A method for rapidly collecting high-activity gleditsia sinensis pollen
By employing specific pressure filtration and water absorption drying processes, the problems of low pollen collection efficiency and loss of activity in soapberry have been solved, achieving efficient and rapid pollen collection, which is suitable for soapberry breeding and scientific research.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- GUIZHOU UNIV
- Filing Date
- 2024-07-17
- Publication Date
- 2026-05-12
AI Technical Summary
Existing technologies for collecting soapberry pollen are inefficient, time-consuming, and labor-intensive, and pollen activity is easily lost, making it difficult to meet the needs of modern planting and research.
By employing specific pressure filtration and water absorption drying processes, combined with the control of key parameters in each step, including pollen dissolution, preliminary sieving, pressure filtration, water absorption drying, and vibration crushing and pulverization, we ensure efficient collection and preservation of pollen activity.
It enables efficient and rapid collection of soapberry pollen, improves the quantity and quality of collection, reduces pollen loss, is suitable for large-scale application, and enhances its value for breeding and scientific research.
Smart Images
Figure CN118661636B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of pollen collection technology, and more particularly to a rapid method for collecting highly active soapberry pollen. Background Technology
[0002] The soapberry (Gleditsia sinensis) is a unique, multifunctional, and economically valuable tree species in my country, widely used in afforestation, landscaping, and traditional medicine. However, as a dioecious plant with male and female flowers, it primarily relies on cross-pollination, with the hermaphroditic flowers serving as functional females. This results in low natural pollination efficiency, leading to problems in actual production such as inconsistent varietal quality, poor pod quality and low yield, and declining seed uniformity. Collecting soapberry pollen for assisted pollination is an important way to improve pod yield and quality.
[0003] The pollen of *Gleditsia sinensis* is spherical or elliptical, and its surface may have textures or protrusions. When dispersing, it usually clumps together into small particles, adhering to the anther wall, petals, and flower stalks, making it difficult to collect. Currently, the conventional method for collecting *Gleditsia sinensis* pollen is to manually peel off the anthers, dry them, and collect and preserve them. This method is inefficient, time-consuming, labor-intensive, and yields very little pollen, failing to meet the needs of modern rapid cultivation and research. Furthermore, the pollen is prone to loss of viability during the collection process.
[0004] Therefore, developing a method that can quickly collect soapberry pollen while ensuring its activity is of great significance to the development of soapberry-related industries and scientific research. Summary of the Invention
[0005] To address the aforementioned problems, this invention provides a method for the rapid collection of highly active soapberry pollen. This method achieves rapid collection of highly active soapberry pollen through specific pressure filtration and water absorption drying processes, combined with the control of key parameters in each step.
[0006] The rapid collection method for highly active soapberry pollen of the present invention includes the following steps:
[0007] S1. Pollen dissolution: Collect male inflorescences of Gleditsia sinensis that are in full bloom and shedding pollen, put them into ultrapure water and stir slowly, repeat 3 times, combine the solutions to obtain an aqueous solution A containing pollen.
[0008] S2. Preliminary sieving: The pollen-containing aqueous solution is sieved to remove large pieces of inflorescence residue, petals, broken anthers and other impurities, to obtain pollen aqueous solution B;
[0009] S3. Pressure filtration: The pollen aqueous solution B is subjected to pressure filtration to obtain pollen;
[0010] S4. Water absorption and drying: Remove excess moisture from the pollen, and then dry it once at a temperature of 28°C and a humidity of 60% to 80% to obtain a solid pollen cake. Then, the solid pollen cake is appropriately dispersed, and the dispersed pollen is dried a second time at a temperature of 28°C and a humidity of 30%.
[0011] S5. Vibration and Powdering: After the dried pollen is vibrated and crushed until it reaches the required powder state, it is stored in a container and preserved under appropriate conditions to complete the rapid collection of soapberry pollen.
[0012] Furthermore, the soap pod is one or more of the following plants belonging to the genus Gleditsia (Gleditsia Linn): Gleditsia sinensis Lam., Gleditsia japonica var. delavayi (Franch.) LC Li, Gleditsia japonica Lodd. ex W. Baxter, Gleditsia fera (Lour.) Merr., and Gleditsia japonica Miq. var. velutina L.C. Li.
[0013] Furthermore, the inflorescences were collected at noon.
