Genetically modified reduced-browning fruit-producing plant and produced fruit thereof, and method of obtaining such
Inventors
Armstrong, John • Lane, William David
Assignees
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Publication Number
US-8563805-B2
Publication Date
2013-10-22
Expiration Date
Abstract
A genetically modified fruit-producing plant, said plant having sufficiently reduced total Polyphenol Oxidase (PPO) activity relative to a wild type of said plant to reduce browning in the fruit of said plant relative to said wild type, wherein the reduced total PPO activity results from a reduction in activity of at least two PPO isoenzymes in said plant relative to said wild type, or a cell, seed, seedling, part, tissue, cell, fruit or progeny of said plant.
Core Innovation
The invention relates to a genetically modified apple plant whose fruit has reduced browning potential relative to wild type. The reduced browning potential results from reduction in expression of genes encoding at least four polyphenol oxidase (PPO) isoenzymes in the apple plant relative to the wild type.
The core technical approach suppresses multiple PPO isoenzymes by providing first, second, third, and fourth heterologous nucleic acid molecules. Each heterologous nucleic acid molecule comprises at least 100 contiguous nucleotides possessing 100% sequence identity to the specified sequence references, encodes a corresponding PPO isoenzyme amino acid sequence, and reduces expression of that PPO isoenzyme.
The invention also defines nucleic acid constructs that include a promoter and the multi-PPO suppressing nucleic acid molecules. One construct arrangement operably links the first, second, third, and fourth nucleic acid molecules to the promoter in sense orientation, while another construct encodes an mRNA capable of forming a stem loop structure with first and second sets separated by a spacer and arranged in antisense orientation.
Claims Coverage
The independent claims cover genetically modified apple plants and related biological materials having reduced fruit browning potential by reducing expression of at least four PPO isoenzymes, and also cover nucleic acid constructs and methods for producing such plants. The coverage includes multiple independent claim families with the principal inventive features focused on multi-isoenzyme PPO gene expression reduction using defined heterologous nucleic acids.
Reduced browning via expression reduction of at least four PPO isoenzymes
A genetically modified apple plant, cell, seed, seedling, part, tissue, fruit, or progeny in which reduced browning potential relative to wild type results from reduction in expression of genes encoding at least four polyphenol oxidase (PPO) isoenzymes, using four heterologous nucleic acid molecules that each encode a respective PPO isoenzyme and reduce expression of that isoenzyme.
Producing reduced-browning apple plants by reducing expression of at least four PPO isoenzymes
A method for producing a genetically modified apple plant having reduced browning potential in fruit relative to wild type by introducing into an apple plant the first, second, third, and fourth heterologous nucleic acid molecules and selecting an apple plant comprising said first, second, third, and fourth heterologous nucleic acid molecules.
Producing reduced-browning apple plants by reducing expression in apple plant cells of at least four PPO isoenzymes
A method for producing a genetically modified apple plant with reduced browning potential in fruit relative to wild type by introducing into an apple plant cell the first, second, third, and fourth heterologous nucleic acid molecules, selecting an apple plant cell comprising all four, and regenerating an apple plant therefrom.
Nucleic acid construct with promoter and four sense-linked PPO suppressing nucleic acids
A nucleic acid construct comprising a promoter and a first, second, third, and fourth heterologous nucleic acid molecule that each encodes a respective PPO isoenzyme and reduces expression of that PPO isoenzyme, where the nucleic acid molecules are operably linked to the promoter in sense orientation.
Stem-loop-forming nucleic acid construct with promoter and antisense PPO suppressing sets separated by a spacer
A nucleic acid construct encoding an mRNA capable of forming a stem loop structure, comprising a promoter, a first set of nucleic acid sequences, a spacer, and a second set of nucleic acid sequences; the first set comprises first through fourth heterologous nucleic acid molecules and the second set comprises the first, second, third, and fourth nucleic acid sequences operably linked to the promoter in antisense orientation.
Across the independent claims, the shared inventive concept is reduced fruit browning potential achieved by reducing expression of at least four PPO isoenzymes using four defined heterologous nucleic acid molecules, with nucleic acid construct embodiments specifying promoter linkage in sense orientation or an mRNA stem-loop structure with antisense orientation separated by a spacer, and with plant production via introducing, selecting, and regenerating from genetically modified apple plant cells.
Stated Advantages
Reduced browning potential of fruit relative to wild type.
Reduced browning potential results from reduction in expression of genes encoding at least four PPO isoenzymes.
Documented Applications
Genetically modified apple plants, including fruit, progeny, and related plant materials, having reduced browning potential relative to wild type.
Methods for producing genetically modified apple plants with reduced browning potential by introducing and selecting multi-PPO suppressing heterologous nucleic acid molecules and regenerating plants from modified apple plant cells.
Nucleic acid constructs encoding and/or forming mRNA with promoter-linked, sense- or antisense-arranged nucleic acid sequences for reducing expression of PPO isoenzymes in apple contexts.
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