Endocarp-specific promoter and uses thereof
Inventors
Callahan, Ann M. • Dardick, Christopher D.
Assignees
US Department of Agriculture USDA
Publication Number
US-12060565-B1
Publication Date
2024-08-13
Expiration Date
2043-03-15
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Abstract
The disclosure relates to polynucleotides comprising an endocarp-specific promoter sequence from the peach peroxidase (POX) gene operably linked to a heterologous nucleotide sequence encoding a product of interest. The polynucleotides can be used to specifically alter endosperm gene expression in fruit. In a transgenic cell, the polynucleotides may block formation of phenylpropanoid pathway lignin-pathway precursors, formation of lignin monomers, and/or polymerization of lignin monomers.
Core Innovation
The invention provides an endocarp-specific promoter, identified from the peach peroxidase (POX) gene, which can be used to produce plants with altered stone tissue without negatively impacting plant growth or fruit production. This promoter sequence is operably linked to heterologous nucleotide sequences encoding products of interest, such as RNA molecules or polypeptides, particularly those involved in the lignin biosynthetic pathway. The polynucleotides comprising this promoter can be included within vectors, host cells, plants, or plant parts to specifically modulate gene expression in the endocarp tissue layer of drupe fruits.
The problem addressed by the invention arises from the presence of hard, lignified endocarp tissue (stone) in stone fruits such as cherries, plums, apricots, and others. This woody tissue hinders the development of seedless fruit and complicates processing, leading to costly pitting and quality control measures. Prior to this invention, no promoters specific to the endocarp tissue had been identified, limiting the ability to genetically modify endocarp properties without affecting other tissues. The invention solves this by identifying and cloning a promoter sequence specific to the endocarp, derived from the peach peroxidase gene, which can drive specific expression of gene constructs to alter lignification and stone formation, potentially softening or eliminating the hard endocarp in stone fruits and related drupes.
Claims Coverage
The patent contains two primary independent claims covering a polynucleotide comprising an endocarp-specific promoter sequence linked to a heterologous nucleotide sequence, and a method for modifying endocarp development in plants using such polynucleotides. The claims emphasize the specificity of the promoter and the targeted modification of lignin-related pathways within endocarp tissue.
Endocarp-specific promoter polynucleotide linked to heterologous nucleotide sequence
A polynucleotide comprising an endocarp-specific promoter sequence at least 99% identical to SEQ ID NO: 1 operably linked to a heterologous nucleotide sequence encoding a product of interest, where the product can be an RNA molecule or polypeptide, including those involved in the lignin biosynthetic pathway or negative regulators of endocarp development and lignification.
Method for modifying endocarp development in fruit
A method comprising transforming a plant cell with the polynucleotide comprising the endocarp-specific promoter, regenerating a plant from the transformed cell, and growing the regenerated plant to produce fruit exhibiting modified endocarp development, including reduced lignin formation, where the plant can be a drupe species.
The independent claims cover the provision of an endocarp-specific promoter sequence used to control heterologous gene expression specifically in endocarp tissue for altering stone development, and a method for using this polynucleotide to genetically modify endocarp properties in fruit-bearing drupe plants.
Stated Advantages
The promoter drives gene expression specifically in the endocarp tissue, allowing targeted modification of stone tissue without adverse effects on other plant tissues or overall plant growth.
Use of this promoter enables production of fruit with modified or reduced lignin formation in the endocarp, facilitating seedless fruit development and easier processing.
The technology has the potential to reduce production costs in processing industries by minimizing stone fragments and improving the quality of processed stone fruit products.
Documented Applications
Use in transforming drupe fruit plants (e.g., plum, peach, nectarine, apricot, date, mango, cherry, almond, coffee, jujube, oil palm, pistachio, damson, olive, coconut, holly fruit, elder fruit, mulberry, blackberry, or raspberry) to alter endocarp development.
Driving expression of RNA hairpins or polypeptides that inhibit lignin biosynthesis pathways, including blocking formation of phenylpropanoid pathway lignin precursors, lignin monomers, or polymerization enzymes, to produce fruit with reduced or modified stone tissue.
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