Method for improving germination rate of hemerocallis citrina baroni seeds
By combining gibberellin and hydrogen peroxide, the problems of low germination rate and uneven germination of daylily seeds were solved, achieving a highly efficient and simple method to improve seed germination rate, which is applicable to the field of agricultural biotechnology.
Patent Information
- Application Number
- CN202511991593.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-26
- Publication Date
- 2026-02-06
AI Technical Summary
Daylily seeds have a low germination rate and uneven germination, and are sensitive to environmental conditions. Traditional treatment methods have limited effectiveness and cannot meet the needs of large-scale production.
By treating daylily seeds with a combination of gibberellin (GA3) and hydrogen peroxide (H2O2), a synergistic effect was achieved by breaking physiological dormancy, promoting embryo cell division, improving seed coat permeability, and activating germination-related enzyme systems.
It significantly improves the germination rate of daylily seeds to 92.67%, with more concentrated and faster germination. The operation is simple, low-cost, and standardized, with strong reproducibility.
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Figure CN121464786A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the field of agricultural biotechnology, in particular to a method for improving the germination rate of daylily seeds. BACKGROUND
[0002] Daylily (Hemerocallis fulva L. Hemerocallis citrina Baroni) is a perennial herb of Hemerocallis genus, which has important edible, medicinal and ornamental values. Currently, the main propagation method in production is division, but it has low propagation coefficient, high seedling cost and easy disease transmission. Seed propagation can significantly improve the propagation efficiency, but it has the following technical obstacles:
[0003] Low germination rate: the germination rate of daylily seeds under natural conditions is only 50.00%, and the dense seed coat is the main factor limiting water permeation and gas exchange. Although the germination rate can be improved to 88.00% after peeling, the operation is complicated and the embryo is easily damaged, which is not suitable for large-scale production. In addition, there may be physiological dormancy mechanism in the seed, resulting in uneven germination.
[0004] High environmental sensitivity: the seed germination has strict requirements on temperature, humidity and other conditions. Although traditional seed soaking treatment (such as 40-80℃ warm water soaking) has certain effect, high temperature above 60℃ can easily produce inhibitory effect, and the germination rate is only 66.00%-75.00%, and the problem of uneven emergence cannot be solved. SUMMARY
[0005] In view of the above technical problems of the prior art, the present application provides a method for improving the germination rate of daylily seeds, which significantly improves the germination rate of daylily seeds by combined treatment of gibberellin (GA3) and hydrogen peroxide (H2O2). The gibberellin breaks the physiological dormancy of the seed, promotes the division of the embryo cells, and at the same time, the hydrogen peroxide moderately oxidizes the seed coat, improves the permeability, and provides active oxygen signal molecules to activate the germination related enzyme system. The two have a synergistic effect through the triple mechanism of "breaking dormancy-promoting permeability-activating".
[0006] To solve the above technical problems, the present application adopts the following technical scheme: A method for improving the germination rate of daylily seeds, comprising the following steps: Soaking the daylily seeds, then adding hydrogen peroxide and gibberellin for combined treatment, and culturing until germination to obtain daylily germinated seeds.
[0007] In a preferred embodiment of the present application, the combined treatment means that the daylily seeds are first treated in hydrogen peroxide, and then treated in gibberellin.
[0008] In a preferred embodiment of the present application, the seed soaking treatment method is to soak the daylily seeds in sterile water at 25-80℃, and the seed soaking treatment time is 24-36h.
[0009] In the preferred embodiment of the present application, the volume concentration of hydrogen peroxide in the hydrogen peroxide and hormone combined treatment is 1%-3%, the mass concentration of gibberellin is 25mg / L-75mg / L, and the total time of the combined treatment is 24h-36h.
[0010] In the preferred embodiment of the present application, the concentration of hydrogen peroxide in the hydrogen peroxide and hormone combined treatment is 1%, and the concentration of gibberellin is 25mg / L.
[0011] In the preferred embodiment of the present application, the culture conditions are as follows: temperature 22℃-25℃, relative humidity 55%-60%, and no light.
[0012] Compared with the prior art, the present application has the beneficial effects that: 1. The method for significantly improving the seed germination rate of daylily by using the combined treatment of gibberellin (GA3) and hydrogen peroxide (H2O2) is simple in operation, low in cost, breaks seed physiological dormancy by using gibberellin, promotes embryo cell division, at the same time, moderately oxidizes seed coat by using hydrogen peroxide, improves permeability, provides active oxygen signal molecules to activate germination related enzyme system, and produces synergistic effect through the three mechanisms of "breaking dormancy-promoting permeability-activating activation", so that the germination rate is greatly improved, and the germination period is significantly shortened.