[0014] Further, the ratio of inflorescence to water in S1 is 10-20 g / L. Preferably, the ratio of inflorescence to water in S1 is 15 g / L.
[0015] Further, the stirring speed in S1 is 100-150 r / min. Preferably, the stirring speed in S1 is 120 r / min.
[0016] Further, the screen mesh size in S2 is 20-80 mesh. Preferably, the screen mesh size in S2 is 50 mesh.
[0017] Further, the pressure filtration described in S3 is carried out in a rubber piston cylinder with a pollen isolation mesh installed at one end. The diameter of the rubber piston cylinder is 10-20 cm, and the height is 30-60 cm. The mesh size is 500-600 mesh. Preferably, the diameter of the rubber piston cylinder is 15 cm and the height is 40 cm. Preferably, when the soapberry is *Gleditsia sinensis* or *Gleditsia australis*, the mesh size is 500 mesh; when the soapberry is *Gleditsia yunnanensis*, *Gleditsia sinensis*, or *Gleditsia velutipes*, the mesh size is 600 mesh.
[0018] Further, the filter is removed when the pollen layer thickness on the isolation screen is 0.5–1.5 mm, thus ending the filtration process. Preferably, the filter is removed when the pollen layer thickness on the isolation screen is 1 mm, thus ending the filtration process.
[0019] Furthermore, the pressure of the pressure filtration described in S3 is 5-6 kPa, and the rubber cylinder is shaken at 6-8 s intervals during the pressurization process.
[0020] Furthermore, the first drying time in S4 is 30 minutes, and the second drying time is 8 minutes.
[0021] Furthermore, the oscillating crushing and pulverizing is carried out in a vortex oscillator with a frequency of 1000-1500 times / min and an oscillation processing time of 1-3 minutes. Preferably, the oscillator frequency is 1000 times / min and the oscillation processing time is 1 minute.
[0022] Furthermore, the pollen is collected in a sealed container containing color-changing silica gel and stored at -80°C.
[0023] Furthermore, the color of the color-changing silica gel changes with humidity.
[0024] Compared with the prior art, the beneficial technical effects of the present invention are as follows:
[0025] This invention enables direct extraction of soapberry pollen from the inflorescence, greatly increasing the amount of soapberry pollen collected.
[0026] This invention improves pollen collection efficiency and reduces pollen loss during collection through a pressure filtration process. It also filters pollen, removing smaller, poorly developed pollen.
[0027] This invention, through a specific drying process, greatly avoids the loss of pollen activity during extraction, enabling the collection of large quantities of viable soapberry pollen in a short time, making it suitable for large-scale pollen collection operations.
[0028] The method for collecting soapberry pollen provided by this invention is simple to operate and easy to implement. It can solve complex problems in soapberry pollen collection, improve the amount and quality of pollen collected, and is applicable to different types of soapberry, thus enhancing its application value in soapberry breeding, planting industry and scientific research. Attached Figure Description
[0029] The present invention will be further described below with reference to the accompanying drawings.
[0030] Figure 1 This is a process flow diagram of the pollen collection method of the present invention. Detailed Implementation
[0031] This invention provides a rapid method for collecting highly active soapberry pollen, specifically including the following steps:
[0032] S1. Pollen dissolution: Collect male inflorescences of Gleditsia sinensis that are in full bloom and shedding pollen, put them into ultrapure water and stir slowly, repeat 3 times, combine the solutions to obtain an aqueous solution A containing pollen.
[0033] S2. Preliminary sieving: The pollen-containing aqueous solution is sieved to remove large pieces of inflorescence residue, petals, broken anthers and other impurities, to obtain pollen aqueous solution B;
[0034] S3. Pressure filtration: The pollen aqueous solution B is subjected to pressure filtration to obtain pollen;
[0035] S4. Water absorption and drying: Remove excess moisture from the pollen, and then dry it once at a temperature of 28°C and a humidity of 60% to 80% to obtain a solid pollen cake. Then, the solid pollen cake is appropriately dispersed, and the dispersed pollen is dried a second time at a temperature of 28°C and a humidity of 30%.
[0036] S5. Vibration and Powdering: After the dried pollen is vibrated and crushed until it reaches the required powder state, it is stored in a container and preserved under appropriate conditions to complete the rapid collection of soapberry pollen.
