Application US20200107510

GENERATION OF NEW VARIETIES OF CANNABIS BY CHEMICAL MUTAGENESIS OF CANNABIS CELL SUSPENSIONS
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US20200107510

1. A method of generating and selecting mutant Cannabis plants through mutagenesis of isolated Cannabis plant cells, comprising: (7) (4)

subjecting plant parts of one or more Cannabis plants to a pectinase treatment to obtain living cells of the one or more Cannabis plants;

suspending the living cells in a mutagenic solution comprising methane sulfonate (EMS) and dimethyl sulfoxide (DMSO) to obtain mutated Cannabis cells;

centrifuging the mutated Cannabis cells to obtain pelleted cells; and

providing the pelleted cells on culture media.

2. The method of claim 1, comprising: (1) (1)

cutting the plant parts pieces having an area of approximately 1 mm²prior to said subjecting the plant parts to the pectinase treatment.

3. The method of claim 2, comprising, after the subjecting and prior to the suspending: (0) (3)

4. The method of claim 1, wherein said subjecting the plant parts to the pectinase treatment comprises: (1) (2)

6. The method of claim 1, comprising: (0) (1)

7. The method of claim 1, wherein the mutagenic solution includes nitrous acid. (1) (0)

9. The method of claim 1, comprising: (0) (1)

10. The method of claim 1, wherein: (1) (2)

13. The method of claim 1, wherein said providing the pelleted cells on culture media comprises: (2) (3)

providing the pelleted cells on a first culture media to obtain individual calli;

subsequently transferring individual calli from the first culture media to a second culture media and incubating the transferred individual calli to obtain plantlets;

subsequently transferring the plantlets to a third culture media.

14. The method of claim 13, wherein the second culture media is a root and shoot culture media or a somatic embryogenesis media. (0)

15. The method of claim 13, comprising: after 4 to 6 inches of growth of the plantlets into plants in the third culture media, transferring the plants to 4-6 inch biodegradable pots containing soil for further growth, the biodegradable pots at least partially composed of peat moss and wood pulp. (1) (0)

16. The method of claim 15, comprising: (1) (2)

after said further growth of the plants, examining variability of the plants for at least one characteristic, comprising at least one of cannabinoid profile, size, branching, height, water stress tolerance, salinity tolerance, temperature tolerance, and flowering time; and

selecting ones of the plants that exhibit improvements over others of the plants for the at least one characteristic for further propagation.

17. The method of claim 16, wherein the at least one characteristic includes a cannabinoid profile, and the cannabinoid profile including a desired range for a ratio of CBD:THC or THC:CBD. (1) (0)

19. A method of generating and selecting mutant Cannabis plants through mutagenesis of isolated Cannabis plant cells, comprising: (1) (8)

subjecting plant parts of Cannabis plants to pectinase treatment;

incubating the pectinase-treated plant parts at a first temperature in a first incubation;

filtering the incubated plant parts to obtain filtrate-containing cells;

centrifuging the filtrate-containing cells in a first centrifuging operation;

suspending the centrifuged filtrate-containing cells in a mutagenic solution that includes methane sulfonate (EMS) and dimethyl sulfoxide (DMSO);

incubating the mutagenic solution at a second temperature after said suspending in a second incubation to obtain mutated Cannabis cells;

centrifuging the mutagenic solution containing the mutated Cannabis cells in a second centrifuging operation to obtain pelleted cells;

providing the pelleted cells in a series of culture media.

20. The method of claim 19, wherein: (0) (2)

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Application US20200107510

GENERATION OF NEW VARIETIES OF CANNABIS BY CHEMICAL MUTAGENESIS OF CANNABIS CELL SUSPENSIONS

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