[0013] 2. The seed germination rate of daylily is systematically improved from 60.00% in the natural state to 92.67% by the combined treatment; the germination uniformity is high: the germination potential and germination index are significantly improved by the combined treatment and other methods, which means that the seed germination is more concentrated and faster, and the seedling growth is more consistent; the operation is standardized and the reproducibility is good: the present application provides specific and quantitative treatment parameters (such as concentration, temperature, and time), forms a standardized process, is easy to popularize and apply, and has strong reproducibility.
[0014] 3. The method for improving the germination rate of daylily seeds described in this invention has a synergistic effect. The discovered combined treatment scheme of gibberellin and low-concentration hydrogen peroxide produces a synergistic effect of "1+1>2", which is a significant technological advancement. First, gibberellin directly participates in breaking physiological dormancy. GA disrupts the balance between abscisic acid and GA in the seed, activates signaling pathways, and induces the synthesis of hydrolytic enzymes such as α-amylase and protease in the endosperm or cotyledons. This further decomposes the starch, protein, and other macromolecules stored in the seed into soluble sugars or amino acids, providing energy and material basis for embryo growth and development. It mainly solves the problem of "physiological dormancy". Hydrogen peroxide mainly solves the problems of physical dormancy and signal transduction. It not only softens the seed coat, but also activates the expression of a series of germination-related genes during seed germination and participates in the GA signal transduction pathway, enhancing the cell's sensitivity to GA. When used in combination, their effects are not simply additive, but rather mutually reinforcing, primarily manifested in the following ways: First, signaling pathways are cross-linked and amplified. H2O2 can oxidize and inhibit negative regulators of the GA signaling pathway or activate positive regulators, thereby sensitizing seed cells to GA. Downstream reactions of GA also produce reactive oxygen species, thus strengthening signal cycling and accelerating the breaking of dormancy. Second, the order and synergistic effect of breaking dormancy: H2O2 acts first, softening the seed coat, increasing oxygen permeability, and acting as an initial signal to "awaken" seed cells, breaking physical dormancy. GA then dominates the biochemical reactions. With physical dormancy broken and cells pre-activated, GA enters the embryo, efficiently initiating the synthesis of hydrolytic enzymes and related genes, promoting the decomposition of stored substances. Third, there is a synergistic effect of energy conversion: the oxygen provided by the decomposition of H2O2 directly promotes aerobic respiration in the embryonic cells, providing immediate energy for the complex biochemical processes induced by GA (such as enzyme synthesis and cell division). Therefore, the combined use of both is not a simple superposition of exogenous substances, but rather an interaction of a series of physical, chemical, and biochemical processes. Attached Figure Description
[0015] Figure 1 The dynamic changes in the germination of daylily seeds in different years.
[0016] Figure 2 This invention relates to the effect of soaking temperature on the germination rate A and germination index B of daylily seeds.
[0017] Figure 3 The present invention relates to the germination rate A and germination index B of daylily seeds treated with hydrogen peroxide.
[0018] Figure 4 This invention relates to the effects of different stratification times on the germination rate (A) and germination index (B) of daylily seeds.
[0019] Figure 5This invention relates to the effects of plant growth regulators on the germination of daylily seeds, wherein A represents the effect of different concentrations of GA3 on the germination of daylily seeds, B represents the effect of different concentrations of 6-BA on the germination of daylily seeds, and C represents the effect of different concentrations of KT on the germination of daylily seeds. Detailed Implementation
[0020] The following detailed description, in conjunction with embodiments of the present invention and accompanying drawings, provides a clear and complete illustration of the technical solutions in these embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.
[0021] It should be noted that all technical terms used in this invention are for the purpose of describing specific embodiments only and are not intended to limit the scope of protection of this invention. Unless otherwise specified, all raw materials, reagents, instruments and equipment used in the following embodiments of this invention can be purchased from the market or prepared by existing methods.
[0022] Example 1 (1) Soak the daylily seeds in water at 25°C. After disinfection, place 50 seeds in a petri dish and add sterile water at the corresponding temperature. Let them cool naturally to room temperature. Set the light incubator to no light (24h), temperature 25°C and humidity 60%.
[0023] (2) Set the gibberellin concentration to 25 mg / L and the hydrogen peroxide concentration to 1%. Set up 3 replicates. For each replicate, select 50 plump and uniformly sized seeds. Place the seeds in the solution of the concentration combination. Soak the seeds in gibberellin and hydrogen peroxide for 12 hours each. Rinse the seeds 3 times with sterile water to remove the residual treatment solution on the seed surface. Place the seeds evenly on a petri dish lined with a layer of filter paper and two layers of gauze. Cultivate in a light incubator until germination, and obtain daylily germination seeds.
