Method for rapidly improving seed germination rate of floating leaf sagittaria sagittifolia
By combining low-temperature preservation and HCl solution treatment with alternating dark and light cultivation, the problem of low seed germination rate of *Sagittaria floribunda* was solved, achieving efficient seed germination and rapid propagation, and providing technical support for the research and production of *Sagittaria floribunda*.
Patent Information
- Application Number
- CN202310773380.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-06-28
- Publication Date
- 2025-12-30
- Estimated Expiration
- 2043-06-28
AI Technical Summary
The seeds of *Sagittaria floribunda* have a low germination rate and a long germination time, making it difficult to meet the needs of scientific research and production. Moreover, the germination rate is extremely low under natural conditions.
After preserving the seeds at low temperatures, they were sterilized with NaClO solution, then treated with HCl solution for a specific time, and then cultured under alternating dark and light conditions, with 12 hours of darkness and 12 hours of light, at temperatures of 20℃ and 25℃, while keeping the seeds moist.
It significantly improved the germination rate of Sagittaria floribunda seeds, shortened the germination time, and achieved a germination rate of over 90%, reducing the seedling cycle, improving seed utilization efficiency, and providing technical support for scientific research and production.
Smart Images
Figure GDA0004463434230000041 
Figure GDA0004463434230000051 
Figure GDA0004463434230000061
Abstract
Description
Technical Field
[0001] This invention belongs to the field of aquatic plant cultivation technology, specifically relating to a method for rapidly increasing the germination rate of floating-leaved arrowhead seeds. Background Technology
[0002] Sagittaria natans Pall., a perennial aquatic floating-leaved herbaceous plant belonging to the genus Sagittaria in the family Alismataceae, is particularly susceptible to ecological and environmental impacts. Its distribution and population size in my country are declining, and it is on the verge of extinction (Dai Can, Tang Luying. RAPD analysis of genetic diversity of the endangered plant Sagittaria natans [J]. Amino Acids and Biological Resources, 2005(01):6-9.). It has been listed in the "National Second-Class Protected Plant List" and can be queried through the China Rare and Endangered Plants Information System (http: / / www.iplant.cn / rep / protlist).
[0003] Sagittaria floribunda primarily reproduces asexually, but can also be propagated by seeds. Seed germination is a fundamental life stage in which plants begin to grow and is a prerequisite for plant growth and development, playing a crucial role in the plant life cycle (Mihalte L, Sestras RE, Feszt G. Methods to improve the germination rate of cactus seeds [J]. Bulgarian Journal of Agricultural Sciences, 2011, 17(3): 288-295.). At the same time, seed germination is affected by many external factors, such as storage conditions and environmental factors, as well as by the characteristics of the seeds themselves, such as seed dormancy characteristics, maturity, water content, plumpness, and vigor (Koornneef M, Leónie Bentsink, Hilhorst H. Seed dormancy and germination [J]. Latest Views in Plant Biology, 2002, 5(1): 33-36. DOI: 10.1016 / S1369-5266(01)00219-9.). More importantly, different species exhibit varying seed dormancy and germination characteristics. Therefore, different seeds should be given appropriate methods to break dormancy and improve germination rates. Currently, there are no reported studies on the germination of *Sagittaria floribunda* seeds. Observations under natural conditions have revealed an extremely low germination rate, while seed viability testing shows high seed activity, indicating that dormancy cannot be broken under natural conditions. To address this issue, the applicant has conducted long-term research on *Sagittaria floribunda* seed germination, providing a method to significantly improve the germination rate and solve the problem of low germination rates, and even germination occurring the following year, in research and production. Summary of the Invention
[0004] The purpose of this invention is to provide a method for rapidly increasing the germination rate of *Sagittaria buergeriana* seeds. This method effectively improves the germination rate of *Sagittaria buergeriana* seeds, significantly shortens the germination time, and ensures consistent germination times. It meets the requirements of scientific research and production, and solves the problem of low germination rates or even germination every other year in scientific research and production. This lays a technical foundation for subsequent research on *Sagittaria buergeriana*. At the same time, it can also provide a reference for research on rapid in vitro propagation of other aquatic plants.
[0005] To solve the above problems, the technical solution adopted by the present invention is as follows:
[0006] A method for rapidly increasing the germination rate of arrowhead seeds includes the following steps:
[0007] (1) Seed collection and storage: During the seed maturity period, the seeds are bagged with gauze bags. After the seeds mature, the gauze bags are collected and taken back to the laboratory to be cleaned. The cleaned seeds are placed in a seed storage bottle filled with tap water and stored in a refrigerator at low temperature.
