阅读说明：本技术 一种工业大麻植株的高效雌化方法 (Efficient estrualization method of industrial hemp plants ) 是由 李骏 刘祥忠 汤小苏 于 2020-12-30 设计创作，主要内容包括：本发明公开了一种工业大麻植株的高效雌化方法,包括：使用纳米银胶体喷雾处理工业大麻雌株诱导其转换为雌雄同株,分化出雄性花蕾,产生可育花粉,然后自由授粉,结籽得到全雌性工业大麻种子。本发明采用纳米银胶体对植株进行诱导,将雌性大麻植株转化为雌雄同株大麻,强制雌株产生可育花粉来对其他雌株进行授粉,可以得到96％以上的纯雌性(或称全雌性)工业大麻种子。所需胶体银溶液量较少,诱导成本降低、程序简化,诱导雌化成功率高且稳定。(The invention discloses a high-efficiency estrualization method of industrial hemp plants, which comprises the following steps: the industrial hemp female plant is treated by nano silver colloid spray to induce the industrial hemp female plant to be converted into hermaphrodite, male flower buds are differentiated to generate fertile pollen, and then the fertile pollen is freely pollinated and seeded to obtain the all-female industrial hemp seed. The invention adopts nano silver colloid to induce plants, converts female hemp plants into hermaphrodite hemp plants, forces the female plants to generate fertile pollen to pollinate other female plants, and can obtain more than 96 percent of pure female (or called full female) industrial hemp seeds. The required colloidal silver solution is less, the induction cost is reduced, the procedure is simplified, and the success rate of induction estrification is high and stable.)

1. A high-efficiency estrualization method of industrial hemp plants, characterized by comprising: the industrial hemp female plant is treated by nano silver colloid spray to induce the industrial hemp female plant to be converted into hermaphrodite, male flower buds are differentiated to generate fertile pollen, and then the fertile pollen is freely pollinated and seeded to obtain the all-female industrial hemp seed.

2. The method for high-efficiency estrification of industrial hemp plants according to claim 1, wherein the particle size of the nano silver particles in the nano silver colloid is not more than 50 nm.

3. The efficient estrification method of industrial hemp plants according to claim 1, wherein the mass concentration of nano silver in the nano silver colloid is 15-50 mg/L.

4. The method for high-efficiency estrification of industrial hemp plants according to claim 1, wherein the nano silver colloid is obtained by electrolysis of elemental silver in water;

the voltage of the electrolysis is 9-12V.

5. A method for efficient estrification of industrial hemp plants according to any one of claims 1 to 4, comprising the steps of:

(1) carrying out nano silver colloid spraying treatment on the industrial hemp female plant continuously every day, taking spraying moisture without liquid dripping as a standard, and adjusting the photoperiod to 12 hours, and lighting for 12 hours and darkness, so that the industrial hemp female plant is converted into a hermaphrodite plant;

(2) stopping spraying, continuously culturing for 3-4 weeks, differentiating mature male flower buds from the marijuana of the same male and female plants, generating fertile pollen, and carrying out seed setting after free pollination to obtain the all-female industrial marijuana seeds.

6. The method for the efficient estrification of industrial hemp plants according to claim 5, wherein in step (1), the industrial hemp female plants are obtained by seed cultivation or cloning;

the cloning is to clone the mother plant by using a cutting branch.

7. The method for high-efficiency estrification of industrial hemp plants according to claim 5, wherein in step (1), the nano silver colloid spray treatment is performed 2-3 times per day.

8. The method for high-efficiency estrification of industrial hemp plants according to claim 5, wherein in step (1), the nano silver colloid spray treatment is continued for 5-15 days or until the plant development is completed.

9. The method for the efficient estrification of industrial hemp plants according to claim 5, wherein the total amount of nano silver colloid sprayed per female industrial hemp plant during the spraying treatment period in step (1) is not more than 500 mL.

Technical Field

The invention relates to the technical field of agricultural planting, in particular to a high-efficiency estrualization method of industrial hemp plants.

