Method for treating polygonum cuspidatum seeds

Abstract

The invention relates to the technical field of biological seedling raising, and discloses a method for treating polygonum cuspidatum seeds and a method for breaking dormancy and promoting germination of the polygonum cuspidatum seeds. The method for treating the polygonum cuspidatum seeds comprises the following steps: (1) performing ultraviolet radiation on the polygonum cuspidatum seeds; (2) performing circulating freezing treatment on the polygonum cuspidatum seeds subjected to ultraviolet radiation; and (3) mixing the polygonum cuspidatum seeds subjected to circulating freezing treatment with a carbendazim powder solution, and then taking out and airing. The polygonum cuspidatum seeds treated by the method are subjected to germination culture, the germination rate and germination potential of the polygonum cuspidatum seeds can be greatly improved, the seedling culture success rate is further improved, and meanwhile, the treatment method is simple and easy to implement.

Description

Technical Field

[0001] This invention relates to the field of biological seedling technology, specifically to a method for treating Polygonum cuspidatum seeds and a method for germinating and cultivating Polygonum cuspidatum seeds. Background Technology

[0002] Polygonum cuspidatum, as a traditional Chinese medicine, has a long history of medicinal use. It was first recorded in the "Mingyi Bielu" and listed as a medium-grade medicine. Its dried rhizome and root have the effects of promoting diuresis and relieving jaundice, clearing heat and detoxifying, dispersing blood stasis and relieving pain, and relieving cough and resolving phlegm. It is included in the "Pharmacopoeia of the People's Republic of China" and is a commonly used medicinal material in traditional Chinese medicine clinical treatment of damp-heat jaundice, rheumatic arthralgia and other diseases. It is also the core raw material of many Chinese patent medicines. In recent years, with the in-depth development of Polygonum cuspidatum extract in the fields of medicine and health care products, its market demand has continued to surge. However, Polygonum cuspidatum seeds have a very low germination rate due to seed dormancy (physical dormancy and physiological dormancy). Physical dormancy is mainly manifested in the following three aspects: (1) Hard and dense seed coat: Polygonum cuspidatum achenes have a 3-angled hard shell. The seed coat is keratinized, impermeable to water and air, which hinders the entry of water and oxygen, and the embryo cannot absorb water and swell. (2) Influence of persistent perianth: Seeds are stored with persistent perianth, which further reduces permeability. The perianth needs to be removed before sowing. (3) Mechanical resistance: The seed coat has high mechanical strength, and the radicle is difficult to break through. Physiological dormancy (endogenous inhibition of the embryo) is mainly manifested in the following two aspects: (1) Embryo is not fully matured: Mature seed embryos are morphologically mature, but physiologically immature. The endogenous abscisic acid (ABA) content is high and the gibberellin (GA) is insufficient, which inhibits germination. (2) Phototropism: Polygonum cuspidatum seeds are light-loving seeds. The germination rate is significantly reduced in the dark. Light is an important condition for breaking physiological dormancy.

[0003] In existing technologies, methods for breaking dormancy in Polygonum cuspidatum seeds mainly involve soaking in warm water or treating with plant hormones, which have limitations in improving the germination rate. Therefore, this invention provides a new method for treating Polygonum cuspidatum seeds, which can further improve the germination rate and germination potential, increase the seedling success rate, and is simple to implement. Summary of the Invention

[0004] The purpose of this invention is to overcome the problems of low germination rate and germination potential of Polygonum cuspidatum seeds and low seedling success rate in the prior art, and to provide a method for treating Polygonum cuspidatum seeds. This method is simple and easy to implement. The germination rate and germination potential of Polygonum cuspidatum seeds treated by this method can be greatly improved, thereby improving the seedling success rate.

