Plants having increased oil quality

Inventors

Marks, Michael DavidChopra, RatanFolstad, NicoleWyse, Donald L.Sedbrook, John C.

Assignees

Illinois State UniversityUniversity of Minnesota System

Publication Number

US-12221612-B2

Publication Date

2025-02-11

Expiration Date

2039-02-12

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Abstract

This document relates to methods and materials for generating oilseed (e.g., pennycress) plants that have oil with reduced levels of polyunsaturated fatty acids (PUFAs) and/or increased levels of oleic acid. For example, oilseed plants having reduced expression levels of one or more polypeptides involved in fatty acid biosynthesis (e.g., fatty acid desaturase 2 (FAD2) and reduced oleate desaturation 1 (ROD1)), as well as methods and materials for making and using such oilseed plants are provided.

Core Innovation

This invention provides methods and materials for generating oilseed plants, such as pennycress, that produce oils with reduced levels of polyunsaturated fatty acids (PUFAs) and/or increased levels of oleic acid. For example, the invention describes pennycress plants in which the expression or activity of polypeptides involved in fatty acid biosynthesis—particularly fatty acid desaturase 2 (FAD2) and reduced oleate desaturation 1 (ROD1)—is reduced due to specific genetic modifications. These modifications may include substitutions, insertions, deletions, or frameshift mutations in relevant genes.

The problem addressed is that natural fatty acid profiles of wild pennycress strains make pennycress oil inedible for humans and suboptimal for both biofuel and food production. By altering the genetic makeup of pennycress plants to reduce PUFA content and/or increase oleic acid levels, the invention improves oil quality, making it more suitable for a range of applications and comparable to oils like canola and olive oil.

The invention encompasses specific genetic modifications in the FAD2 gene, ROD1 gene, and optionally the FAE1 gene, using, for example, site-specific genome editing techniques. These engineered pennycress plants, or their seeds, exhibit significant changes in their oil fatty acid compositions: enhanced oleic acid content and reduced PUFAs, with some lines also exhibiting reduced or eliminated erucic acid. The document details representative nucleotide and amino acid sequences, methods of mutagenesis (such as EMS treatment and CRISPR-Cas9), and the resulting improvements in seed oil composition.

Claims Coverage

The independent claim of the patent covers a genetically modified pennycress plant with specific characteristics relating to fatty acid composition through targeted modifications of the ROD1 gene.

Loss-of-function modification in ROD1 gene in pennycress

A pennycress plant with a genome comprising a modified reduced oleate desaturation 1 (ROD1) gene. The ROD1 gene harbors a loss-of-function modification effective to knock out the expression and/or activity of ROD1 polypeptides. This modification is specifically defined as being effective in reducing levels of polyunsaturated fatty acids (PUFAs) and/or increasing levels of oleic acid in the plant, compared to a pennycress plant lacking the modification. The unmodified coding sequence of the ROD1 gene encodes a ROD1 polypeptide having the sequence of SEQ ID NO:10.

The claim coverage centers on pennycress plants genetically modified to include a loss-of-function in the ROD1 gene, resulting in reduced PUFAs and/or increased oleic acid in the oil.

Stated Advantages

The invention enables the production of oilseed plants with improved oil quality, offering reduced levels of PUFAs and/or increased levels of oleic acid compared to wild type plants.

Oil with reduced PUFAs provides increased stability, such as an extended shelf life and a wider temperature range for use, including at elevated temperatures.

High oleic acid levels are desirable for human food sources and as a feedstock, as well as for the production of various bio-based products.

These oilseed plants can be grown between other major crops (e.g., corn and soybean) without displacing existing crops, maximizing land use efficiency.

The improved oil compositions make pennycress oil comparable to high-value oils such as canola, olive oil, and high oleic sunflower.

Documented Applications

Production of biofuels, including biodiesel and bio jet fuel, from pennycress oil with reduced PUFA and/or increased oleic acid content.

Use of oil from modified pennycress plants in the creation of bioproducts, such as biopolymers for bioplastics.

Application of the oil as animal feed supplements.

Use as edible oil for human consumption.

Use of oil with reduced PUFAs in high-temperature applications, such as food fryers or as lubricants for mechanical systems.

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