Transgenic plants that overexpress a R2R3-MYB transcription factor gene and methods for producing the same
Inventors
Berkowitz, Gerald • Haiden, Samuel • Ma, Yi
Assignees
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Publication Number
US-12103950-B2
Publication Date
2024-10-01
Expiration Date
Abstract
The present disclosure provides compositions and methods related to genetically engineered plants. In particular, the present disclosure provides novel compositions for transgenic cannabis plants that overexpress a R2R3-MYB transcription factor gene and methods related to the genetic regulation of the development of a glandular trichome in plants.
Core Innovation
The invention relates to transgenic or genetically engineered plants and plant parts comprising a R2R3-MYB transcription factor gene, specifically MIXTA/CsMIXTA (SEQ ID NO:1), expressed under control of an operably linked heterologous promoter or homologous promoter. The recombinant nucleic acid molecules include a nucleotide sequence encoding the R2R3-MYB transcription factor having the nucleic acid sequence of SEQ ID NO:1 and a nucleotide sequence encoding the promoter operably linked to the transcription factor coding sequence. The described embodiments further include expression vectors, host cells, and chimeric genes integrated into the genome of a genetically engineered plant or plant cell.
The invention addresses increasing or upregulating trichome formation in a plant or plant cell by transforming and growing the plant or plant cell under conditions allowing expression of the SEQ ID NO:1 R2R3-MYB transcription factor. The stated functional outcome is that expression of the R2R3-MYB transcription factor increases or upregulates trichome formation compared to a wildtype or control plant. In Cannabis sativa, the document also states increased production or accumulation of cannabinoid compounds, including A9-tetrahydrocannabinol (THC), cannabichromene (CBC), cannabidiol (CBD), and combinations.
The described embodiments include assessments indicating transcription factor activity and nuclear localization, as well as phenotypic increases in glandular trichomes. CsMIXTA expression patterns in female flower development are described as correlating with cannabinoid biosynthetic genes/levels. In Nicotiana tabacum, ectopic expression is described as increasing glandular trichome density and branching, and the results are quantified by microscopy and qPCR. These observations are used in support of the claimed increase in glandular trichome formation and, for Cannabis sativa, cannabinoid accumulation compared to wildtype or control.
Claims Coverage
The independent claims cover four principal categories: recombinant nucleic acids, genetically engineered plants/plant cells, methods of increasing or upregulating trichome formation, and methods of increasing cannabinoid formation. Across these categories, the core inventive features are the SEQ ID NO:1 R2R3-MYB transcription factor operably linked to homologous or heterologous promoter sequences, chimeric genome integration in plants, and the functional outcomes of increased trichome formation and increased cannabinoid formation in Cannabis sativa contexts.
Promoter-linked SEQ ID NO:1 R2R3-MYB transcription factor nucleic acid
A recombinant nucleic acid molecule comprising a nucleotide sequence encoding a R2R3-MYB transcription factor from Cannabis sativa having the nucleic acid sequence of SEQ ID NO:1, and a nucleotide sequence encoding a heterologous promoter, wherein the nucleotide sequence encoding the R2R3-MYB transcription factor is operably linked to the nucleotide sequence encoding the heterologous promoter.
Chimeric genome-integrated SEQ ID NO:1 R2R3-MYB transcription factor
A genetically engineered plant or plant cell comprising a chimeric gene integrated into its genome, the chimeric gene comprising a nucleotide sequence encoding a R2R3-MYB transcription factor and having the nucleic acid sequence of SEQ ID NO: 1 operably linked to homologous or heterologous promoter.
Transformation and expression to increase or upregulate trichome formation
A method of increasing or upregulating trichome formation in a plant or plant cell, comprising transforming a plant or plant cell with a nucleotide sequence that encodes a R2R3-MYB transcription factor having the nucleic acid sequence of SEQ ID NO:1, and growing the plant or plant cell under conditions which allow for expression of SEQ ID NO: 1, wherein expression increases or upregulates trichome formation in the plant when compared to a wildtype or control plant.
Transformation and expression to increase cannabinoid formation via trichome upregulation
A method of increasing cannabinoid formation in a plant or plant cell, comprising transforming a cannabinoid producing plant or plant cell with a nucleotide sequence that encodes a R2R3-MYB transcription factor having the nucleic acid sequence of SEQ ID NO: 1, and growing the plant or plant cell under conditions which allow for expression of the R2R3-MYB transcription factor wherein expression increases or upregulates trichome formation and increases or upregulates formation of a cannabinoid in the plant or plant cell.
Overall, the claims primarily concentrate on recombinant nucleic acid constructs and chimeric genome-integrated plants that express a SEQ ID NO:1 R2R3-MYB transcription factor from Cannabis sativa under operable homologous or heterologous promoter control, with functional results stated as increased trichome formation and, in cannabinoid-producing contexts, increased cannabinoid formation.
Stated Advantages
Increases or upregulates trichome formation compared to a wildtype or control plant.
Increases or upregulates formation of cannabinoids in a plant or plant cell.
In Cannabis sativa, increased production or accumulation of A9-tetrahydrocannabinol (THC), cannabichromene (CBC), cannabidiol (CBD), or combinations compared to a wildtype or control Cannabis sativa plant.
Documented Applications
Genetically engineered Cannabis sativa plants and plant parts with increased cannabinoid accumulation, including increased A9-tetrahydrocannabinol (THC), cannabichromene (CBC), and cannabidiol (CBD) compared to wildtype or control.
Increasing or upregulating trichome formation in a plant or plant cell by expressing the SEQ ID NO:1 R2R3-MYB transcription factor.
Increasing cannabinoid formation in a cannabinoid producing plant or plant cell, where expression increases or upregulates trichome formation and increases or upregulates cannabinoid formation.
Ectopic expression in Nicotiana tabacum is described as increasing glandular trichome density and branching, with quantified phenotyping and expression analyses.
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