[0005] To achieve the above objectives, the present invention provides a method for processing Polygonum cuspidatum seeds, the method comprising the following steps: (1) Expose the seeds of Polygonum cuspidatum to ultraviolet light; (2) Cyclic freezing treatment was performed on the seeds of Polygonum cuspidatum after ultraviolet irradiation; (3) Mix the frozen Polygonum cuspidatum seeds with a solution of polyoxin powder, and then take them out and air dry.

[0006] Preferably, in step (1), the irradiation conditions include: a wavelength of 100-350 nm and an intensity of 50-150 μW / cm. 2 The time is 2-8 hours.

[0007] Preferably, in step (1), the irradiation conditions include: a wavelength of 280-320 nm and an intensity of 75-120 μW / cm². 2 The time is 3-6 hours.

[0008] Preferably, in step (2), the number of cyclic freezing treatments is 2-5; more preferably 2-4.

[0009] Preferably, in step (2), the freezing conditions in the cyclic freezing process include: a temperature of -60 to -20°C and a time of 10-30 hours.

[0010] Preferably, in step (2), the thawing conditions in the cyclic freezing process include: a temperature of 20-40°C and a time of 2-6 hours.

[0011] Preferably, in step (2), the thawing method in the cyclic freezing process is water bath thawing.

[0012] Preferably, in step (3), the weight ratio of carbendazim powder to water in the carbendazim powder solution is 1g:500-800mL.

[0013] Preferably, in step (3), the weight ratio of the Polygonum cuspidatum seeds to the volume of the carbendazim powder solution is 1 kg: 3-5 L.

[0014] Preferably, in step (3), the mixing conditions include: a temperature of 15-40°C and a time of 1-30 min.

[0015] The second aspect of the present invention provides a method for germinating and culturing Polygonum cuspidatum seeds, the method comprising: treating Polygonum cuspidatum seeds according to the above method, and then germinating and culturing the treated Polygonum cuspidatum seeds.

[0016] Compared with the prior art, the technical solution of the present invention has the following advantages: The method for treating Polygonum cuspidatum seeds described in this invention is simple and easy to implement. Furthermore, the method described in this invention can effectively break the dormancy state of Polygonum cuspidatum seeds, thereby greatly improving the germination rate and germination potential, and thus increasing the seedling success rate. Detailed Implementation

[0017] The following provides a detailed description of specific embodiments of the present invention. It should be understood that the specific embodiments described herein are for illustrative and explanatory purposes only and are not intended to limit the scope of the invention.

[0018] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0019] This invention provides a method for processing Polygonum cuspidatum seeds, the method comprising the following steps: (1) Expose the seeds of Polygonum cuspidatum to ultraviolet light; (2) Cyclic freezing treatment was performed on the seeds of Polygonum cuspidatum after ultraviolet irradiation; (3) Mix the frozen Polygonum cuspidatum seeds with a solution of polyoxin powder, and then take them out and air dry.

[0020] In the method for processing Polygonum cuspidatum seeds described in this invention, the Polygonum cuspidatum seeds are mature seeds that are 2-3 years old, healthy, plump, undamaged, free from mold, and free from pests and diseases, and the purity of the Polygonum cuspidatum seeds is ≥95% and the cleanliness is ≥98%.

[0021] In the method for treating Polygonum cuspidatum seeds described in this invention, in step (1), the Polygonum cuspidatum seeds are subjected to ultraviolet irradiation, which significantly enhances the activity of key hydrolytic enzymes such as α-amylase in the seeds, thereby increasing the energy supply rate required for seed germination; in step (2), the Polygonum cuspidatum seeds after ultraviolet irradiation are subjected to cyclic freezing treatment, which causes microcracks in the seed coat of Polygonum cuspidatum due to physical stress caused by drastic temperature changes, thereby improving water permeability and air permeability and breaking dormancy; in step (3), the Polygonum cuspidatum seeds after cyclic freezing treatment are mixed with a fungicide powder solution, and then taken out and dried, which disinfects and sterilizes the seeds, further reducing the interference of pests and pathogens, and then taken out and dried; based on the above steps, the dormancy state of Polygonum cuspidatum seeds can be broken, the germination rate and germination potential of Polygonum cuspidatum seeds can be improved, thereby improving the seedling success rate of Polygonum cuspidatum.

