EP3720275A1 - Propyl cannabinoid hemp plants, methods of producing and methods of using them - Google Patents
Propyl cannabinoid hemp plants, methods of producing and methods of using themInfo
- Publication number
- EP3720275A1 EP3720275A1 EP18885114.1A EP18885114A EP3720275A1 EP 3720275 A1 EP3720275 A1 EP 3720275A1 EP 18885114 A EP18885114 A EP 18885114A EP 3720275 A1 EP3720275 A1 EP 3720275A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- cannabis
- plant
- terpene
- content
- weight
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
- A01H6/28—Cannabaceae, e.g. cannabis
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
- A01H5/02—Flowers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/01—Hydrocarbons
- A61K31/015—Hydrocarbons carbocyclic
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
- A61K31/05—Phenols
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
Definitions
- the invention relates to Specialty Cannabis hemp plants with low ⁇ 9 -tetrahydrocannabinol and high propyl cannabinoid contents, as well as compositions mimicking the cannabinoid and Terpene Profiles of said plants, and methods for making and using said cannabis plants and compositions.
- Cannabis is a genus of flowering plants that includes at least three species, Cannabis sativa, Cannabis indica, and Cannabis ruderalis as determined by plant phenotypes and secondary metabolite profiles.
- cannabis nomenclature is often used incorrectly or interchangeably.
- Cannabis literature can be found referring to all cannabis varieties as “sativas” or all cannabinoid-producing plants as "indicas.” Indeed the promiscuous crosses of indoor cannabis breeding programs have made it difficult to distinguish varieties; with most cannabis being sold in the United States, having features of both sativa and indica species.
- Hemp also known as industrial hemp, is a type of cannabis plant grown specifically for the industrial uses of its derived products.
- hemp has been defined as any cannabis plant that has no more than three-tenths of one percent (i.e., 0.3%) concentration of THC.
- Several European countries similarly define hemp as a cannabis plant that has no more than two- tenths of one percent (i.e., 0.2%) concentration of THC.
- Hemp lines with the requisite low quantities of THC have traditionally been produced by selectively breeding plants to either not express the THCA synthase gene, or alternatively, to not produce cannabinoids at all. These breeding approaches result in hemp plants with low cannabinoid diversity, and reduced medicinal properties. Moreover, many hemp plants with null or unexpressed THCA synthase enzymes continue to produce low levels of THC, which can sometimes exceed legal limits if permitted to go past maturity, resulting in total loss of a crop. The Colorado Department of Agriculture for example, requires growers to destroy their entire crop, if even one plant tests higher than 0.3% (8 CCR 1203-23 5.53).
- plants, plant parts, plant tissues and plant cells are produced to contain novel and useful combinations of cannabinoids with improved recreational and medicinal effects.
- the Specialty Cannabis plants, plant parts, plant tissues and plant cells of the present disclosure comprise no more than 0.3% THC content, while also accumulating propyl cannabinoids.
- the present disclosure teaches, a cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part, tissue, or cell thereof, which is capable of producing a female inflorescence, said inflorescence comprising: a) a functional B D allele; b) a propyl cannabinoid max content of at least 1.0% by weight; c) a tetrahydrocannabinol (THC max) content of no more than 0.3% by weight, wherein the contents of all cannabinoids are measured by high performance liquid chromatography (HPLC) and calculated based on dry weight of the inflorescence.
- HPLC high performance liquid chromatography
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part, tissue, or cell thereof of the present disclosure is represented by a representative sample of seed producing said plant that has been deposited under NCIMB Nos. 43258, 43259, and 43260.
- the present disclosure teaches a terpene producing, diploid cannabis hemp plant cell from a female inflorescence (i) a cannabis hemp plant, (ii) an asexual clone of the plant, or (iii) a part of the plant, wherein said cannabis hemp plant, asexual clone of the plant or part of the plant produces the female inflorescence, said inflorescence comprising: a) a functional B D allele; b) a propyl cannabinoid max content of at least 1.0% by weight; c) a tetrahydrocannabinol (THC max) content of no more than 0.3% by weight, wherein the contents of all cannabinoids are measured by high performance liquid chromatography (HPLC) and calculated based on dry weight of the inflorescence; wherein and wherein samples of seed that produce plants comprising a), b), and c) have been deposited under NCIMB Nos. 43258, 432
- the present disclosure teaches a dry sinsemilla cannabis inflorescence comprising: a) a B D allele; b) a propyl cannabinoid max content of at least 1.0% by weight; c) a tetrahydrocannabinol (THC max) content of no more than 0.3% by weight, wherein the contents of all cannabinoids are measured by high performance liquid chromatography (HPLC) and calculated based on dry weight of the inflorescence; wherein and wherein samples of seed that produce plants comprising a), b), and c) are obtainable from seed deposited under NCIMB Nos. 43258, 43259, and 43260.
- HPLC high performance liquid chromatography
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof does not comprise a functional ⁇ allele.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprises a B D /B D genotype.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprises a BO/B D genotype.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprises a terpene oil content greater than about 1.0% by weight; wherein the terpene oil content is the additive content of terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene as measured by GC-FID and calculated based on dry weight of the inflorescence.
- the terpene oil content is the additive content of terpinolene, alpha phellandrene, beta ocimene, carene,
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprises a terpene oil content greater than about 1.5% by weight.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprises a terpene oil content greater than about 2.0% by weight.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprises a propyl cannabinoid max content of at least 2% by weight.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprises a propyl cannabinoid max content of at least 3% by weight.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprises a THC max content of no more than 0.2% by weight.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprises a THC max content of no more than 0.1% by weight.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprises a THC max content of no more than 0.01% by weight.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprises a THC max content of no more than 0.00% by weight.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprises a Terpene Profile in which myrcene is not the dominant terpene; wherein the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene.
- the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is terpinolene.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is alpha phellandrene.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is beta ocimene.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is carene.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is limonene.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is gamma terpinene.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is alpha pinene.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is alpha terpinene.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is beta pinene.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is fenchol.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is camphene.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is alpha terpineol.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is alpha humulene.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is beta caryophyllene.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is linalool.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile wherein the first or second most abundant terpene in the Terpene Profile is caryophyllene oxide.
- the cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part (e.g., inflorescence), tissue, or cell thereof comprise a Terpene Profile in which myrcene is the first or second most abundant terpene in the Terpene Profile; wherein the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene.
- the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene
- the present disclosure teaches a method of producing a cannabis extract, said method comprising the steps of: contacting the inflorescence of Specialty Cannabis hemp with a solvent, thereby producing a cannabis extract.
- the method of producing a cannabis extract comprises heating said extract, thereby decarboxylating at least 70% of the cannabinoid content of the extract.
- the method of producing a cannabis extract comprises winterizing said extract.
- the present disclosure teaches a cannabis extract from the Specialty Cannabis hemp plants of the present disclosure.
- the cannabis extract is selected from the group consisting of kief, hashish, bubble hash, solvent reduced oils, sludges, e-juice, and tinctures.
- the present disclosure teaches a cannabis extract comprising greater than 10% propyl cannabinoid max content, greater than 10% terpene oil content, and less than 1% THC max content as measured by HPLC and based on weight of the extract.
- the present disclosure teaches a method of breeding cannabis hemp plants with high propyl cannabinoid content max and low THC max content, said method comprising: (i) making a cross between a first cannabis hemp plant of the Specialty Cannabis hemp, and a second cannabis plant to produce an Fl plant (ii) harvesting the resulting seed; (iii) growing said seed; and (iv) selecting for high propyl cannabinoid content max and low THC max content; wherein the resulting selected cannabis hemp plant comprises at least 1.0% propyl cannabinoid max content by weight, and no more than 0.3% THC max content by weight.
- the present disclosure teaches a method of producing cannabis hemp plants with high propyl cannabinoid content max and low THC max content, said method comprising: (i) obtaining a cannabis seed, or cutting from a first cannabis hemp plant of any one of the Specialty Cannabis hemp of the present disclosure, (ii) placing said cannabis seed or cutting in an environment conducive to plant growth; (iii) allowing said cannabis seed or cutting to produce a new cannabis plant; (iv) selecting for high propyl cannabinoid content max and low THC max content; wherein the resulting selected cannabis hemp plant is comprises at least 1.0% propyl cannabinoid max content by weight, and no more than 0.3% THC max content by weight.
- the present disclosure teaches a cannabis hemp female inflorescence, said inflorescence comprising: a) a functional B D allele; b) a propyl cannabinoid max content of at least 1.0% by weight; c) a tetrahydrocannabinol (THC max) content of no more than 0.3% by weight, wherein the contents of all cannabinoids are measured by high performance liquid chromatography (HPLC) and calculated based on dry weight of the inflorescence.
- HPLC high performance liquid chromatography
- the present disclosure teaches a cannabis hemp female inflorescence, said inflorescence comprising: a) a functional B D allele; b) a propyl cannabinoid max content of at least 1.0% by weight; c) a tetrahydrocannabinol (THC max) content of no more than 0.3% by weight, wherein the contents of all cannabinoids are measured by high performance liquid chromatography (HPLC) and calculated based on dry weight of the inflorescence, wherein a representative sample of seed producing plants with said inflorescence has been deposited under NCIMB Nos. 43258, 43259, and 43260.
- HPLC high performance liquid chromatography
- the present disclosure teaches a terpene producing, diploid cannabis hemp plant cell from a female inflorescence (i) a cannabis hemp plant, (ii) an asexual clone of the plant, or (iii) a part of the plant, wherein said cannabis hemp plant, asexual clone of the plant or part of the plant produces the female inflorescence, said inflorescence comprising: a) a functional B D allele; b) a propyl cannabinoid max content of at least 1.0% by weight; c) a tetrahydrocannabinol (THC max) content of no more than 0.3% by weight, wherein the contents of all cannabinoids are measured by high performance liquid chromatography (HPLC) and calculated based on dry weight of the inflorescence; wherein and wherein samples of seed that produce plants comprising a), b), and c) have been deposited under NCIMB Nos. 43258, 432
- the present disclosure teaches a dry, non-viable (i) cannabis hemp plant or (ii) part thereof, wherein said cannabis hemp plant or part thereof, comprises at least a portion of a female inflorescence, said inflorescence comprising: a) a B D allele; b) a propyl cannabinoid max content of at least 1.0% by weight; c) a tetrahydrocannabinol (THC max) content of no more than 0.3% by weight, wherein the contents of all cannabinoids are measured by high performance liquid chromatography (HPLC) and calculated based on dry weight of the inflorescence, wherein samples of seed that produce plants comprising a), b), and c) have been deposited under NCIMB Nos. 43258, 43259, and 43260.
- HPLC high performance liquid chromatography
- the present disclosure teaches a composition
- a composition comprising: a) a propyl cannabinoid max content of at least 20% by weight; b) a cannabidiol (CBD max) content of at least 10% by weight; and c) a tetrahydrocannabinol (THC max) content of no more than 10% by weight; wherein the contents of all cannabinoids are measured by high performance liquid chromatography (HPLC) and calculated based on weight of the composition.
- HPLC high performance liquid chromatography
- the present disclosure teaches a composition comprising a terpene oil content greater than about 10% by weight; wherein the terpene oil content is the additive content of terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene as measured by GC-FID and calculated based on weight of the composition.
- the terpene oil content is the additive content of terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fen
- the present disclosure teaches a composition, wherein the composition comprises a terpene oil content greater than about 15% by weight.
- the present disclosure teaches a composition, wherein the composition comprises a terpene oil content greater than about 20.0% by weight.
- the present disclosure teaches a composition, wherein the composition comprises a propyl cannabinoid max content of at least 30% by weight. [0061 ] In some embodiments, the present disclosure teaches a composition, wherein the composition comprises a propyl cannabinoid max content of at least 40% by weight.
- the present disclosure teaches a composition, wherein the composition comprises a propyl cannabinoid max content of at least 50% by weight.
- the present disclosure teaches a composition, wherein the composition comprises a propyl THC max content of no more than 0.5% by weight.
- the present disclosure teaches a composition, wherein the composition comprises a propyl THC max content of no more than 0.3% by weight.
- the present disclosure teaches a composition, wherein the composition comprises a propyl THC max content of no more than 0.2% by weight.
- the present disclosure teaches a composition, wherein the composition comprises a Terpene Profile in which myrcene is not the dominant terpene; wherein the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene.
- the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is terpinolene.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is alpha phellandrene.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is beta ocimene.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is carene.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is limonene.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is gamma terpinene. [ 0073] In some embodiments, the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is alpha pinene.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is alpha terpinene.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is beta pinene.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is fenchol.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is camphene.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is alpha terpineol.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is alpha humulene.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is beta caryophyllene.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is linalool.
- the present disclosure teaches a composition, wherein the first or second most abundant terpene in the Terpene Profile is caryophyllene oxide.
- the present disclosure teaches a composition, wherein the composition comprises a Terpene Profile in which myrcene is the first or second most abundant terpene in the Terpene Profile; wherein the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene.
- the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene
- Figure 1A-B Depicts the current model biosynthetic pathway for several major cannabinoids.
- Figure 1A- Geranyl pyrophosphate (GPP) and olivetolic acid (OA) are condensed by the geranyl pyrophosphate olivetolate geranyl transferase (GOT) to form cannabigerolic acid (CBGA).
- GPP and divarinic acid are condensed by GOT to form cannabigerovarinic acid (CBGVA).
- Figure IB- CBGA or CBGVA is transformed to: (1) THCA/THCVA by THCA synthase, (2) CBCA/CBCVA by CBCA synthase, or (3) CBDA/CBDVA by CBDA synthase.
- Figure 2 Depicts a sample questionnaire used for volunteer trials of Specialty Cannabis and cannabinoid compositions of the present disclosure. This questionnaire will be provided to volunteers with each cannabis blend sample or cannabinoid composition to measure the effects of the sample when administered.
- Figure 3 Depicts the breeding scheme for the O3.S2.01xO9.S1.01' high propyl cannabinoid Specialty Cannabis hemp line.
- Lines THV01 and CBD05.S1-P24 were crossed to produce Fl line V24.
- V24 was selfed to produce sibling lines V24.S1.N5 and V24.S1.03.
- Line V24.S1.03 was selfed to produce O3.S2.01, which was used in a later cross.
- Lines V24.S1.N5 and V24.S1.03 were crossed to produce Fl line O3.N5.09. O3.N5.09 was then selfed to produce 09. SI.01. Line 09.
- THC and total propyl cannabinoid content of the progeny and parental lines is provided in parenthesis under each named plant as a weight percentage based on the dry weight of the inflorescence. This breeding scheme produced novel plants with THC content below 0.3% and high propyl cannabinoid content.