[0024] Example 2 (1) Soak the daylily seeds in water at 40°C. After disinfection, place 50 seeds in a petri dish, add sterile water at the corresponding temperature, and let them cool naturally to room temperature. Set the light incubator to no light (24h), temperature 25°C, and humidity 60%.
[0025] (2) Set the gibberellin concentration to 25 mg / L and the hydrogen peroxide concentration to 1%. Set up 3 replicates. For each replicate, select 50 plump and uniformly sized seeds. Place the seeds in the solution of the concentration combination. Soak the seeds in gibberellin and hydrogen peroxide for 12 hours each. Rinse the seeds 3 times with sterile water to remove the residual treatment solution on the seed surface. Place the seeds evenly on a petri dish lined with a layer of filter paper and two layers of gauze. Cultivate in a light incubator until germination, and obtain daylily germination seeds.
[0026] Example 3 (1) Soak the daylily seeds in water at 60°C. After disinfection, place 50 seeds in a petri dish, add sterile water at the corresponding temperature, and let them cool naturally to room temperature. Set the light incubator to no light (24h), temperature 25°C, and humidity 60%.
[0027] (2) Set the gibberellin concentration to 25 mg / L and the hydrogen peroxide concentration to 1%. Set up 3 replicates. For each replicate, select 50 plump and uniformly sized seeds. Place the seeds in the solution of the concentration combination. Soak the seeds in gibberellin and hydrogen peroxide for 12 hours each. Rinse the seeds 3 times with sterile water to remove the residual treatment solution on the seed surface. Place the seeds evenly on a petri dish lined with a layer of filter paper and two layers of gauze. Cultivate in a light incubator until germination, and obtain daylily germination seeds.
[0028] Example 4 (1) Soak the daylily seeds in water at 80°C. After disinfection, place 50 seeds in a petri dish and add sterile water at the appropriate temperature. Let them cool naturally to room temperature. Set the light incubator to no light (24h), temperature 25°C and humidity 60%.
[0029] (2) Set the gibberellin concentration to 25 mg / L and the hydrogen peroxide concentration to 1%. Set up 3 replicates. For each replicate, select 50 plump and uniformly sized seeds. Place the seeds in the solution of the concentration combination. Soak the seeds in gibberellin and hydrogen peroxide for 12 hours each. Rinse the seeds 3 times with sterile water to remove the residual treatment solution on the seed surface. Place the seeds evenly on a petri dish lined with a layer of filter paper and two layers of gauze. Cultivate in a light incubator until germination, and obtain daylily germination seeds.
[0030] Example 5 (1) Soak the daylily seeds in water at 25°C. After disinfection, place 50 seeds in a petri dish and add sterile water at the corresponding temperature. Let them cool naturally to room temperature. Set the light incubator to no light (24h), temperature 25°C and humidity 60%.
[0031] (2) The gibberellin concentration was set at 25 mg / L and the hydrogen peroxide concentration was 3%. Three replicates were set. In each replicate, 50 plump and uniformly sized seeds were selected. The seeds were placed in the solution of the concentration combination. After soaking in gibberellin and hydrogen peroxide for 12 hours each, the seeds were rinsed three times with sterile water to remove the residual treatment solution on the seed surface. The seeds were then evenly placed on a petri dish lined with a layer of filter paper and two layers of gauze and cultured in a light incubator until germination, thus obtaining daylily germination seeds.
[0032] Example 6 (1) Soak the daylily seeds in water at 25°C. After disinfection, place 50 seeds in a petri dish and add sterile water at the corresponding temperature. Let them cool naturally to room temperature. Set the light incubator to no light (24h), temperature 25°C and humidity 60%.
[0033] (2) The gibberellin concentration was set at 25 mg / L and the hydrogen peroxide concentration was 6%. Three replicates were set. In each replicate, 50 plump and uniformly sized seeds were selected. The seeds were placed in the solution of the concentration combination. After soaking in gibberellin and hydrogen peroxide for 12 hours each, the seeds were rinsed three times with sterile water to remove the residual treatment solution on the seed surface. The seeds were then evenly placed on a petri dish lined with a layer of filter paper and two layers of gauze and cultured in a light incubator until germination, thus obtaining daylily germination seeds.
[0034] Example 7 (1) Soak the daylily seeds in water at 25°C. After disinfection, place 50 seeds in a petri dish and add sterile water at the corresponding temperature. Let them cool naturally to room temperature. Set the light incubator to no light (24h), temperature 25°C and humidity 60%.