[0037] In one embodiment, the soapberry is one or more of the following plants belonging to the genus Gleditsia: Gleditsia sinensis Lam., Gleditsia japonica var. delavayi (Franch.) LCLi, Gleditsia japonica Lodd. ex W. Baxter, Gleditsia fera (Lour.) Merr., and Gleditsia japonica Miq. var. velutina LCLi.
[0038] In one embodiment, the inflorescences are collected at noon to ensure that the male inflorescences are in optimal pollen-shedding condition, maximizing the amount and quality of pollen collected.
[0039] In one embodiment, the ratio of inflorescence to water in S1 is 10-20 g / L.
[0040] In one embodiment, the ratio of inflorescence to water in S1 is 15 g / L.
[0041] This invention ensures that ultrapure water completely covers the inflorescence by limiting the ratio of inflorescence to water, allowing the pollen to fully dissolve and disperse in the water, facilitating subsequent extraction steps. At the same time, it avoids problems such as prolonged filtration time, increased extraction time, and reduced extraction efficiency and pollen viability caused by excessive solution.
[0042] In one embodiment, the stirring speed in S1 is 100-150 r / min.
[0043] In one embodiment, the stirring speed in S1 is 120 r / min.
[0044] This invention limits the stirring speed to ensure that the pollen and water are fully mixed while maintaining the activity and integrity of the pollen. Through multiple stirrings and water changes, pollen can be extracted from the inflorescence to the maximum extent, ensuring that impurities and residues in the pollen are effectively removed, thereby improving the purity and activity of the final pollen.
[0045] In one embodiment, the screen size in S2 is 20 to 80 mesh.
[0046] In one embodiment, the screen size in S2 is 50 mesh.
[0047] This invention, by limiting the size of the sieve in the initial sieving process, can effectively remove large-sized impurities from the solution, such as inflorescence fragments and petals, thereby obtaining a relatively clean pollen aqueous solution. The purpose of this step is to improve the purity and quality of the pollen aqueous solution, providing a good foundation for subsequent processing and use, and avoiding pollen loss while ensuring efficient impurity removal.
[0048] In one embodiment, the pressure filtration described in S3 is carried out in a rubber piston cylinder with a pollen isolation net installed at one end. The diameter of the rubber piston cylinder is 10-20 cm and the height is 30-60 cm. The specification of the isolation net is 500-600 mesh.
[0049] In one embodiment, the diameter of the rubber piston cylinder is 15cm and the height is 40cm.
[0050] This invention does not strictly limit the specific equipment used for pressure filtration. The reason for recommending a rubber piston cylinder is that this device has a simple structure, is easy to operate, simplifies the pressure filtration process, reduces costs, and improves efficiency. This invention does not strictly limit the specifications of the rubber piston cylinder; the specific specifications can be selected according to the actual pollen extraction requirements. In principle, it should be able to hold sufficient pollen aqueous solution while effectively isolating pollen particles. However, during pressurization, the rubber cylinder should be intermittently agitated as needed to avoid clogging.
[0051] In one embodiment, when the soapberry is soapberry or South China soapberry, the mesh size of the isolation net is 500 mesh; when the soapberry is Yunnan soapberry, mountain soapberry, or downy soapberry, the mesh size of the isolation net is 600 mesh.
[0052] This invention selects pollen separation nets with different mesh sizes for different types of soapberry pollen to adapt to the characteristics of different varieties of soapberry pollen, thereby further improving the efficiency and accuracy of separation.
[0053] In one embodiment, the pollen layer on the isolation net is removed when its thickness is 0.5–1.5 mm, thus ending the filtration process.
[0054] In one embodiment, the pollen layer on the isolation net is removed when its thickness reaches 1 mm, thus ending the filtration process.
[0055] In one embodiment, the pressure of the pressurized filter in S3 is 5-6 kPa, and the rubber cylinder is shaken at 6-8 s intervals during the pressurization process.
[0056] This invention strictly limits the pressure of the pressurized filtration process. Appropriate pressure effectively propels the pollen aqueous solution through the pollen separator, ensuring that most pollen particles are separated and collected on the separator. Excessive or insufficient pressure can affect the separation effect; too high a pressure may damage the separator or crush the pollen, while too low a pressure may result in incomplete separation, failing to achieve the desired pollen screening effect and affecting pollen quality and activity. Furthermore, appropriate pressure combined with a specific mesh size screen can prevent damage to the rubber piston cylinder or separator due to excessive force, protecting the long-term use and safe operation of the equipment. Finally, by limiting the pressure, the pressurized filtration process can be more stably controlled, ensuring consistency and repeatability in each operation, and improving the efficiency and stability of the entire collection process.