[0008] (2) Seed treatment: The seeds that have been treated at low temperature for a certain period of time are taken out, disinfected with 2% NaClO solution for 10 min, and then washed with distilled water several times. A portion of the seeds are randomly selected and seed activity is measured with 1% TTC solution. The seed activity rate is between 78% and 80%. The germination rate will be calculated based on the detected activity rate. The remaining seeds are treated with HCl solution for 0.5 h to 8 h, and then washed repeatedly with distilled water for later use.
[0009] (3) Germination culture: Place the seeds treated in step (2) evenly in a petri dish lined with a layer of filter paper, add a certain amount of distilled water to completely moisten the seed surface, cover the petri dish, and place it in an incubator for culture. Set the germination conditions as 12h darkness at 20℃ and 12h light at 25℃, and repeat the alternation. Add distilled water regularly to keep the seeds moist.
[0010] Furthermore, the seeds are subjected to low-temperature treatment for more than 6 months in step (2).
[0011] Furthermore, in step (2), the effective concentration of the HCl solution is 1%-30%, with an optimal concentration of 10%; the optimal treatment time is 5.5-6.5h.
[0012] Compared with the prior art, the technical effects of the present invention are as follows:
[0013] 1. The dormancy of floating-leaf arrowhead seeds is a major obstacle in breeding and seedling cultivation. This invention effectively solves the technical difficulty of breaking the dormancy of floating-leaf arrowhead seeds and solves the technical problems of low germination rate and slow germination of floating-leaf arrowhead seeds. Through this technical measure, the germination rate of floating-leaf arrowhead seeds is high (over 90%).
[0014] 2. The method of this invention is simple and feasible, low in cost, controllable in process, and free from secondary pollution. It overcomes the technical bottleneck of extremely low germination rate and long germination cycle of floating arrowhead seeds under natural conditions. The germination rate can reach more than 90% within one month, which greatly improves the germination rate and reduces the seedling cycle.
[0015] 3. It effectively improved the seed utilization efficiency of *Sagittaria floribunda* and enabled its rapid propagation. For scientific breeding work aimed at obtaining as many offspring plants as possible, the germination rate is undoubtedly very important, laying a technical foundation for subsequent research on *Sagittaria floribunda*. At the same time, it can also provide a reference for the research on rapid in vitro propagation of other aquatic plants. Attached Figure Description
[0016] Figure 1 This is a schematic diagram of seed germination in experimental group 1 of Example 1 of the present invention (38 days);
[0017] Figure 2 This is a schematic diagram of seed germination in experimental group 2 of Example 2 of the present invention (38 days);
[0018] Figure 3 This is a schematic diagram of the seed germination status (38 days) in experimental group 3 of Example 3 of the present invention;
[0019] Figure 4 This is a schematic diagram of seed germination in Comparative Example 1 of Embodiment 4 of the present invention (38 days);
[0020] Figure 5 This is a schematic diagram of seed germination in Comparative Example 2 of Embodiment 5 of the present invention (38 days);
[0021] Figure 6 This is a schematic diagram of the seed germination of Comparative Example 3 in Example 6 of the present invention (38 days);
[0022] Figure 7 This is a schematic diagram of seeds stained with 1% TTC (activity assay: viable seeds are stained red, otherwise they are inactive). Detailed Implementation
[0023] The technical solution of the present invention will be further described below with reference to the embodiments: Among them, embodiments 1-6 are comparisons of methods using HCl solution of the same concentration for different times, KOH solution treatment, KNO3 solution treatment and no treatment respectively; embodiments 7 and 8 are comparisons of methods using HCl solution of different concentrations and HCl solution of the same concentration for different treatment times.
[0024] Example 1: Experimental group 1 adopted a method to improve the germination rate of Sagittaria buergeriana seeds, the steps of which are as follows:
[0025] (1) Seed collection and storage: During the seed maturity period, the seeds are bagged with gauze bags. After the seeds mature, the gauze bags are collected and brought back to the laboratory to be cleaned. The cleaned seeds are placed in a seed storage bottle filled with tap water and stored in a refrigerator at 4°C.
[0026] (2) Seed treatment: Seeds stored for more than 6 months were taken out, disinfected with 2% NaClO solution for 10 min, and then washed with distilled water 3 times. A portion was randomly selected for seed activity determination. The seed activity rate was found to be about 78.5%, which is high, but some seeds were not viable. The germination rate was calculated based on the detected activity rate. The remaining seeds were treated with 10% HCl solution for 6 h, and then washed repeatedly with distilled water for later use.
