Industrial hemp male attractant agent, preparation method and application thereof

Abstract

The present application relates to the technical field of agricultural application, in particular to a kind of industrial hemp male luring agent and its preparation method and application.The present application uses the common chemical reagent alpha-amino isobutyric acid (AIB) and gibberellin GA3 as effective component and prepares male luring agent, compared with existing male luring agent silver thiosulfate (STS), raw material is more easily obtained, cost is more inexpensive, use is safer, effect is more optimal, has the potential for large-scale production practice.Using the male luring agent proposed in the present application to spray the inflorescence parts of industrial hemp female plants at the beginning of flowering can induce female plants to be male, and produce fertile pollen.The pollen produced by the induced male plants is used to pollinate the inflorescences of normal female plants, which can produce seeds and obtain all-female seeds, which are used for industrial hemp female seed production.

Description

Technical Field

[0001] This invention relates to the fields of agricultural cultivation technology and crop propagation technology, specifically to an industrial hemp male-inducing agent, its preparation method, and its application. Background Technology

[0002] marijuana( Cannabis sativa Cannabis (L.), also known as hemp, fire hemp, thread hemp, and cold hemp, is a species of hemp belonging to the genus Cannabis in the family Cannabinaceae, order Urticales, class Magnoliopsida. Cannabis An annual herbaceous plant. Industrial hemp is a cultivated type developed at the beginning of this century to maximize profits and minimize harms. It refers to hemp varieties bred through genetic modification that contain less than 0.3% tetrahydrocannabinol (THC), a hallucinogenic and addictive substance, and have no drug-related value. Due to its safety, versatility, and significant economic value, industrial hemp has shown broad development prospects in recent years.

[0003] Industrial hemp is a typical dioecious crop, meaning male and female plants are not distinguishable from each other at the seed level (the male-to-female ratio is generally 1:1). It is usually only after sowing and reaching the early reproductive growth stage that male and female plants can be visually identified. The economic value of male and female plants differs; male plants yield and produce significantly higher quality fiber than female plants. In production, if the goal is fiber for textiles, male hemp is preferred for harvesting; if the goal is for edible seeds or medicinal use of flowers and leaves, female hemp is preferred. The medicinal component cannabidiol (CBD) in female plants is much higher than in male plants, but the CBD content decreases by 60% after pollination. To obtain higher CBD content, it is necessary to manually identify the sex of the industrial hemp before pollination and remove male plants. In production aimed at seed use and CBD medicinal purposes, an all-female population is the optimal choice.

[0004] Currently, a feasible method for propagating all-female industrial hemp seeds is to use chemical induction to induce female plants to produce fertile X-type male pollen. This induced pollen is then used to pollinate normal female plants in the same population, resulting in all-female (XX-type) seeds. Most industrial hemp varieties bred in my country are dioecious. However, the main obstacles limiting the use of dioecious or female varieties for all-female seed production and large-scale promotion are low production efficiency, complex processes, and a lack of environmental friendliness. Currently, the traditional male-inducing agent—silver thiosulfate (STS)—is sprayed to induce male flowers. This agent and method have drawbacks such as complex preparation, difficult storage (requiring darkness and low temperatures), high plant mortality, and environmental unfriendliness. Therefore, further research and development of a scientific, efficient, environmentally friendly, and scalable male-inducing agent and technology is crucial. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention uses α-aminoisobutyric acid (AIB) combined with gibberellin GA3 as the active ingredient to form a male-inducing agent to induce maleization in female plants and produce fertile pollen.

[0006] To achieve the above objectives, the present invention provides the following technical solution: In a first aspect, the present invention provides an industrial hemp male-inducing agent, wherein the active ingredients of the male-inducing agent are α-aminoisobutyric acid and gibberellin GA3; The final concentration of α-aminoisobutyric acid in the androgen inducer is 1-4 g / L, more preferably 1 g / L, 2 g / L, 3 g / L, or 4 g / L, and preferably 4 g / L; the final concentration of gibberellin GA3 in the androgen inducer is 25 mg / L.

[0007] Secondly, the present invention provides a method for preparing the aforementioned industrial hemp male-inducing agent, comprising the following steps: Prepare an aqueous solution of 1-4 g / L α-aminoisobutyric acid; Gibberellin GA3 was added to the aqueous solution of α-aminoisobutyric acid to prepare a mixed solution with a final concentration of 25 mg / L of gibberellin GA3, thus obtaining the industrial hemp male-inducing agent.

[0008] Thirdly, this invention proposes the application of the male-inducing agent in inducing maleization of female industrial hemp plants, producing fertile pollen, and preparing all-female seeds of industrial hemp.