[0057] In one embodiment, the first drying time in S4 is 30 minutes, and the second drying time is 8 minutes.
[0058] This invention employs a two-stage drying process, with strictly controlled temperature and humidity. Inconsistent process parameters during the two drying stages can lead to incomplete or excessive drying, both of which negatively impact pollen preservation and subsequent application. Strict control of drying conditions ensures appropriate pollen moisture content, maintaining its optimal preservation state. Furthermore, as a biological material, pollen's activity and biological characteristics are significantly affected by drying conditions. Appropriate humidity and temperature conditions maintain the pollen's natural state, preventing damage or inactivation during drying, thus ensuring the reliability and accuracy of the pollen in experiments or applications. Excessive drying can lead to pollen pulverization or damage, affecting its effectiveness in experiments. By strictly controlling drying time and conditions, such quality loss can be effectively avoided, ensuring the integrity and stability of the pollen.
[0059] In one embodiment, the oscillating crushing and pulverizing is carried out in a vortex oscillator with a frequency of 1000-1500 times / minute and an oscillation processing time of 1-3 minutes.
[0060] In one embodiment, the oscillator frequency is 1000 times / minute and the oscillation processing time is 1 minute.
[0061] Generally, this invention does not strictly limit the type of oscillator, but a vortex oscillator is recommended to provide uniform and powerful vibration. This method provides uniform and powerful vibration, effectively breaking down the initially crushed pollen into the desired powder state, and ensuring that the pollen is evenly stressed during oscillation, avoiding localized over-crushing or under-crushing. Furthermore, the oscillator frequency directly affects the pollen crushing effect and the final powder state. Selecting a frequency range of 1000-1500 Hz ensures effective crushing while avoiding over-crushing that could lead to pollen loss or alteration of its biological characteristics, effectively and uniformly crushing the pollen into a powder state suitable for subsequent experiments or applications. Simultaneously, during the oscillation and pulverization process, tracing with tracing paper is necessary to provide an anti-static surface, reducing the impact of static electricity on the pollen, preventing pollen adhesion to the container walls, and improving pollen collection and processing efficiency and quality. Finally, during oscillation, the pollen crushing state should be checked every 20 seconds to ensure the pollen reaches the desired powder state while avoiding over-processing. This periodic inspection can be adjusted according to the actual situation to ensure that the pollen is broken up evenly and properly, maintaining its activity and stability in experiments or applications.
[0062] In one embodiment, the pollen is collected in a sealed container containing color-changing silica gel and stored at -80°C.
[0063] In one embodiment, the color-changing silica gel changes color with changes in humidity.
[0064] This invention tests the pollen preservation effect by placing a certain amount of silica gel in a pollen preservation container, regularly checking for color changes in the silica gel, and replacing it promptly. This effectively ensures the long-term preservation and usability of the pollen. Furthermore, the hygroscopic nature of silica gel allows it to absorb moisture from the air, maintaining a dry environment within the container, which also contributes to the long-term preservation of the pollen.
[0065] The technical solution provided by the present invention will be further described below with reference to the embodiments.
[0066] Example 1
[0067] A rapid method for collecting highly active soapberry pollen, comprising the following steps:
[0068] S1. Pollen Dissolution: At noon during the peak flowering period, collect male inflorescences of Gleditsia sinensis Lam. that are shedding pollen and place them in a stirrer containing ultrapure water, ensuring that the ratio of inflorescence to water is 15 g / L and that the water volume is sufficient to completely cover the inflorescence; stir slowly at 120 r / min for 1 min. After stirring, transfer the pollen-containing aqueous solution to a new container and replace it with fresh ultrapure water. Repeat this process 3 times to obtain pollen-containing aqueous solution A.
[0069] S2. Preliminary sieving: Use a 50-mesh sieve to filter the pollen-containing aqueous solution A to remove large pieces of inflorescence residue, petals, broken anthers and other impurities, to obtain pollen aqueous solution B.
