# US 11,299,744 B2 · Transgenic plants expressing type 2C protein phosphatase abscisic acid (PP2CABA) proteins and uses thereof

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- Number: US 11,299,744 B2
- Document ID: US11299744B2
- Status: Granted
- Kind: B2
- Application number: 15/838,702
- Filed: 2017-12-12
- Published: 2018-06-14
- Granted: 2022-04-12
- Earliest claimed: 2016-12-12
- Assignees: Academia Sinica
- Applicants: Academia Sinica
- Inventors: Su-May Yu; Chun-Hsien Lu; Tuan-Hua David Ho; Shuen-Fang Lo
- Web page: https://patentgrep.com/patent/US11299744B2

## CPC

- C12N 15/8273: {for drought, cold, salt resistance}
- C12N 9/16: acting on ester bonds (3.1)
- C12N 15/82: for plant cells {, e.g. plant artificial chromosomes (PACs)}
- C12N 15/8255: {involving lignin biosynthesis}
- C12N 15/8261: {with agronomic (input) traits, e.g. crop yield}
- C12N 15/8271: {for stress resistance, e.g. heavy metal resistance}
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The present disclosure relates to transgenic plants that over-express PP2CABA and methods of using such for enhancing osmotic stress tolerance.

## Claims

### Claim 1

A method of improving growth, stress tolerance and root architecture of a plant, comprising:
(a) transforming plant cells with a DNA construct comprising an exogenous nucleic acid operably linked to a heterologous promoter, wherein the exogenous nucleic acid encodes short form type 2C protein phosphatase abscisic acid (PP2CABA) protein consisting of the amino acid sequence as set forth in SEQ ID NO: 4, to obtain transformed plant cells overexpressing said short form PP2CABA protein, and wherein said DNA construct and said exogenous nucleic acid do not encode long form PP2CABA protein having the amino acid sequence set forth in SEQ ID NO: 2;
(b) growing said transformed plant cells obtained in step (a) to regenerate a plurality of transgenic plants overexpressing said short form PP2CABA protein; and
(c) selecting a transgenic plant from said plurality of transgenic plants regenerated in step (b) that exhibits a lower lateral roots (LR) to primary roots (PR) ratio and a higher tolerance to abiotic osmotic or drought stress, as compared to (i) a control plant of the same species lacking said DNA construct and grown under identical growth conditions, and (ii) a transgenic plant overexpressing said long form PP2CABA protein and grown under identical growth conditions.

### Claim 2 (depends on 1)

The method of claim 1, wherein the exogenous nucleic acid encoding short form PP2CABA protein comprises the nucleic acid sequence as set forth in SEQ ID NO: 3, which does not comprise the nucleic acid sequence as set forth in SEQ ID NO: 1.

### Claim 3 (depends on 1)

The method of claim 1, wherein the heterologous promoter is selected from the group consisting of a constitutive promoter, a tissue-specific promoter, a developmental stage-specific promoter, and a promoter inducible by biotic or abiotic stress.

### Claim 4 (depends on 1)

The method of claim 1, wherein the heterologous promoter is a constitutive promoter selected from the group consisting of a maize ubiquitin (Ubi) promoter, a rice actin (Act1) promoter, and a cauliflower mosaic virus 35S (CaMV35S) promoter.

### Claim 5 (depends on 1)

The method of claim 1, wherein the heterologous promoter is a tissue-specific promoter selected from the group consisting of a rice glutelin (GluB) promoter, a rubisco small subunit (rbcS) promoter, and a maize zein gene promoter.

### Claim 6 (depends on 1)

The method of claim 1, wherein the heterologous promoter is a developmental stage-specific promoter selected from the group consisting of a rice alpha-amylase (α-Amylase) promoter, and a rice glycine rich RNA binding protein (GRRP-A1) promoter.

### Claim 7 (depends on 1)

The method of claim 1, wherein the heterologous promoter is a promoter inducible by biotic or abiotic stress, which is selected from the group consisting of an Arabidopsis rd29A promoter, an Arabidopsis corl SA promoter, an Arabidopsis kinl promoter, an Arabidopsis heat-shock factor (HSF) promoter, an Arabidopsis C-repeat-binding factor (CBF1) promoter, an Arabidopsis dehydration-responsive element binding protein (DREB1A) promoter, a rice HVA1 promoter, a rice HVA22 promoter, a rice PP2CABA promoter, an alcohol dehydrogenase (Adh) promoter, an ethanol-inducible promoter, an alpha-amylase promoter, and a synthetic ABRC321 promoter.