Producing method of <i>Cannabis sativa </i>L. seed

Abstract

The present invention relates to a method for producing Cannabis sativa L. seed, and more particularly, to a method for producing Cannabis sativa L. seed for efficiently cultivating a low-narcotic Cannabis variety, and when Cannabis sativa L. seeds produced by the production method are used, it is possible to cultivate Cannabis female plants where female flowers are formed, and it is possible to improve the quantity of Cannabis female flowers. Further, when a Cannabis extract is prepared using the Cannabis female flowers, it is possible to prepare a low-narcotic Cannabis extract.

Claims

1. A method for producing Cannabis sativa L. seed capable of cultivating Cannabis female plants, the method comprising: spraying a first fertilizer composition on soil before sowing Cannabis sativa L. seeds; sowing Cannabis sativa L. seeds after tilling and preparing the soil on which the first fertilizer composition is sprayed; inducing the formation of budding flowers by providing Cannabis sativa L. germinated from the Cannabis sativa L. seeds with light, wherein the light is provided under a photosynthetic photon flux density (PPFD) condition of 15 to 20 μMm.sup.−2s.sup.−1 for 5 to 16 hours for 15 to 30 days: and then providing light under a photosynthetic photon flux density (PFFD) condition of 30 to 40 μMm.sup.−2s.sup.−1 for 5 to 16 hours for 15 to 30 days; inducing the formation of male budding flowers in Cannabis female plants by removing all Cannabis sativa L. male plants and retaining female plants where male budding flowers are formed by spraying a second fertilizer composition on Cannabis sativa L. female plants; after the second fertilizer composition was sprayed the light is provided under a photosynthetic photon flux density (PPFD) condition of 10 μMm.sup.−2s.sup.−1 for 10 to 12 hours and; producing seeds by fertilizing pollen in the male budding flowers formed in the Cannabis female plants with female flowers.

2. The method of claim 1, wherein the first fertilizer composition comprises a nitrogen fertilizer, a phosphate fertilizer, a potassium fertilizer, and porous silica.

3. The method of claim 1, wherein the second fertilizer composition comprises a nitrogen fertilizer, a phosphate fertilizer, a potassium fertilizer, and porous silica, and the porous silica is a product in which a mixture of solvent, surfactant aminoethoxyvinyl glycine (AVG), α-aminoisobutyric acid (AIB), and an aqueous silver chloride (AgCl) solution is adsorbed.

4. The method of claim 1, wherein the female flowers are used as a Cannabis extract by harvesting flowers and flower petals, and the Cannabis extract is low narcotic.

5. The method of claim 4, wherein the low narcotic comprises tetrahydrocannabinol (THC) in an amount of 0.5 wt % or less in the ingredients of the total extract.

6. The method of claim 4, wherein the low narcotic comprises cannabidiol (CBD) in an amount of 0.5 wt % or more in the ingredients of the total extract.

Description

DETAILED DESCRIPTION

(1) Hereinafter, the Examples of the present invention will be described in detail such that a person skilled in the art to which the present invention pertains can easily carry out the present invention. However, the present invention can be implemented in various different forms, and is not limited to the Examples described herein.

(2) Production of Cannabis sativa L. Seeds Using Fertilizer Composition

(3) Production of Cannabis sativa L. Seeds

(4) Cannabis sativa L. seeds were produced using an IH3 variety known as a low-narcotic variety.

(5) Cannabis sativa L. seeds were sown and cultivated in a glass greenhouse around March. Cannabis sativa L. seeds were sown at an interval of 50 cm in the glass greenhouse. Before sowing, a first fertilizer composition was sprayed on the soil, the soil was tilled and prepared, and then 4 to 5 seeds were each hill-seeded. After sowing, when Cannabis sativa L. was germinated, all seedlings were thinned out while only one healthy seedling was retained.

(6) When seeds were sown and Cannabis sativa L. germinated, light was provided under a photosynthetic photon flux density (PPFD) condition of 20 μMm.sup.−2s.sup.−1 for 10 hours to 12 hours daily.

(7) When cannabis flower buds were formed, male plants were all removed and all thinned out while retaining only female plants.

(8) Thereafter, the formation of male flowers in cannabis female plants was induced by spraying a second fertilizer composition. Even after the second fertilizer composition was sprayed, the condition under which light was provided was maintained in the same manner as described above.

(9) When it was confirmed that male budding flowers were formed in the cannabis female plant and male flowers were blooming, natural fertilization was induced by operating a large electric fan in the glass greenhouse to blow pollen of male flowers by wind.

(10) After the natural fertilization was sufficiently induced, seeds could be confirmed by the unaided eye. When the seeds were sufficiently browned and could be collected, the seeds were collected, and then dried. Thereafter, Cannabis sativa L. seeds were produced by shaking out and harvesting seeds.

