CN113615497B - Method for reducing tetrahydrocannabinol content in hemp plants - Google Patents
Method for reducing tetrahydrocannabinol content in hemp plants Download PDFInfo
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- CN113615497B CN113615497B CN202110980946.9A CN202110980946A CN113615497B CN 113615497 B CN113615497 B CN 113615497B CN 202110980946 A CN202110980946 A CN 202110980946A CN 113615497 B CN113615497 B CN 113615497B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C21/00—Methods of fertilising, sowing or planting
- A01C21/005—Following a specific plan, e.g. pattern
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
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Abstract
The invention discloses a method for reducing the content of tetrahydrocannabinol in hemp plants, which is to spray nano graphene dispersion liquid on the hemp plants after the flowering phase. According to the method for reducing the content of the tetrahydrocannabinol in the hemp plants, the nano graphene dispersion liquid is sprayed on the hemp plants after the flowering phase, so that the content of the tetrahydrocannabinol in the flowers and the leaves is obviously reduced and is lower than 0.3%, and the method meets the relevant legal regulations. The hemp plant sprayed with the nano graphene dispersion liquid has no obvious reduction in the biological yield of the flower and leaf, and the flower and leaf yield tends to increase in a proper concentration range, mainly because the nano graphene dispersion liquid is a two-dimensional nano material consisting of carbon atoms, the photosynthetic utilization rate of the plant can be enhanced, the plant growth is promoted, and the flower and leaf yield is increased.
Description
Technical Field
The invention relates to the technical field of agriculture, in particular to a method for reducing the content of tetrahydrocannabinol in hemp plants.
Background
Hemp (Cannabis sativa L.) is a traditional commercial crop, has a long history of cultivation, and is mainly applied to a plurality of fields such as textile, paper making, medicine and the like. Cannabinoids (cannabinoids) are a class of secondary metabolic natural products containing alkyl and monoterpene groups that are unique to the cannabis plant, and currently, 120 kinds of cannabinoids have been isolated, mainly Tetrahydrocannabinol (THC), Cannabidiol (CBD), Cannabinol (CBN), Cannabigerol (CBG), and the like. Research finds that CBD has the functions of blocking adverse effects of certain polyphenols on human nervous systems, blocking breast cancer metastasis, treating epilepsy, resisting rheumatoid arthritis, resisting insomnia and other physiological activities, and has good effect on treating multiple sclerosis. However, THC has hallucinogenic addiction, so that cannabis also becomes a well-known drug source plant. In order to make efficient and safe use of hemp plants, the european union, canada and the china Yunnan province, etc., stipulate in the form of law that hemp plants with a THC content of < 0.3% are industrial hemp. The industrial hemp has no drug utilization value and is popularized by adopting a licensed planting mode. Therefore, the THC content in the hemp plants is effectively reduced to be lower than 0.3 percent, and the method has important significance for the popularization and the utilization of the hemp plants and the guarantee of public safety.
The cannabinoid substances are genetically controlled and mainly exist in glandular hairs of marijuana female plant leaves, calyx and the like, and the content of the cannabinoid substances is related to the number of glandular hairs, the structure of glandular hairs and the like. Researchers can reduce the THC content in the flowers and leaves of the female marijuana plant by a conventional breeding means or a molecular breeding means. However, conventional breeding has a long breeding period, and suitable materials are required to be searched for as parents, while the current molecular breeding technology of marihuana is still imperfect. Cannabinoid accumulation is affected by external environmental conditions in addition to the developmental processes of cannabis plants. Researches show that the total content of cannabinoids in the marihuana leaves is influenced by various environmental factors such as illumination intensity, environmental temperature, soil fertilizer, insect infection, exogenous additives and the like. Studies by Jade (2016) et al found that treatment of hemp plants with three layers of shading, excessive nitrogen fertilizer and severe drought reduced the THC content during the full bloom and initial fruit stages, but shading was difficult to adapt to large-scale planting, excessive nitrogen fertilizer could cause severe environmental pollution, and severe drought reduced plant biomass. Therefore, the important significance of finding a fast and easily realized technical means for reducing the THC content in the hemp plants is great.
