A red oil tamarix breeding artificial pollination regulator, preparation process and use method

Abstract

This invention discloses an artificial pollination regulator for red oil toon seed breeding, its preparation process, and its application method. The regulator comprises component A and component B. Component A includes gibberellin GA3, D-glucose, and disodium hydrogen phosphate buffer; component B includes 24-epibrassinolide, potassium dihydrogen phosphate, and citrate buffer. This invention precisely matches the physiological needs of the initial flowering and full-blown flowering stages through a cyclical time chain of "component A-component B-component A-component B," combined with optimized regulator components, increasing pollination success rate by 30%-40%, and significantly improving seed weight per thousand seeds, plumpness, and germination rate, directly solving the technical bottleneck of unstable natural pollination and large quality fluctuations. The dissolution process ensures the uniformity of the regulator, and combined with pH stability verification, seed quality parameter fluctuations are controlled within ±2%, increasing rapid growth by 30%-40%, improving seed quality stability, and increasing annual yield per plant from 1.5 kg to 3.5 kg. The use of a high-pressure sprayer with an extended high-pressure spray boom reduces manual labor and costs.

Description

Technical Field

[0001] This invention relates to the field of red oil toon breeding technology, specifically to an artificial pollination regulator for red oil toon breeding, its preparation process, and its application method. Background Technology

[0002] Red toon, a native Chinese species of economic forest tree, is rich in vitamins, flavonoids, and essential amino acids in its tender shoots, giving it high nutritional value. However, in recent years, due to a sharp decline in wild red toon populations, it has been upgraded from a third-level protected plant to a second-level protected plant. To save this endangered plant, attention needs to be paid to its propagation and breeding.

[0003] As a tall tree species, traditional sexual reproduction relies on natural pollination or artificial intervention. However, natural pollination depends on wind and insect pollination, which is significantly affected by environmental temperature and humidity, such as the optimal pollination temperature of 18-25℃, humidity of 60%-70%, and the activity cycle of pollinating insects. Existing artificial pollination requires collecting male pollen during the peak flowering period, which necessitates manual climbing of the tree to collect pollen with a pollen collector. After drying and screening, the pollen is sprayed onto the stigma of the female flower using an artificial pollinator. This operation is dangerous, requires 2-3 skilled workers to cooperate, takes 2-3 hours per tree, and needs to be repeated multiple times, resulting in high labor costs. Therefore, it is difficult to meet the needs of large-scale selective breeding.

[0004] According to monitoring data from the Ministry of Agriculture and Rural Affairs in 2025, the seeds of traditionally sexually propagated red toon from Shandong production areas have significant quality defects: the average seed weight is only 0.008-0.010g (thousand-seed weight 8-10g), the plumpness rate is 38%-45%, and the germination rate is 40%-55%. Under natural pollination, the pollination success rate is reduced by 15%-20% due to the decrease in wild populations and pollinating insects. A 2009 study by the Toona sinensis Research Institute of Shanyang County, Shaanxi Province, pointed out that the germination rate of seeds under traditional methods fluctuates within ±10%. The Toona sinensis Research Institute of Shanyang County, Shaanxi Province, in its "Series of Techniques for Year-Round Cultivation of Toona sinensis: 1. Biological Characteristics and Seedling Propagation of Toona sinensis" (China Fruits and Vegetables, 2009(9):49-51), recorded that the seed plumpness rate of seeds naturally pollinated using traditional methods is only 40%-50%, while artificial pollination can increase it to 50%-60%, but the germination rate still fluctuates within 45%-60%, with limited improvement in rapid growth. Moreover, the operation is difficult and costly, making it difficult to promote and apply. Summary of the Invention

[0005] To address the technical deficiencies in existing red oil toon breeding processes, this invention employs a three-tiered technical system: optimized regulator components, synergistic acid-base environment control, and precise spatiotemporal application of pesticides. This system enables precise regulation of the pollination process during flowering, thereby improving pollination success rates and seed quality stability. The technical solution adopted is as follows: the regulator includes component A (initial flowering initiator): gibberellin GA 315mg / L, D-glucose 8g / L, and disodium hydrogen phosphate buffer 0.015mol / L.

