Control of phenotype in plants
Inventors
Boddupalli, Sekhar • Boukharov, Andrey • STAMLER, Rio • Li, Zhongsen • TREMBLAY, Arianne • SCHAUER, Stephen • Tiwari, Shiv B. • Salmeron, John
Assignees
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Publication Number
US-11555197-B2
Publication Date
2023-01-17
Expiration Date
Abstract
The invention provides a compositions and methods for controlling phenotypic traits in plants. Genes of interest are placed under the control of a gene switch to allow inducible control or expression of a gene of interest “on-demand” by treatment of the plant with a chemical ligand.
Core Innovation
The invention provides one or more vectors or polynucleotides comprising a VGEvy gene switch that induces expression of a nucleic acid controlling one or more traits in a plant in the presence of a chemical ligand. The VGEvy gene switch comprises a transactivation domain, a DNA-binding domain, and a ligand-binding domain having an amino acid sequence of SEQ ID NO:2. The regulatory element is operably connected 5′ of the trait-controlling nucleic acid, such that the gene switch induces trait expression when the ligand is present.
The plant traits that are controlled include flowering and floral characteristics, resistance and protection traits, and traits related to nutrition or exogenous gene expression. The document further describes control of flowering using nucleic acids that encode full-length polypeptides or biologically active fragments of flowering-related genes, including Gigantea (GI), Nuclear Factor Y (NFY), Constans (CO), protein farnesyltransferase (PFT), Agamous-Like 15 (AGL15), Flowering Locus T (FT), Twin Sister of FT (TSF), FD (FD bZIP transcription factor), APETALA1 (AP1), SOC1, AGL24, LEAFY (LFY), and SPL3.
Activation of the VGEvy gene switch is described using non-steroidal diacylhydrazine ligands, including methoxyfenozide and diacylhydrazine, and the ligand system is described in the context of chemically inducible gene switches for on-demand trait control. The document also discusses additional switch systems beyond the VGEvy/EcR approach, including rapamycin/FKBP-FRB, tetracycline/Lac, cumate, riboswitch, and biotin-based gene switches, together with selection of regulatory elements and promoters that connect the DNA-binding domain to expression of the trait-controlling nucleic acid.
Claims Coverage
The independent claim covers one or more vectors or polynucleotides comprising a VGEvy gene switch, a trait-controlling nucleic acid, and a regulatory element operably connected 5′ of that trait-controlling nucleic acid, wherein the switch induces expression in the presence of a chemical ligand. The cited dependent refinements specify trait scope, a flowering gene panel, ligand identity, regulatory element architecture, and integration into the plant genome.
VGEvy gene switch induces trait expression in presence of chemical ligand
A vector or polynucleotide comprising a polynucleotide encoding a VGEvy gene switch including a transactivation domain, a DNA-binding domain, and a ligand-binding domain having an amino acid sequence of SEQ ID NO:2; a nucleic acid which controls one or more traits in a plant; and a regulatory element operably connected 5′ of the nucleic acid, wherein the VGEvy gene switch induces expression in the presence of a chemical ligand.
Regulatory element with recognition site and promoter for VGEvy
The vectors or polynucleotides comprise a regulatory element that includes a recognition site for the DNA-binding domain of a VGEvy gene switch and a promoter.
Nucleic acid controlling one or more traits including flowering, resistance, and nutrition-related traits
The nucleic acid controls one or more traits in a plant, wherein the traits include flowering-, drought-, herbicide-, pest-, and disease-resistance characteristics, floral color or aroma intensity, accumulation of specific nutrients, taste, aroma, color, nutritional value, or expression of an exogenous gene.
Flowering control by encoding specified flowering gene polypeptides or active fragments
The nucleic acid which controls one or more traits in a plant comprises encoding a full-length polypeptide or a biologically active fragment of one or more of Gigantea (GI), Nuclear Factor Y (NFY), Constans (CO), protein farnesyltransferase (PFT), Agamous-Like 15 (AGL15), Flowering Locus T (FT), Twin Sister of FT (TSF), FD (bZIP transcription factor), APETALA1 (AP1), SOC1, AGAMOUS-LIKE 24 (AGL24), Leafy (LFY), and SPL3.
Chemical ligand activation using methoxyfenozide or diacylhydrazine
The VGEvy gene switch is activated by a chemical ligand selected from methoxyfenozide or diacylhydrazine.
Integration into plant genome
The plant further includes one or more vectors or polynucleotides integrated into the plant genome.
Overall, the claim coverage centers on vectors or polynucleotides that combine a VGEvy gene switch with a 5′ regulatory element to drive expression of a trait-controlling nucleic acid upon chemical ligand presence. Dependent refinements narrow the trait categories, specify the ligand identity, define the regulatory element architecture, and require integration into the plant genome.
Stated Advantages
External temporal control of gene expression in plants based on ligand-dependent induction of transcription.
Ligand selection supports control over gene-switch activation using different ligand classes depending on gene-switch type.
Inducible control enables trait modulation including flowering control and disease resistance via inducible antimicrobial or immune genes.
Documented Applications
Controlled flowering in plants using nucleic acids and regulatory elements operably connected 5′ of trait control.
Disease resistance in plants through inducible expression of antimicrobial or immune genes.
Trait modulation in plants including flowering and other trait categories controlled by ligand-activated VGEvy gene switch systems.
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