Disclosure of Invention
The invention provides a method for reducing the content of tetrahydrocannabinol in hemp plants, which aims to solve the technical problem that the content of tetrahydrocannabinol in the hemp plants cannot be reduced quickly and easily in the prior art.
The technical scheme adopted by the invention is as follows:
a method for reducing tetrahydrocannabinol content in hemp plants comprises spraying nano-graphene dispersion liquid on the hemp plants after the flowering phase.
Further, the nano graphene dispersion liquid comprises a nano graphene oxide dispersion liquid and/or a reduced nano graphene dispersion liquid.
Further, the concentration of the nano graphene dispersion liquid is 10 mg/L-100 mg/L.
Further, the nano graphene dispersion liquid is sprayed on the hemp plants 7 to 28 days after the flowering period.
Further, 20-30 mL of nano graphene dispersion liquid is sprayed on each hemp plant.
Further, the nano graphene dispersion liquid is sprayed once every 1-3 days, and the spraying times are 5-7 times.
Further, before the nano graphene dispersion liquid is sprayed on the hemp plants after entering the flowering phase, the method further comprises the following steps:
s1, culturing the hemp plants for 16-20 h according to the daily illumination duration;
s2, culturing according to the daily illumination time of 10-12 h until entering the flowering phase.
Further, in step S1, the marijuana plants with the plant height of 13 cm-23 cm are cultured for 16 h-20 h according to the daily illumination time, the day and night temperature is 25-30 ℃, and the marijuana plants are cultured and grown until the plant height is 40 cm-50 cm.
Further, after the nano graphene dispersion liquid is sprayed on the hemp plants after the flowering phase, the method further comprises the following steps: continuously culturing for 14-56 days under the environment that the temperature of day and night is 22-28 ℃.
The invention has the following beneficial effects:
according to the method for reducing the content of the tetrahydrocannabinol in the hemp plants, the nano graphene dispersion liquid is sprayed on the hemp plants after the flowering phase, so that the content of the tetrahydrocannabinol in the flowers and the leaves is obviously reduced and is lower than 0.3%, and the method meets the relevant legal regulations. The hemp plant sprayed with the nano graphene dispersion liquid has no obvious reduction in the biological yield of the flower and leaf, and the flower and leaf yield tends to increase in a proper concentration range, mainly because the nano graphene dispersion liquid is a two-dimensional nano material consisting of carbon atoms, the photosynthetic utilization rate of the plant can be enhanced, the plant growth is promoted, and the flower and leaf yield is increased.
The method for reducing the content of the tetrahydrocannabinol in the hemp plants is simple to operate, is environment-friendly, greatly reduces the content of the tetrahydrocannabinol in the hemp plants while not obviously reducing the biomass of the plants, and is easy to realize large-area application.
In addition to the above-described objects, features and advantages, the present invention has other objects, features and advantages. The present invention will be described in further detail below.
Detailed Description
It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present invention will be described in detail with reference to examples.
According to the method for reducing the content of tetrahydrocannabinol in the hemp plants, the nano-graphene dispersion liquid is sprayed on the hemp plants after the flowering phase.
According to the method for reducing the content of the tetrahydrocannabinol in the hemp plants, the nano graphene dispersion liquid is sprayed on the hemp plants after the flowering phase, so that the content of the tetrahydrocannabinol in the flowers and the leaves is obviously reduced and is lower than 0.3%, and the method meets the relevant legal regulations. The hemp plant sprayed with the nano graphene dispersion liquid has no obvious reduction in the biological yield of the flower and leaf, and the flower and leaf yield tends to increase in a proper concentration range, mainly because the nano graphene dispersion liquid is a two-dimensional nano material consisting of carbon atoms, the photosynthetic utilization rate of the plant can be enhanced, the plant growth is promoted, and the flower and leaf yield is increased.
