US-20260125693-A1 - GENETICALLY ENGINEERED VEGETABLES

Abstract

The present disclosure relates at least in part to genetic engineering of vegetable (e.g., tomato) plants to, for example, produce heme, a molecule contributing to a meat-like flavor, and to the incorporation of male sterility in these plants to facilitate the production of F1 hybrid seeds. Described herein, in some embodiments, are methods for constructing and introducing into such plants genetic constructs containing both a heme gene and a male sterility gene, the resulting plants, and the use of these plants in hybrid seed production. The present disclosure also provides at least in part methods for enhancing lycopene content in plants, such as tomato plants, either through genetic modification or selective breeding. While introducing key genes like Psy is effective, adding multiple genes or knocking out competing pathways can lead to more substantial increases in lycopene levels. Enhanced lycopene content can provide added health benefits, including potential cancer protection, making these plants, such as tomatoes, particularly valuable in the context of functional foods. The plants may be of varieties with high lycopene content combined with genetic modifications as provided herein. The present disclosure offers a comprehensive solution for producing plants (e.g., tomatoes) with increased nutritional value.

Inventors

• Gabriella Rothberg
• Jonathan M. Rothberg

Assignees

• Impossible Vines Inc.

Dates

Publication Date

20260507

Application Date

20251031

Claims (20)

1. 1 . A genetic construct or set of genetic constructs comprising a first promoter operably linked to a first coding sequence and a second promoter operably linked to a second coding sequence, wherein the first coding sequence encodes a heme-producing gene, and wherein the second coding sequence encodes a male-sterility gene.
2. 2 . The genetic construct or set of genetic constructs of claim 1 , wherein the genetic construct comprises the nucleotide sequence of SEQ ID NO: 3 or SEQ ID NO: 4.
3. 3 . The genetic construct or set of genetic constructs of claim 1 , wherein the heme-producing gene is a leghemoglobin gene derived from Glycine max.
4. 4 . The genetic construct or set of genetic constructs of claim 3 , wherein the leghemoglobin gene comprises the nucleotide sequence of SEQ ID NO: 5.
5. 5 . The genetic construct or set of genetic constructs of claim 1 , wherein the male-sterility gene is a barnase gene derived from Bacillus amyloliquefaciens.
6. 6 . The genetic construct or set of genetic constructs of claim 5 , wherein the barnase gene comprises the nucleotide sequence of SEQ ID NO: 6.
7. 7 . The genetic construct or set of genetic constructs of claim 1 , wherein the first promoter and/or the second promoter is a constitutive promoter.
8. 8 . The genetic construct or set of genetic constructs of claim 7 , wherein the constitutive promoter comprises the nucleotide sequence of SEQ ID NO: 9.
9. 9 . The genetic construct or set of genetic constructs of claim 1 , wherein the first promoter and/or the second promoter is a plant promoter.
10. 10 . The genetic construct or set of genetic constructs of claim 9 , wherein the plant promoter comprises the nucleotide sequence of any one of SEQ ID NOs: 10-12.
11. 11 . The genetic construct or set of genetic constructs of claim 1 further comprising a terminator sequence, wherein the terminator sequence comprises the nucleotide sequence of SEQ ID NO: 13 or SEQ ID NO: 14.
12. 12 . The genetic construct or set of genetic constructs of claim 1 further comprising an antibiotic resistance gene.
13. 13 . The genetic construct or set of genetic constructs of claim 12 , wherein the antibiotic resistance gene is a hygromycin resistance gene.
14. 14 . The genetic construct or set of genetic constructs of claim 13 , wherein the hygromycin resistance gene comprises the nucleotide sequence of SEQ ID NO: 7.
15. 15 . The genetic construct or set of genetic constructs of claim 1 further comprising a first flanking sequence and a second flanking sequence, wherein the first flanking sequence comprises the nucleotide sequence of SEQ ID NO: 15 and wherein the second flanking sequence comprises the nucleotide sequence of SEQ ID NO: 16.
16. 16 . A plant cell, comprising the genetic construct or set of genetic constructs of claim 1 .
17. 17 . The plant cell of claim 16 , wherein the plant cell is tomato plant cell or an eggplant plant cell.
18. 18 . A method, the method comprising transforming a plant cell with the genetic construct or set of genetic constructs of claim 1 .
19. 19 . The method of claim 18 , wherein transforming is via Agrobacterium -mediated transformation.
20. 20 . The method of claim 18 , wherein the plant cell is tomato plant cell or an eggplant plant cell.

