Insect resistant genes and methods of use
Inventors
Garbers, Amanda Marie • Lehtinen, Duane • CHOUGULE, NANASAHEB • Beyerlein, Aaron • Zaitseva, Jelena • Eberle, Timothy • Ding, Lei
Assignees
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Publication Number
US-12241075-B2
Publication Date
2025-03-04
Expiration Date
Abstract
Compositions and methods for conferring pesticidal activity to bacteria, plants, plant cells, tissues and seeds are provided. Compositions comprising a coding sequence for a toxin polypeptide are provided. The coding sequences can be used in DNA constructs or expression cassettes for transformation and expression in plants and bacteria. Compositions also comprise transformed bacteria, plants, plant cells, tissues, and seeds. In particular, isolated toxin nucleic acid molecules are provided. Additionally, amino acid sequences corresponding to the polynucleotides are encompassed, and antibodies specifically binding to those amino acid sequences. In particular, the present invention provides for isolated nucleic acid molecules comprising nucleotide sequences encoding the amino acid sequence shown in any of SEQ ID NO: 16 to 30, or the nucleotide sequence set forth in any of SEQ ID NO: 1 to 15, as well as variants and fragments thereof.
Core Innovation
The invention relates to insect resistant genes and methods of use, including recombinant pesticidal/toxin genes derived from Axmi486. The document describes nucleotide sequences and corresponding amino acid sequences, including Axmi486 nucleic acids (SEQ ID NOs 1–15) and Axmi486 proteins (SEQ ID NOs 16–30), and includes Axmi486 variants with encoded pesticidal activity for pest control.
A central aspect is the use of recombinant nucleic acid molecules and recombinant polypeptides comprising pesticidal activity associated with Axmi486, including nucleic acid sequences selected from SEQ ID NO:10 and sequences encoding the Axmi486 polypeptide sequence of SEQ ID NO:25. The document also describes modifications in Axmi486 polypeptides at specified positions that provide increased resistance against Southern armyworm (SAW), while maintaining activity against other target insects.
The invention addresses insect pests and provides approaches for engineered bacteria and transgenic plants that incorporate Axmi486 sequence variants. The described genetic constructs support pest control by killing or protecting against pest taxa, with particular emphasis on increased resistance or mortality against Southern armyworm species while retaining pesticidal activity against other listed pests. The document also describes use for producing or selling commodity plant products containing detectable sequence/protein content.
Claims Coverage
The partial claim set includes multiple independent claims with four independently claimed subject-matter categories: recombinant nucleic acids, modified Axmi486 encoding sequences for increased SAW resistance, recombinant pesticidal polypeptides, and plants/plant-cell embodiments, plus independent methods for protecting plants and increasing yield. Across the independent claims, the inventive scope is anchored by pesticidal activity sequences selected from SEQ ID NO:10 and a sequence encoding the Axmi486 amino acid sequence, and by engineered Axmi486 modifications at specified positions for enhanced SAW resistance.
Recombinant nucleic acid molecule encoding pesticidal activity
A recombinant nucleic acid molecule comprising a nucleotide sequence encoding an amino acid sequence having pesticidal activity, wherein said nucleotide sequence is selected from the group consisting of: (a) the nucleotide sequence set forth in SEQ ID NO: 10; and (b) a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 25.
Axmi486 encoding sequence with modified amino acid positions for increased SAW resistance
A recombinant nucleic acid sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:25, encoding an Axmi486 polypeptide having a modified amino acid corresponding to positions 60, 96, 97, 100 and 275, wherein the polypeptide confers increased resistance against Southern armyworm (SAW) as compared to a control not comprising said modifications.
Recombinant pesticidal polypeptide defined by SEQ ID NO:25
A recombinant polypeptide with pesticidal activity comprising the amino acid sequence of SEQ ID NO:25.
Plant or plant cell with stably incorporated pesticidal DNA construct
A plant or plant cell having stably incorporated into its genome a DNA construct comprising a nucleotide sequence that encodes a protein having pesticidal activity, wherein said nucleotide sequence is selected from the group consisting of: (a) the nucleotide sequence set forth in SEQ ID NO:10; and (b) a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:25.
Method for protecting a plant by expressing a pesticidal polypeptide
A method for protecting a plant from a pest, comprising expressing in a plant or cell thereof a nucleotide sequence that encodes a pesticidal polypeptide, wherein said nucleotide sequence is selected from the group consisting of: (a) the nucleotide sequence set forth in SEQ ID NO:10; and (b) a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:25.
Method for increasing yield in a plant using a stably incorporated pesticidal construct
A method for increasing yield in a plant comprising growing in a field a plant or a seed thereof having stably incorporated into its genome a DNA construct comprising a nucleotide sequence that encodes a protein having pesticidal activity, wherein said nucleotide sequence is selected from the group consisting of: (a) the nucleotide sequence set forth in SEQ ID NO:10; and (b) a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:25, wherein said field is infested with a pest against which said polypeptide has pesticidal activity.
Across the independent claims, the patent centers on nucleic acids and polypeptides comprising pesticidal activity tied to Axmi486 sequences selected from SEQ ID NO:10 and SEQ ID NO:25, and on plants and plant cells with stably incorporated DNA constructs encoding those pesticidal proteins. Separate independent protection and yield-increasing methods require expression of the specified pesticidal sequences in plants and fields infested with targeted pests, with claim 2 specifically emphasizing Axmi486 modifications at positions 60, 96, 97, 100 and 275 to confer increased resistance against SAW compared to controls.
Stated Advantages
Engineered Axmi486 variants increase resistance or mortality against Southern armyworm (SAW) compared to a control not comprising the modifications.
The modified Axmi486 variants maintain activity against other target insects including Plutella xylostella, Anticarsia gemmatalis, Diatraea spp., and Heliothis/Helicoverpa species.
Provides methods of killing or protecting plants against listed pest taxa, including lepidopteran, hemipteran, coleopteran, dipteran, and nematode pests.
Provides a method for increasing yield in a plant by growing plants or seeds with stably incorporated pesticidal DNA constructs in pest-infested fields.
Provides commodity plant products containing detectable sequence/protein.
Documented Applications
Engineered bacteria and transgenic plants used for pest control, including protection against Southern armyworm and other listed insect pests.
Use of modified Axmi486 variants to increase resistance or mortality against Southern armyworm (SAW) while maintaining activity against other target insects.
Methods for protecting a plant from a pest by expressing a nucleotide sequence encoding a pesticidal polypeptide in a plant or plant cell.
Methods for increasing yield in a plant by growing in a field a plant or seed having a stably incorporated DNA construct encoding a pesticidal protein, where the field is infested with a pest against which the polypeptide has pesticidal activity.
Production and sale of commodity plant products comprising seeds or grain, including whole or processed forms, containing detectable sequence/protein content.
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