[0009] Fourthly, this invention proposes a method for applying an industrial hemp male-inducing agent, specifically: spraying the male-inducing agent at the initial flowering stage of the female industrial hemp plant (i.e., the initial stage of stigma appearance); Alternatively, an industrial hemp male-attracting agent containing a surfactant can be sprayed, wherein the surfactant accounts for 0.01% of the volume of the male-attracting agent.

[0010] Furthermore, the spraying area is the main stem inflorescence region and the lateral branch inflorescence region of the female industrial hemp plant during the initial flowering stage.

[0011] Furthermore, during spraying, apply 5–10 mL of the male-inducing agent to each main stem or lateral branch until the corresponding flower spike area is completely wetted.

[0012] Furthermore, the spraying frequency is once every 7 days, and a total of 3 sprays are generally applied.

[0013] About 30 days after the first spraying, male buds can be seen appearing at the sprayed area, and the number of male buds gradually increases, they open and shed pollen.

[0014] Compared with the prior art, the present invention has the following advantages: This invention utilizes common chemical reagents α-aminoisobutyric acid (AIB) and gibberellin (GA3) as active ingredients to prepare a male-inducing agent. Compared with the currently used male-inducing agent silver thiosulfate (STS), the raw materials are more readily available, the cost is lower, the use is safer, and the effect is superior, making it potential for large-scale production. Spraying this male-inducing agent on the flower spikes of female industrial hemp plants at the initial flowering stage can convert over 80% of the female plants into male plants, producing fertile pollen. In industrial hemp seed production, using this male-inducing agent to induce masculinization in treated rows, and then using the induced pollen to pollinate untreated female plants, can achieve the production of all-female seeds. Attached Figure Description

[0015] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0016] Figure 1 This image shows a photograph of the flower spike when the stigma of a female industrial hemp plant begins to appear at the initial flowering stage, as shown in an embodiment of the present invention. Figure 2 This image shows a photograph of a small number of male buds appearing 30 days after spraying a male-inducing agent onto the flower spikes of female industrial hemp plants in an embodiment of the present invention. Figure 3 This image shows a photograph of a large number of male buds appearing on the female hemp plants 45 days after spraying them with the male-inducing agent, as described in an embodiment of the present invention. Figure 4 The diagram shows the pollen activity generated after male buds were induced by agents 1, 2, and 3 in an embodiment of the present invention. Figure 5 The diagram shows the pollen activity produced after male buds were induced by control reagents 2 and 3 in the comparative examples of this invention. Figure 6 The diagram shows the pollen activity produced after male buds were induced by control reagents 4, 5, and 6 in the comparative examples of this invention. Figure 7 The diagram shows flower spikes induced by agents 1, 2, and 3 in embodiments of the present invention. Figure 8 The diagram shows flower spikes induced by control reagent 1, control reagent 2, and control reagent 3 in the comparative examples of the present invention. Figure 9 The diagram shows flower spikes induced by control agents 4, 5, and 6 in the comparative examples of the present invention. Detailed Implementation

[0017] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0018] Example 1 The industrial hemp male-inducing agent proposed in this invention is prepared using α-aminoisobutyric acid as the active ingredient. The specific process is as follows: Take 1g, 2g, and 4g of α-aminoisobutyric acid (AIB) powder respectively and dissolve them in 1L of water and stir well. Take 25mg of GA3 powder and dissolve it in 0.5mL of anhydrous ethanol and shake well. The dissolved GA3 powder was added to aqueous solutions of α-aminoisobutyric acid at mass-volume ratios of 1 g / L, 2 g / L, and 4 g / L, respectively, to obtain androgen-inducing agents.

[0019] It should be noted that the male enhancement drug solution is best prepared and used immediately, and the GA3 solution should be refrigerated if used the next day.

[0020] To ensure the duration of action of male enhancement agents, when using male enhancement agents, you can add 0.01% of Tween-80 (polyoxyethylene sorbitan monooleate) to the male enhancement agent and stir well before use.

[0021] 0.01% (0.01% of the total volume of the male-inducing agent) of Tween-80 was added to the 1g / L, 2g / L and 4g / L male-inducing agents prepared in this embodiment, respectively, and the resulting agents were named Agent 1, Agent 2 and Agent 3.

[0022] Comparative Example 1 A low-concentration silver thiosulfate (STS) solution was used as a chemical inducer (CN104335895A) to induce cannabis production. The specific preparation method is as follows: STS solutions of 0.5 mmol / L (refer to Example 4 of the patent), 1 mmol / L (refer to Example 3 of the patent), and 2 mmol / L (refer to Example 1 of the patent) were prepared according to the method disclosed in the patent with publication number CN104335895A. The obtained STS solutions were respectively designated as control reagent 1, control reagent 2, and control reagent 3.