The main role of tetrahydrocannabinol is not to increase the adaptability of the plant itself under abiotic environmental stresses. The content of tetrahydrocannabinol in the top flower leaves at the initial stage of the hemp female plant is highest, so that the reproductive growth process of the hemp plant is presumed to play a role in defending against pest and disease attack by accumulating the tetrahydrocannabinol. Graphene, as a nano-environment foreign object, may stimulate the defense system of plants and reduce the tetrahydrocannabinol content.
The method for reducing the content of the tetrahydrocannabinol in the hemp plants is simple to operate, is environment-friendly, greatly reduces the content of the tetrahydrocannabinol in the hemp plants while not obviously reducing the biomass of the plants, and is easy to realize large-area application.
The hemp plants are clonal seedling groups with consistent genetic background and are robust stock plants of a hemp strain DMG245 which is scientific research material of a hemp research institute quality resource team of the Chinese academy of agricultural sciences.
In this embodiment, the nano graphene dispersion liquid includes a nano graphene oxide dispersion liquid or a reduced nano graphene dispersion liquid.
In this embodiment, the concentration of the nano-graphene dispersion liquid is 10mg/L to 100 mg/L. The concentration of the nano graphene dispersion liquid is lower than 10mg/L, the effect of reducing the content of the tetrahydrocannabinol in the hemp plants is not obvious, the concentration of the nano graphene dispersion liquid is higher than 100mg/L, the graphene is easy to agglomerate in the dispersion liquid and cannot enter plant cells, and the effect of reducing the content of the tetrahydrocannabinol is not obvious.
In this example, the nano-graphene dispersion was sprayed on hemp plants 7 to 28 days after the flowering phase. The nano graphene dispersion liquid is sprayed on the hemp plants which enter the flowering phase for 7 to 28 days, is the initial flowering phase of the plants, is the early stage of accumulation of the tetrahydrocannabinol, is sensitive to stimulation of environmental foreign matters, and is easily influenced, so that the accumulation of the tetrahydrocannabinol is obviously reduced by spraying the nano graphene dispersion liquid on the hemp plants which enter the flowering phase for 7 to 28 days.
In this example, 20-30 mL of the nano-graphene dispersion liquid was sprayed to each hemp plant. Because the biological yield of the plants is high and the plant yield is lower than 20mL, the whole plant of the plant cannot be sprayed in place, and the treatment effect is influenced; and above 30mL would cause unnecessary waste. In addition, the THC content cannot be obviously reduced by further increasing the spraying amount, so that 20-30 mL of the spraying agent is better sprayed to each hemp plant.
In the embodiment, the nano graphene dispersion liquid is sprayed every 1-3 days, and the spraying times are 5-7 times. The spraying frequency is mainly influenced by the length of the flowering phase. 5-7 times, and 1-3 days are provided for each time, so that the nano graphene dispersion liquid is sprayed on the whole flowering phase of the hemp plants, and the accumulation of tetrahydrocannabinol in the flowering phase is effectively controlled, namely the spraying is finished in the flowering phase.
In this embodiment, before the nano graphene dispersion liquid is sprayed on the hemp plants after entering the flowering phase, the method further comprises:
s1, culturing the hemp plants according to the daily illumination time length of 16-20 h;
s2, culturing the seedlings according to the daily illumination time length of 10-12 h until the seedlings enter the flowering phase.
The accumulation of cannabinoids is influenced by the growth process of the hemp plants and the external environment conditions, the growth and development of the hemp plants are greatly influenced by illumination, the nutrition and the reproduction of the hemp plants are fast under the long-day condition, and once the illumination duration is changed, the cannabinoids can enter the flowering phase to start the reproduction and the growth. Moreover, the accumulation of THC varies with the different developmental stages of the plant.
In the embodiment, in step S1, the hemp plants with plant height of 13cm to 23cm are cultured for 16h to 20h according to the daily illumination time, the day and night temperature is 25 ℃ to 30 ℃, and the hemp plants are cultured and grown until the plant height is 40cm to 50 cm.
