CN112655718B

CN112655718B - Industrial hemp female plant male chemical agent and method

Info

Publication number: CN112655718B
Application number: CN202011486049.4A
Authority: CN
Inventors: 李亚玲; 李亚滨; 刘国杰
Original assignee: Fujian Sanan Sino Science Photobiotech Co Ltd
Current assignee: Fujian Sanan Sino Science Photobiotech Co Ltd
Priority date: 2020-12-16
Filing date: 2020-12-16
Publication date: 2022-01-04
Anticipated expiration: 2040-12-16
Also published as: CN112655718A

Abstract

The invention discloses an industrial hemp female plant androgenetic agent and a preparation method thereof.

Description

Industrial hemp female plant male chemical agent and method

Technical Field

The invention relates to the technical field of industrial hemp planting, in particular to an industrial hemp female plant male chemical agent and a method.

Background

Cannabis sativa L, Cannabis sativa of Cannabaceae, Cannabis of Cannabaceae, annual herbaceous plant, named as Yamamai seedling, Linum usitatissimum, Sesamum indicum, Himalayan teasel root, Cannabis sativa L. In China, hemp has been used for thousands of years, and widely relates to the fields of textile, building materials, medicines, foods and the like.

With the development of industrial hemp industry, the cultivation of new varieties of industrial hemp is an urgent need. The production and preservation of high-quality industrial hemp seeds are a key part in the breeding and research of medicinal industrial hemp. The active medicinal components of the marijuana hermaphrodite plant are accumulated in the female plant.

In the traditional industrial hemp breeding method, the male and female seeds are half in each half, and the seeds need to wait until the plants grow to the breeding period and are distinguished by flower buds, so that the problems of slow breeding process and high planting and maintenance cost can be caused by a long period.

Disclosure of Invention

In view of the above, the present invention aims to provide an industrial hemp female plant androgenetic agent which induces the male plant of hemp to generate pure female pollen, thereby obtaining pure female offspring and solving the above problems.

The invention adopts the specific technical scheme that:

an industrial hemp female plant male chemical agent comprises the following components according to molar concentration: 0.3-1.8mmol/L silver thiosulfate and 0-22mmol/L potassium dihydrogen phosphate.

Correspondingly, the invention also provides a method for the androgenesis of the industrial hemp female plant, which is to spray a female plant androgenetic agent on the flowering branches of the industrial hemp female plant, wherein the female plant androgenetic agent comprises the following components in molar concentration: 0.3-1.8mmol/L silver thiosulfate and 0-22mmol/L potassium dihydrogen phosphate.

Preferably, the female plant androgenizing agent is used in an amount of 10-20mL per flowering branch.

More preferably, the female-plant androgenic agent is administered once every 1 to 7 days at a frequency of 8 to 32 consecutive times.

More preferably, the time for the onset of the female plant androgenization agent is 2 to 3 days before the plant switches from the vegetative to the reproductive phase.

The invention has the beneficial effects that:

1. the method for preparing the pure female seeds does not need a sex distinguishing step, shortens the breeding time and greatly reduces the cultivation cost.

2. Compared with the existing single stamina-promoting medicament, the silver thiosulfate and monopotassium phosphate compound medicament provided by the invention has the advantages that the compound formula weakens the side effect of phytotoxicity, the normal growth and development of plants are not influenced, the activity of pure female pollen is obviously improved, and the average pollen activity is 10% higher than that of the single stamina-promoting medicament.

Drawings

FIG. 1 shows the weight of pollen in the unit inflorescence sample of each treatment group in example 1;

FIG. 2 shows the germination rates of pollen samples of the treatment groups of example 1;

FIG. 3 shows the plant heights of the treatment groups in example 2;

FIG. 4 shows the stem thickness of each treatment group in example 2;

FIG. 5 shows the specific leaf weight of each treatment group in example 2;

FIG. 6 shows chlorophyll for each treatment group in example 2;

FIG. 7 shows the moisture content of the leaves of each treatment group of example 2;

FIG. 8 shows the pollen weight of an individual plant after drying harvested for each treatment group in example 2;

FIG. 9 shows pollen germination rates for each treatment group in example 2.

Detailed Description

The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto, and various substitutions and alterations can be made without departing from the technical idea of the present invention as described above, according to the common technical knowledge and the conventional means in the field.

Example 1

1. Experimental materials:

a) plant material: robust D, F, 4; u2 medicinal marijuana cutting plant.

b) The culture medium is coconut husk: 1, vermiculite: 1.

c) the drug is an AR grade chemical agent.

2. The experimental method comprises the following steps:

2.1 parameter settings

2.1.1 dosing onset change:

a) starting point A: 5 weeks old (the day of cutting is calculated as the starting point);

b) starting point B: the flower bud is less than 2cm, and the branch of inflorescence is 0.5cm long.