[0006] Component B (flowering growth promoter): 24-epibrassinolide 0.3 g / L, potassium dihydrogen phosphate 2.0 g / L, citrate buffer 0.01 mol / L.

[0007] The preparation of component A includes: weighing 0.015 g of gibberellin GA3 using a precision balance, transferring it to a 100 mL beaker, adding 50 mL of deionized water, and stirring magnetically until completely dissolved. GA3 must be handled in the dark; the container should be wrapped with aluminum foil during weighing. Weigh 8.00 g of D-glucose and add it to the above gibberellin GA3 solution, continuing to stir until completely dissolved. Measure 1000 mL of 0.015 mol / L disodium hydrogen phosphate buffer and slowly add it to the above solution while stirring. Continue until the total volume of the component solution reaches 1 L, with a pH of 5.8-6.0 after preparation.

[0008] The preparation of component B includes: weighing 0.30 g of 24-epibrassinolide using a precision balance, transferring it to a 100 mL beaker, adding 50 mL of deionized water, and magnetically stirring until completely dissolved. This reagent must be handled in the dark, and the container should be wrapped with aluminum foil during weighing. Weighing 2.0 g of potassium dihydrogen phosphate and adding it to the above 24-epibrassinolide solution, continuing to stir until completely dissolved. Slowly adding 1000 mL of 0.01 mol / L citrate buffer to the above solution while stirring. The total volume of the component solution reaches 1 L, thus obtaining the component B solution. The pH of the prepared solution is 5.5-5.8.

[0009] Component A of this invention is a flowering initiator, in which gibberellin (GA) is a classic plant growth regulator. In plant physiology, gibberellins have been proven to promote cell elongation, break dormancy, induce flowering, and promote pollen tube growth. Its use at the initial flowering stage aims to utilize its role in promoting pollen tube germination and elongation, laying the foundation for subsequent fertilization.

[0010] D-glucose, as an energy substance and carbon source that plants can directly utilize, provides the necessary energy support for the rapid metabolism of flowers and the development of reproductive organs during the initial flowering stage.

[0011] Disodium hydrogen phosphate buffer maintains the pH of the drug solution in a slightly acidic range of 5.8-6.0, which is close to the pH of the extracellular environment of most plants. This helps maintain the stability of active ingredients such as gibberellin and promotes the absorption of the drug solution by plant tissues.

[0012] Component B is a flowering growth promoter, in which 24-epibrassinolide is a highly effective, broad-spectrum plant growth regulator belonging to the brassinolide class. Its physiological effects include promoting cell division and elongation, improving photosynthetic efficiency, enhancing stress resistance, and significantly promoting pollen germination and fertilization. Use during peak flowering aims to maximize its effect on promoting reproductive cell activity and fertilization success.

[0013] Potassium dihydrogen phosphate is a high-quality phosphorus and potassium compound fertilizer. Phosphorus is an important component of energy substances (such as ATP) and nucleic acids, and is crucial for flowering, fertilization, and seed formation; potassium participates in regulating osmotic pressure and enzyme activity, can enhance plant resistance, and promote the transport of nutrients to reproductive organs.

[0014] Citrate buffer is also used to stabilize drug solutions, creating a suitable and stable microenvironment to ensure the effectiveness of active ingredients.

[0015] When spraying the A and B component solutions, a specific spraying sequence should be adopted, following a cyclical time chain of "A component - B component - A component - B component". Spray component A on the first day of the initial flowering stage, spray component B on the fourth day of full bloom, and after an 8-day interval, spray component A on the ninth day of the initial flowering stage, and spray component B on the twelfth day of full bloom.

[0016] Spraying settings: Spraying should be carried out between 6-8 am or 5-7 pm, with a water consumption of 35L per mu, a nozzle pressure of 2.5MPa, and a spray width of 1.2m.

[0017] The spraying equipment is a standard "high-pressure sprayer + extended high-pressure spray boom".

[0018] This invention still uses manual operation, but its advantage is that it allows for observation of the growth and flowering of the mother tree during critical periods, and allows for fertilization and maintenance of the mother tree in advance.

[0019] Initial flowering stage (first spray of component A): When the flower bud opening rate is 10%-20%, the reproductive organs of the flower begin to become active. Spraying component A, which is based on gibberellin, at this time aims to "start" the germination of pollen tubes and the development of ovules, preparing for the upcoming large-scale flowering and pollination.