Background

Cannabidiol (hereinafter referred to as CBD) is a main chemical component in medicinal plant cannabis sativa, is a non-addictive component in cannabis sativa, has pharmacological effects of spasm resistance, anxiety resistance, inflammation resistance and the like, and is drawing more and more attention in various fields of medical use, consumption, food, new materials and the like.

Cannabidiol is mainly enriched in flowers and leaves of female plants, and the content of cannabidiol in male plants is extremely low; to improve the efficiency and productivity of industrial extraction of CBD, it is desirable that industrial cannabis for cannabidiol extraction be whole female. The existing industrial hemp in China is a male and female heterostrain, 50 percent of seeds generated by the industrial hemp are male and 50 percent of seeds generated by the industrial hemp are female under natural conditions, and then male strains need to be artificially selected and removed during seed production. When industrial hemp is planted in large quantities in the field, a large amount of time, field and resources are consumed for managing the plants. If one or two male plants are missed during the screening process, the female plants in the whole hemp field are pollinated, and the content of cannabidiol in the female plants is seriously reduced. In addition, the flowers and leaves of the all-female variety have large yield increase amplitude and regular characters; the all-female variety does not produce seeds, and the potential safety hazard of private seed reservation is also avoided. Therefore, for planting industrial hemp for extracting cannabidiol, from the aspects of production cost, economic benefit, supervision and the like, all female seeds need to be adopted as much as possible to obtain all female plants.

By applying the feminization technology, female plants are forced to generate fertile pollen to pollinate other female plants, so that pure female (or named as full female) industrial hemp seeds can be obtained, and the full female industrial hemp population is obtained when the seeds are planted. The technology can reduce the cost of screening male plants, eliminate the possibility of accidental pollination in the field, reduce the cost of time, field, manpower, resources and the like, improve the economic benefit and is one of the key technologies for the development of the industry.

Patent specification with publication number CN104335895A discloses a method for inducing pure female industrial hemp, which is a chemical induction method, specifically, Silver Thiosulfate (STS) solution and method are adopted to carry out 2 times of spray treatment on the top shoot of main stem and side branch of female plant, and remove male plant, induce male bud on female plant, convert female plant into hermaphrodite plant, and seed after free pollination, the seed is the full female industrial hemp seed. And then, intensively planting the obtained all-female seeds, performing secondary chemical induction, and obtaining the all-female industrial hemp seeds without emasculation. The implementation effect can reach 38% -85% of female plants to become the same male and female beads, and the later generation is the all-female industrial hemp seeds. The above patent technology adopts silver thiosulfate solution to induce plants. However, the method can only achieve the transformation efficiency of the same male and female plants of up to 85 percent, the male flowers account for most, two times of induction treatment are needed before and after the transformation, the full-female industrial hemp seeds can be propagated, the transformation efficiency is not high enough, the procedure is complicated, and the planting cost is increased. In addition, silver thiosulfate is extremely toxic, the used residual liquid needs to be recovered and specially treated, and the silver thiosulfate is forbidden to be used in Europe at present, and the acquisition cost is high.

KR1020040060528A discloses a method for producing female seeds by inducing male flowers on male and female stamens, wherein silver nitrate (AgNO) is added to the female hemp during vegetative growth₃) The solution is sprayed onto female cannabis, and the female cannabis is treated under short day conditions to induce male flowers in the female cannabis. However, silver nitrate is also a toxic substance to the human body.

Disclosure of Invention

Aiming at the defects in the field, the invention provides a high-efficiency estrification method of industrial hemp plants, which adopts nano silver colloid to induce the plants, converts the female hemp plants into hermaphrodite hemp, forces the female plants to generate fertile pollen to pollinate other female plants, and can obtain more than 96% of pure female (or called full female) industrial hemp seeds. The required colloidal silver solution is less, the induction cost is reduced, the procedure is simplified, and the success rate of induction estrification is high and stable.