- FIG. 4 Depicts the breeding scheme for the '03. S2.16x09. S 1.01' high propyl cannabinoid Specialty Cannabis hemp line. Lines THV01 and CBD05.S1-P24 were crossed to produce Fl line V24. V24 was selfed to produce sibling lines V24.S1.N5 and V24.S1.03. Line V24. S 1.03 was selfed to produce 03. S2.16, which was used in a later cross. Lines V24. S 1.N5 and V24.S1.03 were crossed to produce Fl line O3.N5.09. O3.N5.09 was then selfed to produce 09. SI.01. Line 09.
- V24 was selfed to produce sibling lines V24.S1.N5 and V24.S1.03. Lines V24.S1.N5 and V24.S1.03 were crossed to produce Fl sibling lines O3.N5.09 and 03.N5.12. O3.N5.09 was then selfed to produce O9.S1.01. Line O3.N5.09 was also crossed with 03.N5.12 to produce line 012.09.10. Line 09. SI.01 was then crossed to 012.09.10 to produce final progeny line O12.09.10x09.S1.01. The THC and total propyl cannabinoid content of the progeny and parental lines is provided in parenthesis under each named plant as a weight percentage based on the dry weight of the inflorescence. This breeding scheme produced novel plants with THC content below 0.3% and high propyl cannabinoid content.
- FIG. 6 Depicts the breeding scheme for the 'V24.Sl.P09xO9.S1.01 high propyl cannabinoid Specialty Cannabis hemp line.
- Lines THV01 and CBD05.S1-P24 were crossed to produce Fl line V24.
- V24 was selfed to produce sibling lines V24.S1.P09, V24.S1.N5, and V24.S1.03.
- Lines V24.S1.N5 and V24.S1.03 were crossed to produce Fl line O3.N5.09.
- O3.N5.09 was then selfed to produce O9.S1.01.
- Line O9.S1.01 was then crossed to V24.S1.P09 to produce final progeny V24.Sl.P09xO9.S1.01.
- the THC and total propyl cannabinoid content of the progeny and parental lines is provided in parenthesis under each named plant as a weight percentage based on the dry weight of the inflorescence.
- This breeding scheme produced novel plants with THC content below 0.3% and high propyl cannabinoid content.
- FIG. 7 Depicts the breeding scheme for the 'V24.S2.26xO9.S1.01 ' high propyl cannabinoid Specialty Cannabis hemp line.
- Lines THVOl and CBD05.S1-P24 were crossed to produce Fl line V24.
- V24 was selfed to produce sibling lines V24.S1.24, V24.S1.N5, and V24.S1.03.
- V24.S1.24 was separately selfed to produce V24.S2.26.
- Lines V24.S1.N5 and V24.S1.03 were crossed to produce Fl line O3.N5.09. O3.N5.09 was then selfed to produce O9.S1.01.
- Line O9.S1.01 was then crossed to V24.S2.26 to produce final progeny V24.S2.26xO9.S1.01 line.
- the THC and total propyl cannabinoid content of the progeny and parental lines is provided in parenthesis under each named plant as a weight percentage based on the dry weight of the inflorescence.
- This breeding scheme produced novel plants with THC content below 0.3% and high propyl cannabinoid content.
- Figure 8. Depicts the breeding scheme for the 'O9.S1.01xO9.S1.01' high propyl cannabinoid Specialty Cannabis hemp line. Lines THV01 and CBD05.S1-P24 were crossed to produce Fl line V24.
- V24 was selfed to produce sibling lines V24.S1.N5 and V24.S1.03. Lines V24.S1.N5 and V24.S1.03 were crossed to produce Fl line O3.N5.09. O3.N5.09 was then selfed to produce 09. S 1.01. Line 09. SI.01 was then selfed again to produce final progeny 09. SI.01x09. SI.01 line.
- the THC and total propyl cannabinoid content of the progeny and parental lines is provided in parenthesis under each named plant as a weight percentage based on the dry weight of the inflorescence. This breeding scheme produced novel plants with THC content below 0.3% and high propyl cannabinoid content
- the term "about” refers to plus or minus 10% of the referenced number.
- reference to an absolute content of a particular terpene of "about 1%” means that that terpene can be present at any amount ranging from 0.9% to 1.1% content by weight.
- the instant specification will often refer to content by weight of various compounds and mixtures (e.g., individual volatiles, terpenes and cannabinoids, or defined sets thereof, such as terpene oil content, cannabinoid content, propyl cannabinoid content).
- Persons having skill in the art will understand the context under which the weight content is being used, and will thus recognize the appropriate frame (i.e. denominator) to use when expressing such a content.
- weight contents from cannabis inflorescences are reported in terms of dry weight of the inflorescence (i.e. by calculating weight of the compound of interest divided by dry weight of inflorescence, multiplied by 100).
- Weight contents of extracts or other compositions is based on the weight of the extract or composition, respectively (i.e., by calculating weight of the compound of interest divided by weight of the composition, multiplied by 100).
- the content of a compound or mixture in an extract or edible will be expressed in terms of absolute weight (e.g., a brownie with 500 mg of THC).
- the invention provides cannabis plants.
- plant refers to plants in the genus of Cannabis and plants derived thereof. Such as cannabis plants produced via asexual reproduction, tissue culture, and via seed production.
- plant part refers to any part of a plant including but not limited to the embryo, shoot, root, stem, seed, stipule, leaf, petal, flower, inflorescence, bud, ovule, bract, trichome, branch, petiole, internode, bark, pubescence, tiller, rhizome, frond, blade, ovule, pollen, stamen, and the like.
- Plant parts may also include certain extracts such as kief or hash, which includes cannabis trichomes or glands. In some embodiments, plant part should also be interpreted as referring to individual cells derived from the plant.
- plant cell refers to any totipotent plant cell from a cannabis plant.
- Plant cells of the present disclosure include cells from a cannabis plant shoot, root, stem, seed, stipule, leaf, petal, inflorescence, bud, ovule, bract, trichome, petiole, internode.
- the disclosed plant cell is from a cannabis trichome.
- the term dominant refers to a terpene that is the most abundant in the Terpene Profile either in absolute content as a percentage by dry weight, or in relative content as a percentage of the Terpene Profile.
- a or “an” refers to one or more of that entity; for example, "a gene” refers to one or more genes or at least one gene.
- a gene refers to one or more genes or at least one gene.
- the terms “a” (or “an”), “one or more” and “at least one” are used interchangeably herein.
- reference to “an element” by the indefinite article “a” or “an” does not exclude the possibility that more than one of the elements is present, unless the context clearly requires that there is one and only one of the elements.
- the term a plant may refer to more than one plants.
- a "landrace” refers to a local variety of a domesticated plant species that has developed largely by natural processes, by adaptation to the natural and cultural environment in which it lives. The development of a landrace may also involve some selection by humans but it differs from a formal breed that has been selectively bred deliberately to conform to a particular formal, purebred standard of traits.
- the International Code of Zoological Nomenclature defines rank, in the nomenclatural sense, as the level, for nomenclatural purposes, of a taxon in a taxonomic hierarchy (e.g., all families are for nomenclatural purposes at the same rank, which lies between superfamily and subfamily). While somewhat arbitrary, there are seven main ranks defined by the international nomenclature codes: kingdom, phylum/division, class, order, family, genus, and species. Further taxonomic hierarchies used in this invention are described below.
- the invention provides plant cultivars.
- the term “cultivar” means a group of similar plants that by structural features and performance (i.e., morphological and physiological characteristics) can be identified from other varieties within the same species.
- the term “cultivar” variously refers to a variety, strain or race of plant that has been produced by horticultural or agronomic techniques and is not normally found in wild populations. The terms cultivar, variety, strain and race are often used interchangeably by plant breeders, agronomists and farmers.
- variable means a plant grouping within a single botanical taxon of the lowest known rank, which grouping, irrespective of whether the conditions for the grant of a breeder's right are fully met, can be i) defined by the expression of the characteristics resulting from a given genotype or combination of genotypes, ii) distinguished from any other plant grouping by the expression of at least one of the said characteristics and iii) considered as a unit with regard to its suitability for being propagated unchanged.
- the invention provides methods for obtaining plant lines.
- the term "line” is used broadly to include, but is not limited to, a group of plants vegetatively propagated from a single parent plant, via tissue culture techniques or a group of inbred plants which are genetically very similar due to descent from a common parent(s).
- a plant is said to "belong” to a particular line if it (a) is a primary transformant (T0) plant regenerated from material of that line; (b) has a pedigree comprised of a T0 plant of that line; or (c) is genetically very similar due to common ancestry (e.g., via inbreeding or selfing).
- T0 primary transformant
- the term “pedigree” denotes the lineage of a plant, e.g. in terms of the sexual crosses affected such that a gene or a combination of genes, in heterozygous (hemizygous) or homozygous condition, imparts a desired trait to the plant.
- inbreeding refers to the production of offspring via the mating between relatives.
- the plants resulting from the inbreeding process are referred to herein as “inbred plants” or “inbreds.”
- LOQ refers to the limit of quantitation for Gas Chromatography (GC) and High Performance Liquid Chromatography (HPLC) measurements.
- secondary metabolites refers to organic compounds that are not directly involved in the normal growth, development, or reproduction of an organism. In other words, loss of secondary metabolites does not result in immediate death of said organism.
- single allele converted plant refers to those plants that are developed by a plant breeding technique called backcrossing wherein essentially all of the desired morphological and physiological characteristics of an inbred are recovered in addition to the single allele transferred into the inbred via the backcrossing technique.
- sample includes a sample from a plant, a plant part, a plant cell, an extract, or a composition, or from a transmission vector, or a soil, water or air sample.
- the invention provides offspring.
- the term "offspring” refers to any plant resulting as progeny from a vegetative or sexual reproduction from one or more parent plants or descendants thereof.
- an offspring plant may be obtained by cloning or selfing of a parent plant or by crossing two parent plants and include sellings as well as the Fl or F2 or still further generations.
- An F1 is a first-generation offspring produced from parents at least one of which is used for the first time as donor of a trait, while offspring of second generation (F2) or subsequent generations (F3, F4, etc.) are specimens produced from sellings of F1's, F2's etc.
- An F1 may thus be (and usually is) a hybrid resulting from a cross between two true breeding parents (true-breeding is homozygous for a trait), while an F2 may be (and usually is) an offspring resulting from self-pollination of said Fl hybrids.
- the invention provides methods for crossing a first plant with a second plant.
- crossing refers to the process by which the pollen of one flower on one plant is applied (artificially or naturally) to the ovule (stigma) of a flower on another plant.
- Backcrossing is a process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (Fl), back to one of the parents of the hybrid progeny. Backcrossing can be used to introduce one or more single locus conversions from one genetic background into another.
- Fl first generation hybrid
- the present invention provides methods for obtaining plant genotypes comprising recombinant genes.
- genotype refers to the genetic makeup of an individual cell, cell culture, tissue, organism (e.g., a plant), or group of organisms.
- the present invention provides homozygotes.
- homozygote refers to an individual cell or plant having the same alleles at one or more loci.
- the present invention provides homozygous plants.
- homozygous refers to the presence of identical alleles at one or more loci in homologous chromosomal segments.
- the present invention provides hemizygotes.
- hemizygotes or “hemizygous” refers to a cell, tissue, organism or plant in which a gene is present only once in a genotype, as a gene in a haploid cell or organism, a sex-linked gene in the heterogametic sex, or a gene in a segment of chromosome in a diploid cell or organism where its partner segment has been deleted.
- the present invention provides heterozygotes.
- heterozygote and “heterozygous” refer to a diploid or polyploid individual cell or plant having different alleles (forms of a given gene) present at least at one locus.
- the cell or organism is heterozygous for the gene of interest that is under control of the synthetic regulatory element.
- the invention provides self-pollination populations.
- self- crossing means the pollen of one flower on one plant is applied (artificially or naturally) to the ovule (stigma) of the same or a different flower on the same plant.
- the invention provides ovules and pollens of plants.
- ovule refers to the female gametophyte
- polyen means the male gametophyte
- the invention provides methods for obtaining plants comprising recombinant genes through transformation.
- transformation refers to the transfer of nucleic acid (i. e. , a nucleotide polymer) into a cell.
- genetic transformation refers to the transfer and incorporation of DNA, especially recombinant DNA, into a cell.
- the invention provides transformants comprising recombinant genes.
- the term "transformant” refers to a cell, tissue or organism that has undergone transformation.
- the original transformant is designated as “T0” or “T0.”
- Selling the T0 produces a first transformed generation designated as "F1" or "T1.”
- cannabinoid profile refers to the detectable (i.e., "non-trace") cannabinoids present in a sample, such as in cannabis inflorescence material, or a composition.
- references to plants with novel or diverse cannabinoid profiles in this document refers to plants with novel combinations or levels of cannabinoids within a single sample. The level at which cannabinoids can be detected will vary slightly depending on the techniques used, and the cannabinoid being tested. For the purposes of this disclosure, cannabinoid levels below 1.0% will be considered “trace” amounts.
- propyl cannabinoids refers to cannabinoids with propyl (i.e., C3) carbon tails.
- Propyl cannabinoids include, but are not limited to, CBGVA, THCVA, CBDVA, CBCVA, and their decarboxylated variants.
- pentyl cannabinoids refers to cannabinoids with pentyl (i.e., C5) carbon tails.
- Pentyl cannabinoids include, but are not limited to CBGA, THCA, CBDA, CBCA, and their decarboxylated variants.
- propyl cannabinoid content refers to the additive content of the propyl cannabinoids, as measured by dry weight of the inflorescence, or the composition comprising the propyl cannabinoid.
- propyl cannabinoid max content refers to the additive content of the potential decarboxylated propyl cannabinoids (as converted by formulas provided in this disclosure). This term is meant to indicate the quantity of propyl cannabinoid content that would be present if all the propyl cannabinoids were decarboxylated. Unless indicated otherwise, the terms “propyl cannabinoid content” and “propyl cannabinoid max content” are used interchangeably.
- low THC content refers to THC contents of no more than 0.3% THC.
- the present disclosure refers to BT, BD, or B0 alleles.
- BT allele or “ ⁇ allele” refers to a gene coding for a THCA synthase enzyme.
- BD allele or “B D allele” refers to a gene coding for a CBDA synthase enzyme.
- B0 allele or “Bo allele” refers to a gene coding for a null THCA or CBDA synthase enzyme.
- a BT allele containing cannabis plant would be expected to accumulate THCA/THCVA
- a BD allele containing cannabis plant would be expected to accumulate CBDA/CBDVA.
- Plants with no functional ⁇ alleles, such as plants with B D B D or B D B 0 genotypes can, in many instances, accumulate low quantities of THC. This THC accumulation can sometimes go beyond 0.3%, causing the plants to no longer be categorized as hemp. Plants of the present disclosure cure this problem, in part by shifting the flux of THC synthesis to THCV.