Generally, light changes are initiated at 5 weeks of age, entering the flowering phase of the flowering phase, followed by significant flower bud differentiation at about 1-2 weeks — i.e., A and B are expected to differ by about 2 weeks, i.e., 14 days.

2.1.2 androgenic agent (GA3& STS) comparison;

a) GA3 gibberellin

b) STS silver thiosulfate complex

2.1.3 dosing regimen (concentration/cycle) changes;

2.2 groups (3 repeats per group)

2.2.1 control group (mode used in preliminary experiments) (treatment group 1), administration starting point A, STS 0.3mmol/L, once daily, 32 times of continuous administration;

2.2.2GA3 group (treatment group 2), starting point B for administration, GA30.3mmol/L, once per week, for 8 consecutive administrations;

2.2.3 STS 1 group (treatment group 3), administration starting point B, STS 0.3mmol/L, once daily, 32 consecutive administrations;

2.2.4 STS 2 group (treatment group 4), starting point B of administration, STS 1.8mmol/L, once every 3 days, 8 times of continuous administration;

2.2.5 STS 3 group (treatment group 5), starting point B of administration, STS 1.8mmol/L, once per week, 8 consecutive administrations.

2.3, environmental control:

2.3.1 temperature: 20 +/-2 DEG C

2.3.2 relative humidity: 50 to 70 percent

2.3.3 illumination:

a) and (3) growing period: water-cooled lamp, spectrum in growth period, 200-;

b) and (3) promoting flowering period: water-cooled lamp, spectrum in growth period, 400-;

c) and (3) flowering period: water-cooled lamp, flowering spectrum, 500-.

2.3.4 nutrient solution:

a) nutrient solution in the growth period: EC 2.2 ms/cm; pH 5.5-6.0

b) Nutrient solution for flowering phase: EC 2.5 ms/cm; pH 5.5-6.0

2.3.5 plant types: in the initial state, 4 primary branches and 8-10 secondary branches are controlled, the difference of plant heights is less than 2cm, and the difference of stem thicknesses is less than 0.5 mm; after the inflorescence is obviously differentiated, the plant type is not trimmed.

2.4 pollen harvest standard:

2.4.1 in the inflorescence of the main branch, most male flowers are cracked, 1-2 male flowers are completely opened, and then the male flowers can be harvested;

2.4.2 sampling: 3 portions of each plant were continued for 10cm inflorescences starting from the top. Manually separating male flowers, placing in a parchment paper bag, sealing, and drying in an oven at 25 + -1 deg.C;

2.4.3 drying the male flowers, sieving the dried male flowers with a 50-mesh sieve, and separating to obtain pollen;

2.4.4 notes: after harvesting one male flower or separating one pollen, the gloves need to be replaced, and the male flower is sprayed in a wind and disinfected by alcohol.

3. And (4) comparing and analyzing results:

3.1 time to appearance of Male flowers

TABLE 1 castration experiments first time male flowering

The result shows that for STS androgenic agents, the administration starting point has no great influence on the emergence time of male flowers, and the male flowers appear when three varieties are administered for 16-17 days; the frequency of administration is approximately high, the relatively earlier the male flowers appear. Drug concentrations did not show a significant effect on the time spent in males.

And GA₃The time spent by the agent is the latest and does not take effect on F, presumably for too long an effect period, whose flowering phase has ended before the agent takes effect.

3.2 the weight of each treated pollen is shown in figure 1: FIG. 1 shows a unit inflorescence which is 10cm in length from the top;

data are presented as (Average ± SE); by means of Duncan analysis, pairwise comparison between the same species can be achieved, the marks of different letters are remarkably different, and p is less than 0.05.

In experiment group 2, the F variety did not produce male flowers, so pollen was not collected.

As can be seen from FIG. 1, the different varieties do not respond consistently to different treatment regimes.

For variety D, treatment 4 had the most pollen amount, and there was no significant difference between the other groups;

for the 4 varieties, there was no significant difference in pollen amount between groups;

for the F species, except treatment 2, pollen amounts 4>5>1 ═ 3, i.e. high concentrations with short dosing intervals, produced the greatest amount of pollen.

Overall, treatment 4 produced relatively the most pollen.

3.3 pollen viability for each treatment is shown in FIG. 2:

Since treatment group 2 performed specifically, only 3 plants produced viable pollen throughout the experimental group and could not be statistically analyzed, and were listed individually as follows:

table 2 pollen sample viability of treatment group 2

As can be seen from FIG. 2 and Table 2, each treatment group produced effective pollen, but the different varieties were not sensitive to the agent, wherein treatment group 2 (gibberellin) did not produce pollen for the F variety.

For the D variety, the pollen viability of the treatment 4 is relatively highest, but has no significant difference compared with the pollen viability of the treatment 3, and has no significant difference among other groups;

for the 4 varieties, the pollen viability of the treatment 3 is relatively lowest, but has no significant difference compared with 5, and has no significant difference among other groups; and 1>3, the two groups differ only in the origin of administration;

for the F variety, there was no significant difference in pollen viability among the other groups except treatment 2.