[0020] Peak Flowering Period Promotion (First Spray of Component B): The peak flowering period, when the flower bud opening rate reaches 50%-70%, is a critical window for pollination and fertilization. Spraying Component B, with 24-epibrassinolide as its core, at this time aims to "strongly promote" pollen germination, pollen tube elongation, and fertilization processes, directly affecting the pollination success rate.

[0021] Supplementing during the initial flowering period (second spray of component A): Spray component A again on the 9th day of the initial flowering period. This is intended to supplement late-blooming flowers or those whose first treatment was insufficient, ensuring that there is sufficient "start-up" support throughout the entire transition from initial flowering to full bloom.

[0022] Consolidation during peak flowering (second spray of component B): Spray component B again in the later stage of peak flowering to consolidate fertilization effect and provide continuous nutrition (phosphorus and potassium) and growth regulation support for the development of ovaries and ovules that have been successfully fertilized in the early stage, which helps to reduce sterility and improve seed plumpness.

[0023] This invention constructs a closed-loop regulatory chain of "initiation-promotion-supplementation-consolidation". Components A and B are complementary in function, with component A focusing on initiating development and component B focusing on promoting fertilization and nutrition. Through precise timing of spraying, exogenous chemical intervention is synchronized with the plant's internal reproductive and developmental rhythm, thereby systematically improving the efficiency and quality of the entire process from pollen germination to seed maturation.

[0024] The beneficial effects of this invention are as follows: By precisely matching the physiological needs of the initial flowering stage and the full flowering stage through the "A component-B component-A component-B component" cyclical time sequence chain, combined with the optimization of regulator components, the pollination success rate is improved, and the thousand-seed weight, plumpness rate and germination rate of seeds are significantly improved, directly solving the technical bottleneck of unstable natural pollination and large quality fluctuations.

[0025] Using a mobile "high-pressure sprayer + extended high-pressure spray boom" for spraying avoids the dangerous operation of manually climbing tall trees, reducing the operation time per tree from 2-3 hours to 0.5 hours, reducing labor costs by 50%, and reducing the difficulty and cost of manual operation.

[0026] By standardizing the formulation process and application parameters, the fluctuation of seed quality indicators is reduced, the rapid growth rate is increased by 30%-40%, and the annual yield per plant is increased from 1.5 kg to 3.5 kg, significantly improving the stability of seed quality and yield. Attached Figure Description

[0027] Figure 1 A timeline diagram for spraying the flowering period regulator for red oil toon. Detailed Implementation

[0028] Example 1

[0029] This invention relates to an artificial pollination regulator for red oil toon seed breeding, its preparation process, and its usage. The regulator comprises component A: gibberellin GA3 15 mg / L (accurate to 0.01 mg), D-glucose 8 g / L, and disodium hydrogen phosphate buffer 0.015 mol / L. The gibberellin GA3 is transferred to a 100 mL beaker, and 50 mL of deionized water is added. The mixture is magnetically stirred until completely dissolved (approximately 5 minutes), avoiding light. When weighing, the container is wrapped with aluminum foil. D-glucose is then added, and stirring continues until completely dissolved (approximately 10 minutes). Finally, disodium hydrogen phosphate buffer is added. 1000 mL of the 0.015 mol / L disodium hydrogen phosphate buffer is slowly added while stirring until the total volume of the solution reaches 1 L. The pH of the prepared solution is 5.9.

[0030] Component B: 0.3 g / L 24-epibrassinolide (accurate to 0.01 mg), 2.0 g / L potassium dihydrogen phosphate, 0.01 mol / L citrate buffer. Transfer 24-epibrassinolide to a 100 mL beaker, add 50 mL of deionized water, and stir magnetically until completely dissolved (approximately 5 minutes). This reagent must be handled in the dark; wrap the container with aluminum foil when weighing. Add potassium dihydrogen phosphate and continue stirring until completely dissolved (approximately 10 minutes). Finally, add citrate buffer: Measure 1000 mL of 0.01 mol / L citrate buffer and add it slowly while stirring. Continue until the total volume of this component solution reaches 1 L, and the pH of the prepared solution is 5.8.