[0070] S3. Pressure filtration: The obtained pollen aqueous solution B is poured into a rubber piston cylinder with a diameter of 15cm and a height of 40cm. A 500-mesh pollen isolation screen is installed at one end of the cylinder. Push the piston to increase the pressure, so that the pressure inside the tube is 5kPa. At this time, a large number of pollen particles are attached to the pollen isolation screen, and the filtrate contains only a small amount of underdeveloped and smaller pollen.
[0071] S4. Absorption and Drying: When the pollen layer attached to the pollen isolation net is 1mm thick, remove the net and place it on absorbent paper to remove excess moisture. Then, place the net in an incubator at 28℃ and 65% humidity for a first drying of 30 minutes. Use the pollen isolation net to completely wrap the solid pollen cake formed after drying, and gently crush the pollen cake by hand to disperse the lumps of pollen. Pay attention to uniform pressure and avoid over-crushing to ensure that the pollen cake does not fall apart during the crushing process. After most of the pollen is dispersed, place it in an incubator at 28℃ and 30% humidity for a second drying of 8 minutes.
[0072] S5. Shaking and Powdering: Take a 50mL centrifuge tube, and use tracing paper to tightly adhere to the inner wall of the plastic tube. Scrape the dried pollen from the separating mesh into the centrifuge tube lined with tracing paper. Fill the tube with 10wt% water-absorbing silica gel beads. Shake the centrifuge tube using a vortex shaker at a frequency of 1000 times / min for 1 minute. To avoid over-crushing the pollen, check the pollen fragmentation status every 20 seconds until the pollen reaches the desired powder state.
[0073] Example 2
[0074] A rapid method for collecting highly active soapberry pollen, comprising the following steps:
[0075] S1. Pollen Dissolution: At noon during the peak flowering period, collect male inflorescences of Gleditsia sinensis Lam. that are shedding pollen and place them in a stirrer containing ultrapure water, ensuring that the ratio of inflorescence to water is 15 g / L and that the water volume is sufficient to completely cover the inflorescence; stir slowly at 120 r / min for 1 min. After stirring, transfer the pollen-containing aqueous solution to a new container and replace it with fresh ultrapure water. Repeat this process 3 times to obtain pollen-containing aqueous solution A.
[0076] S2. Preliminary sieving: Use a 50-mesh sieve to filter the pollen-containing aqueous solution A to remove large pieces of inflorescence residue, petals, broken anthers and other impurities, to obtain pollen aqueous solution B.
[0077] S3. Pressure filtration: The obtained pollen aqueous solution B is poured into a rubber piston cylinder with a diameter of 15cm and a height of 40cm. A 500-mesh pollen isolation screen is installed at one end of the cylinder. Push the piston to increase the pressure, so that the pressure inside the tube is 5kPa. At this time, a large number of pollen particles are attached to the pollen isolation screen, and the filtrate contains only a small amount of underdeveloped and smaller pollen.
[0078] S4. Absorption and Drying: When the pollen layer attached to the pollen isolation net is 1mm thick, remove the net and place it on absorbent paper to remove excess moisture. Then, place the net in an incubator at 28℃ and 60% humidity for a first drying of 30 minutes. Use the pollen isolation net to completely wrap the solid pollen cake formed after drying, and gently crush the pollen cake by hand to disperse the lumps of pollen. Pay attention to uniform force and avoid over-crushing to ensure that the pollen cake does not fall apart during the crushing process. After most of the pollen is dispersed, place it in an incubator at 28℃ and 30% humidity for a second drying of 8 minutes.
[0079] S5. Shaking and Powdering: Take a 50mL centrifuge tube, and use tracing paper to tightly adhere to the inner wall of the plastic tube. Scrape the dried pollen from the separating mesh into the centrifuge tube lined with tracing paper. Fill the tube with 10wt% water-absorbing silica gel beads of pollen weight. Shake the centrifuge tube using a vortex shaker at a frequency of 1000 times / min for 1 minute. To avoid over-shaping the pollen, check the pollen crushing status every 20 seconds until the pollen reaches the desired powder state.