(11) Difference in growth of Cannabis sativa L. according to difference in first fertilizer composition

(12) 1. Preparation of Agar Extract

(13) After an extraction solvent distilled water was added to an agar (Gelidium amansii) extract sample at a ratio of 1:10 (w:v) and then the agar extract sample was completely immersed, extraction was performed repeatedly three times at 80° C. under reflux for each 3 hours. The liquid extract was filtered with a Whatman No. 2 filter paper. A powder sample was prepared by concentrating the filtrate at 60° C. under reduced pressure and used for the experiment.

(14) 2. Preparation of Glasswort Extract

(15) A glasswort extract was prepared using the same method as in the agar extract.

(16) 3. Preparation of Fine Powder Silica

(17) A fine powder silica was prepared so as to have an average particle diameter of 5 to 10 μm by grinding silica, and was immersed in a mixed extract in which the agar extract and the glasswort extract were mixed at a weight ratio of 1:1 to immerse the mixed extract in the fine powder silica.

(18) 4. Preparation of Fertilizer Composition

(19) A fertilizer composition was prepared by mixing 50 parts by weight of a phosphate fertilizer, 50 parts by weight of a potassium fertilizer, and 60 parts by weight of fine powder silica with 100 parts by weight of a nitrogen fertilizer.

(20) 5. Difference in Growth of Cannabis sativa L. According to Use of Fertilizer Composition

(21) The differences in growth of Cannabis sativa L. were compared using Cannabis sativa L. seeds in which only female plants collected in the previous experiment were cultivated.

(22) Before sowing Cannabis sativa L. seeds, i) 10 kg of the fertilizer composition was sprayed, the soil was tilled and prepared, and then Cannabis sativa L. seeds were sown ii) for comparison, 10 kg of the nitrogen fertilizer was sprayed, the soil was tilled and prepared, and then Cannabis sativa L. seeds were sown iii) for comparison, 10 kg of a fertilizer composition obtained by removing only fine powder silica from the above fertilizer composition was sprayed, the soil was tilled and prepared, and then Cannabis sativa L. seeds were sown

(23) After the Cannabis sativa L. seeds were sown under the conditions of i) to iii), the differences in growth were compared by measuring the lengths of Cannabis sativa L. under the same growth conditions.

(24) After the Cannabis sativa L. was cultivated for approximately 2 months, the lengths of the Cannabis sativa L. were compared. For confirmation of sample sizes, 10 seedlings of Cannabis sativa L. were each used, and the lengths of Cannabis sativa L. were measured on average.

(25) According to the measurement result, in the case of i) fertilizer, it was confirmed that the average length of the Cannabis sativa L. was 1.7 m, and in the case of ii) fertilizer, the average length of the Cannabis sativa L. was approximately 1.2 m, and in the case of iii) fertilizer, it was confirmed that the average length of the Cannabis sativa L. was 1.3 m.

(26) It was confirmed that substantially the same degree of growth of Cannabis sativa L. was confirmed except for i) fertilizer, and i) fertilizer promoted the growth of Cannabis sativa L., so that a better degree of growth of Cannabis sativa L. could be confirmed.

(27) Preparation of Second Fertilizer Composition

(28) Ethanol, AVG, AIB, an aqueous silver chloride (AgCl) solution, and Tween 80 were mixed and stirred so as to be uniformly mixed using a stirrer, and then the resulting mixture was prepared as an ethylene inhibitor.

(29) TABLE-US-00001 TABLE 1 ES1 ES2 ES3 ES4 ES5 Ethanol 100 100 100 100 100 AVG 20 30 40 50 60 AIB 20 30 40 50 60 AgCl 5 10 12 15 20 Tween 80 10 10 10 10 10 (Unit parts by weight)

(30) Porous silica powder was immersed in ES1 to ES5, the porous silica powder particles were sufficiently impregnated with ES1 to ES5, respectively, and then 50 parts by weight of a phosphate fertilizer, 50 parts by weight of a potassium fertilizer, and 60 parts by weight of fine powder silica were mixed with 100 parts by weight of a nitrogen fertilizer, thereby preparing a second fertilizer composition.

(31) Evaluation of Formation of Male Flowers

(32) The second fertilizer composition for ES1 to ES5 was sprayed on cannabis female plants in each section, and it was confirmed whether male flowers were formed.

(33) It was confirmed by the unaided eye whether male budding flowers were formed. Considering that a plurality of budding flowers was formed at one cannabis plant, the number of male budding flowers was confirmed, a case where 0 and less than 5 male budding flowers were formed, a case where 5 to 10 male budding flowers were confirmed, and a case where more than 10 male budding flowers were formed were marked with X, Δ, and ∘, respectively. The results are shown in the following Table 2.

