Sorghum-derived transcription regulatory elements predominantly active in root hair cells and uses thereof
Inventors
Baerson, Scott R. • Pan, Zhiqiang • Polashock, James J
Assignees
US Department of Agriculture USDA
Publication Number
US-10815491-B2
Publication Date
2020-10-27
Expiration Date
2035-05-20
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Abstract
Transcription regulatory elements, namely promoter and terminator sequences, obtained from Sorghum bicolor that drive RNA transcription predominately in root hair cells are described, as well as cassettes, expression vectors, and genetically modified plants containing these transcription regulatory elements. The genetically modified plants can be gymnosperms, dicots, or monocots. Methods of directing transcription of a heterologous polynucleotide under control of these transcription regulatory elements in a genetically modified plant's root hair cells are also provided.
Core Innovation
The invention relates to transcription regulatory elements, specifically promoters and terminators, derived from Sorghum bicolor that predominantly drive RNA transcription in root hair cells. These elements are incorporated into cassettes and expression vectors to generate genetically altered plants, including gymnosperms, dicots, and monocots, which selectively express heterologous polynucleotides in root hair cells.
The problem addressed is the need for promoters that direct strong, root hair-specific expression of heterologous genes to improve traits related to nutrient uptake, disease resistance, and other root-associated functions. Existing promoters often lack the required specificity or expression levels in root hair cells, limiting efficient genetic manipulation of this key cell type.
This invention provides DNA sequences for promoters (SEQ ID NOs: 1, 3, 5, 7, 12, 16, 20, and 24) and terminators (SEQ ID NOs: 2, 4, 6, 8, 13, 17, 21, and 25), or sequences with at least 95% identity thereto, that are predominantly or selectively active in root hair cells. The constructs enable transcription of heterologous genes in root hair cells to confer improvements such as enhanced disease resistance, nutrient and water uptake, bioremediation, and allelochemical production without adverse effects caused by non-cell specific expression.
Claims Coverage
The patent claims cover 30 claims with a focus on cassettes, genetically altered plants, plant parts, methods of transcription regulation, and production of proteins or RNA using specific promoter and terminator sequences. The main inventive features include root hair cell-active promoters and terminators derived from Sorghum bicolor, their use in genetically altered plants, and methods for selective expression in root hair cells.
Promoter sequences for root hair-specific transcription regulation
A cassette comprising a promoter selected from SEQ ID NO: 1, 3, or 5 that regulates transcription of a heterologous polynucleotide (encoding protein or RNA) predominantly in plant root hair cells.
Incorporation of terminators linked to the heterologous polynucleotide
The cassette optionally includes a terminator selected from SEQ ID NO: 2, 4, 6, or 8 or sequences at least 95% identical thereto, operably linked to the 3′ end of the heterologous polynucleotide to facilitate transcription termination.
Genetically altered plants and plant material containing the cassette
Genetically altered plants, seeds, pollen, cells, tissues, and progeny comprising the cassette with the root hair-specific promoter and heterologous polynucleotide, optionally including the terminator, capable of expressing desired traits in root hair cells.
Methods of selectively directing transcription to root hair cells
Methods involving introducing the cassette into plant cells by transformation or introgression to produce genetically altered plants where the heterologous polynucleotide is predominantly transcribed in root hair cells, applicable to gymnosperms, monocots, and dicots.
Methods of producing proteins or RNA of interest in root hair cells
Methods for producing a protein or RNA predominantly in root hair cells by introducing the cassette with the specified promoter and heterologous polynucleotide, selecting altered plants and allowing growth to express the product selectively in root hair cells.
The claims comprehensively cover the composition of cassettes with Sorghum bicolor-derived root hair-specific promoters and terminators, genetically altered plants and plant materials containing these cassettes, and methods for achieving selective gene expression and protein or RNA production in root hair cells across various plant species.
Stated Advantages
The transcription regulatory elements direct accurate and high-level expression specifically in root hair cells, enabling targeted genetic modification.
Root hair-specific promoters reduce adverse effects such as potential crop yield reductions caused by non-cell-specific expression of inhibitory gene products.
The promoters deliver higher levels of recombinant gene products in root hairs than previously characterized promoters.
These elements facilitate the production of phenotypes improving disease resistance, nutrient uptake, stress tolerance, beneficial microbial interactions, and other desirable traits related to the plant root-soil interface.
Documented Applications
Expression of heterologous DNA in genetically altered plants to promote colonization of beneficial rhizosphere-associated microorganisms.
Enhancement of nutrient and water uptake through transporter or channel proteins expressed in root hair cells.
Improvement of nitrogen fixation efficiency in leguminous crops by expression in root hair cells.
Facilitation of bioremediation by proteins expressed in root hair cells.
Inhibition of colonization by soil-borne parasites such as nematodes.
Reduction of competition from neighboring plants via allelochemical production in root hair cells.
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