[0022] In a preferred embodiment, in order to improve the germination rate and germination potential of Polygonum cuspidatum seeds while reducing damage from ultraviolet light irradiation, the irradiation conditions in step (1) include: a wavelength of 100-350 nm and an intensity of 50-150 μW / cm². 2 The time is 2-8 hours; more preferably, the wavelength is 280-320 nm and the intensity is 75-120 μW / cm. 2The treatment time is 3-6 hours; based on ultraviolet irradiation, the activity of key hydrolytic enzymes such as α-amylase is significantly enhanced, thereby increasing the energy supply rate required for the germination of Polygonum cuspidatum seeds.

[0023] In a preferred embodiment, in order to further improve the germination rate and germination potential of Polygonum cuspidatum seeds, the number of cycles of freezing treatment in step (2) is 2-5; preferably 2-4. Based on this cycle of freezing treatment, due to the drastic temperature change, the seed coat of Polygonum cuspidatum is made to produce microcracks through physical stress, thereby improving water permeability and air permeability, thus breaking dormancy and improving the germination rate and germination potential of Polygonum cuspidatum seeds, thereby improving the seedling success rate of Polygonum cuspidatum.

[0024] In a preferred embodiment, in step (2), the freezing conditions in the cyclic freezing process include: a temperature of -60 to -20°C and a time of 10 to 30 hours; preferably, the temperature is -50 to -30°C and the time is 12 to 24 hours.

[0025] In a preferred embodiment, in step (2), the thawing conditions in the cyclic freezing process include: a temperature of 20-40°C and a time of 2-6 hours.

[0026] In one specific implementation, in order to further improve the germination rate and germination potential of Polygonum cuspidatum seeds, in step (2), the thawing method in the cyclic freezing treatment is water bath thawing.

[0027] In a further preferred embodiment, step (2) further includes: taking out the Polygonum cuspidatum seeds after the last cycle of thawing and drying them; preferably, air-drying them.

[0028] In the method for treating Polygonum cuspidatum seeds described in this invention, in step (3), the carbendazim powder is a fungicide commonly used in the field, and there are no special requirements for its specific brand. In a specific embodiment, the brand of the carbendazim powder can be one or more of Dafusheng 50% carbendazim wettable powder and Guoguang 50% carbendazim wettable powder.

[0029] In a preferred embodiment, in order to improve the sterilization effect while ensuring a high germination rate and germination potential, in step (3), the weight ratio of carbendazim powder to water in the carbendazim powder solution is 1g:500-800mL; specifically, it can be 1g:500mL, 1g:550mL, 1g:600mL, 1g:650mL, 1g:700mL, 1g:750mL or 1g:800mL.

[0030] In a preferred embodiment, in order to further improve the sterilization effect while ensuring a high germination rate and germination potential, in step (3), the weight ratio of the amount of Polygonum cuspidatum seeds to the volume of the carbendazim powder solution is 1 kg: 3-5 L.

[0031] In a preferred embodiment, the mixing conditions in step (3) include a temperature of 15-40°C and a time of 1-30 min.

[0032] The second aspect of the present invention provides a method for germinating and culturing Polygonum cuspidatum seeds, the method comprising: treating Polygonum cuspidatum seeds according to the above method, and then germinating and culturing the treated Polygonum cuspidatum seeds.

[0033] In one specific embodiment, the germination culture method is as follows: a moist filter paper is placed in a petri dish, the treated Polygonum cuspidatum seeds are placed in the petri dish, and then the petri dish is placed in an incubator with constant temperature, humidity and light for culture. Sterile water is added daily to keep the petri dish moist but not waterlogged.