A method for efficient estrualization of industrial hemp plants, comprising: the industrial hemp female plant is treated by nano silver colloid spray to induce the industrial hemp female plant to be converted into hermaphrodite, male flower buds are differentiated to generate fertile pollen, and then the fertile pollen is freely pollinated and seeded to obtain the all-female industrial hemp seed.

The present invention stimulates the production of male flowers in crops by reducing the level of ethylene in plant tissues or inhibiting the production of ethylene. The invention adopts nano silver colloid as ethylene inhibitor, and the ethylene inhibitor solution is sprayed on female plants to stimulate the female plants to produce male anther. Nano silver colloids are the simplest and most effective method. The nano silver colloid adopted by the invention has no toxicity and no corrosiveness, meets the requirements of environmental protection and sustainable development, and has the advantages of simple configuration, low cost and simple induction process.

Preferably, the particle size of the nano silver particles in the nano silver colloid is not more than 50nm, and the nano silver colloid in the particle size range is beneficial to further improving the inductivity.

Preferably, the mass concentration of the nano silver in the nano silver colloid is 15-50 mg/L, and the nano silver colloid with the concentration lower than 15mg/L can cause the generated male anther to contain a small amount of sterile pollen.

The nano silver colloid can be purchased directly or prepared by self. The existing common preparation methods of the nano-silver colloid comprise methods such as a chemical reduction method, a microwave-assisted method and an in-situ method (see preparation and characterization of the nano-silver colloid and application thereof in life science, which are published in 2016 (chemistry of life), volume 36, pages 185 to 192 of life science of Chennan et al), but raw materials and equipment involved in the methods are complex, and the methods are not suitable for large-scale agriculture. Preferably, the nano silver colloid is obtained by electrolyzing a silver simple substance in water. The silver simple substance can be a silver block with the purity of not less than 99 percent, and can also be simply replaced by a silver wire with the purity of not less than 99 percent, silver solder and the like. The nano silver particles generated by silver elementary substances such as electrolytic silver blocks and the like can be kept in a suspended state in water, and the silver particles cannot be removed by a common filtering means.

More preferably, the voltage of the electrolysis is 9-12V. Higher voltages may also be used, but may present safety concerns. Specifically, a constant voltage power supply or a battery can be used for supplying the voltage.

Still more preferably, the method for preparing the nano silver colloid comprises the steps of:

(A) taking 2 crocodile clips, and respectively clamping a silver block;

(B) welding the 2 alligator clips to one ends of 2 leads respectively, wherein the other ends of the 2 leads are connected with the positive electrode and the negative electrode of a power supply respectively;

(C) immersing the silver blocks clamped by the 2 alligator clamps in distilled water, and ensuring that the clamped 2 silver blocks are not contacted with each other;

(D) and (3) turning on a power supply, moving away the silver block after 15-25 min, detecting the mass concentration of the nano silver in the obtained nano silver colloid, removing black silver oxide on the surface of the silver block if the mass concentration of the nano silver in the obtained nano silver colloid is lower than 15mg/L, putting the silver block into the obtained nano silver colloid again, continuing electrifying, and detecting again every 1-10 min until the mass concentration of the nano silver in the obtained nano silver colloid is 15-50 mg/L.

After the preparation of the nano silver colloid is completed, removing black silver oxide on the surface of the silver block, and storing the silver block in a box for the next use; the obtained nano silver colloid is stored in a brown glass bottle in a dark place and is placed in a cool place. Prohibited from being stored in the refrigerator.

Preferably, the high-efficiency estrification method of industrial hemp plants comprises the following steps:

(1) carrying out nano silver colloid spraying treatment on the industrial hemp female plant continuously every day, taking spraying moisture without liquid dripping as a standard, and adjusting the photoperiod to 12 hours, and lighting for 12 hours and darkness, so that the industrial hemp female plant is converted into a hermaphrodite plant;

(2) stopping spraying, continuously culturing for 3-4 weeks, differentiating mature male flower buds from the marijuana of the same male and female plants, generating fertile pollen, and carrying out seed setting after free pollination to obtain the all-female industrial marijuana seeds.