Description

CROSS REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority under 35 U.S.C. § 119 (e) to U.S. Provisional Application No. 63/715,207, filed Nov. 1, 2024, the entire contents of which are herein incorporated by reference. REFERENCE TO AN ELECTRONIC SEQUENCE LISTING The contents of the electronic sequence listing (1063470000US02-SEQ-KVC.xml; Size: 77,300 bytes; and Date of Creation: Oct. 31, 2025) are herein incorporated by reference in their entirety. FIELD The present disclosure generally relates, at least in part, to the field of plant biotechnology and genetic engineering. More specifically, it relates to the production of genetically engineered vegetable plants, such as tomato (Solanum lycopersicum) plants and eggplant (Solanum melongena) plants, engineered to express a protein to enhance iron content and contribute to a meat-like flavor, optionally combined with a male sterility gene to facilitate F1 hybrid seed production. The present disclosure also relates to increasing lycopene production, such as in tomato plants, to, in some embodiments, enhance its health benefits and antioxidant properties. SUMMARY Vegetables are widely recognized for their nutritional and health benefits, and the global demand for plant-based foods has continued to increase in response to growing consumer interest in sustainability and wellness. Tomatoes are among the most extensively cultivated vegetables and serve as an important dietary source of vitamins and antioxidants. In particular, the naturally occurring carotenoid, lycopene, imparts the characteristic red color and has health-promoting properties, including potential roles in reducing cancer risk and supporting cardiovascular and skin health. Iron and associated carrier molecules, such as heme, contribute to the characteristic meat-like flavor of many foods. The iron content in tomatoes, however, is inherently low, limiting their contribution to dietary iron intake. Advances in plant genetic engineering can enable the precise enhancement of nutritional and sensory compounds in crops, offering an efficient and sustainable means to improve food quality, reduce agricultural inputs, and align with global trends towards plant-based diets. Accordingly, developing genetically improved tomato plant varieties that exhibit enhanced lycopene accumulation and/or elevated iron levels provides a promising opportunity to deliver greater nutritional benefits to the world's expanding population. Aspects of the present disclosure relate, at least in part, to genetically engineered plants (e.g., vegetable plants such as tomato plants and eggplant plants) that can exhibit enhanced nutritional and/or sensory properties through the introduction of genes coding for heme production and, optionally, male sterility to enable controlled F1 hybrid seed production. The present disclosure further encompasses strategies for enhancing lycopene biosynthesis through genetic modification to yield plants (e.g., vegetable plants such as tomato plants) that can produce fruit (e.g., tomatoes) with improved antioxidant capacity. Aspects of the present disclosure further provide genetically engineered vegetable plants that combine enhanced nutritional composition with improved hybridization efficiency. In some embodiments, the present disclosure relates to tomatoes or eggplants engineered to express a protein to produce heme, an iron-carrier molecule contributing to a meat-like flavor, while also introducing a male-sterility gene to facilitate efficient and controlled F1 hybrid seeds. The integration of these two traits enables the development of hybrid tomato or eggplant varieties that deliver desirable flavors without the need for manual emasculation during breeding. Aspects of the present disclosure provides methods for enhancing lycopene biosynthesis in vegetable fruit, such as tomato fruit, through the overexpression of key enzymes involved in its biosynthesis, such as phytoene synthase (Psy). In some embodiments, the present disclosure relates to the use of gene-editing technologies, such as CRISPR/Cas9-mediated technology, to downregulate competing pathways, thereby reducing precursor flux toward lycopene production and achieving elevated antioxidant content. The present disclosure thus can integrate flavor enhancement, nutritional fortification, and reproductive control into a single transgenic platform. This combination allows for sustainable, high-value vegetable cultivation that aligns with consumer demands for nutritious, plant-based foods, with improved sensory traits such as meatier tastes. Furthermore, the present disclosure provides the corresponding genetic constructs, transformation methods, and resulting plants, seeds, and fruits produced thereby. Accordingly, aspects of the present disclosure relate to a genetic construct or set of genetic constructs comprising a first promoter operably linked to a first coding sequence and, optionally, a second promoter opera