[0034] The following examples further illustrate the method for processing Polygonum cuspidatum seeds and the method for germinating and cultivating Polygonum cuspidatum seeds according to the present invention. These examples are implemented based on the technical solution of the present invention, providing detailed implementation methods and specific operating procedures; however, the scope of protection of the present invention is not limited to the following examples.

[0035] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods in the art. Unless otherwise specified, the experimental materials used in the following embodiments are commercially available.

[0036] Example 1 (1) A wavelength of 305 nm and an intensity of 100 μW / cm were used. 2 One kilogram of Japanese knotweed seeds were irradiated with ultraviolet light for 3 hours. (2) The seeds of Polygonum cuspidatum after ultraviolet irradiation were subjected to cyclic freezing treatment for 3 cycles. In each cycle of freezing treatment, the freezing temperature was -40℃ and the time was 18h. The water bath thawing temperature was 35℃ and the time was 3h. After the cycle of freezing treatment was completed, the seeds of Polygonum cuspidatum were air-dried. (3) Soak 1 kg of Polygonum cuspidatum seeds after cyclic freezing treatment in 4 L of carbendazim powder solution (the weight ratio of carbendazim powder to water volume is 1 g: 500 mL solution) at 30 °C for 5 min, and then take them out and air dry.

[0037] Example 2 (1) A wavelength of 296 nm and an intensity of 90 μW / cm were used. 2 One kilogram of Japanese knotweed seeds were irradiated with ultraviolet light for 5 hours. (2) The seeds of Polygonum cuspidatum after ultraviolet irradiation were subjected to cyclic freezing treatment for 4 cycles. In each cycle of freezing treatment, the freezing temperature was -50℃ and the time was 15h. The water bath thawing temperature was 30℃ and the time was 3h. After the cycle of freezing treatment was completed, the seeds of Polygonum cuspidatum were air-dried. (3) Soak 1 kg of Polygonum cuspidatum seeds after cyclic freezing treatment in 5 L of carbendazim powder solution (the weight of carbendazim powder to the volume of water is 1 g: 500 mL solution) at a temperature of 35 °C for 5 min, and then take them out and air dry.

[0038] Example 3 The procedure was carried out in accordance with Example 1, except that in step (1), the wavelength of the ultraviolet irradiation was 350 nm, i.e.: (1) A wavelength of 350 nm and an intensity of 100 μW / cm were used. 2 One kilogram of Japanese knotweed seeds were irradiated with ultraviolet light for 3 hours. (2) The seeds of Polygonum cuspidatum after ultraviolet irradiation were subjected to cyclic freezing treatment for 3 cycles. In each cycle of freezing treatment, the freezing temperature was -40℃ and the time was 18h. The water bath thawing temperature was 35℃ and the time was 3h. After the cycle of freezing treatment was completed, the seeds of Polygonum cuspidatum were air-dried. (3) Soak 1 kg of Polygonum cuspidatum seeds after cyclic freezing treatment in 4 L of carbendazim powder solution (the weight ratio of carbendazim powder to water volume is 1 g: 500 mL solution) at 30 °C for 5 min, and then take them out and air dry.

[0039] Example 4 The procedure was carried out in accordance with Example 1, except that in step (1), the intensity of the ultraviolet irradiation was 50 μW / cm. 2 ,Right now: (1) A wavelength of 305 nm and an intensity of 50 μW / cm were used. 2 One kilogram of Japanese knotweed seeds were irradiated with ultraviolet light for 3 hours. (2) The seeds of Polygonum cuspidatum after ultraviolet irradiation were subjected to cyclic freezing treatment for 3 cycles. In each cycle of freezing treatment, the freezing temperature was -40℃ and the time was 18h. The water bath thawing temperature was 35℃ and the time was 3h. After the cycle of freezing treatment was completed, the seeds of Polygonum cuspidatum were air-dried. (3) Soak 1 kg of Polygonum cuspidatum seeds after cyclic freezing treatment in 4 L of carbendazim powder solution (the weight ratio of carbendazim powder to water volume is 1 g: 500 mL solution) at 30 °C for 5 min, and then take them out and air dry.