[0027] (3) Germination culture: Place the seeds treated in step (2) evenly in a petri dish lined with a layer of filter paper, add a certain amount of distilled water to completely moisten the seed surface, cover the petri dish, and place it in an incubator for culture. Set the germination conditions as 12h darkness at 20℃ and 12h light at 25℃, and repeat the alternation. Replenish distilled water in time to keep the seeds moist.
[0028] Example 2: Experimental group 2 adopted a method to improve the germination rate of Sagittaria buergeriana seeds, the steps of which are as follows:
[0029] (1) Seed collection and preservation: Same as in Example 1;
[0030] (2) Seed treatment: Seeds stored for more than 6 months were taken out, disinfected with 2% NaClO solution for 10 min, and then washed with distilled water 3 times. A portion was randomly selected for seed activity determination. The seed activity rate was found to be about 79.5%, which is relatively high, but some seeds were not viable. The germination rate will be calculated based on the detected activity rate. The remaining seeds were treated with 10% HCl solution for 5.5 h, and then washed repeatedly with distilled water for later use.
[0031] (3) Germination culture: Same as in Example 1.
[0032] Example 3: Experimental group 3 adopted a method to improve the germination rate of Sagittaria buergeriana seeds, the steps of which are as follows:
[0033] (1) Seed collection and preservation: Same as in Example 1;
[0034] (2) Seed treatment: Seeds stored for more than 6 months were taken out, disinfected with 2% NaClO solution for 10 min, and then washed with distilled water 3 times. A portion was randomly selected for seed activity determination. The seed activity rate was found to be about 78%, which is high, but some seeds were not viable. The germination rate will be calculated based on the detected activity rate. The remaining seeds were treated with 10% HCl solution for 6.5 h, and then washed repeatedly with distilled water for later use.
[0035] (3) Germination culture: Same as in Example 1.
[0036] Example 4: Comparative Example 1 uses a method to improve the germination rate of Sagittaria buergeriana seeds, the steps of which are as follows:
[0037] (1) Seed collection and preservation: Same as in Example 1;
[0038] (2) Seed treatment: Take out the seeds that have been stored for more than 6 months, disinfect them with 2% NaClO solution for 10 minutes, and then wash them repeatedly with distilled water. Randomly take a portion for seed activity test. The seed activity rate was found to be about 79%, which is high, but some seeds have no activity. The germination rate will be converted with the detected activity rate. Soak the remaining seeds in distilled water for later use.
[0039] (3) Germination culture: Same as in Example 1.
[0040] Example 5: Comparative Example 2 uses a method to improve the germination rate of Sagittaria buergeriana seeds, the steps of which are as follows:
[0041] (1) Seed collection and preservation: Same as in Example 1;
[0042] (2) Seed treatment: Seeds stored for more than 6 months were taken out, disinfected with 2% NaClO solution for 10 minutes, and then repeatedly washed with distilled water. A portion was randomly selected for seed activity testing. The seed activity rate was found to be about 78.5%, which is relatively high, but some seeds were not viable. The germination rate will be calculated based on the detected activity rate. The seeds were soaked in 5%-30% KOH solution for 1-8 hours, and then repeatedly washed with distilled water before use.
[0043] (3) Germination culture: Same as in Example 1.
[0044] Example 6: Comparative Example 3 uses a method to improve the germination rate of Sagittaria buergeriana seeds, the steps of which are as follows:
[0045] (1) Seed collection and preservation: Same as in Example 1;
[0046] (2) Seed treatment: Take out the seeds that have been stored for more than 6 months, disinfect them with 2% NaClO solution for 10 minutes, and then wash them repeatedly with distilled water. Randomly take a portion for seed activity determination. The seed activity rate was found to be about 79%, which is high, but some seeds have no activity. The germination rate will be calculated based on the detected activity rate. The remaining seeds are soaked in 0.1-10.0 g / L KNO3 solution for 1-8 hours, and then washed repeatedly with distilled water for later use; (3) Germination culture: Same as in Example 1.
[0047] Experimental conclusions of Examples 1-6: Effects of the above methods on the germination of Sagittaria buergeriana seeds.
[0048] The seed germination rates of experimental groups 1-3 and control groups 1-3 in Examples 1-6 above were statistically analyzed, and the results are shown in Table 1 below:
[0049] Table 1. Seed germination status after treatment using the methods in Examples 1-6
[0050]
[0051] As can be seen from Table 1, under the premise that other experimental conditions remain unchanged, the HCl solution treatment has the best effect, with a high germination rate of Sagittaria buergeriana seeds (over 90%) and a short germination time, with a large number of seeds germinating on the first day, which is significantly superior to other methods.
