Methods for producing hybrid seed

Inactive Publication Date: 2007-08-23
MONSANTO TECH LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0018] In another aspect, this invention provides a method for producing a hybrid seed, including the steps of: (a) providing a first parent plant including a transgenic plant grown from a first transgenic plant cell containing in its genome a first recombinant DNA construct that transcribes to RNA including (i) at least one exogenous miRNA recognition site recognizable by a first mature miRNA, and (ii) a first messenger RNA encoding a protein imparting tolerance to a first herbicide, wherein the first mature miRNA is specifically expressed in male reproductive tissue of the first parent plant, thereby specifically suppressing expression of the protein in the male reproductive tissue, and wherein male sterility of the first parent plant is inducible by application of the first herbicide to the first parent plant; (b) providing a second parent plant including a transgenic plant grown from a second transgenic plant cell containing in its genome

Problems solved by technology

A major limitation in the production of hybrid seed for many crop species is the lack of simple, reliable and economical methods of generating sterility in at least one parent (especially the male parent, to result in male-sterility while leaving female gametes intact and accessible for pollination by a suitable pollen donor).
In some species, this is a straightforward although labor-intensive and therefore expensive process (e.g., detasselling in maize).
In other species, such physical emasculation is difficult because of the plant's anatomy.
How

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Examples

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Example

Example 1

[0189] This illustrates non-limiting examples of recombinant DNA constructs for suppression of at least one target gene; these constructs, when designed to include a miRNA recognition site, allow expression of the gene suppression element in specific tissues.

[0190]FIG. 1A schematically depicts non-limiting examples of recombinant DNA constructs of the invention for suppression of at least one target gene. These constructs include at least one first gene suppression element (“GSE” or “GSE1”) for suppressing at least one first target gene, wherein the first gene suppression element is embedded in an intron flanked on one or on both sides by non-protein-coding DNA. These constructs utilize an intron (in many embodiments, an intron derived from a 5′ untranslated region or an expression-enhancing intron is preferred) to deliver a gene suppression element without requiring the presence of any protein-coding exons (coding sequence). The constructs can optionally include at least...

Example

Example 2

[0194] This example describes one non-limiting method of determining the potential usefulness of a miRNA's recognition site, by determining the miRNA's expression pattern. Knowledge of the spatial or temporal distribution of a given miRNA's expression is useful, e.g., in designing recombinant constructs to be expressed in a spatially or temporally specific manner. This example discloses mature miRNA expression patterns in maize and provides sequences of recognition sites for these miRNAs that are suitable for inclusion in recombinant DNA constructs useful in maize and other plants.

[0195] Total RNA was isolated from LH244 maize plants using Trizol (Invitrogen, Carlsbad, Calif.). Seven developmental stages were used, including roots and shoot meristems from germinating seedlings, juvenile (V1 to V2) and adult leaves (V7 to V8), stalk internode, tassel before shedding, and immature (approximately 1″) ears. Five micrograms total RNA was resolved on 17% PAGE-Urea as described ...

Example

Example 3

[0197] This non-limiting example describes recombinant DNA constructs of the invention, useful for suppressing expression of a target RNA in a specific cell of or derived from a multicellular eukaryote such as a plant cell or an animal cell, and methods for their use. The constructs include a promoter operably linked to DNA that transcribes to RNA including at least one exogenous miRNA recognition site recognizable by a mature miRNA expressed in a specific cell of a multicellular eukaryote, and target RNA to be suppressed in the specific cell, wherein said target RNA is to be expressed in cells of the multicellular eukaryote other than the specific cell.

[0198] Strong constitutive promoters that are expressed in nearly all plant cells have been identified (e.g., CaMC 35S, OsAct), but strong spatially specific (cell- or tissue-specific) and temporally specific promoters have been less well characterized. To limit target RNA or transgene expression to a specific cell or tiss...

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Abstract

This invention provides methods for producing a non-natural hybrid seed. Also disclosed are specific miRNAs and miRNA recognition sites useful for conferring inducible sterility on a crop plant, and recombinant DNA construct including such exogenous miRNA recognition sites.

Description

PRIORITY CLAIMS AND REFERENCE TO RELATED APPLICATIONS [0001] This application claims benefit of priority to U.S. Provisional Patent Applications No. 60 / 726,106, filed on 13 Oct. 2005, and 60 / 836,246, filed on 7 Aug. 2006, which are incorporated by reference in their entirety herein.INCORPORATION OF SEQUENCE LISTINGS [0002] The sequence listing that is contained in the file named “38-21(54232)C.rpt” which is 70 kilobytes (measured in operating system MS-Windows), was created on 19 Sep. 2006, and is located in computer readable form on a compact disk (CD-R), is filed herewith and incorporated herein by reference. The sequence listings contained in the files “38-21(54232)A.rpt” (file size of 61 kilobytes, recorded on 12 Oct. 2005, and filed with U.S. Provisional Application 60 / 726,106 on 13 Oct. 2005) and “38-21(54232)B.rpt” (file size of 68 kilobytes, recorded on 7 Aug. 2006, and filed with U.S. Provisional Application 60 / 836,246 on 7 Aug. 2006) are incorporated by reference in their ...

Claims

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Application Information

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IPC IPC(8): A01H5/00C12N15/82A01H5/10
CPCC12N15/8218C12N15/8287C12N15/8289A01H5/10A01H1/02A01H5/00C12N15/829C12N15/8271C12N15/8279C12N15/8286C12N15/8274C12N15/8222C12N15/8231
Inventor ALLEN, EDWARDSGILBERTSON, LARRYHOUMARD, NANCYHUANG, SHIHSHIEHIVASHUTA, SERGEYROBERTS, JAMES
Owner MONSANTO TECH LLC
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