[0040] Example 5 The implementation is carried out in accordance with Example 1, except that in step (2), the number of cyclic freezing cycles is 5, that is: (1) A wavelength of 305 nm and an intensity of 100 μW / cm were used. 2 One kilogram of Japanese knotweed seeds were irradiated with ultraviolet light for 3 hours. (2) The seeds of Polygonum cuspidatum after ultraviolet irradiation were subjected to cyclic freezing treatment for 5 cycles. In each cycle of freezing treatment, the freezing temperature was -40℃ and the time was 18h. The water bath thawing temperature was 35℃ and the time was 3h. After the cycle of freezing treatment was completed, the seeds of Polygonum cuspidatum were air-dried. (3) Soak 1 kg of Polygonum cuspidatum seeds after cyclic freezing treatment in 4 L of carbendazim powder solution (the weight ratio of carbendazim powder to water volume is 1 g: 500 mL solution) at 30 °C for 5 min, and then take them out and air dry.

[0041] Example 6 The procedure was carried out in accordance with Example 1, except that in step (2), the freezing temperature during the cyclic freezing process was -60°C, i.e.: (1) A wavelength of 305 nm and an intensity of 100 μW / cm were used. 2 One kilogram of Japanese knotweed seeds were irradiated with ultraviolet light for 3 hours. (2) The seeds of Polygonum cuspidatum after ultraviolet irradiation were subjected to cyclic freezing treatment for 3 cycles. In each cycle of freezing treatment, the freezing temperature was -60℃ and the time was 18h. The water bath thawing temperature was 35℃ and the time was 3h. After the cycle of freezing treatment was completed, the seeds of Polygonum cuspidatum were air-dried. (3) Soak 1 kg of Polygonum cuspidatum seeds after cyclic freezing treatment in 4 L of carbendazim powder solution (the weight ratio of carbendazim powder to water volume is 1 g: 500 mL solution) at 30 °C for 5 min, and then take them out and air dry.

[0042] Example 7 The method described in Example 1 is followed, except that in step (2), the thawing method during cyclic freezing is room temperature thawing, without water bath thawing, i.e.: (1) A wavelength of 305 nm and an intensity of 100 μW / cm were used. 2 One kilogram of Japanese knotweed seeds were irradiated with ultraviolet light for 3 hours. (2) The seeds of Polygonum cuspidatum after ultraviolet irradiation were subjected to cyclic freezing treatment for 3 cycles. In each cycle of freezing treatment, the freezing temperature was -40℃ and the time was 18h. The thawing temperature was 35℃ and the time was 3h. After the cycle of freezing treatment was completed, the seeds of Polygonum cuspidatum were air-dried. (3) Soak 1 kg of Polygonum cuspidatum seeds after cyclic freezing treatment in 4 L of carbendazim powder solution (the weight ratio of carbendazim powder to water volume is 1 g: 500 mL solution) at 30 °C for 5 min, and then take them out and air dry.

[0043] Comparative Example 1 The implementation is carried out in accordance with Example 1, except that step (1) is omitted, i.e.: (1) 1 kg of Polygonum cuspidatum seeds were subjected to cyclic freezing treatment for 3 cycles. In each cycle of freezing treatment, the freezing temperature was -40℃ and the time was 18h. The water bath thawing temperature was 35℃ and the time was 3h. After the cyclic freezing treatment was completed, the Polygonum cuspidatum seeds were air-dried. (2) Soak 1 kg of Polygonum cuspidatum seeds after cyclic freezing treatment in 4 L of carbendazim powder solution (the weight of carbendazim powder to the volume of water is 1 g: 500 mL solution) at a temperature of 30 °C for 5 min, and then take them out and air dry.

