Low glucosinolate pennycress meal and methods of making
Inventors
Ulmasov, Tim • Sedbrook, John C. • Marks, Michael David • McGinn, Michaela G. • Chopra, Ratan • Jarvis, Brice Allen
Assignees
Illinois State University • CoverCress Inc • University of Minnesota System
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Publication Number
US-10988772-B2
Publication Date
2021-04-27
Expiration Date
Abstract
Pennycress (Thlaspi arvense) seed, seed lots, seed meal, and compositions with reduced glucosinolate content as well as plants that yield such seed, seed lots, seed meal, and compositions are provided. Methods of making and using the pennycress plants and/or seed that provide such seed, seed lots, seed meal, and compositions are also provided.
Core Innovation
The invention relates to pennycress seed that contains reduced sinigrin, specifically having less than 30 micromoles sinigrin per gram by dry weight. The document describes seed lots and pennycress seed meal, including defatted and non-defatted meal, where compositions are characterized by sinigrin content below multiple thresholds.
The reduced sinigrin trait is tied to genetic suppression or loss-of-function affecting sinigrin biosynthesis, transport, hydrolysis, and/or regulatory genes, with resulting functional outcomes reflected in reduced transcripts and/or reduced enzymatic or transport activity. In particular, the document describes pennycress seeds that carry at least one loss-of-function mutation in an endogenous pennycress gene encoding the polypeptide of SEQ ID NO: 3 or an allelic variant thereof, where the loss-of-function mutation reduces expression of the polypeptide or reduces 2-oxoglutarate-dependent dioxygenase activity of the polypeptide.
The document also describes pennycress seeds that comprise at least one transgene or genome rearrangement that suppresses expression of an endogenous pennycress gene encoding the polypeptide of SEQ ID NO: 3 or an allelic variant thereof. Allelic variants of the SEQ ID NO: 3 polypeptide are defined by at least 95% sequence identity to SEQ ID NO: 3, and the described genetic changes are validated by observed genotype–phenotype relationships, including reductions in measured sinigrin levels using both NIRS and wet-lab assays, and by co-segregation results in edited and mutagenized lines.
Claims Coverage
The document includes one independent claim, directed to a pennycress seed with defined sinigrin content and specified genetic suppression of a target pennycress polypeptide (SEQ ID NO: 3). Dependent claims primarily tighten the sinigrin thresholds and broaden alternative endogenous genetic targets and/or define a defatted seed meal production use case with further sinigrin limits.
Low-sinigrin pennycress seed with defined SEQ ID NO: 3 genetic suppression
A pennycress seed having less than 30 micromoles sinigrin per gram by dry weight, wherein the seed comprises at least one loss-of-function mutation in an endogenous pennycress gene encoding the polypeptide of SEQ ID NO: 3 (or an allelic variant) that reduces expression or reduces 2-oxoglutarate-dependent dioxygenase activity; or at least one transgene or genome rearrangement that suppresses expression of the endogenous pennycress gene encoding the polypeptide of SEQ ID NO: 3 (or an allelic variant), with allelic variants having at least 95% sequence identity to SEQ ID NO: 3.
Further reduced sinigrin threshold for pennycress seed
A pennycress seed characterized by containing 1 to 15 micromoles of sinigrin per gram by dry weight.
Functional loss-of-function requirement for SEQ ID NO: 3 polypeptide
A pennycress seed comprising at least one loss-of-function mutation in a pennycress gene encoding the polypeptide of SEQ ID NO: 3 (or an allelic variant), wherein the loss-of-function mutation reduces polypeptide expression or reduces 2-oxoglutarate-dependent dioxygenase activity of the polypeptide.
Alternative endogenous targets linked to SEQ ID NO: 1 and SEQ ID NO: 2
A pennycress seed comprising at least one loss-of-function mutation in an endogenous pennycress coding sequence or gene specified by SEQ ID NO: 1 or SEQ ID NO: 2, or an allelic variant with at least 95% sequence identity to the respective SEQ ID.
Defatted pennycress seed meal with low sinigrin via solvent extraction and separation
A method for producing defatted pennycress seed meal having less than 30 micromoles sinigrin per gram by dry weight by solvent-extracting a pennycress seed lot and separating the extracted seed meal from the solvent.
Further reduced sinigrin threshold for defatted seed meal
The method of producing defatted pennycress seed meal further requires that the defatted seed meal contains less than 15 micromoles of sinigrin per gram (dry weight).
Across the independent claim and its dependents, the claimed inventive concept centers on pennycress with defined low sinigrin content achieved by genetic suppression or loss-of-function targeting the polypeptide of SEQ ID NO: 3 (with specified functional consequences and an allelic identity criterion), with optional tightening of sinigrin thresholds, alternative endogenous targets linked to SEQ ID NO: 1 and SEQ ID NO: 2, and an additional method claim for producing defatted seed meal with further sinigrin limits.
Stated Advantages
Documented Applications
Animal feed use of low-glucosinolate pennycress seed and seed meal compositions having reduced sinigrin levels.
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