- BT, BD, and B0 alleles are detectable through direct sequencing (Onofri et al., 2015 "Sequence heterogeneity of Cannabidiolic- and tetrahydrocannabinolic acid-synthase in Cannabis sativa L. and its relationship with chemical phenotype" Phytochemistry Vol 116 pgs 57-68). Persons having skill in the art can also determine the presence of a BT, BD, or homozygous B0 alleles by studying the cannabinoid profile of the plant. BT alleles result in the accumulation of THCA and/or THCV A, while BD alleles result in the accumulation of CBDA and/or CBDVA.
- genotype at the B allele can be assessed by analyzing the cannabinoid profile of cannabis tissue.
- the term "functional ⁇ allele” or “functional B D allele” refers to an allele that results in the cannabis plant accumulating greater than 1.5% THC max +THCV max or greater than 1.5% CBD max + CBDV max, respectively. Cannabinoid accumulation below this level is typically attributed to residual activity of otherwise “null” alleles.
- references to cannabinoids in a plant, plant part, extract, or composition of the present disclosure should be understood as references to both the acidic and decarboxylated versions of the compound (e.g., THCmax as determined by the conversion guidelines described in this document, and understood by those skilled in the art).
- references to high THC contents of a cannabis plant in this disclosure should be understood as referencing to the combined THC and THCA content.
- THCmax and THC max are interchangeably used in this document. This is true for all other cannabinoids discussed in this document.
- winterizing or “winterization” refers to the process by which plant lipids and waxes are removed from a cannabis extract. Persons have skill in the art will immediately recognize how to winterize an extract. Briefly, winterization is the dissolving the cannabis extract into a polar solvent (most commonly ethanol) at sub-zero temperatures. Doing so separates the waxes and lipids from the oil, forcing them to collect at the top of the mixture for easy filtration/collection. Typically, winterization is conducted by mixing ethanol and hash oil into a container and placing it into a sub-zero freezer.
- a polar solvent most commonly ethanol
- the term "maturity,” “harvest maturity,” or “floral maturity” refers to the developmental stage at which a cannabis plant is ready for harvest. Persons having skill in the art will recognize maturity based on the plant's morphologies. Cannabis plants are considered to be at harvest maturity when fan leaves begin to yellow, and when inflorescences begin to take on a 'frosted' appearance, as trichomes develop on calyxes and lower portions of bracts. If bracts and inflorescent parts turn overly yellow and/or if the 'frosted' appearance is visible from afar, this could indicate the plant is beyond maturity. The color of trichomes can also be used to determine maturity.
- Trichomes from cannabis plants first look small and clear, but gradually enlarge, and progressively become 'milkier' and opaque with continued maturation, finally displaying a desiccated appearance and amber color.
- harvest maturity is defined as the time period between the enlarged clear trichome developmental stage and the opaque/milky trichome developmental stage.
- Amber trichomes in cannabis plants are, in some embodiments, an indication of overly mature trichomes.
- the present disclosure uses the terms "maturity,” “harvest maturity,” and “floral maturity” interchangeably. Unless otherwise noted, all cannabinoid and terpene values of cannabis plants discussed in this document refer to the level of those compounds present in a cannabis inflorescence at harvest maturity.
- Tepene Profile is defined as the absolute and relative values of 17 of the most expressed terpenes in the Specialty Cannabis hemp and compositions of the present disclosure: terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene.
- a survey of the terpene profiles of several cannabis varieties has found that these terpenes express at high enough levels so as to have their own pharmacological effects and also to act in synergy with cannabinoids.
- Tepene Essential Oil or “Terpene Essential Oil Content” refers to the additive contents of all the terpenes in the Terpene Profile, as measured by weight of the dry inflorescence or cannabinoid composition.
- Cannabis is an annual, dioecious, flowering herb. Its leaves are typically palmately compound or digitate, with serrated leaflets. Cannabis normally has imperfect flowers, with staminate "male” and pistillate “female” flowers occurring on separate plants. It is not unusual, however, for individual plants to separately bear both male and female flowers (i.e., have monoecious plants). Although monoecious plants are often referred to as "hermaphrodites,” true hermaphrodites (which are less common in cannabis) bear staminate and pistillate structures on individual flowers, whereas monoecious plants bear male and female flowers at different locations on the same plant.
- the life cycle of cannabis varies with each variety but can be generally summarized into germination, vegetative growth, and reproductive stages. Because of heavy breeding and selection by humans, most cannabis seeds have lost dormancy mechanisms and do not require any pre- treatments or winterization to induce germination (See Clarke, RC et al. "Cannabis: Evolution and Ethnobotany” University of California Press 2013). Seeds placed in viable growth conditions are expected to germinate in about 3 to 7 days. The first true leaves of a cannabis plant contain a single leaflet, with subsequent leaves developing in opposite formation, with increasing number of leaflets. Leaflets can be narrow or broad depending on the morphology of the plant grown. Cannabis plants are normally allowed to grow vegetatively for the first 4 to 8 weeks.
- cannabis plants can grow up to 2.5 inches a day, and are capable of reaching heights of 20 feet or more.
- Indoor growth pruning techniques tend to limit cannabis size through careful pruning of apical or side shoots.
- Cannabis has long been used for drug and industrial purposes, including fiber (hemp), for seed and seed oils, for medicinal purposes, and as a recreational drug.
- Industrial fiber hemp products are made from cannabis plants selected to produce an abundance of fiber.
- hemp varieties of Cannabis have been bred to produce minimal levels of THC, the principal psychoactive constituent responsible for the psychoactivity associated with marijuana.
- Marijuana varieties of Cannabis on the other hand typically refer to plants that have been bred to produce high levels of THC and other secondary metabolites, including other cannabinoids and terpenes.
- marijuana cannabis strains used as a drug and industrial hemp both derive from the Cannabis family and contain trace amounts or more of the psychoactive component tetrahydrocannabinol (THC), they are distinct strains with unique phytochemical compositions and uses. Hemp typically has lower concentrations of THC and higher concentrations of cannabidiol (CBD), which decreases or eliminates the psychoactive effects of the plant.
- CBD cannabidiol
- the legality of industrial hemp varies widely between countries. Some governments regulate the concentration of THC and permit only hemp that is bred with an especially low THC content.
- Industrial hemp can be further subdivided into the category of fiber hemp, and resinous hemp.
- Fiber Hemp is used to make a variety of commercial and industrial products including rope, clothes, food, paper, textiles, plastics, insulation and biofuel.
- the bast fibers can be used to make textiles that are 100% hemp, but they are commonly blended with other organic fibers such as flax, cotton or silk, to make woven fabrics for apparel and furnishings.
- the inner two fibers of the plant are more woody and typically have industrial applications, such as mulch, animal bedding and litter.
- drying hemp oil from the seeds becomes solid and can be used in the manufacture of oil-based paints, in creams as a moisturizing agent, for cooking, and in plastics.
- Hemp seeds have been used in bird feed mix as well. Also, more than 95% of hemp seed sold in the European Union was used in animal and bird feed according to the 2013 research data. Thus, the hemp seed can be used for animal and bird feed.
- Resinous hemp refers to cannabis plants that meet the low THC requirements of hemp regulatory programs described above, but which are bred for the production of non-THC cannabinoids.
- the cannabinoids produced from resinous hemp have become a popular source of medical and dietary supplement products.
- the most popular dietary supplement produced from resinous hemp is CBD oil and related products.
- the first genome sequence of Cannabis which is estimated to be 820 Mb in size, was published in 2011 by a team of Canadian scientists (van Bakel et al, "The draft genome and transcriptome of Cannabis sativa" Genome Biology 12:R102).
- Cannabis varieties will flower without the need for external stimuli, most varieties have an absolute requirement for inductive photoperiods in the form of short days or long nights to induce fertile flowering.
- the first sign of flowering in cannabis is the appearance of undifferentiated flower primordial along the main stem of the nodes. At this stage, the sex of the plants are still not distinguishable. As the flower primordia continue to develop, female (pistillate), and male (staminate) flowers can be distinguished.
- the plants and inflorescences of the present disclosure are seedless, sinsemilla. In some embodiments, the plants and inflorescences of the present disclosure are unpollinated.
- Cannabis plants produce a unique family of terpeno-phenolic compounds called cannabinoids.
- Cannabinoids, terpenoids, and other compounds are secreted by glandular trichomes that occur most abundantly on the floral calyxes and bracts of female plants.
- CBD cannabidiol
- THC ⁇ 9- tetrahydrocannabinol
- Cannabinoids accumulate at the highest levels in the trichomes of cannabis inflorescences. However, cannabinoids have been detected in nearly all cannabis organs (see John K. Hemphill et al, "Cannabinoid Content of Individual Plant Organs From Different Geographical Strains of Cannabis Sativa L.” Journal of Natural Products, Vol 43, No. 1 Jan-Feb, 1980). Applicant has similarly detected terpenes in non-inflorescence parts of cannabis plants. Thus, in some embodiments, the plant cells of the present disclosure are terpene and cannabinoid producing cells.
- Cannabinoids are the most studied group of secondary metabolites in cannabis. Most exist in two forms, as acids and in neutral (decarboxylated) forms.
- the acid form is designated by an “A” at the end of its acronym (i.e. THC A).
- THC A The phytocannabinoids are synthesized in the plant as acid forms, and while some decarboxylation does occur in the plant, it increases significantly post- harvest and the kinetics increase at high temperatures. (Sanchez and Verpoorte 2008).
- the biologically active forms for human consumption are the neutral forms.
- cannabinoids in their acid forms can be converted to their non-acidic forms through a process called decarboxylation.
- Decarboxylation is usually achieved by (optionally) thorough drying of the plant material followed by heating it, often by either combustion, vaporization, or heating or baking in an oven.
- Cannabinoid compositions can similarly be decarboxylated by being exposed to heat.
- the total measured content of acid cannabinoid variants forms should be adjusted to account for the loss of the carboxyl group. In some embodiments, this adjustment can be made by multiplying the molar content of the acidic cannabinoid forms by the molecular weight of the corresponding decarboxylated cannabinoid. Other shorthand conversions are also available for quickly converting acidic cannabinoid content to active cannabinoid content.
- THCmax (THCA ⁇ 0.877) + THC.
- references to cannabinoids in a plant, plant part, extract, or composition of the present disclosure includes both the acidic and decarboxylated versions of the compound (e.g., THCmax as determined by the conversion guidelines described above, and understood by those skilled in the art).
- References to a cannabinoid content (however it is measured) in a claim should be understood as representing theoretical maximums of decarboxylated "active" cannabinoid contents, plus converted contents of acidic versions of the same cannabinoid, unless otherwise indicated.
- the cannabinoids in the Specialty Cannabis plants, plant parts, extracts and compositions of the present disclosure include, but are not limited to, A9-Tetrahydrocannabinol (A9-THC), A8-Tetrahydrocannabinol (A8-THC), Cannabichromene (CBC), Cannabicyclol (CBL), Cannabidiol (CBD), Cannabielsoin (CBE), Cannabigerol (CBG), Cannabinidiol (CBND), Cannabinol (CBN), Cannabitriol (CBT), and their propyl homologs, including, but are not limited to cannabidivarin (CBDV), A9-Tetrahydrocannabivarin (THCV), cannabichromevarin (CBCV), and cannabigerovarin (CBGV), and their acidic variants.
- A9-Tetrahydrocannabinol A9-THC
- Non-THC cannabinoids include one or more of THCV, CBD, CBDV, CBC, CBCV, CBN, CBG, and ⁇ 8THC (a.k.a. D8THC) cannabinoids, and their acidic variants.
- non-THC cannabinoids include propyl THCVA and THCV.
- THC is the principal psychoactive constituent (or cannabinoid) of the cannabis plant.
- the initially synthesized and accumulated form in plant is THC acid (THCA).
- THC has mild to moderate analgesic effects, and cannabis can be used to treat pain by altering transmitter release on dorsal root ganglion of the spinal cord and in the periaqueductal gray. Other effects include relaxation, alteration of visual, auditory, and olfactory senses, fatigue, and appetite stimulation. THC has marked antiemetic properties, and may also reduce aggression in certain subjects (Hoaken (2003). "Drugs of abuse and the elicitation of human aggressive behavior”. Addictive Behaviors 28: 1533-1554).
- THC The pharmacological actions of THC result from its partial agonist activity at the cannabinoid receptor CB1, located mainly in the central nervous system, and the CB2 receptor, mainly expressed in cells of the immune system (Pertwee, 2006, "The pharmacology of cannabinoid receptors and their ligands: An overview”. International Journal of Obesity 30: SI 3- S18.)
- the psychoactive effects of THC are primarily mediated by its activation of CBlG-protein coupled receptors, which result in a decrease in the concentration of the second messenger molecule cAMP through inhibition of adenylate cyclase (Elphick et al., 2001, "The neurobiology and evolution of cannabinoid signaling".
- THC has an anticholinesterase action, which may implicate it as a potential treatment for Alzheimer's and Myasthenia (Eubanks et al., 2006, "A Molecular Link Between the Active Component of Marijuana and Alzheimer's Disease Pathology”. Molecular Pharmaceutics 3 (6): 773-7.)
- THC occurs mainly as tetrahydrocannabinolic acid (THCA, 2-COOH-THC).
- THCA tetrahydrocannabinolic acid
- 2-COOH-THC tetrahydrocannabinolic acid
- Geranyl pyrophosphate and olivetolic acid react, catalyzed by an enzyme to produce cannabigerolic acid, which is cyclized by the enzyme THC acid synthase to give THCA.
- THCA is decarboxylated producing THC.
- the pathway for THCA biosynthesis is similar to that which produces the bitter acid humulone in hops.
- THC variants include:
- CBD is a cannabinoid found in cannabis. Cannabidiol has displayed sedative effects in animal tests (Pickens, 1981, "Sedative activity of cannabis in relation to its delta' -trans- tetrahydrocannabinol and cannabidiol content". Br. J. Pharmacol. 72 (4): 649-56). Some research, however, indicates that CBD can increase alertness, and attenuate the memory-impairing effect of THC.
- CBD reduces growth of aggressive human breast cancer cells in vitro and reduces their invasiveness (McAllister et al., 2007, “Cannabidiol as a novel inhibitor of Id-1 gene expression in aggressive breast cancer cells”. Mol. Cancer Ther. 6 (11): 2921-7.)
- Cannabis produces CBD-carboxylic acid through the same metabolic pathway as THC, until the last step, where CBDA synthase performs catalysis instead of THCA synthase.
- CBDA synthase performs catalysis instead of THCA synthase.
- CBG is a non-psychoactive cannabinoid found in the Cannabis genus of plants.
- Cannabigerol is found in higher concentrations in hemp rather than in varieties of Cannabis cultivated for high THC content and their corresponding psychoactive properties.