[0023] Add 25 mg / L of GA3 to STS solutions of 0.5 mmol / L, 1 mmol / L, and 2 mmol / L respectively. The resulting solutions are designated as Control Drug 4, Control Drug 5, and Control Drug 6, respectively.

[0024] Field application: In the field, seeds of 'Yunma Female No. 1' were directly sown. The female industrial hemp plants were observed to begin flowering, i.e., when the stigmas began to appear (e.g.) Figure 1 (As shown) The agents prepared in Example 1 and Comparative Example 1 were sprayed on the main stem and lateral branches three times, once every 7 days. The leaves and tips of the main stem and lateral branches were sprayed until the leaf surface was completely wetted but no liquid dripped (approximately 5-10 mL). A photograph of the stigma one month after spraying is shown. Figure 2 As shown, male flower buds have already appeared. A photo of the stigma 45 days after spraying the pesticide is shown. Figure 3 As shown, the spikelet area exhibits a large number of male buds.

[0025] It should be noted that, to avoid interference between different agents, each concentration should be sprayed with a separate sprayer. Otherwise, spray the control (with water) first, then spray the low, medium, and high concentrations separately, and clean the sprayer each time before using the next concentration. The best time to spray is throughout the day on a cloudy day, or before 10:00 AM or after 3:00 PM on a sunny day.

[0026] The effects of different agents on the male induction rate and pollen activity of 'Yunma Female No. 1' plants in Example 1 and Comparative Example 1 are shown in Table 1: Table 1

[0027] The male induction rate of industrial hemp refers to the proportion of female plants that develop male characteristics or male plants induced by a certain treatment method (such as the use of exogenous agents). In this embodiment, the male induction rate is calculated based on the ratio of the number of male plants induced after treatment to the number of female plants before treatment.

[0028] Microscopic images of the effects of different pesticides on the pollen activity of 'Yunma Female No. 1' are shown below. Figure 4 , Figure 5 and Figure 6 As shown, where, Figure 4 and Figure 7 The images show pollen activity and inflorescence patterns after induction with pesticides 1, 2, and 3, respectively. Figure 5 The images show pollen activity after induction by spraying control agent 2 and control agent 3, respectively. No pollen was collected in the group sprayed with control agent 1 (poor pollen dispersal), so there are no related pollen activity images. Figure 8 The images show flower spikes after induction by spraying control agent 1, control agent 2, and control agent 3, respectively. Figure 6 and Figure 9 The following diagrams show pollen activity and inflorescence spikes after induction with control agents 4, 5, and 6, respectively. Figures 7-9 The concentration of the drug treatment in each graph ranges from low to high, for example... Figure 7The first row contains AIB 1 g / L + GA 3 25 mg / L, the second row contains AIB 2 g / L + GA 3 25 mg / L, and the third row contains AIB 4 g / L + GA 3 25 mg / L. As can be seen from Table 1, the agent 3 proposed in this invention exhibits the best male-inducing rate and pollen activity for cannabis.

[0029] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An industrial hemp male-inducing agent, characterized in that, The active ingredients of the male-inducing agent are α-aminoisobutyric acid and gibberellin GA3; The final concentration of α-aminoisobutyric acid in the androgen inducer is 1-4 g / L, and the final concentration of gibberellin GA3 in the androgen inducer is 25 mg / L.

2. A method for preparing an industrial hemp male-inducing agent, characterized in that, Includes the following steps: Prepare an aqueous solution of 1-4 g / L α-aminoisobutyric acid; Gibberellin GA3 was added to the aqueous solution of α-aminoisobutyric acid to prepare a male-inducing agent with a final concentration of 25 mg / L of gibberellin GA3.

3. The application of the male-inducing agent according to claim 1 or the male-inducing agent prepared according to the method of claim 2 in the masculinization of female industrial hemp plants, the production of fertile pollen, and the production of all-female seeds of industrial hemp.

4. A method for applying an industrial hemp male-inducing agent, characterized in that, During the initial flowering stage of female industrial hemp plants, spray the male-inducing agent as described in claim 1; Alternatively, an industrial hemp male-attracting agent containing a surfactant can be sprayed, wherein the surfactant accounts for 0.01% of the volume of the male-attracting agent.

5. The method according to claim 4, characterized in that, The surfactant is Tween-80.

6. The method according to claim 4, characterized in that, The spraying area includes the main stem and lateral branch flower spike areas of the female industrial hemp plants during the initial flowering stage.

7. The method according to claim 4, characterized in that, When spraying, spray 5-10 mL of the male-inducing agent on each main stem or lateral branch until the corresponding flower spike area is completely wetted.

8. The method according to claim 4, characterized in that, The spraying frequency is once every 7 days, for a total of 3 sprays.

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