[0031] The spraying process utilizes a Yamaha gasoline engine, a high-pressure sprayer, and an extended high-pressure spray boom. For example... Figure 1 As shown, the spraying schedule is as follows: spray component A on the first day of initial flowering, spray component B on the fourth day of full bloom, and after an 8-day interval, spray component A again on the ninth day of initial flowering, and then spray component B on the twelfth day of full bloom. Spray between 6-8 am or 5-7 pm, using 35L of water per acre, with a nozzle pressure of 2.5MPa and a spray width of 1.2m.

[0032] To verify the technical effectiveness of this invention, a ten-randomized block design was used in the field. The control group was sprayed with plain water, while the experimental group used the technical solution of this invention. The results are shown in Table 1.

[0033] Table 1

[0034]

[0035] After comparison, the experimental group had an average seed weight of 0.017-0.019g, a plumpness rate of 85-90%, and a germination rate of 90-95%; the control group had an average seed weight of about 0.009g, a plumpness rate of about 41%, and a germination rate of about 42%.

[0036] The annual yield per plant in the experimental group was around 3.5 kg, which was about 133% higher than the yield of 1.5 kg in the control group.

[0037] Through experimental verification, this technical solution has achieved precise control over the pollination of red oil toon flowers by clearly defining the components, proportions, process steps, application timing and parameters.

[0038] While the specific embodiments of the present invention have been described in detail above, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention, and modifications or variations without creative effort are still within the protection scope of the present invention.

Claims

1. A pollination regulator for breeding red oil toon, characterized in that: The regulator comprises component A and component B; component A comprises gibberellin GA3 15 mg / L, D-glucose 8 g / L, and disodium hydrogen phosphate buffer 0.015 mol / L; component B comprises 24-epibrassinolide 0.3 g / L, potassium dihydrogen phosphate 2.0 g / L, and citrate buffer 0.01 mol / L.

2. The preparation process of the artificial pollination regulator for red oil toon seed breeding as described in claim 1, characterized in that: The preparation of component A includes: weighing 0.015 g of gibberellin GA3 and transferring it to a 100 ml beaker, adding 50 ml of deionized water, and stirring magnetically until completely dissolved; weighing 8 g of D-glucose and adding it to the above gibberellin GA3 solution, stirring until completely dissolved; and slowly adding 1000 ml of 0.015 mol / L disodium hydrogen phosphate buffer to the above solution to make the total volume of the component solution 1 L. The preparation of component B includes: weighing 0.3 g of 24-epibrassinolide and transferring it to a 100 ml beaker, adding 50 ml of deionized water, and stirring magnetically until completely dissolved; weighing 2.0 g of potassium dihydrogen phosphate and adding it to the above 24-epibrassinolide solution, and stirring until completely dissolved; taking 1000 ml of 0.01 mol / L citrate buffer and slowly adding it to the above solution to make the total volume of the component solution 1 L.

3. The preparation process of the artificial pollination regulator for red oil toon seed breeding as described in claim 2, characterized in that: The gibberellin GA3 and 24-epibrassinolide must be handled in the dark, and the container should be wrapped with aluminum foil when weighing.

4. The preparation process of the artificial pollination regulator for red oil toon seed breeding as described in claim 2, characterized in that: The pH value of the solution prepared with component A is 5.8-6.0, and the pH value of the solution prepared with component B is 5.5-5.

8.

5. The preparation process of the artificial pollination regulator for red oil toon seed breeding as described in claim 2, characterized in that: The prepared solutions of components A and B are stored in brown glass bottles to protect them from light.

6. The method of using the artificial pollination regulator for red oil toon seed breeding as described in claim 1 or 2, characterized in that: Spray component A on the first day of the initial flowering period of the red cedar, and spray component B on the fourth day of full bloom. After an 8-day interval, spray component A on the ninth day of the initial flowering period and spray component B on the twelfth day of full bloom.

7. The method of using the artificial pollination regulator for red oil toon seed breeding as described in claim 6, characterized in that: Spraying should be carried out between 6-8 am or 5-7 pm, using 35L of water per mu, with a nozzle pressure of 2.5MPa and a spray width of 1.2m.

8. The method of using the artificial pollination regulator for red oil toon seed breeding as described in claim 6, characterized in that: The spraying equipment uses a high-pressure sprayer and an extended high-pressure spray boom.

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