- Cannabigerol has been found to act as a high affinity a2-adrenergic receptor agonist, moderate affinity 5-HT1 A receptor antagonist, and low affinity CB1 receptor antagonist. It also binds to the CB2 receptor.
- Cannabigerol has been shown to relieve intraocular pressure, which may be of benefit in the treatment of glaucoma (Craig et al.
- CBG variants include:
- CBN is a mildly to non-psychoactive substance cannabinoid found in Cannabis sativa and Cannabis indica/afghanica. It is also a metabolite of tetrahydrocannabinol (THC). CBN acts as a weak agonist of the CB1 and CB2 receptors, with lower affinity in comparison to THC.
- THC tetrahydrocannabinol
- CBC bears structural similarity to the other natural cannabinoids, including tetrahydrocannabinol, tetrahydrocannabivarin, cannabidiol, and cannabinol, among others. Evidence has suggested that it may play a role in the anti-inflammatory and anti-viral effects of cannabis, and may contribute to the overall analgesic effects of cannabis.
- Non-limiting examples of CBC variants include:
- Cannabivarin also known as cannabivarol or CBV, is a non-psychoactive cannabinoid found in minor amounts in the hemp plant Cannabis sativa. It is an analog of cannabinol (CBN) with the side chain shortened by two methylene bridges (-CH2-). CBV is an oxidation product of tetrahydrocannabivarin (THCV, THV).
- CBDV is a non-psychoactive cannabinoid found in Cannabis. It is a homolog of cannabidiol (CBD), with the side-chain shortened by two methylene bridges (CH2 units). Cannabidivarin has been found reduce the number and severity of seizures in animal models (US Pat Application 13/075,873). Plants with relatively high levels of CBDV have been reported in feral populations of C. indica (
- THCV tetrahydrocannabinol
- THC tetrahydrocannabinol
- THCV tetrahydrocannabinol
- This terpeno-phenolic compound is found naturally in Cannabis, sometimes in significant amounts.
- THCV has been shown to be a CB1 receptor antagonist, i.e. it blocks the effects of THC.
- Tetrahydrocannabinol has been shown to increase metabolism, help weight loss and lower cholesterol in animal models (US Pat Application 11/667,860)
- Cannabicyclol is a non-psychotomimetic cannabinoid found in the Cannabis species.
- CBL is a degradative product like cannabinol. Light converts cannabichromene to CBL.
- Non-limiting examples of CBL variants include:
- Non-limiting examples of CBT variants include:
- Non-limiting examples of CBE variants include:
- cannabinoids synthesis More details of cannabinoids synthesis and the properties and uses of these cannabinoids are described in Russo (2011, Taming THC: potential cannabis synergy and phytocannabinoid- terpenoid entourage effects, British Journal of Pharmacology, 163 : 1344-1364), Russo et al. (2006, A tale of two cannabinoids: the therapeutic rationale for combining tetrahydrocannabinol and cannabidiol, Medical Hypothesis, 2006, 66:234-246), Celia et al.
- phenolic precursors such as geranyl pyrophosphate (GPP) and polyketide, olivetolic acid (OA) are condensed by geranyl pyrophosphate olivetolate geranyl transferase (GOT) to form cannabigerolic acid (CBGA).
- GPP and divarinic acid can be condensed by GOT to form cannabigerovarinic acid (CBGVA).
- CBGA or CBGVA are considered to be the "primary cannabinoids" from which others can be produced.
- CBGA/CBGVA is quickly transformed in plants into, for example: (1) CBCA/CBCVA by CBCA synthase; (2) THCA/THCVA by THCA synthase; or (3) CBDA/CBDVA by CBDA synthase.
- CBCA/CBCVA by CBCA synthase
- THCA/THCVA by THCA synthase
- CBDA/CBDVA by CBDA synthase.
- cannabis plants can be categorized into THC-CBD chemotypes based on the state of the B locus BT BT (THC producing, chemotype I), BD BD (CBD producing, chemotype ⁇ II), and BT/BD (producing both THC and CBD, chemotype II). Additional information on the genetic regulation of cannabinoids can be found in de Meijer et al. I, II, II ⁇ , and IV (I: 2003, Genetics, 163:335-346; II: 2005, Euphytica, 145: 189-198; ⁇ II: 2009, Euphytica, 165:293-311; and IV: 2009, Euphytica, 168:95-112).
- the BT and BD alleles are known, and can be easily detected using methods known to those skilled in the art, including Northerns, PCR, sequencing, or Westerns.
- a representative sequence of THCA synthase is available at GenBank ID AB057805.1.
- a representative sequence of the CBDA synthase is available at GenBank ID AB292682.1.
- the specialty plants and compositions of the present disclosure comprise novel Terpene Profiles.
- Terpenes are a large and diverse class of organic compounds, produced by a variety of plants. They are often strong smelling and thus may have had a protective function.
- Terpenes are derived biosynthetically from units of isoprene, which has the molecular formula C 5 H 8 .
- the basic molecular formulae of terpenes are multiples of ( C 5 H 8 ) n where n is the number of linked isoprene units.
- the isoprene units may be linked together "head to tail" to form linear chains or they may be arranged to form rings.
- Non-limiting examples of terpenes include Hemiterpenes, Monoterpenes, Sesquiterpenes, Diterpenes, Sesterterpenes, Triterpenes, Sesquarterpenes, Tetraterpenes, Polyterpenes, and Norisoprenoids.
- cannabis In addition to cannabinoids, cannabis also produces over 120 different terpenes (Russo 2011, Taming THC: potential cannabis synergy and phytocannabinoid-terpenoid entourage effects, British Journal of Pharmacology, 163 : 1344- 1364). Within the context and verbiage of this document the terms 'terpenoid' and 'terpene' are used interchangeably.
- Cannabinoids are odorless, so terpenoids are responsible for the unique odor of cannabis, and each variety has a slightly different profile that can potentially be used as a tool for identification of different varieties or geographical origins of samples (Hillig 2004. "A chemotaxonomic analysis of terpenoid variation in Cannabis” Biochem System and Ecology 875- 891). Indeed, recent studies have concluded that terpene production in cannabis plants is strongly inherited, and is little influenced by environmental factors. (Casano et al 2011. "Variations in terpene profiles of different strains of Cannabis sativa” Acta Horticulturae 925:115-121).
- Terpenes also provide a unique and complex organoleptic profile for each variety that is appreciated by both novice users and connoisseurs.
- Critical differences between many popular commercial cannabis strains can be largely attributed to differences in terpene profiles, which provide each line with their distinctive aroma and pharmacological effects.
- the popular "cookies" strain of cannabis is noted by its myrcene and limonene.
- myrcene and limonene In addition to many circulatory and muscular effects, some terpenes interact with neurological receptors. A few terpenes produced by cannabis plants also bind weakly to cannabinoid receptors.
- Terpenoids can alter the permeability of cell membranes and allow in either more or less THC, while other terpenes can affect serotonin and dopamine chemistry as neurotransmitters.
- Terpenoids are lipophilic, and can interact with lipid membranes, ion channels, a variety of different receptors (including both G- protein coupled odorant and neurotransmitter receptors), and enzymes. Some are capable of absorption through human skin and passing the blood brain barrier.
- terpenes are considered to be pharmacologically relevant when present in concentrations of at least 0.05% in plant material (Hazekamp and Fischedick 2010. "Metabolic fingerprinting of Cannabis sativa L., cannabinoids and terpenoids for chemotaxonomic and drug standardization purposes" Phytochemistry 2058-73; Russo 2011, Taming THC: potential cannabis synergy and phytocannabinoid-terpenoid entourage effects, British Journal of Pharmacology, 163:1344-1364).
- THC potential cannabis synergy and phytocannabinoid-terpenoid entourage effects
- Terpenoids can be extracted from the plant material by steam distillation (giving you essential oil) or vaporization, however the yield varies greatly by plant tissue, type of extraction, age of material, and other variables (McPartland and Russo 2001 "Cannabis and Cannabis Extracts: Greater Than the Sum of Their Parts?" Hayworth Press).
- the present disclosure teaches methods for extracting cannabinoids and terpenes.
- Other methods for producing reproducible and quantifiable cannabinoid and terpene measurements are known to persons having skill in the art.
- the yield of terpenoids in cannabis inflorescences is less than 2% by weight on analysis; however, it is thought that they may comprise up to 10% of the trichome content. A few of the most recognized terpenes and non-terpene volatiles in cannabis are discussed below.
- D-Limonene also known as limonene
- limonene is a monoterpenoid that is widely distributed in nature and often associated with citrus. It has strong anxiolytic properties in both mice and humans, apparently increasing serotonin and dopamine in mouse brain. D-limonene has potent antidepressant activity when inhaled. It is also under investigation for a variety of different cancer treatments, with some focus on its hepatic metabolite, perillic acid.
- ⁇ -Myrcene also known as myrcene, is a monoterpenoid also found in cannabis, and has a variety of pharmacological effects. It is often associated with a sweet fruit like taste. It reduces inflammation, aids sleep, and blocks hepatic carcinogenesis, as well as acting as an analgesic and muscle relaxant in mice.
- D-Linalool also known as linalool
- linalool is a monoterpenoid with very well-known anxiolytic effects. It is often associated with lavender, and frequented used in aromatherapy for its sedative impact. It acts as a local anesthetic and helps to prevent scarring from burns, is anti-nociceptive in mice, and shows anti-glutamatergic and anticonvulsant activity.
- ⁇ -Pinene is a monoterpene common in nature, also with a plethora of effects on mammals and humans. It acts as an acetylcholinesterase inhibitor, which aids memory and counteracts the short-term memory loss associated with ⁇ 9 -THC intoxication, is an effective antibiotic agent, and shows some activity against MRS A.
- a-pinene is a bronchodilator in humans and has anti-inflammatory properties via the prostaglandin E-l pathway (Russo 2011, Taming THC: potential cannabis synergy and phytocannabinoid-terpenoid entourage effects, British Journal of Pharmacology, 163: 1344-1364).
- ⁇ -Caryophyllene is often the most predominant sesquiterpenoid in cannabis. It is less volatile than the monoterpenoids, thus it is found in higher concentrations in material that has been processed by heat to aid in decarboxylation. It is very interesting in that it is a selective full agonist at the CB 2 receptor, which makes it the only phytocannabinoid found outside the cannabis genus. In addition, it has anti-inflammatory and gastric cytoprotective properties, and may even have antimalarial activity.
- Caryophyllene oxide is another sesquiterpenoid found in cannabis, which has antifungal and anti-platelet aggregation properties. As an aside, it is also the molecule that drug-sniffing dogs are trained to find (Russo 2011, Taming THC: potential cannabis synergy and phytocannabinoid- terpenoid entourage effects, British Journal of Pharmacology, 163: 1344-1364).
- Nerolidol is a sesquiterpene that is often found in citrus peels that exhibits a range of interesting properties. It acts as a sedative, inhibits fungal growth, and has potent anti-malarial and antileishmanial activity. It also alleviated colon adenomas in rats (Russo 2011, Taming THC: potential cannabis synergy and phytocannabinoid-terpenoid entourage effects, British Journal of Pharmacology, 163:1344-1364). Phytol is a diterpene often found in cannabis extracts. It is a degradation product of chlorophyll and tocopherol. It increases GABA expression and therefore could be responsible the relaxing effects of green tea and wild lettuce.
- Table 2- A non-limiting list of the medical effects of some of the most common terpenes found in cannabis
- the absolute cannabinoid and terpene contents of a plant are calculated based on weight of cannabinoid or terpene present in a sample divided by the dried weight of the dried trimmed inflorescence.
- Dried inflorescences refer to harvested inflorescence tissue dried to ⁇ 10% moisture level.
- terpene and cannabinoid contents are further adjusted to account for any remaining moisture content, by removing the weight of any remaining moisture from the measured weight of the inflorescence.
- Moisture content of a flower can be determined by a variety of analytical methods. Persons having skill in the art will be familiar with methods for measuring moisture content.
- the present disclosure teaches the use of FTIR analysis for calculating moisture content of inflorescences.
- the present disclosure teaches the use of additional drying steps in desiccant chambers to calculate remaining moisture contents.
- trimmed inflorescence refers to inflorescences with sun (sugar) leaves cut off such that only the calyx and reproductive buds remain. Trimming can be performed manually, through careful manicuring of harvested tissue, or via automated mechanical methods.
- the present disclosure also teaches methods of pre-screening grown seeds for specific cannabinoid contents.
- the types of cannabinoids produced by a cannabis inflorescence can also be determined in the field via thin layer chromatography (TLC) analysis (see “Cannabis Inflorescence & Leaf QC” from The American Herbal Pharmacopeia 2013).
- the present disclosure will often refer to Specialty Cannabis comprising a selected cannabinoid or terpene content. In some instances, the present disclosure will refer to Specialty Cannabis that produces inflorescences comprising a selected cannabinoid or terpene content. It will be understood that both of these statements are interchangeable, and that references to the cannabinoid or terpene contents of a Specialty Cannabis refer to the contents of the inflorescences those plants produce.
- the present disclosure provides Specialty Cannabis with novel cannabinoid profiles.
- the Specialty Cannabis of the present disclosure qualifies as industrial hemp.
- the presently disclosed inventions are based in part on the instant inventors' discovery that resinous hemp plants could be bred to produce high (non-trace) quantities of propyl cannabinoids, while also accumulating no more than 0.3% THC. The novelty of the presently disclosed invention is discussed in more detail below.
- GW Pharmaceuticals has developed and patented a cannabis variety that does not produce any THC (US 9,035,130), and could thus address the harvest maturity concerns of traditional hemp. This plant however, has been bred to comprise a "cannabinoid knockout factor" that blocks cannabinoid biosynthesis entirely. Thus, the plant patented by GW pharmaceuticals would only be helpful for fiber production, and would not serve the same purpose as the Specialty Cannabis hemp lines of the present disclosure, which also accumulate high levels of cannabinoids, including CBD, CBG, and propyl cannabinoids.
- the Specialty Cannabis plants of the present disclosure address the limitations of traditional resinous hemp lines. Without wishing to be bound by any theory, the present inventors believe that the presently disclosed Specialty Cannabis plant comprise one or more alleles redirecting cannabinoid flux to propyl (C3) cannabinoids. This genetic feature prevents the accumulation of THC, thus ensuring that the Specialty Cannabis plants of the present disclosure qualify as hemp under U.S. and foreign law. This is particularly important for large crops, where harvest can several days, or weeks to complete.
- the Specialty Cannabis hemp lines of the present disclosure solve this problem, reducing production costs, and ensuring that the grower will remain within the law.
- the presently disclosed Specialty Cannabis lines are also beneficial to consumers, who now gain access to additional non-THC cannabinoids in their hemp products.
- the Specialty Cannabis plants of the present disclosure represent a new category of cannabis plants in which high (non-trace) levels of propyl cannabinoids accumulate with no more than 0.3% THC.
