Compositions and methods for improving crop yields through trait stacking

Patent metadata

Field	Value
Publication number	US-12385060-B2
Application number	US-202017634381-A
Country	US
Kind code	B2
Filing date	Aug 13, 2020
Priority date	Aug 14, 2019
Publication date	Aug 12, 2025
Grant date	Aug 12, 2025

Abstract

The present disclosure provides modified, transgenic, or genome edited/mutated corn plants that are semi-dwarf and have one or more improved ear traits and yield relative to a control plant, such as increased ear area, increased ear volume, increased single kernel weight, increased ear fresh weight, increased ear diameter, increased ear length, increased yield, increased kernels per ear, decreased ear tip void, and decreased ear void. The modified, transgenic, or genome edited/mutated corn plants comprise a transgene encoding one or more CONSTANS (CO) or CONSTANS-like (COL) polypeptides and a transgene encoding one or more MADS-box polypeptides, and can also have a reduced expression of one or more GA20 or GA3 oxidase genes. Also provided are methods for producing the modified, transgenic, or genome edited/mutated corn plants.

First claim

The invention claimed is:

1. A modified corn plant or a plant part thereof comprising 1) a first recombinant expression cassette comprising a transcribable DNA sequence encoding a non-coding RNA for suppression of one or more gibberellic acid 20 (GA20) oxidase genes, wherein the non-coding RNA comprises a targeting sequence that is (a) at least 95% complementary to at least 19 consecutive nucleotides of a first mRNA molecule encoding a first endogenous GA20 oxidase protein that has at least 95% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 9, and (b) at least 95% complementary to at least 19 consecutive nucleotides of a second mRNA molecule encoding a second endogenous GA20 oxidase protein that has at least 95% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 15, 2) a second recombinant expression cassette comprising a first DNA sequence encoding a CONSTANS-like (COL) polypeptide that has at least 95% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 168, and 3) a third recombinant expression cassette comprising a second DNA sequence encoding a MADS-box polypeptide that has at least 95% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 176; wherein the modified corn plant is semi-dwarf and has one or more improved ear traits, relative to a control corn plant that does not have the first, second and third recombinant expression cassettes.

2. The modified corn plant or plant part thereof of claim 1 , wherein the first, second and third recombinant expression cassettes are stably integrated into the genome of the corn plant or plant part thereof.

3. The modified corn plant or plant part thereof of claim 1 , wherein the transcribable DNA sequence comprises a sequence that is at least 90% identical or complementary to at least 15 consecutive nucleotides of SEQ ID NO: 39, 53, or 55.

4. The modified corn plant or plant part thereof claim 1 , wherein the first DNA sequence of the second recombinant expression cassette comprises a sequence that is at least 90% identical to SEQ ID NO: 169.

5. The modified corn plant or plant part thereof of claim 1 , wherein the second DNA sequence of the third recombinant expression cassette comprises a sequence that is at least 90% identical to SEQ ID NO: 177.

6. The modified corn plant or plant part thereof of claim 1 , wherein the expression level of an endogenous GA20 oxidase gene is reduced or eliminated in the modified corn plant or plant part thereof.

7. The modified corn plant or plant part thereof of claim 1 , wherein the transcribable DNA sequence encoding the non-coding RNA for suppression of the one or more GA20 oxidase genes is operably linked to a heterologous plant-expressible promoter.

8. The modified corn plant or plant part thereof of claim 7 , wherein the heterologous plant-expressible promoter:
i. is a vascular promoter;
ii. is selected from the group consisting of a sucrose synthase promoter, a sucrose transporter promoter, a Sh1 promoter, Commelina yellow mottle virus (CoYMV) promoter, a wheat dwarf geminivirus (WDV) large intergenic region (LIR) promoter, a maize streak geminivirus (MSV) coat polypeptide (CP) promoter, a rice yellow stripe 1 (YS1)-like promoter, a rice yellow stripe 2 (OsYSL2) promoter, and a combination thereof;
iii. comprises a DNA sequence that is at least 95% identical to one or more of SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70 or SEQ ID NO: 71;
iv. is a rice tungro bacilliform virus (RTBV) promoter; or
v. comprises a DNA sequence that is at least 95% identical to one or more of SEQ ID NO: 65 or SEQ ID NO: 66.