In the step (1), the industrial hemp female strain can be obtained by a seed cultivation or cloning method.

The cloning is preferably carried out by using a cutting branch to clone a mother plant, and the specific steps can be as follows: selecting a female industrial hemp plant needing to maintain the character, cloning, cleaning and disinfecting tools, cutting off branches with the length of 10-20 cm at an angle of 45 degrees below a newly grown branch, and immediately putting into water; removing the large leaves at the bottom, immersing the cut into the cloning gel for 2-3 minutes, and then placing the cut into a nutrient solution (rooting solution) for rapid rooting; finally, the branches are placed in a small pot of moist soil and the activity is restored after one or two days.

If the plant is cultured from the seed stage instead of being directly cloned, the nano silver colloid needs to be sprayed after the plant shows the character, so as to ensure that the plant is female.

In the step (1), preferably, the nano-silver colloid spraying treatment is performed for 2-3 times per day.

Preferably, in the step (1), the nano silver colloid spraying treatment lasts for 5-15 days or until the plant development is completed. Generally, the spraying treatment is continued for about 2 weeks, and according to different environments, the spraying is sometimes only needed for 5-10 days, and the spraying can also be carried out until the plant development is completed, so that the hermaphrodite plants are ensured.

In the process of spraying and inducing the nano-silver colloid, the plant characters can be observed by using a magnifying glass, when the female hemp plants develop, the male pollen sacs can be developed instead of the female calyces and the female pistils, and the same male and female plants can be realized.

And (2) in the whole spraying treatment period in the step (1), the total dosage of the nano silver colloid sprayed by each industrial hemp female plant is not more than 500 mL.

In the high-efficiency estrualization method of industrial hemp plants, an independent cultivation space needs to be manufactured, and the phenomenon that the whole hemp field is destroyed due to accidental pollination of other plants is avoided.

The all-female industrial hemp seeds obtained by the invention are planted in a concentrated way, and a large amount of female hemp plants can be obtained.

It should be noted that hemp plants sprayed with nano-silver colloid cannot be used for production, but the produced pollen or seeds are safe and usable. Therefore, it is preferable to produce pollen from marijuana female plants by using a cloning method, so that the plants can be clearly distinguished from other plants to prevent the other plants from being polluted by spraying nano silver colloid.

Compared with the prior art, the invention has the main advantages that:

1) more than 96% of the obtained industrial hemp seeds are female;

2) only one induction is needed, and the induction operation process is simple;

3) the nano silver colloid is non-toxic and environment-friendly, the cost is low no matter the nano silver colloid is purchased or prepared by oneself, and a simple and easy nano silver colloid preparation method is used during the self-preparation.

Drawings

FIG. 1 is a schematic structural diagram of an apparatus for preparing nano silver colloid according to an embodiment;

FIG. 2 is a photograph of a nano silver colloid spray treatment in example;

FIG. 3 is a photograph of the hermaphrodite strains induced in the examples.

Detailed Description

The invention is further described with reference to the following drawings and specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The following examples are conducted under conditions not specified, usually according to conventional conditions, or according to conditions recommended by the manufacturer.

As shown in fig. 1, this example prepares nano silver colloid according to the following method:

(A) taking 2 crocodile clips, and respectively clamping a silver block, wherein the purity of the silver block is not less than 99%;

(B) welding the 2 alligator clips to one ends of 2 leads respectively, wherein the other ends of the 2 leads are connected with the positive electrode and the negative electrode of a power supply respectively;

(C) adding 3/4 cups of distilled water into a glass beaker or other glass containers, and immersing the silver blocks clamped by the 2 alligator clamps into the distilled water, wherein the silver blocks are respectively positioned on one side of the glass cup and ensure that the 2 clamped silver blocks are not contacted with each other;