[0052] Example 7: A method for improving the germination rate of Sagittaria buergeriana seeds, the steps of which are as follows:
[0053] (1) Seed collection and preservation: Same as in Example 1;
[0054] (2) Seed treatment: Seeds stored for more than 6 months were taken out, disinfected with 2% NaClO solution for 10 min, and then washed with distilled water 3 times. A portion of the seeds were randomly selected for seed activity determination. The seed activity was found to be about 80%, which is high, but some seeds were not viable. The germination rate will be calculated based on the detected activity rate. The remaining disinfected seeds were randomly divided into groups. Each group of seeds was treated with distilled water (CK) and HCl solutions of different concentrations (1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%) for 6 h, and then washed with distilled water multiple times before use.
[0055] (3) Germination culture: The seeds treated in step (2) are evenly placed in each group in a petri dish lined with a layer of filter paper, a certain amount of distilled water is added, the petri dish is covered and placed in an incubator for culture. The germination conditions are set as 12 hours in darkness at 20°C and 12 hours in light at 25°C, and the conditions are repeated. Distilled water is added in time to keep the seeds moist.
[0056] The results are as follows (Table 2):
[0057] Table 2 Seed germination after treatment with HCl solutions of different concentrations.
[0058]
[0059] As shown in Table 2, 1%-10% HCl solutions were effective after 6 hours of treatment, with 10% HCl solution showing the best results. The germination rate of Sagittaria floribunda seeds reached as high as 98%, and the germination time was short, with a large number of seeds germinating on the first day. In contrast, the 15%-30% HCl solutions were not effective, indicating that higher concentrations are not necessarily better.
[0060] Example 8: A method for improving the germination rate of Sagittaria buergeriana seeds, the steps of which are as follows:
[0061] (1) Seed collection and preservation: Same as in Example 1;
[0062] (2) Seed treatment: Seeds stored for more than 6 months were taken out, disinfected with 2% NaClO solution for 10 min, and then washed with distilled water 3 times. A portion of the seeds were randomly selected for seed activity determination. The seed activity was found to be about 79%, which is relatively high, but some seeds were not viable. The germination rate will be calculated based on the detected activity rate. The remaining disinfected seeds were randomly divided into groups. Each group of seeds was treated with distilled water (CK) and 10% HCl solution for different durations (0.5 h, 1 h, 2 h, 4 h, 5.5 h, 6 h, 6.5 h, 8 h), and then washed repeatedly with distilled water for later use.
[0063] (3) Germination culture: The seeds treated in step (2) are evenly placed in each group in a petri dish lined with a layer of filter paper, a certain amount of distilled water is added, the petri dish is covered and placed in an incubator for culture. The germination conditions are set as 12 hours in darkness at 20°C and 12 hours in light at 25°C, and the conditions are repeated. Distilled water is added in time to keep the seeds moist.
[0064] The results are as follows (Table 3):
[0065] Table 3. Effects of different treatment durations with HCl solution on seed germination of *Sagittaria floribunda*.
[0066]
[0067] As can be seen from Table 3, treatment with 10% HCl solution for 6 hours yielded the best results, with a germination rate of up to 98% for Sagittaria buergeriana seeds. Moreover, the germination time was short, with a large number of seeds germinating on the first day.
Claims
1. A method of increasing the seed germination rate of Sagittaria cuneata, characterized by, Comprising the following steps: (1) Seed collection and preservation: before the seed maturation period, the seeds are bagged with a cotton bag, and after the seeds mature, the cotton bag is collected, cleaned and placed in a seed storage bottle with tap water, and stored at 4°C in low temperature; (2) Seed treatment: the seeds treated for more than 6 months are taken out, disinfected with 2% NaClO solution for 10 minutes, then washed with distilled water several times, treated with 1%-10% HCl solution for 0.5-6h, and then washed repeatedly with distilled water; 15%-30% treatment effect is not good, and the higher the concentration, the better the treatment effect; (3) Germination culture: the seeds treated in step (2) are evenly placed in a culture dish with a layer of filter paper, a certain amount of distilled water is added, the culture dish cover is covered, and the culture dish is placed in a culture box, and the germination conditions are set as 12h darkness, 20°C, 12h light, 25°C, alternating repeatedly, and distilled water is added regularly to keep the seeds moist.
2. The method for improving seed germination rate of floating-heart according to claim 1, wherein, The effective concentration of HCl solution in step (2) is 10%, and the treatment time is 5.5-6.5h.