[0035] (2) Set the gibberellin concentration to 50 mg / L and the hydrogen peroxide concentration to 1%. Set up 3 replicates. For each replicate, select 50 plump and uniformly sized seeds. Place the seeds in the solution of the concentration combination. Soak the seeds in gibberellin and hydrogen peroxide for 12 hours each. Rinse the seeds 3 times with sterile water to remove the residual treatment solution on the seed surface. Place the seeds evenly on a petri dish lined with a layer of filter paper and two layers of gauze. Cultivate in a light incubator until germination, and obtain daylily germination seeds.
[0036] Example 8 (1) Soak the daylily seeds in water at 25°C. After disinfection, place 50 seeds in a petri dish and add sterile water at the corresponding temperature. Let them cool naturally to room temperature. Set the light incubator to no light (24h), temperature 25°C and humidity 60%.
[0037] (2) Set the gibberellin concentration to 50 mg / L and the hydrogen peroxide concentration to 3%, set 3 replicates, and select 50 plump and uniformly sized seeds for each replicate. Place the seeds in the solution of the concentration combination, soak them in gibberellin and hydrogen peroxide for 12 hours each, rinse the seeds 3 times with sterile water to remove the residual treatment solution on the seed surface, and place them evenly on a petri dish with a layer of filter paper and two layers of gauze. Cultivate in a light incubator until germination, and obtain daylily germination seeds.
[0038] Example 9 (1) Soak the daylily seeds in water at 25°C. After disinfection, place 50 seeds in a petri dish and add sterile water at the corresponding temperature. Let them cool naturally to room temperature. Set the light incubator to no light (24h), temperature 25°C and humidity 60%.
[0039] (2) Set the gibberellin concentration to 50 mg / L and the hydrogen peroxide concentration to 6%. Set up 3 replicates. In each replicate, select 50 plump and uniformly sized seeds. Place the seeds in the solution of the concentration combination. Soak the seeds in gibberellin and hydrogen peroxide for 12 hours each. Rinse the seeds 3 times with sterile water to remove the residual treatment solution on the seed surface. Place the seeds evenly on a petri dish with a layer of filter paper and two layers of gauze. Cultivate in a light incubator until germination to obtain daylily germination seeds.
[0040] Example 10 (1) Soak the daylily seeds in water at 25°C. After disinfection, place 50 seeds in a petri dish and add sterile water at the corresponding temperature. Let them cool naturally to room temperature. Set the light incubator to no light (24h), temperature 25°C and humidity 60%.
[0041] (2) Set the gibberellin concentration to 75 mg / L and the hydrogen peroxide concentration to 1%. Set up 3 replicates. For each replicate, select 50 plump and uniformly sized seeds. Place the seeds in the solution of the concentration combination. Soak the seeds in gibberellin and hydrogen peroxide for 12 hours each. Rinse the seeds 3 times with sterile water to remove the residual treatment solution on the seed surface. Place the seeds evenly on a petri dish lined with a layer of filter paper and two layers of gauze. Cultivate in a light incubator until germination, and obtain daylily germination seeds.
[0042] Example 11 (1) Soak the daylily seeds in water at 25°C. After disinfection, place 50 seeds in a petri dish and add sterile water at the corresponding temperature. Let them cool naturally to room temperature. Set the light incubator to no light (24h), temperature 25°C and humidity 60%.
[0043] (2) The gibberellin concentration was set at 75 mg / L and the hydrogen peroxide concentration was 3%. Three replicates were set. In each replicate, 50 plump and uniformly sized seeds were selected. The seeds were placed in the solution of the concentration combination. After soaking in gibberellin and hydrogen peroxide for 12 hours each, the seeds were rinsed three times with sterile water to remove the residual treatment solution on the seed surface. The seeds were then evenly placed on a petri dish lined with one layer of filter paper and two layers of gauze and cultured in a light incubator until germination, thus obtaining daylily germination seeds.
[0044] Example 12 (1) Soak the daylily seeds in water at 25°C. After disinfection, place 50 seeds in a petri dish and add sterile water at the corresponding temperature. Let them cool naturally to room temperature. Set the light incubator to no light (24h), temperature 25°C and humidity 60%.
[0045] (2) Set the gibberellin concentration to 75 mg / L and the hydrogen peroxide concentration to 6%. Set up 3 replicates. For each replicate, select 50 plump and uniformly sized seeds. Place the seeds in the solution of the concentration combination. Soak the seeds in gibberellin and hydrogen peroxide for 12 hours each. Rinse the seeds 3 times with sterile water to remove the residual treatment solution on the seed surface. Place the seeds evenly on a petri dish lined with a layer of filter paper and two layers of gauze. Cultivate in a light incubator until germination, and obtain daylily germination seeds.