In this embodiment, after the nano graphene dispersion liquid is sprayed on the hemp plant after entering the flowering phase, the method further includes: continuously culturing for 14-56 days in the environment of day and night temperature of 22-28 ℃.
Examples
The hemp plant is from a robust stock plant of hemp strain DMG245, a scientific research material of germplasm resource team of China academy of agricultural sciences.
The nano graphene oxide dispersion liquid and the reduced nano graphene dispersion liquid are purchased from Beijing Solaibao science and technology Limited, and the solvent is water.
Example 1
Carrying out culture growth on a DMG245 clone seedling hemp plant group with consistent genetic background and average plant height of 15cm under the conditions that the daily illumination time length is 18h and the temperature is 26 +/-1℃, and adjusting the daily illumination time length to 10h after the hemp plant grows to the average plant height of 40cm, namely entering the flowering phase;
after the hemp plants enter the flowering period for 14 days, respectively spraying the hemp plants with nano graphene oxide dispersion solutions with the concentrations of 0mg/L, 10mg/L, 50mg/L and 100mg/L, wherein the spraying amount is 30 mL/plant every two days, and the spraying is carried out for 5 times in total. Culturing in a greenhouse at day and night temperature of 26 + -1 deg.C, periodically watering and fertilizing, and repeating for 3 times. After spraying, the hemp plants are continuously cultured for 21 days, and then samples are taken to determine the flower and leaf yield of the hemp plants and the content of tetrahydrocannabinol in the flower and leaves.
Example 2
Carrying out culture growth on a DMG245 clone seedling hemp plant group with consistent genetic background and average plant height of 20cm under the conditions that the daily illumination time is 16h and the temperature is 26 +/-1 ℃ and adjusting the daily illumination time to 12h when the hemp plant grows to the average plant height of 50cm, namely entering the flowering phase;
after the hemp plants enter the flowering period for 21 days, spraying the hemp plants with prototype nanometer graphene dispersion solutions with the concentrations of 0mg/L, 10mg/L, 50mg/L and 100mg/L respectively, wherein the spraying amount is 25 mL/plant every two days, and the spraying is carried out for 4 times. Culturing in a greenhouse at day and night temperature of 26 + -1 deg.C, periodically watering and fertilizing, and repeating for 3 times. After spraying, the hemp plants are continuously cultured for 35 days, and then sampling is carried out to determine the flower and leaf yield of the hemp plants and the tetrahydrocannabinol content in the flower and leaves.
Example 3
Carrying out culture growth on a DMG245 clone seedling hemp plant population with consistent genetic background and average plant height of 15cm under the conditions that the daily illumination time is 18h and the temperature is 26 +/-1 ℃ until the hemp plant grows to about 50cm, and adjusting the daily illumination time to 12h, namely entering the flowering phase;
after the hemp plants enter the flowering phase for 7 days, the nano graphene oxide dispersion liquid with the concentration of 0mg/L, 10mg/L, 50mg/L and 100mg/L is respectively used for spraying the hemp plants, and the nano graphene oxide dispersion liquid is sprayed once every two days, wherein the spraying amount is 20 mL/plant every time, and the spraying is carried out for 6 times in total. Culturing in a greenhouse at day and night temperature of 26 + -1 deg.C, periodically watering and fertilizing, and repeating for 3 times. After spraying, the hemp plants are continuously cultured for 14 days, and then samples are taken to determine the flower and leaf yield of the hemp plants and the content of tetrahydrocannabinol in the flower and leaves.
Randomly taking 5-15 cm flowers and leaves at the top end of the plant, putting the flowers and leaves into a mesh bag, and drying in the shade. According to the detection regulation recommended in the local standard of Yunnan province, namely 'variety type of industrial hemp' (DB53/295.1-2009), the THC content of the dried flowers and leaves in the shade is detected by a High Performance Liquid Chromatography (HPLC) method. The data are expressed as mean ± sd of 3 replicate groups of plants.