Overall, treatment group 4 was the most viable relative pollen.

In summary, treatment group 4(STS 1.8mmol/L, once every 3 days, with 8 consecutive administrations) was best tested in this experiment.

Example 2

1. Experimental materials:

a) plant material: robust D, F, 4; u2 medicinal marijuana cutting plant.

b) The culture medium is coconut husk: 1, vermiculite: 1.

c) the drug is an AR grade chemical agent.

2. The experimental method comprises the following steps:

2.1 fixed Condition

2.1.1 ambient conditions

2.1.2 Pre-dose plant Pre-treatment

Broad-spectrum bactericide (carbendazim, which is administered according to the recommended dose of commercial medicament) is sprayed on plants to prevent fungal infection.

2.1.3 time of administration

The plants are about 6 weeks old 3 days before flowering.

2.1.4 reshaping mode

2.1.5 time to harvest

Most flowers are yellow green to yellow, most flowers are slightly cracked, and a small part of flowers are completely cracked, so that the pollen is beneficial to dispersing.

2.1.6 pollen drying

Drying by silica gel: separating single flower, and packaging into parchment paper bags; alternately stacking 3 layers with allochroic silicagel layer by layer, and drying at 20 deg.C under the condition that relative humidity is less than or equal to 50% for 12 hr; pollen is separated by a 50-mesh nylon screen; weighing; packaging with parchment paper, vacuum sealing, and storing at-20 deg.C in dark.

2.2 grouping (4 replicates per group)

3. Results and analysis of the experiments

3.1 plant growth index Change

The initial growth indexes of each group of plants are measured on the 1 st day of the test: the plant height, stem thickness, specific leaf weight, chlorophyll and leaf water content have no obvious difference basically, which indicates that the growth vigor of each group of plants is uniform and consistent before the test is started.

After all the test groups were administered, the growth index of each group was measured as shown in FIGS. 3 to 7 below.

Data are presented as (Average ± SD); compared pairwise between groups by Duncan analysis, p is less than 0.001.

As can be seen from FIGS. 3-7, there was no significant difference between the groups in terms of specific leaf weight and leaf water content.

On the morphological data of plant height and stem thickness, the difference between groups is very significant. Generally 9-16 is higher than 1-8.

3.2 the weight of each treated pollen is shown in FIG. 8: data are presented as (Average ± SD); the second group (best formula for phase i test) was used as the control group by LSD analysis, indicating p <0.001, p <0.01, p <0.05, and no difference was noted.

As can be seen from fig. 8, there was a large difference in pollen weight between the groups, with

groups

6, 7, and 8 being significantly better than the control, and

groups

5 and 15 being significantly better than the control, while groups 1 (method provided in literature), 9, 11, and 13 were comparable to the control, and the other groups had poor pollen yield.

3.3 pollen viability

The pollen viability was expressed by measuring the pollen germination rate, and the data is expressed as (Average ± SD) as shown in fig. 9; the second group (best formula for phase i test) was used as the control group by LSD analysis, indicating p <0.001, p <0.01, p <0.05, and no difference was noted.

As can be seen from fig. 9, pollen viability was significantly better in

groups

9 and 16 than in the control, and 1, 4, 6, 7, 8, 10, 12, 13, 14 was comparable to the control.

The invention aims to screen a better androgenesis method and obtain more pollen with high vigor. Thus, considering the combination of pollen yield and vigor,

groups

4, 6, 7, 8, 9 are more potential formulations. Whereas 9 groups were best in terms of pollen appearance. In summary, the results of this experiment suggest that 9 groups (1.8mmol/L silver thiosulfate +22mmol/L potassium dihydrogen phosphate) are the best combination.

Although the embodiments have been described, once the basic inventive concept is obtained, other variations and modifications of these embodiments can be made by those skilled in the art, so that the above embodiments are only examples of the present invention, and not intended to limit the scope of the present invention, and all equivalent structures or equivalent processes using the contents of the present specification and drawings, or any other related technical fields, which are directly or indirectly applied thereto, are included in the scope of the present invention.

Claims

1. A method for the emasculation of industrial hemp female plants is characterized in that a female plant emasculation medicament is sprayed on flower branches of industrial hemp female plants, and the female plant emasculation medicament comprises the following components in molar concentration: 1.8mmol/L silver thiosulfate and 22mmol/L potassium dihydrogen phosphate.

2. The method for the androgenesis of industrial cannabis sativa according to claim 1, wherein the amount of the female plant androgenesis agent is 10-20mL per flowering branch.

3. The method for masculinizing industrial marijuana plants according to claim 2, wherein the frequency of administration of the masculinizing agent is once every 1 to 7 days and 8 to 32 times of continuous administration.

4. The method for masculinizing industrial hemp plants according to claim 3, wherein the time for the onset of the masculinizing agent is 2 to 3 days before the plant is shifted from the vegetative growth phase to the reproductive growth phase.