[0046] Comparative Example 1 Fifty intact seeds were allowed to absorb moisture for 24 hours and then placed directly in a light incubator for germination testing. The light incubator was set to a dark environment (24 hours), a temperature of 25°C, and a humidity of 60%.
[0047] Comparative Example 2 Fifty seeds were sterilized and placed in petri dishes. Sterile water at 25°C was added to each dish, and the dishes were allowed to cool naturally to room temperature. After absorbing water for 24 hours, the seeds were placed in a light incubator for germination testing.
[0048] Comparative Example 3 Fifty seeds were sterilized and placed in petri dishes. Sterile water at 40°C was added to each dish, and the dishes were allowed to cool naturally to room temperature. After absorbing water for 24 hours, the seeds were placed in a light incubator for germination testing.
[0049] Comparative Example 4 Fifty seeds were sterilized and placed in petri dishes. Sterile water at 60°C was added to each dish. The dishes were allowed to cool naturally to room temperature and absorb water for 24 hours before being placed in a light incubator for germination testing.
[0050] Comparative Example 5 Fifty seeds were sterilized and placed in petri dishes. Sterile water at 80°C was added to each dish, and the dishes were allowed to cool naturally to room temperature. After absorbing water for 24 hours, the seeds were placed in a light incubator for germination testing.
[0051] Comparative Example 6 Fifty seeds were soaked in a 1% hydrogen peroxide solution for 24 hours, rinsed three times with sterile water, and evenly dispersed on a petri dish lined with one layer of filter paper and two layers of gauze. The seeds were then cultured in a light incubator. Seeds treated directly with distilled water served as a germination control. The process was repeated three times.
[0052] Comparative Example 7 Fifty seeds were soaked in a 3% hydrogen peroxide solution for 24 hours, rinsed three times with sterile water, and evenly dispersed on a petri dish lined with one layer of filter paper and two layers of gauze. The seeds were then cultured in a light incubator. Seeds treated directly with distilled water served as a germination control. The process was repeated three times.
[0053] Comparative Example 8 Fifty seeds were soaked in a 6% hydrogen peroxide solution for 24 hours, rinsed three times with sterile water, and evenly dispersed on a petri dish lined with one layer of filter paper and two layers of gauze. The seeds were then cultured in a light incubator. Seeds treated directly with distilled water served as a germination control. The process was repeated three times.
[0054] Comparative Example 9 Fifty seeds were soaked in a 9% hydrogen peroxide solution for 24 hours, rinsed three times with sterile water, and evenly dispersed on a petri dish lined with one layer of filter paper and two layers of gauze. The seeds were then cultured in a light incubator. Seeds treated directly with distilled water served as a germination control. The process was repeated three times.
[0055] Comparative Example 10 (1) Disinfect the stratified seeds with 0.1% KMnO4 solution for 30 minutes, rinse with clean water, and then mix the seeds with sterilized river sand in a ratio of 1:3. The moisture content of the river sand should be 40%, that is, it should be able to be formed into a ball when squeezed in the hand and crumble when released.
[0056] (2) After stratification, the seeds were placed in a 4℃ refrigerator for indoor stratification. After 24 hours of absorbing water, 50 seeds were taken out for germination test. The germination status of the seeds was recorded and repeated 3 times.
[0057] Comparative Example 11 (1) Disinfect the stratified seeds with 0.1% KMnO4 solution for 30 minutes, rinse with clean water, and then mix the seeds with sterilized river sand in a ratio of 1:3. The moisture content of the river sand should be 40%, that is, it should be able to be formed into a ball when squeezed in the hand and crumble when released.
[0058] (2) Place the stratified seeds in a 4℃ refrigerator for indoor stratification. During stratification, add water and mix to maintain the moisture of the seed sand. After 15 days, take out 50 seeds for germination test and record the germination status of the seeds. Repeat 3 times.
[0059] Comparative Example 12 (1) Disinfect the stratified seeds with 0.1% KMnO4 solution for 30 minutes, rinse with clean water, and then mix the seeds with sterilized river sand in a ratio of 1:3. The moisture content of the river sand should be 40%, that is, it should be able to be formed into a ball when squeezed in the hand and crumble when released.
[0060] (2) After stratification, the seeds were placed in a 4℃ refrigerator for indoor stratification. During stratification, water was added and mixed to maintain the moisture of the seed sand. After 30 days, 50 seeds were taken out for germination test and the germination status of the seeds was recorded. The test was repeated 3 times.