The results of sampling measurement in example 1 are shown in table 1.
Table 1 test results of example 1
In table 1, compared with the control group (the concentration of the nano graphene oxide dispersion liquid is 0mg/L), the tetrahydrocannabinol content in the hemp plant leaves is respectively reduced by 65.85%, 31.71% and 68.29% after the nano graphene oxide dispersion liquid of 10mg/L, 50mg/L and 100mg/L is sprayed for 21 days, but the flower and leaf yield of the hemp plants is not obviously reduced, and the flower and leaf yield tends to increase after the nano graphene oxide dispersion liquid of 50mg/L is treated.
The results of sampling measurement of example 2 are shown in table 2.
Table 2 test results of example 2
In table 2, compared with the control group (the concentration of the reduced nano-graphene dispersion liquid is 0mg/L), the tetrahydrocannabinol content in the hemp plant flower leaves is respectively reduced by 56.89%, 50.01% and 62.07% 35 days after the reduced nano-graphene dispersion liquids of 10mg/L, 50mg/L and 100mg/L are sprayed, the tetrahydrocannabinol content of the treated plants meets the legal requirement of not higher than 0.3%, the flower and leaf yield of the hemp plants does not change significantly, and the flower and leaf yield also tends to increase after the reduced nano-graphene dispersion liquid of 50 mg/L.
The results of sampling measurement of example 3 are shown in table 3.
Table 3 test results of example 3
In table 3, compared with the control group (the concentration of the nano graphene oxide dispersion liquid is 0mg/L), the tetrahydrocannabinol content in the hemp plant leaves is respectively reduced by 35%, 20% and 45% after the nano graphene oxide dispersion liquid of 10mg/L, 50mg/L and 100mg/L is sprayed for 21 days. And the flower and leaf yield tends to increase after the reduced nano graphene dispersion liquid of 50mg/L is treated.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A method for reducing the content of tetrahydrocannabinol in hemp plants,
spraying the nano graphene dispersion liquid on hemp plants after the flowering phase, wherein the concentration of the nano graphene dispersion liquid is 10-100 mg/L, and the nano graphene dispersion liquid is sprayed on the hemp plants 7-28 days after the flowering phase.
2. The method of reducing tetrahydrocannabinol content in cannabis plants according to claim 1,
the nano graphene dispersion liquid comprises a nano graphene oxide dispersion liquid and/or a reduced nano graphene dispersion liquid.
3. The method of reducing tetrahydrocannabinol content in cannabis plants according to claim 1,
spraying 20-30 mL of nano graphene dispersion liquid to each hemp plant.
4. The method of reducing tetrahydrocannabinol content in cannabis plants according to claim 3,
spraying the nano graphene dispersion liquid once every 1-3 days, wherein the spraying times are 5-7 times.
5. The method of reducing tetrahydrocannabinol content in cannabis plants according to claim 1,
before the nano graphene dispersion liquid is sprayed on the hemp plants after the flowering phase, the method also comprises the following steps:
s1, culturing the hemp plants according to the daily illumination time length of 16-20 h;
s2, culturing the seedlings according to the daily illumination time length of 10-12 h until the seedlings enter the flowering phase.
6. The method of reducing tetrahydrocannabinol content in cannabis plants according to claim 5,
in step S1, culturing the hemp plants with the plant height of 13 cm-23 cm for 16 h-20 h according to the daily illumination time, wherein the day and night temperature is 25-30 ℃, and culturing and growing until the plant height is 40 cm-50 cm.
7. The method of reducing tetrahydrocannabinol content in cannabis plants according to claim 1,
after the nano graphene dispersion liquid is sprayed on the hemp plants after entering the flowering phase, the method further comprises the following steps:
continuously culturing for 14-56 days in the environment of day and night temperature of 22-28 ℃.
8. The method of reducing tetrahydrocannabinol content in cannabis plants according to claim 1,
the tetrahydrocannabinol in the hemp plants sprayed with the nano graphene dispersion liquid is reduced by at least 20%.
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