless otherwise specified, the technical means used in the following embodiments are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following embodiments are commercially available unless otherwise specified.

[0018] The seeds of the *Pterocarya stenoptera* were collected in late October 2024 from six mature seeds of individual plants in two locations: Guanshan National Nature Reserve and Taojiangyuan Nature Reserve in Quannan County, Jiangxi Province. The seeds were washed, dried in the shade, and prepared for use.

[0019] The sand was first sieved through a 2mm sieve, then soaked in a 1g / L carbendazim solution for 1 hour for disinfection, and then rinsed three times with tap water before use. The carbendazim was purchased from Sichuan Runer Technology Co., Ltd.

[0020] Example 1

[0021] I. Experimental Methods.

[0022] 1. The effect of storage methods on seed germination.

[0023] Storage began on November 27, 2024. Each individual plant was used as a replicate, with 6 replicates (6 individual plants). 150 seeds were taken from each individual plant, soaked in water for 24 hours, then disinfected with 75% alcohol for 10 seconds, followed by disinfection with 10% sodium hypochlorite solution for 10 minutes, rinsed 5 times with sterile water, slightly dried, and then individually packaged into seed mesh bags. Three storage methods were employed:

[0024] Natural low-temperature sand storage: Seeds are mixed with moist sand (30% humidity) at a volume ratio of 1:5 and placed into seed bags. All seed bags are then buried in a large outdoor basin of sand. A 20cm layer of sand is laid at the bottom of the basin, followed by the seed bags spread evenly on top and covered with a 15cm layer of sand. A rain shelter is erected over the basin, and artificial water spraying is used to maintain the sand humidity at 30%. During natural low-temperature storage, the average daily maximum temperature is 12.91±1℃, the average daily minimum temperature is 4.12±1℃, the average daily temperature is 8.19±1℃, the average diurnal temperature range is 8.78±1℃, the highest daytime temperature is 21.01℃, and the lowest daytime temperature is -2.1℃. The highest nighttime temperature is 17.15℃, and the lowest nighttime temperature is -2.11℃.

[0025] Low-temperature sand storage: After mixing the seeds with moist sand, put them into seed bags. Bury all the seed bags in a plastic basin filled with moist sand. Place the entire basin in a 4℃ freezer. Cover the basin with a plastic bag to keep it moist and poke small holes for ventilation. Observe it from time to time and keep the sand humidity at 30% by spraying water.

[0026] Constant temperature and humidity storage: Do not mix with wet sand. Wrap all the seed bags containing only seeds with a wet towel and store them in a 4℃ freezer. Observe them from time to time and keep the towel at a certain humidity by spraying water.

[0027] After 87 days of storage, germination rate and radicle length were statistically analyzed on February 22, 2025. Germination was defined as the radicle breaking through the seed coat; the percentage of germinated seeds in each bag out of the initial storage quantity of 150 seeds was the germination rate. From each bag of germinated seeds, the 10 seeds with the longest radicles were selected, and the radicle length of each seed was measured using calipers. The average radicle length of the 10 seeds was calculated.

[0028] To test the effectiveness of seed storage, seedling experiments were conducted using a portion of seeds stored in different ways. On February 22, 2025, 15 healthy, plump seeds with well-developed radicles were selected from each seed bag and sown into 5 small flowerpots, 3 seeds per pot, for a total of 90 pots. The substrate was a 1:1 mixture of potting soil and vermiculite. The potted plants were placed in a greenhouse and watered normally. The germination rate, i.e., the percentage of seeds that sprouted and formed visible seedlings, was measured. Germination rates were recorded three times on days 15, 20, and 25 after sowing.

[0029] 2. The effect of soil substrate on the germination of *Pterocarya stenoptera* seeds.

[0030] On March 12, 2025, a sowing experiment was conducted using seeds that had undergone natural low-temperature sand stratification to germinate in different substrates. Three soil substrate types were used: fine sand, composite nutrient soil, and garden soil. The composite nutrient soil consisted of a 1:1 volume ratio of purchased nutrient soil and vermiculite; the garden soil was soil sourced directly from the nursery. 50-cell pots were used as sowing containers, with three pots sown for each substrate (three replicates). One seed was sown per cell, for a total of 450 seeds in nine pots. The experiment was conducted in a greenhouse with normal watering management. Starting from the 11th day after sowing, the germination rate and the total number of leaves per pot were recorded every five days, for a total of six records. The nutrient soil was purchased from Zhejiang Hongyue Horticulture Co., Ltd., the vermiculite particle size was 3mm-6mm, and the garden soil was collected from the Flower and Nursery Teaching Base of Jiangxi Agricultural University.

[0031] II. Data Processing.

[0032] Statistical analysis was performed using Excel 2016 and IBM SPSS 26.0 software. One-way ANOVA and Duncan's method for multiple comparisons were used to assess differences between different treatments. Data in charts and graphs are presented in the form of "mean ± standard error," with a significance level of 0.05.

