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1. WO1996004392 - TRANSGENIC CEREAL PLANTS

Publication Number WO/1996/004392
Publication Date 15.02.1996
International Application No. PCT/US1995/008977
International Filing Date 26.07.1995
Chapter 2 Demand Filed 29.02.1996
IPC
C12N 15/82 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79Vectors or expression systems specially adapted for eukaryotic hosts
82for plant cells
CPC
C12N 15/8207
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79Vectors or expression systems specially adapted for eukaryotic hosts
82for plant cells ; , e.g. plant artificial chromosomes (PACs)
8201Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
8206by physical or chemical, i.e. non-biological, means, e.g. electroporation, PEG mediated
8207by mechanical means, e.g. microinjection, particle bombardment, silicon whiskers
Applicants
  • PIONEER HI-BRED INTERNATIONAL, INC. [US/US]; 700 Capital Square 400 Locust Street Des Moines, IA 50309, US
Inventors
  • BOWEN, Benjamin, A.; US
  • LOWE, Keith; US
  • ROSS, Margot, C.; US
  • SANDAHL, Gary, A.; US
  • TOMES, Dwight, T.; US
  • SONGSTAD, David, D.; US
  • GORDON-KAMM, William, J.; US
Agents
  • BENT, Stephen, A. ; Foley & Lardner Suite 500 3000 K Street, N.W. Washington, DC 20007-5109, US
Priority Data
08/282,27029.07.1994US
08/483,09107.06.1995US
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) TRANSGENIC CEREAL PLANTS
(FR) CEREALES TRANSGENIQUES
Abstract
(EN)
To obtain a transgenic cereal plant which is stably transformed, an exposed cereal meristem is subjected to biolistic bombardment in order to target non-differentiated meristem cells for transformation. Immature embryos at the early proembryo, mid proembryo, late proembryo, transitional or early coleoptilar stage are harvested for biolistic bombardment. The meristem tissue or cells fated to contribute to the meristem then are manipulated in order to enlarge transgenic sectors, either through selection and/or through effecting a proliferation from the tissue of shoots or multiple meristems $i(per se). The shoot population thus obtained then is screened, by means of a nonlethal enrichment assay, to identify either chimeric sectors that will contribute to germline transmission, or non-sectored, L2 periclinal chimeras that will by definition transmit to progeny. Increased time in culture, under selection, enhances the prospects for sectoral-to-periclinal conversions, and also selects for L1-to-L2 conversions which, through a shift in position, ultimately contribute to the germline. Transgenic sectors also are stabilized during the step of tillering.
(FR)
Pour obtenir une céréale transgénique qui est transformée d'une manière stable, on soumet un méristème de céréale qui a été dégagé, à un bombardement biolistique, en vue d'une transformation des cellules non différenciées du méristème. On recueille des embryons immatures au stade pro-embryonnaire précoce, moyen et tardif, au stade transitionnel ou au stade coléoptilaire précoce pour effectuer le bombardement biolistique. On soumet le tissu du méristème ou des cellules devant contribuer à la formation du méristème à une manipulation pour élargir les secteurs transgéniques, par sélection et/ou en provoquant une prolifération à partir du tissu de pousses ou de méristèmes multiples en eux-mêmes. La population de pousses ainsi obtenue est soumise à une sélection par un test d'enrichissement non létal, pour identifier les secteurs chimères qui vont contribuer à la transmission de la lignée germinale ou des chimères périclinales non sectorisées L2 qui vont, par définition, se transmettre à la progéniture. Une augmentation du temps de culture dans des conditions de sélection augmente les chances de conversions sectorielle en périclinale et également favorise les conversions L1 en L2, qui, par un décalage de position, contribuent en fin de compte à la lignée germinale. Les secteurs transgéniques sont également stabilisés pendant la phase de tallage.
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