[0061] Comparative Example 13 (1) Disinfect the stratified seeds with 0.1% KMnO4 solution for 30 minutes, rinse with clean water, and then mix the seeds with sterilized river sand in a ratio of 1:3. The moisture content of the river sand should be 40%, that is, it should be able to be formed into a ball when squeezed in the hand and crumble when released.
[0062] (2) After stratification, the seeds were placed in a refrigerator at 4°C for indoor stratification. During stratification, water was added and mixed to maintain the moisture of the seed sand. After 45 days, 50 seeds were taken out for germination test and the germination status of the seeds was recorded. The test was repeated 3 times.
[0063] Comparative Example 14 (1) Disinfect the stratified seeds with 0.1% KMnO4 solution for 30 minutes, rinse with clean water, and then mix the seeds with sterilized river sand in a ratio of 1:3. The moisture content of the river sand should be 40%, that is, it should be able to be formed into a ball when squeezed in the hand and crumble when released.
[0064] (2) After stratification, the seeds were placed in a 4℃ refrigerator for indoor stratification. During stratification, water was added and mixed to maintain the moisture of the seed sand. After 60 days, 50 seeds were taken out for germination test and the germination status of the seeds was recorded. The test was repeated 3 times.
[0065] Comparative Example 15 Fifty seeds were soaked in a 25 mg / L gibberellin (GA3) solution for 24 h, then rinsed three times with sterile water, and finally cultured in a light incubator. This process was repeated three times.
[0066] Comparative Example 16 Fifty seeds were soaked in a 50 mg / L gibberellin (GA3) solution for 24 h, then rinsed three times with sterile water, and finally cultured in a light incubator. This process was repeated three times.
[0067] Comparative Example 17 Fifty seeds were soaked in a 75 mg / L gibberellin (GA3) solution for 24 h, then rinsed three times with sterile water, and finally cultured in a light incubator. This process was repeated three times.
[0068] Comparative Example 18 Fifty seeds were soaked in a 100 mg / L solution of 6-benzylaminopurine (6-BA) for 24 h, then rinsed three times with sterile water, and finally cultured in a light incubator. This process was repeated three times.
[0069] Comparative Example 19 Fifty seeds were soaked in a 200 mg / L solution of 6-benzylaminopurine (6-BA) for 24 h, then rinsed three times with sterile water, and finally cultured in a light incubator. This process was repeated three times.
[0070] Comparative Example 20 Fifty seeds were soaked in a 300 mg / L solution of 6-benzylaminopurine (6-BA) for 24 h, then rinsed three times with sterile water, and finally cultured in a light incubator. This process was repeated three times.
[0071] Comparative Example 21 Fifty seeds were soaked in a 10 mg / L kinetin (KT) solution for 24 h, then rinsed three times with sterile water, and finally cultured in a light incubator. This process was repeated three times.
[0072] Comparative Example 22 Fifty seeds were soaked in a 50 mg / L kinetin (KT) solution for 24 h, then rinsed three times with sterile water, and finally cultured in a light incubator. This process was repeated three times.
[0073] Comparative Example 23 Fifty seeds were soaked in a 100 mg / L kinetin (KT) solution for 24 h, then rinsed three times with sterile water, and finally cultured in a light incubator. This process was repeated three times.
[0074] Results Analysis Single treatments have limited effectiveness: Existing technologies using single hormone or single oxidant treatments can partially improve germination, but the germination rate remains low, and there are shortcomings such as poor concentration of germination time and low germination index. Currently, there are no reports on the combined treatment of gibberellin and hydrogen peroxide for daylily seeds. The synergistic mechanism of the two reagents and the optimal concentration ratio are not yet clear, leading to blind spots for producers in the seed treatment process.
[0075] Figure 1 The variation in daylily seed germination rate in different years. Figure 1The results show that under a constant temperature of 25℃, the germination of daylily seeds varied across different years. In 2022 and 2023, the germination rate of daylily seeds increased rapidly at the beginning of the germination test. In 2021, the germination rate increased slowly from day 0 to day 9, then stabilized around day 10, ultimately remaining at a low level with a germination rate of 8.67%. In 2022, the germination rate increased rapidly from day 0 to day 9, reaching a peak of 40.67% from day 10 to day 15. The germination rate of daylily seeds in 2023 was the fastest, peaking around day 15 with a germination rate of 60.00%. Statistical analysis revealed significant differences in the germination rates of daylily seeds from different years (F=85.604, P<0.05). The differences in germination rates for seeds from 2021, 2022, and 2023 were all statistically significant.