- the Specialty Cannabis plants of the present disclosure accumulate no detectable THC content (e.g., less than 0.01% by weight, as measured by HPLC).
- the Specialty Cannabis plants of the present disclosure solve the problems of previously existing hemp lines.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, or 60% total cannabinoids by weight of the dried inflorescence, and all ranges therebetween.
- the Specialty Cannabis of the present disclosure comprise 1-5%, 1-10%, 1-40%, 1-30%, or
- the Specialty Cannabis of the present disclosure produce inflorescences comprising more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, or 60% total cannabinoids by weight of the dried inflorescence while accumulating no more than 0.3% or 0.2% THC content by weight.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% propyl cannabinoids by weight, and all ranges therebetween.
- the Specialty Cannabis of the present disclosure comprise 2%-10%, 3%-30%, or 3%-25% propyl cannabinoids content by weight of the dried inflorescence.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% propyl cannabinoids by weight of the dried inflorescence while accumulating no more than 0.3% or 0.2% THC content by weight.
- the Specialty Cannabis of the present disclosure qualify as industrial hemp under relevant U.S. and European regulations. That is, in some embodiments, the Specialty Cannabis of the present disclosure accumulate no more than 0.3%, or less than 0.2% THC content.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising about 0.00%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, or 0.30% THC by weight of the dried inflorescence, and all ranges therebetween.
- the Specialty Cannabis of the present disclosure comprise 0.00%-0.10%, 0.00%-0.20%, or 0.00%- 0.30% THC content by weight of the dried inflorescence.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising no more than about 0.00%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, or 0.30% THC by weight of the dried inflorescence, while accumulating at least 1% non-THC cannabinoid content by weight.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising no more than about 0.00%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, or 0.30% THC by weight of the dried inflorescence, at any stage in the plant's growth. That is, in some embodiments, the Specialty Cannabis hemp lines of the present disclosure never accumulate greater than 0.3%, or greater than 0.2% THC, even if allowed to develop past harvest maturity.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% CBD by weight of the dried inflorescence, and all ranges therebetween.
- the Specialty Cannabis of the present disclosure comprise 3%-40%, 3%-30%, or 3%-25% CBD content by weight of the dried inflorescence.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% CBD by weight of the dried inflorescence while accumulating no more than 0.3% or 0.2% THC content by weight.
- the Specialty Cannabis of the present disclosure accumulates CBC.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% CBC by weight of the dried inflorescence, and all ranges therebetween.
- the Specialty Cannabis of the present disclosure comprise 3%-40%, 3%-30%, or 3%-25% CBC content by weight of the dried inflorescence.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% CBC by weight of the dried inflorescence while accumulating no more than 0.3% or 0.2% THC content by weight.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% THCV by weight of the dried inflorescence, and all ranges therebetween.
- the Specialty Cannabis of the present disclosure comprise 3%-40%, 3%-30%, or 3%-25% THCV content by weight of the dried inflorescence.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% THCV by weight of the dried inflorescence of the dried inflorescence while accumulating no more than 0.3% or 0.2% THC content by weight.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% CBDV by weight of the dried inflorescence, and all ranges therebetween.
- the Specialty Cannabis of the present disclosure comprise 3%-40%, 3%-30%, or 3%-25% CBDV content by weight of the dried inflorescence.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% CBDV by weight of the dried inflorescence while accumulating no more than 0.3% or 0.2% THC content by weight.
- the Specialty Cannabis of the present disclosure accumulates CBCV.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% CBCV by weight of the dried inflorescence, and all ranges therebetween.
- the Specialty Cannabis of the present disclosure comprise 3%-40%, 3%-30%, or 3%-25% CBCV content by weight of the dried inflorescence.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40% CBCV by weight of the dried inflorescence while accumulating no more than 0.3% or 0.2% THC content by weight.
- the present invention teaches the preference for cannabis plant material with novel Terpene Profiles.
- the Specialty Cannabis of the present disclosure produce inflorescences comprising organoleptically pleasing Terpene Profiles.
- the Specialty Cannabis of the present invention has an absolute content of any one of the 17 terpenes in the Terpene Profile as set forth in Table 3 that is 0%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.4%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.5%, 0.51%, 0.52%, 0.53%,
- the absolute content of any one of the terpenes is between about 0.05% and about 0.85%.
- This paragraph is intended to be read as applying to any specific terpene(s) in a Terpene Profile, such that the name of any one or two or more of these terpenes as specifically referred to elsewhere herein (e.g., linalool) can replace the phrase "any one of the 17 terpenes in the Terpene Profile.”
- the Specialty Cannabis of the present invention has an absolute content of any one of the 17 terpenes in the Terpene Profile that is greater than 0%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.4%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.5%, 0.51%, 0.52%, 0.53%, 0.54%,
- a limonene dominant terpene is used to refer to Terpene Profiles in which limonene is the most abundant terpene in the Terpene Profile (i.e., limonene relative or absolute content is > content of any single one of the 16 other terpenes in the Terpene Profile). Reference to other dominant terpenes is similarly based on said terpene being the most abundant within the Terpene Profile.
- the Specialty Cannabis of the present invention has 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%
- the essential oil content of the Specialty Cannabis varieties of the present invention is between about 0.5% and about 8% by dry weight. In other embodiments the essential oil contents of the Specialty Cannabis varieties of the present invention is between about 1.0% and about 5% by dry weight.
- the Specialty Cannabis of the present invention has greater than 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%,
- the terpene content of the Specialty Cannabis of the present disclosure is described in relative terms as a percentage composition of the total Terpene Profile.
- a Specialty Cannabis with 1.2% absolute terpinolene content and 1.2% limonene content and no other terpenes in the Terpene Profile would said to have 50% terpinolene and 50% limonene relative content.
- the Specialty Cannabis of the present invention has a relative content of any one of the 17 terpenes in the Terpene Profile that is greater than or less than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 7
- the relative content of any one of the terpenes is between 0% and 100%.
- This paragraph is intended to be read as applying to any specific terpene(s) in a Terpene Profile, such that the name of any of one or two or more these terpenes as specifically referred to elsewhere herein (e.g., linalool) can replace the phrase "any one of the 17 terpenes in the Terpene Profile.”
- the Specialty Cannabis of the present disclosure produce female inflorescences. In some embodiments, the Specialty Cannabis of the present disclosure have been feminized to produce female seed. In some embodiments, the supporting seed deposits referenced in the present disclosure are feminized. Persons having skill in the art will be familiar with techniques to feminize cannabis seeds, including breeding through treatment with silver thiosulfate, colloidal silver, hormones, and rodelization method.
- Another important breeding phenotype is flower color.
- the accumulation of anthocyanins, carotenoids, or other color-producing compounds in leaves and flowers of cannabis can have an effect on consumer visual appeal and flavor. Iconic examples of the appeal of color are the popular "Purple Kush”, “Purple Haze”, and “Purple Trainwreck” varieties that express anthocyanins in their late maturation stages to produce dark purple leaves. Color selections can also be based on (but not limited to) unique coloration of stem, leaf, inflorescence, calyx, stamen, trichome bodies and finished products including extracts and hash.
- Cannabis yield is another important factor in breeding.
- Cannabis yield is measured by pounds (lbs), grams (g) or kilograms (Kg) of dried (e.g., -10% moisture) trimmed flowers. Yield can be expressed in terms of yield per plant, yield per watt of light, and yield per square meter of growing area among others. Cannabis yield is also dependent on the growing environment. For example, yields for a particular cannabis strain will vary between outdoor growth long season, outdoor growth short season, or indoor growth. Yield may also be affected by growing conditions such as type of lighting, soil, fertilizer use, size of growing pot, etc.
- the Specialty Cannabis of the present disclosure produce .lg, .2g, .3g, .4g, .5g, .6g, .7g, .8g, .9g, l.Og, l.lg, 1.2g, 1.3g, 1.4g, 1.5g, 1.6g, 1.7g, 1.8g, 1.9g, 2.0g, 2.1g, 2.2g, 2.3g, 2.4g, 2.5g, 2.6g, 2.7g, 2.8g, 2.9g, 3.0g, 3.1g, 3.2g, 3.3g, 3.4g, 3.5g, 3.6g, 3.7g, 3.8g, 3.9g, 4.0g, 4.1g, 4.2g, 4.3g, 4.4g, 4.5g, 4.6g, 4.7g, 4.8g, 4.9g, or 5.0g of dried flowers per watt of light, including all ranges therebetween.
- the Specialty Cannabis of the present invention produces 10g, 15g, 20g, 25g, 30g, 35g, 40g, 45g, 50g, 55g, 60g, 65g, 70g, 75g, 80g, 85g, 90g, 95g, 100g, 105g, 110g, 115g, 120g, 125g, 130g, 135g, 140g, 145g, 150g, 155g, 160g, 165g, 170g, 175g, 180g, 185g, 190g, 195g, 200g, 210g, 220g, 230g, 240g, 250g, 260g, 270g, 280g, 290g, 300g, 310g, 320g, 330g, 340g, 350g, 360g, 370g, 380g, 390g, 400g, 410g, 420g, 430g, 440g, 450g, 460g, 470g, 480g, 490g,
- Root Structure Positive root selection is marked by overall root vigor and adventitious root growth, ease of transplant, rate of root development on clonal propagations, and root shooting from tissue culture samples. Root selections can also be based on resistance to soil and hydroponic pathogens including Pythium.
- Vigor - Selection for plant vigor are marked by tremendous grow rates and robust stem/stalk infrastructure. Often times, selection display morphologies that are very much enlarged compared to sibling progeny.
- Marijuana Botany An Advanced study: The Propagation and Breeding of Distinctive Cannabis by Robert Connell Clarke.
- the present invention also relates to variants, mutants and modifications of the seeds, plant parts and/or whole plants of the cannabis plants of the present invention.
- Variants, mutants and trivial modifications of the seeds, plants, plant parts, plant cells of the present invention can be generated by methods well known and available to one skilled in the art, including but not limited to, mutagenesis (e.g., chemical mutagenesis, radiation mutagenesis, transposon mutagenesis, insertional mutagenesis, signature tagged mutagenesis, site-directed mutagenesis, and natural mutagenesis), knock-outs/knock-ins, antisense and RNA interference.
- mutagenesis e.g., chemical mutagenesis, radiation mutagenesis, transposon mutagenesis, insertional mutagenesis, signature tagged mutagenesis, site-directed mutagenesis, and natural mutagenesis
- knock-outs/knock-ins antisense and RNA interference.
- the present invention also relates to a mutagenized population of the cannabis plants of the present invention, and methods of using such populations.
- the mutagenized population can be used in screening for new cannabis lines that comprise one or more or all of the morphological, physiological, biological, and/or chemical characteristics of cannabis plants of the present invention.
- the new cannabis plants obtained from the screening process comprise one or more or all of the morphological, physiological, biological, and/or chemical characteristics of cannabis plants of the present invention, and one or more additional or different new morphological, physiological, biological, and/or chemical characteristic.
- the mutagenized population of the present invention can be used in Targeting Induced Local Lesions in Genomes ⁇ LLING) screening method, which combines a standard and efficient technique of mutagenesis with a chemical mutagen (e.g., Ethyl methanesulfonate (EMS)) with a sensitive DNA screening-technique that identifies single base mutations (also called point mutations) in a target gene.
- EMS Ethyl methanesulfonate
- TELLING® Detailed description on methods and compositions on TELLING® can be found in Till et al. (Discovery of induced point mutations in maize genes by TILLING, BMC Plant Biology 2004, 4:12), Weil et al., (TILLING in Grass Species, Plant Physiology January 2009 vol. 149 no.
- the plants of the present invention can be used to produce new plant varieties. In some embodiments, the plants are used to develop new, unique and superior varieties or hybrids with desired phenotypes.
- selection methods e.g., molecular marker assisted selection
- breeding methods can be combined with breeding methods to accelerate the process. Additional breeding methods have been known to one of ordinary skill in the art, e.g., methods discussed in Chahal and Gosal (Principles and procedures of plant breeding: biotechnological and conventional approaches, CRC Press, 2002, ISBN 084931321X, 9780849313219), Taji et al.
- Cannabis genome has been sequenced recently (van Bakel et al., The draft genome and transcriptome of Cannabis sativa, Genome Biology, 12(10):R102, 2011). Molecular makers for cannabis plants are described in Datwyler et al.
- molecular markers are designed and made, based on the genome of the plants of the present application.
- the molecular markers are selected from Isozyme Electrophoresis, Restriction Fragment Length Polymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP- PCR), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs). Amplified Fragment Length Polymorphisms (AFLPs), and Simple Sequence Repeats (SSRs) which are also referred to as Microsatellites, etc.
- RFLPs Restriction Fragment Length Polymorphisms
- RAPDs Randomly Amplified Polymorphic DNAs
- AP- PCR Arbitrarily Primed Polymerase Chain Reaction
- DAF Sequence Characterized Amplified Regions
- SCARs Sequence Characterized Amplified Regions
- the molecular markers can be used in molecular marker assisted breeding.
- the molecular markers can be utilized to monitor the transfer of the genetic material.
- the transferred genetic material is a gene of interest, such as genes that contribute to one or more favorable agronomic phenotypes when expressed in a plant cell, a plant part, or a plant.
- said method comprises (i) crossing any one of the plants of the present invention comprising the expression cassette as a donor to a recipient plant line to create a F1 population; (ii) selecting offspring that have expression cassette.
- the offspring can be further selected by testing the expression of the gene of interest.
- the transgenic plant with the expression cassette can serve as a male or female parent in a cross pollination to produce offspring plants, wherein by receiving the transgene from the donor plant, the offspring plants have the expression cassette.
- protoplast fusion can also be used for the transfer of the transgene from a donor plant to a recipient plant.
- Protoplast fusion is an induced or spontaneous union, such as a somatic hybridization, between two or more protoplasts (cells in which the cell walls are removed by enzymatic treatment) to produce a single bi- or multi-nucleate cell.
- the fused cell that may even be obtained with plant species that cannot be interbred in nature is tissue cultured into a hybrid plant exhibiting the desirable combination of traits. More specifically, a first protoplast can be obtained from a plant having the expression cassette.
- a second protoplast can be obtained from a second plant line, optionally from another plant species or variety, preferably from the same plant species or variety, that comprises commercially desirable characteristics, such as, but not limited to disease resistance, insect resistance, valuable grain characteristics (e.g., increased seed weight and/or seed size) etc.
- the protoplasts are then fused using traditional protoplast fusion procedures, which are known in the art to produce the cross.
- embryo rescue may be employed in the transfer of the expression cassette from a donor plant to a recipient plant.
- Embryo rescue can be used as a procedure to isolate embryos from crosses wherein plants fail to produce viable seed.