(34) TABLE-US-00002 TABLE 2 ES1 ES2 ES3 ES4 ES5 Formation of male X Δ ◯ ◯ Δ budding flowers

(35) As shown in Table 2, it was confirmed that the difference in formation of male budding flowers was exhibited according to the difference in content of the ingredient of the second fertilizer composition. In the case of ES1, the effect by the ethylene inhibitor was insignificant, so that it could be confirmed that sex conversion rarely occurred, and in the case of ES2 and ES5, sex conversion occurred, but the degree of conversion was insignificant as compared to other ethylene inhibitors, so that there was a problem in that it was difficult to produce Cannabis sativa L. seeds by natural fertilization.

(36) Production of Cannabis sativa L. Seeds According to Light Conditions and Treatment with Ethylene Inhibitor

(37) Production of Cannabis sativa L. Seeds

(38) Cannabis sativa L. seeds were produced using an IH3 variety known as a low-narcotic variety.

(39) Cannabis sativa L. seeds were sown and cultivated in a glass greenhouse around March. Cannabis sativa L. seeds were sown at an interval of 50 cm in the glass greenhouse. Before sowing, a fertilizer composition was sprayed on the soil, the soil was tilled and prepared, and then 4 to 5 seeds were each hill-seeded. After sowing, when Cannabis sativa L. was germinated, all seedlings were thinned out while only one healthy seedling was retained.

(40) When seeds were sown and Cannabis sativa L. was germinated, light was provided under the same condition as a photosynthetic photon flux density (PPFD) condition of 20 μMm.sup.−2s.sup.−1 for 10 hours to 12 hours daily for approximately 30 days. When cannabis flower buds were formed, male plants were all removed, only female plants were retained, and then light was provided under a PPFD condition of 40 μMm.sup.−2s.sup.−1 for 10 hours to 12 hours daily for 30 days.

(41) Thereafter, the ethylene inhibitor prepared under the same condition as that in the following Table 1 was sprayed on the shoot apex of a cannabis female plant.

(42) After the ethylene inhibitor was sprayed, light was provided under a PPFD condition of 10 μMm.sup.−2s.sup.−1 for 10 hours to 12 hours.

(43) When it was confirmed that male budding flowers were formed in the cannabis female plant on which the ethylene inhibitor was sprayed and male flowers were blooming, natural fertilization was induced by operating a large electric fan in the glass greenhouse to blow pollen of male flowers by wind.

(44) After the natural fertilization was sufficiently induced, seeds could be confirmed by the unaided eye. When the seeds were sufficiently browned and could be collected, the seeds were collected, and then dried. Thereafter, Cannabis sativa L. seeds were produced by shaking out and harvesting seeds.

(45) Preparation of Ethylene Inhibitor

(46) Ethanol, AVG, AIB, an aqueous silver chloride (AgCl) solution, and Tween 80 were mixed and stirred so as to be uniformly mixed using a stirrer, and then the resulting mixture was prepared as an ethylene inhibitor.

(47) TABLE-US-00003 TABLE 3 EE1 EE2 EE3 EE4 EE5 Ethanol 100 100 100 100 100 AVG 20 30 40 50 60 AIB 20 30 40 50 60 AgCl 5 10 12 15 20 Tween 80 10 10 10 10 10 (Unit parts by weight)

(48) Evaluation of Formation of Male Flowers

(49) After the ethylene inhibitor for EE1 to EE5 was each sprayed on 5 seedlings of cannabis female plants, it was confirmed whether male flowers were formed.

(50) It was confirmed by the unaided eye whether male budding flowers were formed. Considering that a plurality of budding flowers was formed at one cannabis plant, the number of male budding flowers was confirmed, a case where 0 and less than 5 male budding flowers were formed, a case where 5 to 10 male budding flowers were confirmed, and a case where more than 10 male budding flowers were formed were marked with X, Δ, and ∘, respectively. The results are shown in the following Table 4.

(51) TABLE-US-00004 TABLE 4 EE1 EE2 EE3 EE4 EE5 Formation of male X ◯ ◯ ◯ Δ budding flowers

(52) As shown in Table 4, it was confirmed that the difference in formation of male budding flowers was exhibited according to the difference in content of the ingredient of the ethylene inhibitor. In the case of EE1, the effect by the ethylene inhibitor was insignificant, so that it could be confirmed that sex conversion rarely occurred, and in the case of EE5, sex conversion occurred, but the degree of conversion was insignificant as compared to other ethylene inhibitors, so that there was a problem in that it was difficult to produce Cannabis sativa L. seeds by natural fertilization.

(53) Difference in Flowering Period Lengths of Female and Male Flowers According to Light Condition

(54) In order to confirm effect of light on the growth of Cannabis sativa L., the lighting conditions were varied, and then the flowering time point of female plants and the flowing time point of male flowers were confirmed, and the difference in flowering periods of female flowers and male flowers was confirmed. During the growth of Cannabis sativa L., the lighting was provided in the same manner as above for 10 to 12 hours daily.