[0076] Figure 2 To investigate the effect of soaking temperature on the germination rate (A) and germination index (B) of daylily seeds, the germination rate of daylily seeds treated at different soaking temperatures showed an increasing trend over time. The germination rate increased slowly from 0-5 days, accelerated from 5-15 days, and gradually stabilized after 15 days. Figure A shows that the germination rate of seeds treated at 60℃ increased rapidly and eventually stabilized at a high level, reaching 81.00%. The germination rate of seeds treated at 80℃ increased slowly in the early stages, and although it increased later, the final germination rate was lower than that of the 60℃ treatment group, at 66.00%. The germination rate trend of the 40℃ treatment was similar to that of the control group (CK). Figure B shows that the germination index of the 60℃ treatment was the highest, at 3.84, significantly higher than that of the 80℃ treatment, but not significantly different from the control group and the 40℃ treatment. This indicates that a soaking temperature of 60℃ has a certain effect on the germination rate and germination index of daylily seeds, while treatment at 80℃ has an inhibitory effect on seed germination.
[0077] As shown in Table 1, the initial germination time was 3.67±0.58 days for the control group (CK, room temperature), 4.67±0.58 days for the 40℃ treatment group, 4.33±2.89 days for the 60℃ treatment group, and 4.67±1.15 days for the 80℃ treatment group. There was no significant difference in initial germination time among the different treatment groups. This indicates that different water temperatures have little effect on the initial germination time of daylily seeds.
[0078] Table 1 shows the germination time of daylily seeds under different soaking temperatures. Different water temperatures had a certain impact on the time required for 50% germination and the final germination time, with significant differences in the final germination time. Compared with the control (room temperature), relatively lower temperatures were more conducive to seed germination, while higher temperature treatments inhibited seed germination.
[0079] Figure 3 To determine the germination rate (A) and germination index (B) of daylily seeds treated with hydrogen peroxide according to this invention, this experiment used hydrogen peroxide to treat the daylily seeds. Figure 3 As shown, with increasing hydrogen peroxide concentration, the germination rate of daylily seeds initially increased and then decreased, reaching its highest point at a 3% hydrogen peroxide concentration with a germination rate of 84.67%. Over time, the germination rate of all treatments showed an upward trend, with the 3% hydrogen peroxide treatment showing the fastest increase and eventually stabilizing at a high level. The germination rates of the 1% and 6% hydrogen peroxide treatments showed similar trends, with germination rates of 70% and 78%, respectively. The 9% hydrogen peroxide treatment showed slow initial growth and a lower final level, with a germination rate of 54%. Compared to the control group (CK), 1%, 3%, and 6% hydrogen peroxide concentrations all promoted daylily seed germination, while the 9% hydrogen peroxide treatment showed a decrease compared to the CK, indicating that increasing hydrogen peroxide concentration inhibits daylily seed germination.
[0080] As shown in Table 2, treatment with hydrogen peroxide at concentrations of 1%, 3%, and 6% promoted the initial germination time of daylily seeds, which were 2.33±1.15 days, 2.67±0.58 days, and 3.00±0.00 days, respectively. However, the 9% concentration treatment inhibited the initial germination time. There was no significant difference in the 50% germination time, indicating that different treatments had a relatively small impact on the time required for seeds to reach 50% germination. Compared to the control group, different concentrations of hydrogen peroxide treatment increased the germination rate of daylily seeds while also prolonging the final germination time.
[0081] Table 2 Germination time of daylily seeds treated with different concentrations of hydrogen peroxide Figure 4 This invention investigates the effects of different stratification times on the germination rate (A) and germination index (B) of daylily seeds. The germination rates varied with stratification time. As the number of stratification days increased, the germination rate initially increased and then decreased. The highest germination rate (90%) was observed at 45 days. Germination rates at other stratification times were 78.67% (15 days), 76.67% (30 days), and 70.00% (60 days), respectively. Compared to the control group (CK), although the germination index did not increase significantly, the germination rate was higher in all cases, indicating that stratification effectively improves the germination rate of daylily seeds.
[0082] As shown in Table 3, different stratification times significantly affected the germination time of daylily seeds. The shorter 15-day stratification treatment delayed the time required for initial germination and 50% germination; the 45-day and 60-day stratification treatments promoted earlier germination and faster attainment of 50% germination, but all stratification treatments delayed the final germination time of the seeds.
[0083] Table 3 Germination time of daylily seeds treated with different stratification times To improve the germination rate of daylily seeds, this experiment used three plant growth regulators to treat the seeds. The results showed that the germination rate was significantly affected by the plant growth regulators.