- the fertilized ovary or immature seed of a plant is tissue cultured to create new plants (see Pierik, 1999, In vitro culture of higher plants, Springer, ISBN 079235267x, 9780792352679, which is incorporated herein by reference in its entirety).
- the recipient plant is an elite line having one or more certain desired traits.
- desired traits include but are not limited to those that result in increased biomass production, production of specific chemicals, increased seed production, improved plant material quality, increased seed oil content, etc. Additional examples of desired traits includes pest resistance, vigor, development time (time to harvest), enhanced nutrient content, novel growth patterns, aromas or colors, salt, heat, drought and cold tolerance, and the like.
- Desired traits also include selectable marker genes (e.g., genes encoding herbicide or antibiotic resistance used only to facilitate detection or selection of transformed cells), hormone biosynthesis genes leading to the production of a plant hormone (e.g., auxins, gibberellins, cytokinins, abscisic acid and ethylene that are used only for selection), or reporter genes (e.g. luciferase, ⁇ -glucuronidase, chloramphenicol acetyl transferase (CAT, etc.).
- the recipient plant can also be a plant with preferred chemical compositions, e.g., compositions preferred for medical use or industrial applications.
- Open-Pollinated Populations The improvement of open-pollinated populations of such crops as rye, many maizes and sugar beets, herbage grasses, legumes such as alfalfa and clover, and tropical tree crops such as cacao, coconuts, oil palm and some rubber, depends essentially upon changing gene-frequencies towards fixation of favorable alleles while maintaining a high (but far from maximal) degree of heterozygosity. Uniformity in such populations is impossible and trueness-to-type in an open-pollinated variety is a statistical feature of the population as a whole, not a characteristic of individual plants. Thus, the heterogeneity of open-pollinated populations contrasts with the homogeneity (or virtually so) of inbred lines, clones and hybrids.
- Population improvement methods fall naturally into two groups, those based on purely phenotypic selection, normally called mass selection, and those based on selection with progeny testing.
- Interpopulation improvement utilizes the concept of open breeding populations; allowing genes to flow from one population to another. Plants in one population (cultivar, strain, ecotype, or any germplasm source) are crossed either naturally (e.g., by wind) or by hand or by bees (commonly Apis mellifera L. or Megachile rotundata F.) with plants from other populations. Selection is applied to improve one (or sometimes both) population(s) by isolating plants with desirable traits from both sources.
- Mass Selection In mass selection, desirable individual plants are chosen, harvested, and the seed composited without progeny testing to produce the following generation. Since selection is based on the maternal parent only, and there is no control over pollination, mass selection amounts to a form of random mating with selection. As stated herein, the purpose of mass selection is to increase the proportion of superior genotypes in the population.
- Synthetics A synthetic variety is produced by crossing inter se a number of genotypes selected for good combining ability in all possible hybrid combinations, with subsequent maintenance of the variety by open pollination. Whether parents are (more or less inbred) seed- propagated lines, as in some sugar beet and beans (Vicia) or clones, as in herbage grasses, clovers and alfalfa, makes no difference in principle. Parents are selected on general combining ability, sometimes by test crosses or topcrosses, more generally by polycrosses. Parental seed lines may be deliberately inbred (e.g. by selfing or sib crossing). However, even if the parents are not deliberately inbred, selection within lines during line maintenance will ensure that some inbreeding occurs. Clonal parents will, of course, remain unchanged and highly heterozygous.
- Pedigreed varieties A pedigreed variety is a superior genotype developed from selection of individual plants out of a segregating population followed by propagation and seed increase of self-pollinated offspring and careful testing of the genotype over several generations. This is an open pollinated method that works well with naturally self-pollinating species. This method can be used in combination with mass selection in variety development. Variations in pedigree and mass selection in combination are the most common methods for generating varieties in self- pollinated crops.
- Hybrids A hybrid is an individual plant resulting from a cross between parents of differing genotypes. Commercial hybrids are now used extensively in many crops, including corn (maize), sorghum, sugar beet, sunflower and broccoli. Hybrids can be formed in a number of different ways, including by crossing two parents directly (single cross hybrids), by crossing a single cross hybrid with another parent (three-way or triple cross hybrids), or by crossing two different hybrids (four- way or double cross hybrids).
- hybrids Strictly speaking, most individuals in an out breeding (i.e., open-pollinated) population are hybrids, but the term is usually reserved for cases in which the parents are individuals whose genomes are sufficiently distinct for them to be recognized as different species or subspecies.
- Hybrids may be fertile or sterile depending on qualitative and/or quantitative differences in the genomes of the two parents.
- Heterosis, or hybrid vigor is usually associated with increased heterozygosity that results in increased vigor of growth, survival, and fertility of hybrids as compared with the parental lines that were used to form the hybrid. Maximum heterosis is usually achieved by crossing two genetically different, highly inbred lines.
- Specialty Cannabis plants of the present invention can be further modified by introducing into the plants one or more transgenes which when expressed lead to desired phenotypes.
- the most common method for the introduction of new genetic material into a plant genome involves the use of living cells of the bacterial pathogen Agrobacterium tumefaciens to literally inject a piece of DNA, called transfer or T-DNA, into individual plant cells (usually following wounding of the tissue) where it is targeted to the plant nucleus for chromosomal integration.
- Agrobacterium-mediated plant transformation involves as a first step the placement of DNA fragments cloned on plasmids into living Agrobacterium cells, which are then subsequently used for transformation into individual plant cells.
- Agrobacterium-mediated plant transformation is thus an indirect plant transformation method.
- Methods of Agrobacterium-mediated plant transformation that involve using vectors with no T-DNA are also well known to those skilled in the art and can have applicability in the present invention. See, for example, U.S. Patent No. 7,250,554, which utilizes P-DNA instead of T-DNA in the transformation vector.
- biolistic bombardment uses ultrafine particles, usually tungsten or gold, that are coated with DNA and then sprayed onto the surface of a plant tissue with sufficient force to cause the particles to penetrate plant cells, including the thick cell wall, membrane and nuclear envelope, but without killing at least some of them (US 5,204,253, US 5,015,580).
- a third direct method uses fibrous forms of metal or ceramic consisting of sharp, porous or hollow needle-like projections that literally impale the cells, and also the nuclear envelope of cells.
- a selection method For efficient plant transformation, a selection method must be employed such that whole plants are regenerated from a single transformed cell and every cell of the transformed plant carries the DNA of interest.
- These methods can employ positive selection, whereby a foreign gene is supplied to a plant cell that allows it to utilize a substrate present in the medium that it otherwise could not use, such as mannose or xylose (for example, refer US 5,767,378; US 5994629). More typically, however, negative selection is used because it is more efficient, utilizing selective agents such as herbicides or antibiotics that either kill or inhibit the growth of nontransformed plant cells and reducing the possibility of chimeras. Resistance genes that are effective against negative selective agents are provided on the introduced foreign DNA used for the plant transformation.
- nptH neomycin phosphotransferase
- herbicides and herbicide resistance genes have been used for transformation purposes, including the bar gene, which confers resistance to the herbicide phosphinothricin (White et al., Nucl Acids Res 18: 1062 (1990), Spencer et al., Theor Appl Genet 79: 625-631(1990), US 4795855, US 5378824 and US 6107549).
- the dhfr gene which confers resistance to the anticancer agent methotrexate, has been used for selection (Bourouis et al., EMBO J. 2(7): 1099-1104 (1983).
- Genes can be introduced in a site directed fashion using homologous recombination. Homologous recombination permits site specific modifications in endogenous genes and thus inherited or acquired mutations may be corrected, and/or novel alterations may be engineered into the genome. Homologous recombination and site-directed integration in plants are discussed in, for example, U.S. Patent Nos. 5,451,513, 5,501,967 and 5,527,695.
- Transgenic plants can now be produced by a variety of different transformation methods including, but not limited to, electroporation; microinjection; microprojectile bombardment, also known as particle acceleration or biolistic bombardment; viral-mediated transformation; and Agrobacterium-mediated transformation. See, for example, U.S. Patent Nos. 5,405,765; 5,472,869; 5,538,877; 5,538,880; 5,550,318; 5,641,664; 5,736,369 and 5,736,369; and International Patent Application Publication Nos.
- Microprojectile bombardment is also known as particle acceleration, biolistic bombardment, and the gene gun (Biolistic® Gene Gun).
- the gene gun is used to shoot pellets that are coated with genes (e.g., for desired traits) into plant seeds or plant tissues in order to get the plant cells to then express the new genes.
- the gene gun uses an actual explosive (.22 caliber blank) to propel the material. Compressed air or steam may also be used as the propellant.
- the Biolistic® Gene Gun was invented in 1983-1984 at Cornell University by John Sanford, Edward Wolf, and Nelson Allen. It and its registered trademark are now owned by E. I. du Pont de Nemours and Company. Most species of plants have been transformed using this method.
- Agrobacterium tumefaciens is a naturally occurring bacterium that is capable of inserting its DNA (genetic information) into plants, resulting in a type of injury to the plant known as crown gall. Most species of plants can now be transformed using this method, including cucurbitaceous species.
- a transgenic plant formed using Agrobacterium transformation methods typically contains a single gene on one chromosome, although multiple copies are possible. Such transgenic plants can be referred to as being hemizygous for the added gene.
- a more accurate name for such a plant is an independent segregant, because each transformed plant represents a unique T-DNA integration event (U. S. Patent No. 6, 156,953).
- a transgene locus is generally characterized by the presence and/or absence of the transgene.
- a heterozygous genotype in which one allele corresponds to the absence of the transgene is also designated hemizygous (U.S. Patent No. 6,008,437).
- Non-limiting examples of methods for transforming cannabis plants and cannabis tissue culture methods are described in Zweger (The Biotechnology of Cannabis sativa, April 2009); MacKinnon (Genetic transformation of Cannabis sativa Linn: a multipurpose fiber crop, doctoral thesis, University of Dundee, Scotland, 2003), MacKinnon et al. (Progress towards transformation of fiber hemp, Scottish Crop Research, 2000), and US 20120311744, each of which is herein incorporated by reference in its entirety for all purposes.
- the transformation can be physical, chemical and/or biological.
- the present disclosure teaches the genetic modification of Specialty Cannabis.
- the Specialty Cannabis of the present disclosure comprise one or more transgenes, or DNA edits.
- the present disclosure teaches transformation of plants (e.g., via agrobacterium, gene gun, or other delivery mechanism).
- the present disclosure teaches gene editing with CRISPR, as disclosed in US Patent Nos 8,697,359, 9,790,490, and US Application No 15/482,603.
- the present disclosure teaches cannabinoid compositions comprising high propyl cannabinoid contents with little to no THC.
- the compositions of the present disclosure are completely derived from cannabis extractions (i.e., all components are derived from the cannabis plant).
- the present disclosure teaches cannabinoid compositions in which only the active cannabinoid and terpene components must be derived from the cannabis plant.
- the present disclosure teaches cannabinoid compositions in which one or more components are derived from sources other than the cannabis plant (e.g., from other organisms, or chemically synthesized).
- the cannabinoid compositions of the present disclosure can, in some embodiments, comprise cannabinoids produced via standard chemical, biochemical, or biocatalytic methods. Persons having skill in the art will be familiar with various synthesis methods, including those of U.S. 9,359,625 and Taura et al. 1996, The Journal of Biological Chemistry, Vol. 271, No. 21, p. 17411-17416.
- compositions of the present disclosure are treated to convert THC to CBD to reduce or eliminate THC content.
- Persons having skill in the art will be familiar with the various methods for converting THC to CBD, including those discussed in US Patent No. 9,259,449.
- the compositions of the present disclosure mimic the cannabinoid and Terpene Profiles of the Specialty Cannabis plants disclosed herein. That is, in some embodiments, the cannabinoid compositions comprise a cannabinoid component with little to no THC, and a terpene component. Thus, in some embodiments, the cannabinoid compositions of the present disclosure comprise no more than 0.3% THC with greater than 1% propyl cannabinoid contents, and at least 1% terpene oil content, as measured by the weight of the composition. Extractions methods designed to preserve cannabinoid and Terpene Profiles are disclosed in other sections of this application.
- the compositions of the present disclosure comprise more concentrated cannabinoid and terpene content than the Specialty Cannabis hemp plants.
- the cannabinoid compositions comprise a CBDV content of greater than 20%, a terpene oil content of greater than 10%, and a THC content of less than 10%, as measured by weight of the composition.
- the present disclosure teaches methods of supplementing cannabis extracts with one or more cannabinoid or terpene to account for any losses of the compounds during the extraction process.
- the present disclosure teaches the formulation of cannabis compositions from individual components (i.e., by mixing individual cannabinoid and terpene components obtained from the same or different sources).
- the cannabinoid compositions of the present disclosure are designed to mimic the organoleptic experience produced by the Specialty Cannabis.
- the cannabinoid compositions of the present disclosure comprise about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 8
- the cannabinoid compositions of the present disclosure comprise more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%
- the cannabinoid compositions of the present disclosure comprise about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 8
- the Cannabinoid compositions of the present disclosure comprise 2%-10%, 3%-30%, or 3%-60% propyl cannabinoids content by weight of the composition.
- the cannabinoid compositions of the present disclosure comprise more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 6
- the cannabinoid compositions of the present disclosure comprise about 0.00%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.30%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% THC by weight of the composition, and all ranges therebetween.
- the cannabinoid compositions of the present disclosure comprise 0.00%-0.10%, 0.00%-0.20%, 0.00%-0.30%, 0.00%-3.00%, or 0.00%-9.00% THC content by weight of the composition.
- the cannabinoid compositions of the present disclosure comprise less than about 0.00%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.30% 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% THC by weight of the composition, while accumulating at least 20% non- THC cannabinoid content by weight of the composition.
- the cannabinoid compositions of the present disclosure comprise about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 8
- the cannabinoid compositions of the present disclosure comprise more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%
- the cannabinoid compositions of the present disclosure comprise about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 8
- the cannabinoid compositions of the present disclosure comprise more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%
- the cannabinoid composition of the present disclosure comprise about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%
- the cannabinoid compositions of the present disclosure comprise more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%,
- the cannabinoid compositions of the present disclosure produce comprise about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%,
- the cannabinoid compositions of the present disclosure comprise 3%-40%, 3%-30%, or 3%-25% CBDV content by weight of the composition.
- the cannabinoid compositions of the present disclosure comprise more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%,
- the cannabinoid compositions of the present disclosure comprise about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 8
- the cannabinoid compositions of the present disclosure comprise more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%,
- the cannabinoid compositions of the present disclosure produce comprise organoleptically pleasing Terpene Profiles.
- the cannabinoid compositions of the present invention has an absolute content of any one of the 17 terpenes in the Terpene Profile that is 0%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.4%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%
- the absolute content of any one of the terpenes is between about 0.05% and about 5%.