9. A seed of the modified corn plant of claim 1 , wherein the seed comprises the first, second, and third recombinant expression cassettes.

10. The seed of claim 9 , wherein a progeny plant grown from the seed is semi-dwarf and has one or more improved ear traits, relative to a control corn plant that does not comprise the first, second, and third recombinant expression cassettes.

11. A commodity or commodity product produced from the seed of claim 9 , comprising the first, second, and third recombinant expression cassettes.

12. A method comprising planting the seed of claim 9 in a growth medium or soil.

13. A plurality of modified corn plants in a field, each modified corn plant comprising the modified corn plant of claim 1 .

14. A method for producing a modified corn plant, the method comprising a method selected from the group consisting of (1)-(17):
(1) a. introducing into a corn cell a first recombinant expression cassette comprising a coding sequence for a COL polypeptide that has at least 95% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 168, wherein the corn cell comprises a second recombinant expression cassette comprising a transcribable DNA sequence encoding a non-coding RNA for suppression of one or more GA20 oxidase genes, wherein the non-coding RNA comprises a targeting sequence that is (a) at least 95% complementary to at least 19 consecutive nucleotides of a first mRNA molecule encoding a first endogenous GA20 oxidase protein that has at least 95% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 9, and (b) at least 95% complementary to at least 19 consecutive nucleotides of a second mRNA molecule encoding a second endogenous GA20 oxidase protein that has at least 95% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 15, and a third recombinant expression cassette comprising a coding sequence for a MADS-box polypeptide that has at least 95% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 176; and
b. regenerating or developing a modified corn plant from the corn cell, wherein the modified corn plant comprises the first, second and third recombinant expression cassettes; and wherein the modified corn plant is semi-dwarf and has one or more improved ear traits, relative to a control corn plant that does not have the first, second and third recombinant expression cassettes;
(2) a. introducing into a corn cell a first recombinant expression cassette comprising a coding sequence for a MADS-box polypeptide that has at least 95% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 176, wherein the corn cell comprises a second recombinant expression cassette comprising a transcribable DNA sequence encoding a non-coding RNA for suppression of one or more GA20 oxidase genes, wherein the non-coding RNA comprises a targeting sequence that is (a) at least 95% complementary to at least 19 consecutive nucleotides of a first mRNA molecule encoding a first endogenous GA20 oxidase protein that has at least 95% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 9, and (b) at least 95% complementary to at least 19 consecutive nucleotides of a second mRNA molecule encoding a second endogenous GA20 oxidase protein that has at least 95% amino acid sequence identity to the amino acid sequence as set forth in SEQ ID NO: 15, and a third recombinant expression cassette comprising a coding sequence for a COL polypeptide that has at least 95% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 168; and
b. regenerating or developing a modified corn plant from the corn cell, wherein the modified corn plant comprises the first, second, and third recombinant expression cassettes; and wherein the modified corn plant is semi-dwarf and has one or more improved ear traits, relative to a control corn plant that does not have the first, second and third recombinant expression cassette

Assignees

• Monsanto Technology Llc

Inventors

• Dietrich Charles R
• Ivleva Natalia
• Slewinski Thomas L

Classifications

• C12N15/827

  Flower development or morphology, e.g. flowering promoting factor [FPF] · CPC title

• C12N15/8262

  involving plant development · CPC title

• C12N15/8218

  Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS] · CPC title

• C12N15/8297 Primary

  Gibberellins; GA3 · CPC title

• A01H6/4684

  Zea mays [maize] · CPC title