US-12618081-B2 - Brown marmorated and harlequin stink bug pheromone enzyme synthesis and uses thereof

Abstract

Described herein are engineered polynucleotides and vectors capable of encoding one or more engineered harlequin and/or brown marmorated stink bug pheromone synthesis enzymes. Also described herein are engineered harlequin and/or brown marmorated stink bug pheromone synthesis enzymes. Also described herein are methods of making modified plants capable of expressing one or more harlequin and/or brown marmorated stink bug pheromone synthesis enzymes.

Inventors

• DOROTHEA THOLL
• JASON LANCASTER

Assignees

• VIRGINIA TECH INTELLECTUAL PROPERTIES, INC.

Dates

Publication Date

20260505

Application Date

20200821

Claims (2)

1. 1 . An engineered polynucleotide comprising: a polynucleotide that is 100% identical to SEQ ID NO: 4; and a heterologous regulatory element operatively coupled to the polynucleotide.
2. 2 . The engineered polynucleotide of claim 1 , wherein the engineered polynucleotide encodes a polypeptide that is 100% identical to SEQ ID NO: 2.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is the U.S. National Stage Application under 35 U.S.C. § 371 of Patent Cooperation Treaty Application No. PCT/US2020/047538, filed on Aug. 21, 2020, entitled “BROWN MARMORATED AND HARLEQUIN STINK BUG PHEROMONE ENZYME SYNTHESIS AND USES THEREOF,” which claims the benefit of and priority to U.S. Provisional Patent Application No. 62/891,148, filed on Aug. 23, 2019, entitled “Production of Stink Bug Pest Aggregation Pheromone Precursors,” the contents of which is incorporated by reference herein in their entireties. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH This invention was made with government support under Grant No. 2016-67013-24759 awarded by USDA National Institute of Food and Agriculture. The government has certain rights in the invention. STATEMENT REGARDING SEQUENCE LISTING This application contains a sequence listing in electronic form as a .txt file entitled “VTIP-0225US2_2025-09-12_ST25”, created on Sep. 12, 2025, and having a size of 96,600 bytes. The content of the sequence listing is herein incorporated by reference in its entirety. TECHNICAL FIELD The subject matter disclosed herein is generally directed to engineered terpene synthesis enzymes, and more particularly those related to those present in insects. BACKGROUND The brown marmorated (Halyomorpha halys) and harlequin (Murgantia histrionica) stink bugs pose a threat to many agriculturally important corps grown throughout the world. The brown marmorated stink bug is very invasive particularly in the U.S., Europe, Eurasia, South America, and eastern Asia. The harlequin stink bug is an important embodiment of cabbage and related crops in the southern half of the United States and other parts of the world with similar climates. If not controlled the harlequin stink bug can easily destroy and entire crop by injuring the host plants by sucking their sap and causing them to wilt and die. Current methods to control these stink bug populations include biological control (e.g. wasps and flies that transmit parasites to the brown marmorated and/or harlequin stink bug) and chemical control. Given the need to find alternatives to chemical control of pests in agriculture, there exists a need for alternative methods for control of pests, such as the brown marmorated and harlequin stink bugs, in agriculture. Citation or identification of any document in this application is not an admission that such a document is available as prior art to the present invention. SUMMARY Described in several example embodiments herein are engineered polynucleotides comprising: one or more polynucleotides that is about 50-100% identical to SEQ ID NO: 3, SEQ ID NO: 4, or both. In certain example embodiments, the engineered polynucleotide encodes a polypeptide that is about 50-100% identical to SEQ ID NO: 1, SEQ ID NO: 2, or both. In certain example embodiments, comprising a polynucleotide capable of encoding a cytochrome P450 enzyme. In certain example embodiments, the cytochrome P450 enzyme is a cytochrome P450 enzyme that is expressed by a brown marmorated stink bug, a harlequin stink bug, or both. In certain example embodiments, the engineered polynucleotide further comprises a polynucleotide capable of encoding an isomerase. In certain example embodiments, the isomerase is an isomerase that is expressed by a brown marmorated stink bug, a harlequin stink bug, or both. Described in several example embodiments herein are engineered polynucleotides capable of encoding two or more of the following polypeptides: (a) a polypeptide that is about 50-100% identical to SEQ ID NO: 1;(b) a polypeptide that is about 50-100% identical to SEQ ID NO: 2;(c) a cytochrome P450 enzyme; and(d) an isomerase. Described in several example embodiments herein are vector systems comprising: one or more polynucleotides of the present description; andoptionally, one or more regulatory elements, wherein one or more of the one or more regulatory elements is operably coupled to the polynucleotide. In certain example embodiments, the regulatory element is a plant-specific regulatory element. In certain example embodiments, the vector system comprises 2 or more vectors, where at least one of the vectors comprises one or more polynucleotides of the present description. Described in several example embodiments herein are engineered stink bug pheromone synthesis systems comprising: one or more TPS polypeptides, wherein each TPS polypeptide is about 50-100% identical to SEQ ID NO: 1, SEQ ID NO: 2, or both; andone or both of a cytochrome P450 polypeptide and an isomerase polypeptide. In certain example embodiments, the system is capable of producing (a) a brown marmorated stink bug pheromone;(b) a brown marmorated stink bug pheromone intermediate;(c) a harlequin stink bug pheromone;(d) a harlequin stink bug pheromone intermediate; or(e) any combination thereof. In certain example embodiments, system is capable of producing (a) a brown marmorated st