(D) and (3) turning on a power supply, controlling the voltage to be 9-12V, moving away the silver block after 20min, detecting the mass concentration of the nano silver in the obtained nano silver colloid to be light gold, if the mass concentration of the nano silver in the obtained nano silver colloid is lower than 15mg/L, removing black silver oxide on the surface of the silver block, putting the silver block into the obtained nano silver colloid again, continuing electrifying, and detecting again every 5min until the mass concentration of the nano silver in the obtained nano silver colloid is 15-50 mg/L. A concentration of less than 15mg/L of nanosilver colloid will result in the resulting male anthers containing a small amount of sterile pollen. The grain diameter of the nano silver particles in the obtained nano silver colloid is not more than 50 nm.

After the preparation of the nano silver colloid is completed, removing black silver oxide on the surface of the silver block, and storing the silver block in a box for the next use; the obtained nano silver colloid is stored in a brown glass bottle in a dark place and is placed in a cool place. Prohibited from being stored in the refrigerator.

The nano silver colloid prepared by the method is used for carrying out the feminization induction on the industrial hemp plant seeds.

An efficient estrualization method of industrial hemp plants, comprising the steps of:

(1) and (4) planting industrial hemp female plants.

The industrial hemp female strain can be obtained by a seed cultivation or cloning method.

The cloning is to clone a mother plant by using a cutting branch, and specifically comprises the following steps: selecting a female industrial hemp plant needing to maintain the character, cloning, cleaning and disinfecting tools, cutting off branches with the length of 10-20 cm at an angle of 45 degrees below a newly grown branch, and immediately putting into water; removing the large leaves at the bottom, immersing the cut into the cloning gel for 2-3 minutes, and then placing the cut into a nutrient solution (rooting solution) for rapid rooting; finally, the branches are placed in a small pot of moist soil and the activity is restored after one or two days.

(2) As shown in fig. 2, the spraying treatment of the nano silver colloid is continuously carried out for 2-3 times per day on the female industrial hemp plants in the planned cultivation area, the spraying is carried out for 2 weeks by taking the standard that the female industrial hemp plants are wet but no liquid drips, the photoperiod is adjusted to 12 hours, the illumination is carried out for 12 hours and the darkness is carried out, the spraying treatment is continuously carried out for 2 weeks, the female industrial hemp plants are induced to be converted into hermaphrodite plants, and the total amount of the nano silver colloid sprayed by the female industrial hemp plants is not more than 500 mL. Plant characters are observed by using a magnifying glass, when the female hemp plants develop, the female calyx and the female pistil are not generated any more, but male anthers develop, and the same male and female plants are realized, as shown in figure 3.

If the plant is cultured from the seed stage instead of being directly cloned, the nano silver colloid needs to be sprayed after the plant shows the character, so as to ensure that the plant is female. According to different environments, only 5-10 days of spraying is needed sometimes; or spraying the mixture until the plant development is completed to ensure that the male and female plants are the same.

In the process, an independent cultivation space needs to be manufactured, and the situation that the whole hemp field is damaged due to accidental pollination of other plants is avoided.

(3) Stopping spraying, continuously culturing for 3-4 weeks, differentiating mature male flower buds from the marijuana of the same male and female plants, generating fertile pollen, and carrying out seed setting after free pollination to obtain the all-female industrial marijuana seeds. The obtained all-female industrial hemp seeds are planted in a concentrated way, and a large number of female hemp plants can be obtained.

It should be noted that hemp plants sprayed with nano-silver colloid cannot be used for production, but the produced pollen or seeds are safe and usable. Therefore, it is preferable to produce pollen from marijuana female plants by using a cloning method, so that the plants can be clearly distinguished from other plants to prevent the other plants from being polluted by spraying nano silver colloid.

Furthermore, it should be understood that various changes and modifications can be made by one skilled in the art after reading the above description of the present invention, and equivalents also fall within the scope of the invention as defined by the appended claims.