Engineering wheat with increased dietary fiber
Inventors
Baltes, Nicholas • Gil Humanes, Javier
Assignees
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Publication Number
US-12012607-B2
Publication Date
2024-06-18
Expiration Date
Abstract
Materials and methods are provided for making plants (e.g., Triticum varieties) with increased levels of dietary fiber, such as by making TALE nuclease-induced mutations in alleles encoding starch branching enzyme IIa (SBEIIa) and starch branching enzyme IIb (SBEIIb).
Core Innovation
The invention engineers a hexaploid Triticum plant, plant part, or plant cell to contain at least three deletion or insertion mutations induced by contact with a transcription activator-like effector (TALE) nuclease. The TALE nuclease is configured to bind a nucleic acid sequence as set forth in SEQ ID NO: 8, and the induced mutations are within the binding site in at least three starch branching enzyme IIa (SBEIIa) alleles endogenous to the plant, plant part, or plant cell in each genome.
The engineered hexaploid Triticum keeps each starch branching enzyme IIb (SBEIIb) allele wild-type while modifying the SBEIIa alleles. The mutant SBEIIa alleles comprise sequences set forth in SEQ ID NO: 11910, 11912, and 11915, respectively, or the sequences set forth in SEQ ID NO: 11910, 11913, and 11915, respectively.
The disclosure further describes generating such mutant Triticum plants by contacting hexaploid Triticum plant cells or plant parts comprising functional SBEII alleles with the TALE nuclease configured to bind to the nucleic acid sequence in SEQ ID NO: 8. The selected plant cell or plant part is grown into a Triticum plant having increased dietary fiber relative to a control where the SBEIIa alleles have not been mutated.
Claims Coverage
The provided excerpt includes three independent claims, covering engineered hexaploid Triticum with specific SBEIIa mutation profiles induced by a TALE nuclease while maintaining wild-type SBEIIb alleles, a Triticum plant with homozygous mutations in endogenous SBEIIa homoeoalleles defined by specific sequence sets, and a method for generating the dietary-fiber-increasing Triticum using TALE nuclease contacting, selection of specified mutation outcomes, and growing to a plant.
TALE-induced multi-allelic SBEIIa binding-site deletions or insertions with wild-type SBEIIb
A hexaploid Triticum plant, plant part, or plant cell comprising at least three deletion or insertion mutations induced by contact with a transcription activator-like effector (TALE) nuclease configured to bind a nucleic acid sequence as set forth in SEQ ID NO: 8, wherein the at least three deletion or insertion mutations are within the binding site of the TALE nuclease in at least three starch branching enzyme IIa (SBEIIa) alleles endogenous to the plant, plant part, or plant cell, in each genome, and wherein each starch branching enzyme IIb (SBEIIb) allele is a wild-type SBEIIb allele, and wherein the Triticum plant, plant part or plant cell has increased levels of dietary fiber as compared to a control hexaploid Triticum plant in which the SBEIIa alleles have not been mutated.
Homozygous SBEIIa homoeoallele mutations defined by SEQ ID NO combinations with increased dietary fiber
A Triticum plant, plant part, or plant cell comprising mutations within three starch branching enzyme Ha (SBEIIa) homoeoalleles endogenous to the plant, plant part, or plant cell, wherein the mutant SBEIIa homoeoalleles comprise the sequences set forth in SEQ ID NO:11910, 11912, and 11915, respectively, or the sequences set forth in SEQ ID NO:11910, 11913, and 11915, respectively, and wherein the Triticum plant, plant part or plant cell has increased levels of dietary fiber as compared to a control hexaploid Triticum plant in which the SBEIIa alleles are wild-type, wherein the plant, plant part, or plant cell contains an A, B, and D genome and the mutations are homozygous mutations in the SBEIIa-A, B and D alleles.
TALE nuclease contacting, selection of binding-site SBEIIa deletions or insertions with wild-type SBEIIb, and growing to a dietary-fiber-increasing plant
A method for generating a Triticum plant that has increased levels of dietary fiber, wherein the method comprises contacting a hexaploid Triticum plant cell or plant part comprising functional SBEII alleles with a TALE nuclease configured to bind to a nucleic acid sequence as set forth in SEQ ID NO: 8; selecting from the plant cell or plant part a plant cell or plant part in which at least one SBEIIa allele in each genome of the plant cell or plant part comprises a deletion or insertion mutation within the binding site of the TALE nuclease and each SBEIIb allele is a wild-type SBEIIb allele; and growing the selected plant cell or plant part into a Triticum plant having increased levels of dietary fiber as compared to a control hexaploid Triticum plant in which the SBEIIa alleles have not been mutated, wherein the mutant SBEIIa alleles comprise the sequences set forth in SEQ ID NO:11910, 11912, and 11915, respectively, or the sequences set forth in SEQ ID NO:11910, 11913, and 11915, respectively, and wherein the selected plant cell or plant part contains an A, B, and D genome and the mutations are homozygous mutations in the SBEIIa-A, B and D alleles.
Across the independent claims, the inventive core is engineering hexaploid Triticum with increased dietary fiber by inducing at least three deletion or insertion mutations within TALE binding sites in endogenous SBEIIa alleles in the A, B, and D genomes while keeping all SBEIIb alleles wild-type. The claims further constrain the mutant SBEIIa alleles to specific SEQ ID NO sequences and, in the method claim, require contacting with a TALE nuclease, selecting for the specified mutation pattern, and growing to a dietary-fiber-increasing Triticum plant.
Stated Advantages
Increased levels of dietary fiber as compared to a control hexaploid Triticum plant in which the SBEIIa alleles have not been mutated.
Documented Applications
No documented applications found
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