(55) The lighting conditions are shown in the following Table 5.

(56) TABLE-US-00005 TABLE 5 Difference in flowering 30 days after After treatment with periods germination 30 days later ethylene inhibitor (day) PE1 20 μMm.sup.−2s.sup.−1PPFD 40 μMm.sup.−2s.sup.−1PPFD 10 μMm.sup.−2s.sup.−1PPFD 1 PE2 20 μMm.sup.−2s.sup.−1PPFD 20 μMm.sup.−2s.sup.−1PPFD 20 μMm.sup.−2s.sup.−1PPFD 16 PE3 40 μMm.sup.−2s.sup.−1PPFD 40 μMm.sup.−2s.sup.−1PPFD 40 μMm.sup.−2s.sup.−1PPFD 20 PE4 10 μMm.sup.−2s.sup.−1PPFD 10 μMm.sup.−2s.sup.−1PPFD 10 μMm.sup.−2s.sup.−1PPFD 15

(57) As shown in Table 5, it was confirmed that according to the difference in lighting conditions, the difference in flowering periods of female flowers and male flowers was exhibited. When the difference in flowering periods is exhibited to be 2 weeks or more, the natural fertilization is impossible, so that it can be said that production of Cannabis sativa L. is greatly affected because the artificial fertilization needs to be performed manually by humans.

(58) Difference in Growth of Cannabis sativa L. According to the Difference in Fertilizers

(59) 1. Production of Agar Extract

(60) After an extraction solvent distilled water was added to an agar (Gelidium amansii) extract sample at a ratio of 1:10 (w:v) and then the agar extract sample was completely immersed, extraction was performed repeatedly three times at 80° C. under reflux for each 3 hours. The liquid extract was filtered with a Whatman No. 2 filter paper. A powder sample was prepared by concentrating the filtrate at 60° C. under reduced pressure and used for the experiment.

(61) 2. Preparation of Glasswort Extract

(62) A glasswort extract was prepared using the same method as in the agar extract.

(63) 3. Preparation of Fine Powder Silica

(64) A fine powder silica was prepared so as to have an average particle diameter of 5 to 10 μm by grinding silica, and was immersed in a mixed extract in which the agar extract and the glasswort extract were mixed at a weight ratio of 1:1 to immerse the mixed extract in the fine powder silica.

(65) 4. Preparation of Fertilizer Composition

(66) A fertilizer composition was prepared by mixing 50 parts by weight of a phosphate fertilizer, 50 parts by weight of a potassium fertilizer, and 60 parts by weight of fine powder silica with 100 parts by weight of a nitrogen fertilizer.

(67) 5. Difference in Growth of Cannabis sativa L. According to Use of Fertilizer Composition

(68) The differences in growth of Cannabis sativa L. were compared using Cannabis sativa L. seeds in which only female plants collected in the previous experiment were cultivated.

(69) Before sowing Cannabis sativa L. seeds, i) 10 kg of the fertilizer composition was sprayed, the soil was tilled and prepared, and then Cannabis sativa L. seeds were sown; ii) for comparison, 10 kg of the nitrogen fertilizer was sprayed, the soil was tilled and prepared, and then Cannabis sativa L. seeds were sown; and iii) for comparison, 10 kg of a fertilizer composition obtained by removing only fine powder silica from the above fertilizer composition was sprayed, the soil was tilled and prepared, and then Cannabis sativa L. seeds were sown.

(70) After the Cannabis sativa L. seeds were sown under the conditions of i) to iii), the differences in growth were compared by measuring the lengths of Cannabis sativa L. under the same growth conditions.

(71) After the Cannabis sativa L. was cultivated for approximately 2 months, the lengths of the Cannabis sativa L. were compared. For confirmation of sample sizes, 10 seedlings of Cannabis sativa L. were each used, and the lengths of Cannabis sativa L. were measured on average.

(72) According to the measurement result, in the case of i) fertilizer, it was confirmed that the average length of the Cannabis sativa L. was 1.7 m, and in the case of ii) fertilizer, the average length of the Cannabis sativa L. was approximately 1.2 m, and in the case of iii) fertilizer, it was confirmed that the average length of the Cannabis sativa L. was 1.3 m.

(73) It was confirmed that substantially the same degree of growth of Cannabis sativa L. was exhibited except for i) fertilizer, and i) fertilizer promoted the growth of Cannabis sativa L., so that a better degree of growth of Cannabis sativa L. could be confirmed.

(74) Although preferred Examples of the present invention have been described in detail hereinabove, the right scope of the present invention is not limited thereto, and it should be understood that many variations and modifications of those skilled in the art using the basic concept of the present invention, which is defined in the following claims, will also fall within the right scope of the present invention.