- This paragraph is intended to be read as applying to any specific terpene(s) in a Terpene Profile, such that the name of any one or two or more of these terpenes as specifically referred to elsewhere herein (e.g., linalool) can replace the phrase "any one of the 17 terpenes in the Terpene Profile.”
- the cannabinoid compositions of the present invention has an absolute content of any one of the 17 terpenes in the Terpene Profile that is greater than 0%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.2%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.3%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.4%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.5%, 0.51%, 0.52%, 0.53%,
- the cannabinoid compositions of the present invention has 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%
- the essential oil content of the cannabinoid compositions of the present invention is between about 0.5% and about 30% by weight of the composition. In other embodiments the essential oil contents of the cannabinoid compositions of the present invention is between about 1.0% and about 25% by weight of the composition.
- the cannabinoid compositions of the present invention has greater than 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%,
- the terpene content of the cannabinoid compositions of the present disclosure is described in relative terms as a percentage composition of the total Terpene Profile.
- a cannabinoid compositions with 1.2% absolute terpinolene content and 1.2% limonene content and no other terpenes in the Terpene Profile would said to have 50% terpinolene and 50% limonene relative content.
- the cannabinoid compositions of the present invention has a relative content of any one of the 17 terpenes in the Terpene Profile that is greater than or less than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 7
- the relative content of any one of the terpenes is between 0% and 100%.
- This paragraph is intended to be read as applying to any specific terpene(s) in a Terpene Profile, such that the name of any of one or two or more these terpenes as specifically referred to elsewhere herein (e.g., linalool) can replace the phrase "any one of the 17 terpenes in the Terpene Profile.”
- additional components are optionally added to the cannabinoid compositions of the present disclosure to improve the taste and/or physical properties of the composition (such as stability, viscosity, appearance of smoke as it is inhaled, etc.).
- additional components include, but are not limited to, sweeteners, natural flavorants, artificial flavorants, colorants, antioxidants, preservatives, chelating agents, viscomodulators, tonicifiers, odorants, opacifiers, suspending agents, binders, thickeners, carriers and mixtures thereof, including, but not limited to, xanthum gum, carboxymethylcellulose, carboxyethylcellulose, hydroxypropylcellulose, methylcellulose, microcrystalline cellulose, starches, dextrins, maltodextrins, other polyols (including sugar alcohols, such as sorbitol, lactitol or mannitol), carbohydrates (e.g., lactose), propylene glycol alginate, gellan gum,
- the cannabinoid compositions of the present disclosure comprise one or more medium chain length triglycerides (MCTs).
- MCTs are triglycerides whose fatty acids have an aliphatic tail of 6-12 carbon atoms.
- the MCT is one or more of caproic acid, caprylic acid, capric acid, lauric acid and mixtures thereof.
- Suitable sources of MCTs are known to those skilled in the art and include, for example, coconut oil and palm kernel oil.
- the cannabinoid compositions of the present disclosure comprise one or more polyesterdiols.
- the polyesterdiol may be a linear two to ten units polymer (also referred to as (ester)2-10 glycol), that is derived from natural or non-natural sources such as vegetables, fruits, bacteria, yeast, algae, or manufactured by chemical processes.
- the polyesterdiol is 1) a polypropylene glycol such as: dipropylene glycol; tripropylene glycol, including tetra-, penta-, hexa-, hepta-, octa-, nona- and decapropylene glycol and other derivatives thereof; 2) a polybutylene glycol such as: dibutylene glycol, tributylene glycol, including terra-, penta-, hexa-, hepta-, octa-, nona- and decabutylene glycol, and other derivatives thereof; 3) also including 2- 10 unit polymers of rare organic ester types such as pentylene, octylene, terpentylene, nonylene, linalylene, isoamylene, isobutylene, geranylene, bornylene, benzylene and allylene, caprylylene, such as, for example, polyis
- the cannabinoid compositions of the present disclosure comprise a linear polyesterdiol selected from the group consisting of: (ethylene)3-10 glycol; (propylene)2-10 glycol; (butylene)2-10 glycol; (pentylene)2-10 glycol; (octylene)2-10 glycol; (terpentylene)2-10 glycol; (nonylene)2-10 glycol; (linalylene)2-10 glycol; (isoamylene)2-10 glycol; (isobutylene)2- 10 glycol; (geranylene)2-10 glycol; (bornylene)2-10 glycol; (benzylene)2-10 glycol; (allylene)2- 10 glycol; and (caprylylene)2-10 glycol; acid or sugar conjugates thereof, and ester or ether or alcohol derivatives thereof.
- a linear polyesterdiol selected from the group consisting of: (ethylene)3-10 glycol; (propylene)2-10 glycol; (butylene)2-10 glycol; (
- the cannabinoid compositions of the present disclosure comprises a carrier selected from the group consisting of: triethylene glycol; tetraethyleneglycol, pentaethylenglycol, hexaethyleneglycol, heptaethyleneglycol, octaethyleneglycol, nonaethylenglycol; decaethylene glycol; dipropylene glycol; tripropylene glycol; tetrapropylene glycol, pentapropylene glycol, hexapropylene glycol, heptapropylene glycol, octapropylene glycol, nonapropylene glycol; decapropylene glycol; dibutylene glycol, tributylene glycol; tetrabutylene glycol, pentabutylene glycol, hexabutylene glycol, heptabutylene glycol, octabutylene glycol, nonabutylene glycol,
- the carrier is selected from the group consisting of borneol, camphor, 1,8-Cineole, citral, geraniol, indomethacin, limonene, linalool, linalyl acetate, ⁇ - myrcene, myrcenol, 1-menthol, menthone, neomenthol, nerol, nerolidol, a-pinene, peppermint oil, pulegone, phytol, terpineol, terpinen-4-ol, thymohydroquinone, thymol, and thymoquinone
- compositions of the present disclosure comprise one or more of propylene glycol, glycerine, vegetable glycerine, and/or water.
- the present disclosure provides for extracts from Specialty Cannabis plants.
- Cannabis extracts or products or the present disclosure include:
- Kief- refers to trichomes collected from cannabis.
- the trichomes of cannabis are the areas of cannabinoid and terpene accumulation. Kief can be gathered from containers where cannabis flowers have been handled. It can be obtained from mechanical separation of the trichomes from inflorescence tissue through methods such as grinding flowers, or collecting and sifting through dust after manicuring or handling cannabis. Kief can be pressed into hashish for convenience or storage.
- Hash- sometimes known as hashish, is often composed of preparations of cannabis trichomes. Hash pressed from kief is often solid.
- Bubble Hash- sometimes called bubble melt hash can take on paste-like properties with varying hardness and pliability. Bubble hash is usually made via water separation in which cannabis material is placed in a cold water bath and stirred for a long time (around 1 hour). Once the mixture settles it can be sifted to collect the hash.
- Solvent reduced oils also sometimes known as hash oil, honey oil, or full melt hash among other names.
- This type of cannabis oil is made by soaking plant material in a chemical solvent. After separating plant material, the solvent can be boiled or evaporated off, leaving the oil behind.
- Butane Hash Oil is produced by passing butane over cannabis and then letting the butane evaporate.
- Budder or Wax is produced through isopropyl extraction of cannabis. The resulting substance is a wax like golden brown paste.
- Another common extraction solvent for creating cannabis oil is C02. Persons having skill in the art will be familiar with C02 extraction techniques and devices, including those disclosed in US 20160279183, US 2015/01505455, US 9,730,911, and US 2018/0000857.
- Tinctures- are alcoholic extracts of cannabis. These are usually made by mixing cannabis material with high proof ethanol and separating out plant material.
- E-juice- are cannabis extracts dissolved in either propylene glycol, vegetable glycerin, or a combination of both. Some E-juice formulations will also include polyethylene glycol and flavorings. E-juice tends to be less viscous than solvent reduced oils and is commonly consumed on e-cigarettes or pen vaporizers.
- Rick Simpson Oil ethanol extractions
- Rick Simpson Oil ethanol extractions
- the extract produced from contacting the cannabis with ethanol is heated so as to decarboxylate the extract.
- the Specialty Cannabis of the present invention is extracted via methods that preserve the cannabinoid and terpenes. In other embodiments, said methods can be used with any cannabis plants.
- the extracts of the present invention are designed to produce products for human or animal consumption via inhalation (via combustion, vaporization and nebulization), buccal absorption within the mouth, oral administration, and topical application delivery methods.
- the present invention teaches an optimized method at which we extract compounds of interest, by extracting at the point when the drying harvested plant has reached 15% water weight, which minimizes the loss of terpenes and plant volatiles of interest. Stems are typically still 'cool' and 'rubbery' from evaporation taking place.
- the chemical extraction of Specialty Cannabis can be accomplished employing polar and non-polar solvents in various phases at varying pressures and temperatures to selectively or comprehensively extract terpenes, cannabinoids and other compounds of flavor, fragrance or pharmacological value for use individually or combination in the formulation of our products.
- the extractions can be shaped and formed into single or multiple dose packages, e.g., dabs, pellets and loads.
- the solvents employed for selective extraction of our cultivars may include water, carbon dioxide, 1,1,1,2-tetrafluoroethane, butane, propane, ethanol, isopropyl alcohol, hexane, and limonene, in combination or series.
- the chemical and mechanical extraction methods of the present invention can be used to produce products that combine chemical extractions with plant parts containing compounds of interest.
- the extracts of the present invention may also be combined with pure compounds of interest to the extractions, e.g. cannabinoids or terpenes to further enhance or modify the resulting formulation's fragrance, flavor or pharmacology.
- the extractions are supplemented with terpenes or cannabinoids to adjust for any loss of those compounds during extraction processes.
- the cannabis extracts of the present invention mimic the chemistry of the cannabis flower material.
- the cannabis extracts of the present invention will about the same cannabinoid and Terpene Profile of the dried flowers of the Specialty Cannabis of the present invention.
- Extracts of the present invention can be used for vaporization, production of e-juice or tincture for e-cigarettes, or for the production of other consumable products such as edibles or topical spreads.
- Cannabis edibles such as candy, brownies, and other foods are a popular method of consuming cannabis for medicinal and recreational purposes.
- the Specialty Cannabis of the present disclosure and/or the cannabinoid compositions of the present disclosure can be used to make cannabis edibles.
- Most cannabis edible recipes begin with the extraction of cannabinoids and terpenes, which are then used as an ingredient in various edible recipes.
- the cannabis extract used to make edibles out of the Specialty Cannabis of the present invention is cannabis butter.
- Cannabis butter is made by melting butter in a container with cannabis and letting it simmer for about half an hour, or until the butter turns green. The butter is then chilled and used in normal recipes.
- Other extraction methods for edibles include extraction into cooking oil, milk, cream, flour (grinding cannabis and blending with flour for baking). Lipid rich extraction mediums/edibles are believed to facilitate absorption of cannabinoids into the blood stream. THC absorbed by the body is converted by the liver into 11 -hydroxy- THC.
- the present disclosure teaches edibles produced from the Specialty Cannabis and/or cannabinoid compositions disclosed herein.
- Plant inflorescence samples were prepared by grinding ⁇ 5 g of dried cannabis flower material in a coffee grinder. From this homogenized material, 1000 ⁇ 20 mg was placed in a 50mL falcon tube with ⁇ 1 g of 2mm beads and 15 niL of working solution. Each sample was placed in the bead beater (1600 MiniG from Spex Sample Prep) and homogenized on high for 6 minutes. Then approximately 2mL of each sample were transferred to 2mL centrifuge vials and centrifuged at 10000 g for 5 minutes. For samples suspected of having higher or lower concentrations of analytes the mass to volume ratio of the extraction could be adjusted. The neat sample was placed in a GC vial for terpene analysis without dilution.
- the supernatant was also diluted with working solution for GC and HPLC analysis.
- a 1:96 dilution provided the appropriate concentration for analysis of cannabinoids present at concentrations above 2.3%, while a 1:6 dilution allowed for analysis of cannabinoids below this level.
- Terpenes were quantified by a method developed on a GC-FID instrument from Perkin Elmer (Waltham, MA). It is recognized among analytical scientists that terpene measurements conducted via HPLC are unreliable, as HPLC is not effective at measuring volatiles, such as terpenes. This method separates and quantifies 17 different terpenoids commonly found in cannabis plant tissue. The terpenoids are each quantified by their own individual calibration curves generated with analytical reference standards (Sigma Aldrich) and all use n-nonane as the internal standard.
- the instrumentation includes a Clarus 680 gas chromatograph (GC) equipped with an autosampler, an Elite-5 column (Perkin Elmer (Waltham, MA), 30 m length, 0.25 mm internal diameter, 0.25 ⁇ thickness film diameter) and a flame ionization detector (FID). Instrument control and data acquisition and analyses was accomplished by TotalChrom software version 1.2.3.4 (Perkin Elmer, Waltham, MA).
- Cannabinoids were quantified by an analytical method developed and run on a Perkin Elmer (Waltham, MA) GC-FID instrument as well. This method was developed to separate six neutral cannabinoids, CBD, CBG, CBN, THC, A8-THC, and CBC. The cannabinoids are each quantified by their own individual calibration curves generated with analytical reference standards (Restek) and all use tricosane as the internal standard. The retention time of THCV was determined by analyzing THV01 (vide infra) by GCMS, however since analytical standards were not available it was "quantified" by referencing the calibration curve for THC.
- Restek analytical reference standards
- the instrumentation includes a Clarus 680 gas chromatograph (GC) equipped with an autosampler, an Elite-lcolumn (Perkin Elmer (Waltham, MA), 30 m length, 0.25 mm internal diameter, 0.25 ⁇ m thickness film diameter) and a flame ionization detector (FID). Instrument control and data acquisition and analyses was accomplished by TotalChrom software version 1.2.3.4 (Perkin Elmer, Waltham, MA).
- the GC-FID cannabinoid assay Due to the very linear nature of the FID detector, the GC-FID cannabinoid assay generally provided satisfactory results over nearly two orders of magnitude (up to 1.0 mg/mL), however in order to use the same calibration solutions and "validation" procedures for both GC and HPLC the range was reduced to that of the HPLC method. Based on the sample mass extracted (500 mg) and a 3x3 mL extraction (low oil samples), a 1 :3 dilution provided quantitation of cannabinoid levels from 0.09-1.35% and the 1:40 dilution from 1.15-18% in the plant matrix.
- HPLC-PDA also known as HPLC-DAD, or simply HPLC
- HPLC-DAD HPLC-DAD
- This method quantifies six neutral cannabinoids (CBD, CBG, CBN, THC, A8-THC, and CBC) as well as the acid cannabinoids THCA, CBDA and CBGA amongst other acidic cannabinoids, based on calibration curves generated with analytical standards and an internal reference standard (ibuprofen).
- HPLC analyses were performed using a Agilent 1290 System (Agilent Technologies, Santa Clara, CA).
