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Methods for producing hybrid seed

A technology for transgenic plants and male sterility, applied in the directions of biochemical equipment and methods, horticultural methods, and botanical equipment and methods, and can solve the problems of chlorosis, insufficient phytotoxic side effects of gametes, and no selectivity of gametes.

Inactive Publication Date: 2016-02-17
MONSANTO TECH LLC
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the levels of glyphosate required to kill most of the male gametes but leave enough female gametes still capable of fertilization often result in wilting or chlorosis of the plants
Therefore, the main disadvantage of using glyphosate as a gameticide is the phytotoxic side effect caused by not being sufficiently selective for gametes, as is generally the case with most chemical gameticides
[0011] Commercial production of hybrid seeds using chemical gametocides is mainly limited by their general lack of selectivity for gametes

Method used

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  • Methods for producing hybrid seed
  • Methods for producing hybrid seed
  • Methods for producing hybrid seed

Examples

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Embodiment 1

[0199] This example sets forth non-limiting examples of recombinant DNA constructs useful for the suppression of at least one target gene; these constructs, when designed to contain miRNA recognition sites, allow expression of gene suppression elements in specific tissues.

[0200] figure 1 A schematically illustrates a non-limiting example of a recombinant DNA construct of the invention useful for suppressing at least one target gene. These constructs comprise at least one first gene suppression element ("GSE" or "GSE1") for the suppression of at least one first target gene, wherein the first gene suppression element is embedded in a region of non-protein-coding DNA. in the introns on either or both sides. These constructs use introns (in many embodiments, preferably those derived from the 5' sequence). The construct may optionally comprise at least one second gene suppression element ("GSE2") for suppressing at least one second target gene, at least one second gene suppre...

Embodiment 2

[0205] This example describes a non-limiting method for determining potentially useful miRNA recognition sites by measuring miRNA expression patterns. Information on the spatial or temporal distribution of expression of a given miRNA can be used, for example, to design recombinant constructs to be expressed in a spatially or temporally specific manner. This example discloses expression patterns of mature miRNAs in maize and provides sequences of recognition sites for these miRNAs suitable for inclusion in recombinant DNA constructs useful in maize and other plants.

[0206] Total RNA was isolated from LH244 maize plants using Trizol (Invitrogen, Carlsbad, CA). Seven developmental stages were used, including root and branch meristems from germinated seedlings, juvenile (V1-V2) and adult leaves (V7-V8), stalk internodes, pre-shedding ears, and immature (approximately 1") ears. 5 μg of total RNA was resolved on 17% PAGE-urea as described in Allen et al. (2004) Nat. Genet., 36: 1...

Embodiment 3

[0212] This non-limiting example describes recombinant DNA constructs of the invention that can be used to inhibit target expression in specific cells of or derived from multicellular eukaryotes, such as plant cells or animal cells. RNA expressions, and methods of their use. The construct comprises a promoter operably linked to DNA transcribed into RNA comprising at least one exogenous miRNA recognition site that is recognized by a mature miRNA expressed in a particular cell of a multicellular eukaryote, and A target RNA that is suppressed in a specific cell, wherein the target RNA is expressed in a cell of a cell-unspecific multicellular eukaryote.

[0213] Strong constitutive promoters expressed in almost all plant cells have been identified (eg CaMC35S, OsAct), but strong spatial (cell or tissue specific) and temporal specific promoters have not been fully characterized. In order to restrict target RNA or transgene expression to specific cell or tissue types in the absence...

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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

[0001] This application is a divisional application of the following applications: filing date: September 20, 2006; application number: 200680046237.7 (PCT / US2006 / 036847); title of invention: same as above. [0002] Priority Claims and References to Related Applications [0003] This application claims priority to U.S. Provisional Patent Application No. 60 / 726,106, filed October 13, 2005, and U.S. Provisional Patent Application No. 60 / 836,246, filed August 7, 2006, which are incorporated herein in their entirety Reference. [0004] Introduction of Sequence Listing [0005] Document "38-21(54232)A.rpt" (file size 61kb, inscribed on 12 October 2005, filed with U.S. provisional application 60 / 726,106 on 13 October 2005), "38-21(54232 )B.rpt" (file size 68kb, inscribed on August 7, 2006, filed with U.S. provisional application 60 / 836,246 on August 7, 2006) and "38-21(54232)C.rpt" (file 70 kb in size, inscribed September 19, 2006, and filed September 20, 2006 with US Provisional A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/06C12N15/82A01G7/06A01H5/10A01H5/08
CPCC12N15/8216C12N15/8217C12N15/8218C12N15/8234C12N15/8238C12N15/8254C12N15/8265C12N15/827C12N15/8275C12N15/8279C12N15/8285C12N15/8286C12N15/8289Y02A40/146
Inventor E.艾伦L.A.吉伯特森N.M.豪马德黄士杰S.I.伊瓦舒塔J.K.罗伯特斯
Owner MONSANTO TECH LLC
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