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-12116585-B2
Publication Date
2024-10-15
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
Genetically modified apple plants are provided with a heterologous nucleic acid molecule that includes two or more nucleic acid sequences corresponding to polyphenol oxidase (PPO) isoenzymes, such that fruit has reduced browning potential relative to fruit of a wild type apple plant. The reduced browning potential results from reduction in expression of two or more genes encoding PPO isoenzymes in the genetically modified apple plant compared to the wild type apple plant.
The heterologous nucleic acid molecule comprises at least 50 contiguous nucleotides of specified sequence identifiers, including at least a second or a fourth nucleic acid sequence, and in embodiments includes at least three of the first, second, third, and fourth sequences. Each included nucleic acid sequence is capable of reducing expression of a corresponding PPO isoenzyme expression target, thereby reducing total PPO activity and associated fruit browning potential.
RNA-based suppression is used to target PPO isoenzyme expression, including transcripts capable of forming an mRNA stem-loop structure, and regulatory and structural features such as operably linked promoters. Example multi-isoenzyme configurations are described using isoenzyme combinations directed to PPO isoenzymes, with evaluation using PPO activity and expression and browning measurements including luminosity change (ΔL) and total color change (ΔE), and field trial observations of reduced bruising and juice browning.
Claims Coverage
The independent claims cover genetically modified apple plants having heterologous nucleic acid molecules that target multiple PPO isoenzyme genes to reduce fruit browning potential relative to wild type. The inventive features are organized around sequence-defined nucleic acid segments, capacity to reduce corresponding PPO isoenzyme expression, and the resulting reduction in expression of multiple PPO genes to produce reduced browning potential; additional dependents tighten the phenotype using an mRNA stem-loop capability and a quantified browning reduction threshold of at least 50%.
Multi-isoenzyme PPO gene expression reduction for reduced fruit browning
A genetically modified apple plant comprising a heterologous nucleic acid molecule where, when two or more of specified nucleic acid sequences are present, each is capable of reducing expression of a corresponding PPO isoenzyme, such that fruit has reduced browning potential relative to fruit of a wild type apple plant, with the reduced browning potential resulting from reduction in expression of two or more genes encoding PPO isoenzymes compared to the wild type apple plant.
Heterologous nucleic acid includes specified PPO isoenzyme sequence segments
The genetically modified apple plant where at least one of the two or more nucleic acid sequences comprises the second or the fourth nucleic acid sequence, and the heterologous nucleic acid molecule comprises at least 50 contiguous nucleotides of SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, and SEQ ID NO:24, such that the presence of required sequences yields reduced browning potential via reduction in expression of corresponding PPO isoenzymes.
Three or more PPO isoenzyme sequences reduce expression of three or more PPO genes
A genetically modified apple plant comprising a heterologous nucleic acid molecule comprising three or more of specified nucleic acid sequences (SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, and SEQ ID NO:24), where when three or more are present each is capable of reducing expression of a corresponding PPO isoenzyme, such that fruit has reduced browning potential relative to fruit of a wild type apple plant, and wherein the reduced browning potential results from reduction in expression of three or more genes encoding PPO isoenzymes compared to the wild type apple plant.
Promoter operably linked to the heterologous nucleic acid and mRNA stem-loop capability
The genetically modified apple plant where the heterologous nucleic acid molecule is operably linked to a promoter and encodes an mRNA molecule capable of forming a stem-loop structure, as part of the construct that achieves reduced browning potential.
Quantified browning reduction relative to wild type
The genetically modified apple plant where fruit browning is reduced by at least 50% compared to the wild type apple plant.
Overall, the independent claims require a genetically modified apple plant with a heterologous nucleic acid molecule containing specified PPO-related sequence segments such that multiple PPO isoenzyme gene expressions are reduced, producing reduced fruit browning potential versus wild type. Coverage is further tightened in dependents by operable promoter linkage, an mRNA stem-loop capable transcript, and a browning reduction threshold of at least 50%.
Stated Advantages
Reduced fruit browning potential relative to fruit of a wild type apple plant.
Reduced expression of two or more or three or more genes encoding polyphenol oxidase (PPO) isoenzymes compared to a wild type apple plant.
Reduced bruising and juice browning as described in field trial evaluations.
Fruit browning reduced by at least 50% compared to a wild type apple plant.
Documented Applications
Fruit juice derived from the fruit of the genetically modified apple plant is used to make a beverage.
Foodstuff comprising the genetically modified apple plant part described in the document.
Plant/product applications in food industries including fresh-cut, juice, beverages, and snack industries.
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