[0084] Figure 5 To investigate the effects of the plant growth regulators of this invention on the germination of daylily seeds, GA3 at four concentrations all improved seed germination rate, with the highest germination rate reaching 90% at a concentration of 50 mg / L. 6-BA significantly affected seed germination rate, with a noticeable effect at a concentration of 200 mg / L, achieving a germination rate of 81.33%. Compared to the control group, KT at a concentration of 10 mg / L also improved seed germination rate to 82.00%, while KT at 50 mg / L and 100 mg / L inhibited seed germination. Among the three hormones used to treat daylily seeds, 50 mg / L GA3 showed the best effect, achieving a germination rate of 90%.
[0085] As shown in Table 4, the initial germination time of GA3 at a concentration of 50 mg / L was 2.67 ± 0.58 days, significantly shorter than that of the control group (CK), indicating that treatment at this concentration promoted earlier seed germination. 6-BA treatment had no significant effect on the initial germination time of daylily seeds, but delayed it at KT concentrations of 50 mg / L and 100 mg / L. All GA3 concentrations prolonged the seed germination time by half, while the 6-BA treatments at concentrations of 100 mg / L, 200 mg / L, and 300 mg / L resulted in germination times of 8.67 ± 1.15 days, 9.00 ± 2.00 days, and 8.00 ± 1.73 days, respectively, shorter than the CK group, indicating that 6-BA could accelerate the germination process to 50%. High concentrations of KT delayed 50% seed germination. Treatment of daylily seeds with different plant growth regulators significantly delayed the final germination time compared to the control group.
[0086] Table 4. Effects of different plant growth regulators on the germination time of daylily seeds. As shown in Table 5, to study the effect of combined treatment with two chemical reagents on seed germination, a total of nine treatments were set up. All different combinations significantly improved the germination rate of daylily seeds. The highest germination rate of 92.67% was achieved when the GA3 concentration was 25 mg / L and the H2O2 concentration was 1%. Overall, the combination of GA3 and H2O2 at low concentrations can improve seed germination rate. Similarly, when the GA3 concentration was 25 mg / L and the H2O2 concentration was 1%, the highest germination potential of 79.33% and the highest germination index of 6.22 were achieved.
[0087] Table 5. Effects of combined GA3 and H2O2 treatment on daylily seed germination. As shown in Table 6, compared to the control group, when the GA3 concentration was 25 mg / L and the H2O2 concentration was 1%, the initial germination time was shortened from 6.33 days to 4.33 days. The time required for 50% germination and the final germination time were similar between the experimental and control groups. Considering the germination rate, germination potential, germination index, and initial germination time, the combined use of GA3 and H2O2 was more effective in improving seed germination rate than using them alone, and the optimal concentration combination was GA3 25 mg / L + H2O2 1%.
[0088] Table 6. Effects of combined GA3 and H2O2 treatment on germination time of daylily seeds. It should be noted that when numerical ranges are involved in this invention, it should be understood that both endpoints of each numerical range and any value between the two endpoints can be selected. Since the steps and methods used are the same as in the embodiments, preferred embodiments are described here to avoid redundancy. Although preferred embodiments of the invention have been described, those skilled in the art, once they understand the basic inventive concept, can make other changes and modifications to these embodiments. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments as well as all changes and modifications falling within the scope of this invention.
[0089] 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 improving the germination rate of daylily seeds, characterized in that, Includes the following steps: Daylily seeds were soaked, then treated with a combination of hydrogen peroxide and gibberellin, and cultured until germination to obtain daylily germination seeds.
2. The method for improving the germination rate of daylily seeds according to claim 1, characterized in that, Combined treatment refers to treatment with hydrogen peroxide first, followed by treatment with gibberellin.
3. The method for improving the germination rate of daylily seeds according to claim 1, characterized in that, In the combined treatment with hydrogen peroxide and hormones, the volume concentration of hydrogen peroxide was 1%-3%, the mass concentration of gibberellin was 25mg / L-75mg / L, and the total treatment time was 24h-36h.
4. The method for improving the germination rate of daylily seeds according to claim 1, characterized in that, In the combined treatment with hydrogen peroxide and hormones, the concentration of hydrogen peroxide was 1% and the concentration of gibberellin was 25 mg / L.
5. The method for improving the germination rate of daylily seeds according to claim 1, characterized in that, The seed soaking treatment method involves soaking daylily seeds in sterile water at 25℃-80℃ for 24h-36h.
6. The method for improving the germination rate of daylily seeds according to claim 1, characterized in that, The cultivation conditions are: temperature 22℃-25℃, relative humidity 55%-60%, and no light.