- the HPLC system comprised G4212A diode array detector, a G1316C temperature controlled column compartment, a G4226A autosampler, and a G4204A quaternary pump. Separation of the cannabinoids was achieved on a Poroshell 120 EC- CI 8 column (2.7 ⁇ , 150mm x 2.1mm i.d, PN 693775-902) with a Poroshell 120 EC-C18 guard column (2.7 ⁇ , 5mm x 2.1mm i.d., PN 821725-911) in place (Agilent Technologies, Santa Clara, CA). Instrument control, data acquisition and integration was achieved with OpenLab CDS ChemStation Rev C.01.06 software (Agilent Technologies, Santa Clara, CA).
- Calibration was achieved by performing a five-point calibration curve (0.016 - 0.25mg/mL for each analyte) followed by linear regression analysis. This analysis was performed with Microsoft Excel (Redmond, WA) software. The calibration curves were generated by injecting each standard in triplicate and the RSDs provided the measure of precision while the absolute accuracy was determined by comparing the concentrations of the standards predicted by the calibration curve to their "known" values determined by dilution ratios. AOAC International standards for accuracy and precision were used as quality guidelines for every calibration. Check standards were run at the start, middle, and end of every analysis, and recalibration was performed when they varied more than +/- 5% of their initial average response.
- the cannabinoid content was quantified by both GC-FID and HPLC.
- the main difference between GC and HPLC is that GC involves thermal stress and mainly resolves analytes by boiling points while HPLC does not involve heat and mainly resolves analytes by polarity.
- This orthogonal approach to analyses is desirable for highly accurate and reproducible results in determining chemotype.
- the first reason is related to the difference between the cannabinoids produced naturally by the plant (the acidic cannabinoids) and those that are bioactive (the neutral cannabinoids). Cannabis biosynthesizes all the cannabinoids in their relatively unstable acidic forms, and these forms are generally not bioactive in the traditional sense.
- THCA neutral cannabinoids
- CBG CBG
- CBC CBC
- CBD CBN
- CBN CBN
- HPLC assay could characterize the naturally occurring chemotypes, but the acidic analytes are not available as standards, so this quantification is approximate and considered for information only.
- the acidic analytes are all quantified by reference to the calibration curve for THCA, and this is not an unreasonable assumption as many of them have approximately the same spectral properties.
- the GC assay is calibrated with analytical standards, but these are the neutral cannabinoids and their formation from the naturally occurring acidic cannabinoids in the GC injector is not quantitative, which complicates exact characterization of the naturally occurring chemotype.
- Method validation is important in establishing that a method is fit for its intended purpose, providing assurance that the results that are reported are precise, accurate, and reflective of the sample.
- Very few labs in the cannabis industry attempt to validate their assays and this fact, combined with inappropriate sampling have resulted in erroneous data for several varieties.
- an abbreviated protocol similar to Single Laboratory Validation (SLV) was carried out.
- the analyte RSDs, recoveries, and precisions at these concentrations satisfied AO AC guidance (based on mg/mL).
- the RSDs for the terpenes at the low, medium, and high concentrations (varied by terpene but generally 0.016, 0.125, and 1.0 mg/mL) were less than 5%, 4%, and 3% respectively.
- the absolute bias for these analytes was generally less than 10%, 4%, and 2%.
- the RSDs for the cannabinoids by both GC and HPLC at the low, medium, and high concentrations were less than 2%, 2%, and 1% respectively.
- the absolute bias for these analytes was generally less than 10%, 2%, and 2%.
- the volunteer trial for higher propyl content hemp will be conducted over 2 weeks. Volunteers will be split into six groups (1-6). Each volunteer in the group will be given two samples (a control and a comparator blend). In this trial, the control comparator blends will be prepared to contain nearly identical levels of a non-propyl cannabinoids (e.g. THC, and/or CBD), but each week the comparator will be formulated so as to include different levels of THCV and/or CBDV added (e.g., either 2%, 5%, or 7.5% CBDV added in).
- a non-propyl cannabinoids e.g. THC, and/or CBD
- Example 3 Volunteer trials using high propyl cannabinoid compositions. (PROPHETIC).
- Volunteer comparison trials will be conducted to determine the effect of increased propyl cannabinoid content in cannabis compositions with no more than 0.3% THC contents. Volunteer trials will be conducted in similar fashion to those of Example 2.
- each volunteer in the group will be given two composition samples (a control and a comparator blend).
- the samples will be provided in single-use e-cigarettes or in tinctures designed to be vaporized or administered to the mucosa/swallowed, respectively.
- the control comparator compositions will be prepared to contain nearly identical levels of a non- propyl cannabinoid (e.g. THC, and/or CBD), but each week the comparator will be formulated so as to include different levels of a propyl cannabinoid added (e.g., either 2%, 5%, or 7.5% CBDV added in).
- One objective of the inventions of the present disclosure was to develop cannabis varieties accumulating high levels of propyl cannabinoids with no more than 0.3% or 0.2% THC content. This goal was achieved through a multi-pronged cannabis breeding program that utilized existing public and proprietary cannabis lines to produce novel cannabis germplasms exhibiting high levels of propyl cannabinoids, with no more than 0.3% or 0.2% THC content across varied genetic and phenotypic backgrounds.
- the cannabinoid profiles of each parental line was determined using HPLC as described in Example 1.
- the resulting measurements of the initial parental lines are summarized in Table 3. All of the initial parental lines exhibited either only trace amounts of propyl cannabinoid content, or accumulated greater than 0.3% THC content.
- Table 3 also reports the cannabinoid contents of several intermediate filial generations generated during each breeding scheme that were used as parents for the final progeny lines.
- One objective of the inventions of the present disclosure was to develop cannabis varieties accumulating high levels of propyl cannabinoids with no more than 0.3% or 0.2% THC content.
- the Specialty Cannabis varieties of the present invention were additionally selected for their ability to produce terpenes that are appealing to patients and that may also provide a pharmacological activity that modifies, enhances or ameliorates the effects of the cannabinoids.
- a secondary objective of the breeding programs of the present disclosure was to produce plants with high terpene oil content and diverse Terpene Profiles.
- F2 seed can further be grown to produce F2 progeny. Selection for desirable phenotypes and/or genotypes can be conducted within the F1, F2, or subsequent progeny since the selections can be maintained (i.e., fixed) via asexual reproduction. Alternatively, the F2 progeny can be crossed among themselves to produce a bulked F3 population from which desired progeny can be selected and/or further generations of crossing can be conducted. Again, selected F2 progeny can be maintained (i.e., fixed) via asexual reproduction. In another embodiment, the resultant F1 progeny can by backcrossed to high propyl cannabinoid parent or a hemp variety to further reinforce the traits of other parent.
- the cannabinoid and terpene components of Specialty Cannabis hemp lines of the present disclosure can be maintained by extracting the cannabinoid and terpene contents.
- the resulting extract will mimic the cannabinoid and terpene contents of the plant/inflorescence.
- extracts of the present disclosure can be stored indefinitely.
- the lines can also be further selected for a specific content of certain other cannabinoids and/or of certain terpenes/terpenoids, and/or for additional phenotypic and genotypic characteristics.
- Desirable phenotypic characteristics include but are not limited to larger plant size (i.e., greater bulk or biomass), higher production of flower buds, larger flowers, more trichomes, shorter plant stature, ability to tolerate lower and/or higher growing temperatures, greater germination percentage, greater seedling vigor, more efficient water usage, disease resistance, pest resistance, and other desirable agronomic and production traits.
- progeny resulting from any selection stage of either the crosses described in Figures 3-8, or any of the described selfing, sib crosses, or backcrossing versions of the breeding regimes of the present invention can be asexually reproduced so as to fix and maintain the desirable content, propyl cannabinoid content, low THC content, the aroma and flavor(s) typical of the desired class, and the other desirable phenotypic and/or genotypic characteristics.
- the resultant selected lines will be designated as Specialty Cannabis Varieties.
- the resultant Specialty Cannabis plants of the present invention also generally have more terpene essential oil content per plant than contemporary hemp varieties. More terpene essential oil per plant means less plant matter is required per treatment/administration, thereby also further minimizing any health risks for medical and recreational cannabis smokers/consumers. This would also further increase production efficiency.
- Example 5 The new Specialty Cannabis varieties created through crosses as described in Example 5 were subjected to cannabinoid and terpene chemical analysis as described in Example 1.
- the resulting breeding schemes of Example 5 produced six separate lines of high propyl cannabinoid Specialty Cannabis hemp germplasm ('O3.S2.01XO9.S1.01,' O3.S2.16XO9.S1.01,' O12.09.10X09.S1.01,' 'V24.S1.P09XO9.S1.01,' 'V24.S2.26XO9.S1.01,' and 'O9.S1.01XO9.S1.01').
- the level of cannabinoids of several plants within each of the high propyl cannabinoid lines was measured by HPLC, and is presented across Tables 5-10. Terpenes for several plants within each of the high propyl cannabinoid lines were measured using GC-FID, and are presented as absolute content measurements based on the percent content by weight of dry inflorescences in Tables 11-13. A summary table of representative plants from each of the high propyl cannabinoid hemp lines is presented in Table 4.
- Cannabinoid containing fractions were decarboxylated at high temperatures (175 C) for a period of about 90 minutes. In addition to decarboxylating, this process also allows for water removal from samples prior to winterization. In the winterization process, the two cannabinoid fractions were pooled and incubated with ethanol at -20 C for a period of at least 24 hours for proper separation of fats and waxes. After incubation, samples were filtered to remove solidified material.
- HPLC high performance liquid chromatography
- a terpene producing, diploid cannabis hemp plant cell from a female inflorescence (i) a cannabis hemp plant, (ii) an asexual clone of the plant, or (iii) a part of the plant, wherein said cannabis hemp plant, asexual clone of the plant or part of the plant produces the female inflorescence, said inflorescence comprising:
- a dry sinsemilla cannabis inflorescence comprising:
- the terpene oil content is the additive content of terpinolene, alpha phellandrene, beta ocimene, caren
- the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pin
- 26. The cannabis hemp plant, or an asexual clone of said cannabis hemp plant, or a plant part, tissue, or cell thereof of embodiment 12, wherein the first or second most abundant terpene in the Terpene Profile is beta caryophyllene.
- the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma
- a method of producing a cannabis extract comprising the steps of: contacting the inflorescence of any one of embodiments 1-29 with a solvent, thereby producing a cannabis extract.
- a method of breeding cannabis hemp plants with high propyl cannabinoid content max and low THC max content comprising:
- the resulting selected cannabis hemp plant comprises at least 1.0% propyl cannabinoid max content by weight, and no more than 0.3% THC max content by weight.
- a method of producing cannabis hemp plants with high propyl cannabinoid content max and low THC max content comprising:
- the resulting selected cannabis hemp plant is comprises at least 1.0% propyl cannabinoid max content by weight, and no more than 0.3% THC max content by weight.
- a cannabis hemp female inflorescence comprising:
- HPLC high performance liquid chromatography
- terpene oil content is the additive content of terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene as measured by GC-FID and calculated based on dry weight of the inflorescence.
- the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camp
- the cannabis hemp female inflorescence of embodiment 46 wherein the first or second most abundant terpene in the Terpene Profile is carene.
- the cannabis hemp female inflorescence of embodiment 46, wherein the first or second most abundant terpene in the Terpene Profile is limonene.
- the cannabis hemp female inflorescence of embodiment 46, wherein the first or second most abundant terpene in the Terpene Profile is beta caryophyllene.
- the cannabis hemp female inflorescence of embodiment 46, wherein the first or second most abundant terpene in the Terpene Profile is linalool.
- the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinen
- a method of producing a cannabis extract comprising the steps of: contacting the inflorescence of any one of embodiments 35-63 with a solvent, thereby producing a cannabis extract.
- a terpene producing, diploid cannabis hemp plant cell from a female inflorescence (i) a cannabis hemp plant, (ii) an asexual clone of the plant, or (iii) a part of the plant, wherein said cannabis hemp plant, asexual clone of the plant or part of the plant produces the female inflorescence, said inflorescence comprising:
- terpene oil content is the additive content of terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene as measured by GC-FID and calculated based on dry weight of the inflorescence.
- the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene
- the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha
- a method of producing a cannabis extract comprising the steps of: contacting the cell of any one of embodiments 67-95 with a solvent, thereby producing a cannabis extract.
- terpene oil content is the additive content of terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene as measured by GC-FID and calculated based on dry weight of the inflorescence.
- the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpin
- the cannabis extract of embodiment 129 wherein said extract is selected from the group consisting of kief, hashish, bubble hash, solvent reduced oils, sludges, e-juice, and tinctures.
- a composition comprising:
- CBD max cannabidiol
- tetrahydrocannabinol THC max
- HPLC high performance liquid chromatography
- composition of embodiment 134 wherein the composition comprises a terpene oil content greater than about 10% by weight;
- terpene oil content is the additive content of terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene as measured by GC-FID and calculated based on weight of the composition.
- composition 141 The composition of any one of embodiments 134-140, wherein the composition comprises a propyl THC max content of no more than 0.5% by weight.
- composition comprises a Terpene Profile in which myrcene is not the dominant terpene; wherein the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is terpinolene.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is alpha phellandrene.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is beta ocimene.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is limonene.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is gamma terpinene.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is alpha pinene.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is alpha terpinene.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is beta pinene.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is fenchol.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is camphene.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is alpha terpineol.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is alpha humulene.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is beta caryophyllene.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is linalool.
- composition of embodiment 144, wherein the first or second most abundant terpene in the Terpene Profile is caryophyllene oxide.
- composition of any one of embodiments 134-143 wherein the composition comprises a Terpene Profile in which myrcene is the first or second most abundant terpene in the Terpene Profile; wherein the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camphene, alpha terpineol, alpha humulene, beta caryophyllene, linalool, caryophyllene oxide, and myrcene.
- the Terpene Profile is defined as terpinolene, alpha phellandrene, beta ocimene, carene, limonene, gamma terpinene, alpha pinene, alpha terpinene, beta pinene, fenchol, camp
Abstract
Description
Claims
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PCT/US2018/064704 WO2019113574A1 (en) | 2017-12-08 | 2018-12-10 | Propyl cannabinoid hemp plants, methods of producing and methods of using them |
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TWI583374B (en) * | 2010-03-30 | 2017-05-21 | Gw伐瑪有限公司 | Use of the phytocannabinoid cannabidivarin (cbdv) in the treatment of epilepsy |
US10792318B2 (en) * | 2013-03-14 | 2020-10-06 | Sc Laboratories, Inc. | Bioactive concentrates and uses thereof |
WO2014145490A2 (en) * | 2013-03-15 | 2014-09-18 | Biotech Institute, Llc | Breeding, production, processing and use of specialty cannabis |
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