[0033] III. Results and Analysis

[0034] 1. Experimental results on the effects of different storage methods on the germination of *Pterocarya stenoptera* seeds.

[0035] Table 1 shows that the germination rates of seeds stored under natural low-temperature sand stratification and constant low-temperature sand stratification were 73.67% and 61.56%, respectively, significantly higher than the 36.44% of those stored under constant low-temperature humidification. After natural low-temperature sand stratification, the radicles were significantly longer than those stored under the other two methods. This indicates that sand stratification is more conducive to the germination and sprouting of *Pterocarya stenoptera* seeds than sandless storage, with natural low-temperature sand stratification being the optimal method.

[0036] In terms of germination rate, the germination rate of seeds stored in natural low-temperature sand was the highest on both the 15th and 20th day after sowing. By the 20th day, the germination rate of seeds stored in natural low-temperature sand had reached over 70%, while the germination rate of seeds stored using the other two methods also reached over 70% by the 25th day, with no significant difference among the three. This indicates that seeds stored in natural low-temperature sand have the fastest germination rate after sowing.

[0037] In summary, from the perspective of seed germination and early seedling emergence, natural low-temperature sand storage is a better method for storing *Pterocarya stenoptera* seeds.

[0038] Table 1: Effects of storage methods on seed germination and emergence of *Pterocarya stenoptera*.

[0039]

[0040] Note: Different letters in the same column represent significant differences between treatments at the 0.05 level, and the same applies to the following table.

[0041] 2. Results of the effects of different soil substrates on the germination of *Pterocarya stenoptera* seeds.

[0042] Table 2 shows that during the six observation periods (days 11-36 post-sowing), there was no significant difference in seed germination rates between the sand and nutrient soil substrates. However, the germination rate of the garden soil substrate was consistently the lowest, with significant differences observed on days 11, 16, 31, and 36 post-sowing. This indicates that using sand and nutrient soil as soil substrates does not significantly affect the germination rate of *Pterocarya stenoptera* seeds in the short term, and both substrates are superior to garden soil. Furthermore, the experiment revealed that compared to germination rate 5, germination rate 6 was lower in the sand and garden soil substrates due to some seedling mortality, while the germination rate in the nutrient soil substrate remained unchanged.

[0043] The overall seedling emergence peak was reached, measured on the 21st day after sowing, by the number of leaves per seedling per hill. The results showed that, if... Figure 1 As shown, the number of leaves per pot was significantly higher in both fine sand and nutrient soil than in garden soil; there was no significant difference between fine sand and nutrient soil, but the number of leaves was slightly higher in nutrient soil.

[0044] In summary, sand and potting soil are superior to garden soil among the three sowing substrates; in the short term, sand and potting soil do not show significant differences in germination rate and leaf number of *Pterocarya stenoptera* seeds.

[0045] Table 2: Effect of sowing substrate on seed germination of *Pterocarya stenoptera*.

[0046]

[0047] Note: Emergence rates of 1 to 6 refer to the emergence rates on days 11, 16, 21, 26, 31, and 36 after sowing, respectively.

[0048] Conclusion: Natural low-temperature stratification of *Pterocarya stenoptera* seeds in moist sand can improve the germination rate and promote rapid seedling emergence. Sand or potting soil can be used as the sowing substrate, but potting soil is preferable in the long run.

[0049] Although preferred embodiments of the invention have been described, those skilled in the art, upon learning the basic inventive concept, can make other changes and modifications to these embodiments. Therefore, the appended claims are intended to be interpreted as including both the preferred embodiments and all changes and modifications falling within the scope of the invention.

[0050] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A method for rapid germination and improved seedling emergence of Acacia mangium seeds, characterized by, The method comprises the following steps: After soaking the seed of Ulmus pumila L. in water for 24 hours, the seed is disinfected, mixed with wet sand, put into a seed bag, buried in wet sand with a humidity of 30%, and a depth of 15 cm, and then stored in a natural low-temperature sand until the germination rate of the seed reaches 73.67±5.31%, thereby obtaining the germinated seed; and the germinated seed is sowed in a seedling raising substrate to raise seedlings, wherein the natural low-temperature condition is that the temperature during the storage is not higher than 21.01℃ during the day, not lower than -2.1℃ during the day, not higher than 17.15℃ at night, and not lower than -2.11℃ at night; The disinfection comprises the following steps: soaking in alcohol with a volume fraction of 75% for 10 seconds, soaking in sodium hypochlorite solution with a volume fraction of 10% for 10 minutes, and finally washing with sterile water for 5 times; The volume ratio of the seed of Ulmus pumila L. to the wet sand in the seed bag is 1:5; The seedling raising substrate is fine sand or a mixed substrate, and the mixed substrate is prepared by mixing nutrient soil and vermiculite at a volume ratio of 1:1; The particle size of the vermiculite is 3mm-6mm.

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