Pioneer Hi-bred International, Inc.

Margit C Ross

William James Gordon-Kamm

Keith S Lowe

Dwight T Tomes

Benjamin A Bowen

Dennis L Bidney

Gary A Sandahl

Laura A Church

Albert Laurence Lu

Christopher Jay Scelonge

Michael D Miller

George J Hoerster

Dorothy A Pierce

Matthew Curtis Harmon

Lijuan Wang

Heather Marie Christensen

David D Songstad

Virginia Crane

William Gordan-Kamm

Kristen Denise Rinehart Krebs

Jeffrey Klever

Bin Cong

Luciano M Jaureguy

Leigh Bangs

Patrea M Hill

Mike Miller

Gary Sandahl

Embodiments disclosed herein relate to the field of plant molecular biology, specifically to DNA constructs for conferring insect resistance to a plant. Embodiments disclosed herein relate to insect resistant corn plant containing event DP-915635-4, and to assays for detecting the presence of event DP-915635-4 in samples and compositions thereof.

Maize event DP-915635-4 and methods for detection thereof

The invention provides improved plant transformation methods. In particular the method provides increased transformation freequency, especially in recalcitrant plants. The method comprises stably transforming a target cell with at least one polynucleotide of interest. The target cell has been previously transformed to stimulate growth of the cell and has gone through at least one cell division.

Methods for enhancing plant transformation frequencies

A method for the production of transgenic plants is provided in which a vector carrying a gene encoding the green fluorescent protein is introduced into cells, the cells are screened for the protein and transformed cells are selected and regenerated. The cellular toxicity of the green fluorescent protein is circumvented by regulating expression of the gene encoding the protein or directing the protein to a subcellular compartment where it is not toxic to the cell. DNA constructs are provided for cell transformation in which the expression of a gene encoding the green fluorescent protein is placed under the control of an inducible promoter. In addition, DNA constructs are provided in which a nucleotide sequence encoding the green fluorescent protein is operably linked to a signal sequence which directs the expressed protein to a subcellular compartment where the protein is not toxic to the cell. Oxidative stress to plant cells transformed with GFP also can be ameliorated by transforming cells with an expression vector comprising genes encoding GFP and an oxygen scavenger enzyme such as superoxide dismutase. The toxicity of GFP in transformed plants can be eliminated by excising the screenable marker gene following detection of transformed cells or sectors. The FLP/FRT system is used in conjunction with GFP as a visible marker for transformation and FRT excision. A nucleotide sequence optimized for expression of the green fluorecent protein in plants is also provided.

Use of the green fluorescent protein as a screenable marker for plant transformation

The invention relates to compositions and methods and for genetically transforming plants. Compositions comprising transformation-enhancing agents and methods of use are provided. The methods involve administering an effective amount of a transformation-enhancing agent to a plant. The compositions and methods of the invention find use in improving transformation efficiency and increasing the embryogenic response of callus.

Transformation-enhancing compositions and methods of use

An improvement in the process of transporting biological materials into living cells by bombarding the cells with microprojectile particles coated with biological material. In the improved process prior to coating of the particles such as tungsten beads for example, with biological material such as DNA, the particles are first pretreated with a strong inorganic acid such as nitric acid.

Pretreatment of microprojectiles prior to using in a particle gun

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 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.

Method for producing transgenic cereal plants

Growing community of inventors

Johnston, IA, United States of America

Margit C Ross