[0044] Comparative Example 2 The implementation is carried out in accordance with Example 1, except that step (2) is omitted, i.e.: (1) A wavelength of 305 nm and an intensity of 100 μW / cm were used. 2 One kilogram of Japanese knotweed seeds were irradiated with ultraviolet light for 3 hours. (2) Soak 1 kg of Japanese knotweed seeds after UV irradiation in 4 L of carbendazim powder solution (the weight of carbendazim powder to the volume of water is 1 g: 500 mL solution). The soaking temperature is 30 °C and the soaking time is 5 min. Then take them out and air dry.

[0045] Test case The treated Polygonum cuspidatum seeds from the examples and comparative examples were subjected to germination culture. The treated Polygonum cuspidatum seeds were placed in petri dishes lined with moist filter paper, and then placed in an incubator at 25±1℃, light intensity 12h / d, and humidity 70% for culture. Sterile water was added daily to keep the petri dishes moist but not waterlogged. The germination rate and germination potential of the Polygonum cuspidatum seeds were statistically analyzed. The germination rate refers to the percentage of normally germinated seeds out of the total number of tested seeds within the specified total germination days (15 days). The germination potential refers to the percentage of normally germinated seeds out of the total number of tested seeds in the early stage of germination (7 days). The results are shown in Table 1. Table 1

[0046] As can be seen from the results in Table 1, treating Polygonum cuspidatum seeds using the method described in this invention and then germinating the treated seeds can greatly improve the germination rate and germination potential of Polygonum cuspidatum seeds, thereby increasing the success rate of Polygonum cuspidatum seedling cultivation. This is of great significance for the cultivation of Polygonum cuspidatum.

[0047] The preferred embodiments of the present invention have been described in detail above; however, the present invention is not limited thereto. Within the scope of the inventive concept, various simple modifications can be made to the technical solutions of the present invention, including combinations of various technical features in any other suitable manner. These simple modifications and combinations should also be considered as the content disclosed in the present invention and are all within the protection scope of the present invention.

Claims

1. A method for treating Polygonum cuspidatum seeds, characterized in that, The method includes the following steps: (1) Expose the seeds of Polygonum cuspidatum to ultraviolet light; (2) Cyclic freezing treatment was performed on the seeds of Polygonum cuspidatum after ultraviolet irradiation; (3) Mix the frozen Polygonum cuspidatum seeds with a solution of polyoxin powder, and then take them out and air dry.

2. The method according to claim 1, characterized in that, In step (1), the irradiation conditions include: wavelength of 100-350 nm and intensity of 50-150 μW / cm. 2 The time is 2-8 hours; Preferably, the irradiation conditions include: a wavelength of 280-320 nm and an intensity of 75-120 μW / cm². 2 The time is 3-6 hours.

3. The method according to claim 1 or 2, characterized in that, In step (2), the number of cyclic freezing processes is 2-5; preferably 2-4.

4. The method according to any one of claims 1-3, characterized in that, In step (2), the freezing conditions in the cyclic freezing process include: a temperature of -60 to -20°C and a time of 10-30 hours.

5. The method according to any one of claims 1-4, characterized in that, In step (2), the thawing conditions in the cyclic freezing process include: a temperature of 20-40℃ and a time of 2-6h.

6. The method according to any one of claims 1-5, characterized in that, In step (2), the thawing method in the cyclic freezing process is water bath thawing.

7. The method according to any one of claims 1-6, characterized in that, In step (3), the weight ratio of carbendazim powder to water in the carbendazim powder solution is 1g:500-800mL.

8. The method according to any one of claims 1-7, characterized in that, In step (3), the weight ratio of the Polygonum cuspidatum seeds to the volume ratio of the carbendazim powder solution is 1 kg: 3-5 L.

9. The method according to any one of claims 1-8, characterized in that, In step (3), the mixing conditions include: a temperature of 15-40℃ and a time of 1-30min.

10. A method for germinating and cultivating Polygonum cuspidatum seeds, characterized in that, The method includes: treating Polygonum cuspidatum seeds according to any one of claims 1-9, and then germinating the treated Polygonum cuspidatum seeds.

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