[0080] Example 3
[0081] A rapid method for collecting highly active soapberry pollen, comprising the following steps:
[0082] S1. Pollen Dissolution: At noon during the peak flowering period, collect male inflorescences of Gleditsia sinensis Lam. that are shedding pollen and place them in a stirrer containing ultrapure water, ensuring that the ratio of inflorescence to water is 15 g / L and that the water volume is sufficient to completely cover the inflorescence; stir slowly at 120 r / min for 1 min. After stirring, transfer the pollen-containing aqueous solution to a new container and replace it with fresh ultrapure water. Repeat this process 3 times to obtain pollen-containing aqueous solution A.
[0083] S2. Preliminary sieving: Use a 50-mesh sieve to filter the pollen-containing aqueous solution A to remove large pieces of inflorescence residue, petals, broken anthers and other impurities, to obtain pollen aqueous solution B.
[0084] S3. Pressure filtration: The obtained pollen aqueous solution B is poured into a rubber piston cylinder with a diameter of 15cm and a height of 40cm. A 500-mesh pollen isolation screen is installed at one end of the cylinder. Push the piston to increase the pressure, so that the pressure inside the tube is 6kPa. At this time, a large number of pollen particles are attached to the pollen isolation screen, and the filtrate contains only a small amount of underdeveloped and smaller pollen.
[0085] S4. Absorption and Drying: When the pollen layer attached to the pollen isolation net is 1mm thick, remove the net and place it on absorbent paper to remove excess moisture. Then, place the net in an incubator at 28℃ and 80% humidity for a first drying of 30 minutes. Use the pollen isolation net to completely wrap the solid pollen cake formed after drying, and gently crush the pollen cake by hand to disperse the lumps of pollen. Pay attention to uniform pressure and avoid over-crushing to ensure that the pollen cake does not fall apart during the crushing process. After most of the pollen is dispersed, place it in an incubator at 28℃ and 30% humidity for a second drying of 8 minutes.
[0086] S5. Shaking and Powdering: Take a 50mL centrifuge tube, and use tracing paper to tightly adhere to the inner wall of the plastic tube. Scrape the dried pollen from the separating mesh into the centrifuge tube lined with tracing paper. Fill the tube with 10wt% water-absorbing silica gel beads of pollen weight. Shake the centrifuge tube using a vortex shaker at a frequency of 1000 times / min for 1 minute. To avoid over-shaping the pollen, check the pollen crushing status every 20 seconds until the pollen reaches the desired powder state.
[0087] Comparative Example 1
[0088] Same as Example 1, except that: drying is performed only once.
[0089] Comparative Example 2
[0090] Same as Example 1, except that only two drying processes are performed.
[0091] Comparative Example 3
[0092] Same as Example 1, except that the primary drying temperature is 30°C and the humidity is 50%.
[0093] Comparative Example 4
[0094] Same as Example 1, except that the secondary drying temperature is 30°C and the humidity is 20%.
[0095] Comparative Example 5
[0096] Same as Example 1, except that: the drying method is as follows: when the pollen layer thickness is 1mm, take out the isolation net, place it on absorbent paper to absorb excess water, spread it on a clean thick paper with a thickness of about 1cm, cover the pollen with a layer of white paper to prevent direct sunlight, and dry the pollen moisture content to an average moisture content of 8% under normal intensity sunlight.
[0097] Comparative Example 6
[0098] Same as Example 1, except that: the drying method is as follows: when the pollen layer thickness is 1mm, take out the isolation net, place it on absorbent paper to absorb excess water, put it in a sealed container, add an appropriate amount of color-changing silica gel to dry the pollen moisture content to an average moisture content of 8%.
[0099] Comparative Example 7
[0100] Similar to Example 1, except that atmospheric pressure filtration was used instead of pressurized filtration. The results showed that atmospheric pressure filtration could not achieve pollen separation, therefore the results of this comparative example were not tested in subsequent performance characterization.
[0101] Comparative Example 8
[0102] Same as Example 1, except that the pressure of the pressurized filter is 8 kPa.
[0103] Comparative Example 9
[0104] Same as Example 1, except that: pollen is collected by manually peeling off the anthers using traditional methods, as follows:
[0105] S1. Pollen collection: Collect a large number of male inflorescences of Gleditsia sinensis in the early morning during the peak flowering period. At this time, the anthers on the inflorescences are pale yellow and have not yet dehisced to release pollen. After bringing the inflorescences back to the laboratory, weigh 150g of the inflorescences and manually remove the anthers with tweezers. Let them dry naturally until the anthers dehisce and release pollen.
