# US 2018/0291387 A1 · Enhanced plants of genus humulus and methods of making and using the same

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- Number: US 2018/0291387 A1
- Document ID: US20180291387A1
- Status: Published application
- Kind: A1
- Application number: 15/950,713
- Filed: 2018-04-11
- Published: 2018-10-11
- Earliest claimed: 2017-04-11
- Assignees: Ebbu, LLC
- Applicants: ebbu, LLC
- Inventors: Robert Fletcher ROSCOW, JR.
- Web page: https://patentgrep.com/patent/US20180291387A1

## CPC

- C12N 15/8223: {Vegetative tissue-specific promoters}
- C12N 15/8243: {involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Disclosed herein is a genetically modified plant of genus humulus not requiring flowering for producing lupulin glands comprising secondary compounds. Provided herein is a new plant of genus humulus having lupulin glands on non-flowering parts of the plant, such as leaves. The disclosed plants have a high mass % of secondary compounds and a high degree of lupulin glands coverage on the surface of the plant. Also disclosed herein are methods of producing secondary compounds from a plant of genus humulus without flowering the plant of genus humulus. For example, the disclosed methods provide for inducing lupulin glands development on a plant of genus humulus without flowering the plant of genus humulus.

## Claims

### Claim 1

A plant of genus humulus, comprising;
lupulin glands on non-flowering parts of the plant of genus humulus;
a vector comprising genetic information; and
wherein the plant of genus humulus comprises a surface area.

### Claim 2 (depends on 1)

The plant of claim 1, wherein the vector is a plasmid comprising a cDNA sequence native to a plant of genus humulus corresponding to lupulin gland induction.

### Claim 3 (depends on 1)

The plant of claim 1, wherein the vector is a purified transformed bacteria comprising a DNA sequence native to a plant of genus humulus, wherein said native DNA sequence corresponds to lupulin gland induction.

### Claim 4 (depends on 1)

The plant of claim 1, comprising lupulin glands on about 25-100% of the surface area of the plant.

### Claim 5 (depends on 4)

The plant of claim 4, comprising lupulin glands on about 50-100% of the surface area of the plant.

### Claim 6 (depends on 4)

The plant of claim 4, comprising lupulin glands on about 70-100% of the surface area of the plant.

### Claim 7 (depends on 1)

The purified transformed bacteria of claim 1, comprising a buffer solution; wherein the vector is suspended in the buffer solution.

### Claim 8 (depends on 1)

The plant of claim 1, comprising 20-80 mass % of secondary compounds of the plant.

### Claim 9 (depends on 8)

The plant of claim 8, comprising about 30-70 mass % of secondary compounds of the plant.

### Claim 10 (depends on 8)

The plant of claim 8, comprising about 40-65 mass % of secondary compounds of the plant.

### Claim 11 (depends on 8)

The plant of claim 8, comprising about 50-60 mass % of secondary compounds of the plant.

### Claim 12

A plant of the genus humulus comprising cDNA fragments corresponding to lupulin gland induction in a plant not of the genus humulus.

### Claim 13 (depends on 12)

The plant of claim 12, wherein the plant not of the genus humulus is a plant of genus Arabidopsis.

### Claim 14

A method of producing secondary compounds, comprising inducing lupulin gland development in a plant of genus humulus.

### Claim 15 (depends on 14)

The method of claim 14, wherein said secondary compounds are chosen from alpha acids, beta acids, essential oils, flavonoids, or terpenes.

### Claim 16 (depends on 15)

The method of claim 15, comprising harvesting at least one alpha acid or at least one terpene during the vegetative growth cycle of the plant of the genus humulus.

### Claim 17 (depends on 14)

The method of claim 14, comprising modifying genetic material of the plant of the genus humulus.

### Claim 18 (depends on 14)

The method of claim 14, comprising introducing non-native DNA sequence into the plant of the genus humulus.

### Claim 19 (depends on 17)

The method of claim 17, comprising overexpressing at least one lupulin gland gene.

### Claim 20 (depends on 19)

The method of claim 19, comprising interfering with expression of at least one lupulin gland gene.

### Claim 21 (depends on 20)

The method of claim 20, wherein the at least one lupulin gland gene is chosen from a bHLH Basic Helix Loop Helix transcription factor, a R2R3 MYB transcription factor, a R3 MYB transcription factor, a WD40 repeat transcription factor/ protein, or a homeodomain transcription factor.