[0106] S2. Preliminary sieving: Use a 50-mesh sieve to sieve the anthers and pollen to collect the pollen;
[0107] S3. Chemical drying: Place the collected pollen in a sealed container, add an appropriate amount of color-changing silica gel, and dry the pollen to an average moisture content of 8%.
[0108] S4. Shaking and Powdering: Scrape the dried pollen into a 50mL centrifuge tube lined with tracing paper. Add absorbent silica gel balls to the tube and shake the centrifuge tube using a vortex shaker at a frequency of 1000 times / min for 1 minute. Check the pollen breakage status every 20 seconds until the pollen is collected in powder form.
[0109] Pollen collection time, pollen collection amount, and pollen viability were measured in Examples 1-3 and Comparative Examples 1-7. Pollen viability was determined using the in vitro germination method, employing a liquid medium of 15% sucrose + 100 mg / L boric acid + 20 mg / L calcium chloride. The results are shown in Table 1.
[0110] Table 1
[0111]
[0112]
[0113] This document uses specific examples to illustrate the principles and implementation methods of the present invention. The descriptions of the above embodiments are only for the purpose of helping to understand the method and core ideas of the present invention. Furthermore, those skilled in the art will recognize that, based on the ideas of the present invention, there will be changes in the specific implementation methods and application scope. Therefore, the content of this specification should not be construed as a limitation of the present invention.
Claims
1. A method for rapid collection of highly active soapberry pollen, characterized in that, Includes the following steps: S1. Pollen dissolution: Collect male inflorescences of Gleditsia sinensis that are in full bloom and shedding pollen, put them into ultrapure water and stir slowly, repeat 3 times, combine the solutions to obtain an aqueous solution A containing pollen. S2. Preliminary sieving: The pollen-containing aqueous solution is sieved to remove impurities, resulting in pollen aqueous solution B; S3. Pressure filtration: The pollen aqueous solution B is subjected to pressure filtration to obtain pollen. The pressure filtration is carried out in a rubber piston cylinder with a pollen isolation net installed at one end. The pressure of the pressure filtration is 5~6 kPa. S4. Water absorption and drying: Remove excess moisture from the pollen, and then dry it once at a temperature of 28°C and a humidity of 60%~80% to obtain a solid pollen cake. Then, the solid pollen cake is appropriately dispersed, and the dispersed pollen is dried a second time at a temperature of 28°C and a humidity of 30%. S5. Vibration and Powdering: After the dried pollen is vibrated and crushed until it reaches the required powder state, it is stored in a container and preserved under appropriate conditions to complete the rapid collection of soapberry pollen.
2. The method according to claim 1, characterized in that, The soapberry species is from the genus *Gnaphalium*. Gleditsia Linn Plant soap pods ( Gleditsia sinensis Lam. ), Yunnan soapberry ( Gleditsiajaponica var.delavayi(Franch.) LCLi ), Mountain soapberry ( Gleditsiajaponica Lodd.ex W.Baxter ), South China soapberry ( Gleditsiafera (Lour.)Merr. ), fluffy soapberry ( Gleditsiajaponica Miq.var.velutina LCLi One or more of them.
3. The method according to claim 1, characterized in that, The ratio of inflorescence to water in S1 is 10~20 g / L.
4. The method according to claim 1, characterized in that, The stirring speed described in S1 is 100~150 r / min.
5. The method according to claim 1, characterized in that, The sieving process described in S2 is carried out using a sieve with a mesh size of 20-80.
6. The method according to claim 1, characterized in that, The pressure filtration described in S3 is carried out in a rubber piston cylinder with a pollen isolation net installed at one end. The diameter of the rubber piston cylinder is 10-20 cm and the height is 30-60 cm. The specification of the isolation net is 500-600 mesh.
7. The method according to claim 1, characterized in that, When the pollen layer on the isolation net is 0.5~1.5 mm thick, remove it and the filtration process ends.
8. The method according to claim 1, characterized in that, During the pressurization process described in S3, the rubber piston cylinder is shaken at intervals of 6-8 seconds.
9. The method according to claim 1, characterized in that, The first drying time in S4 is 30 minutes, and the second drying time is 8 minutes.
10. The method according to claim 1, characterized in that, The oscillation and crushing pulverization is carried out in a vortex oscillator with a frequency of 1000-1500 times / minute and an oscillation processing time of 1-3 minutes.