Pioneer Hi-bred International, Inc.

Her Majesty the Queen in Right of Canada As Represented by Agriculture

Dow Agrosciences LLC

National Research Council of Canada

Sakata Seed Corporation

Her Majesty the Queen in Right of Canada As Represented by the Minister of Agriculture and Agri-food Eastern Cereal & Oilseed, Research Center

Paladin Hybrids, Inc.

Laurian Sylvio Robert

Steven F Fabijanski

Paul G Arnison

Diego Albani

Johann Schernthaner

Steven Fabijanski

Hai Ping Hong

Jean L Gerster

John A Simmonds

Elizabeth Foster

Jas Singh

Stephen Gleddie

Janice Carole Morissette

Tong Wu

Anne M Johnson-Flanagan

The present invention relates to methods to control the spread of recombinant DNA molecules between sexually compatible plants of differing genetic composition. The invention describes the production of transgenic plants that comprise recombinant traits of interest or concern linked to repressible lethal genes. The lethal genes are blocked by the action of repressor molecules produced by the expression of repressor genes located at a different genetic locus. The lethal phenotype is only expressed after the segregation of the repressible lethal gene construct and the repressor gene following meiosis. The present invention may be employed for both open-pollinated and hybrid seed production systems and may be used to maintain genetic purity by blocking unintended introgression of genes from plants devoid of the specific repressor gene. The invention includes methods that impart traits that are desirable for environmentally responsible heterologous protein production, to genetic material used to impart said traits and to new plants and products derived by said methods.

Methods and genetic compositions to limit outcrossing and undesired gene flow in crop plants

This invention is directed to a method for the expression of a gene of interest, or a chimeric or modified gene allowing the localization of a protein, protein fusion, peptide or fragment of interest within the extracellular domain of a floral cell. This method comprises preparing a construct comprising a promoter sequence capable of expressing a gene encoding the protein, protein fusion, peptide, or fragment of interest, within the floral cell; a translated sequence of the protein, protein fusion, peptide, or fragment of interest, which is localized within the extracellular domain of a floral cell; a gene that encodes the protein, protein fusion, peptide, or fragment of interest; and a terminator sequence, and transforming a plant. Plants transformed with such a construct are characterized as having a protein, fragment thereof, or peptide of interest on the surface of a floral cell. Such localized proteins or peptides may be used for the purposes of peptide display, mediating plant sterility, modifying pollen-pistil interactions, altering pollen for insect consumption etc.

Modification of pollen coat protein composition

The present invention relates to novel methods for the transformation of crop species, novel methods for the selection and identification of transformed plant cells and novel methods for recovery of regenerated whole plants. The method also relates to the development of plants with novel traits and plants that contain novel recombinant DNA constructs.

Methods and constructs for plant transformation

A process is described for producing fertile hybrid seed or hybrid seed comprising fertile and sterile seed using male-sterile plants created by employing molecular techniques to manipulate anti-sense gene and other genes that are capable of controlling the production of fertile pollen in plants. These plants are functionally male-sterile plants with pollen from male-fertile plants. Hybrid seed production is simplified and improved by this invention and can be extended to plant crop species for which commercially acceptable hybrid seed production methods are not currently available.

Molecular methods of hybrid seed production

A process is described for producing fertile hybrid seed or hybrid seed comprising fertile and sterile seed using male-sterile plants created by employing molecular techniques to manipulate anti-sense gene and other genes that are capable of controlling the production of fertile pollen in plants. Said plants are functionally male-sterile plants with pollen from male-fertile plants. Hybrid seed production is simplified and improved by this invention and can be extended to plant crop species for which commercially acceptable hybrid seed production methods are not currently available.

A process is described for producing fertile hybrid seed or hybrid seed comprising fertile and sterile seed using male-sterile plants created by employing molecular techniques to manipulate genes that are capable of controlling the production of fertile pollen in plants. Hybrid seed production is simplified and improved by this approach, which can be extended to plant crop species for which commercially acceptable hybrid seed production methods have not been available.

A Brassica sp. genomic clone containing a gene promoter, which directs expression in the tapetum, was isolated. When the promoter was fused with the GUS gene and introduced into B. napus by Agrobacterium mediated transformation, this promoter controlled GUS expression in the tapetum in transgenic B. napus plants. Expression was observed from bud length 2-3 mm, which corresponds to the uninucleate microspore stage, peaked at 4-5 mm which corresponds to the binucleate microspore stage and declined to no detectable level at the open flower stage. There was no GUS expression in other tissues such as root, stem, leaf, sepal, petal or pistil. This promoter will be useful for the temporal and spatial control of gene expression in plants.

Brassica sp. gene promoter highly expressed during tapetum development

A Brassica sp. genomic clone G 363 containing a gene promoter, which directs expression in the stigma, was isolated. Based on microbombardment studies of Brassica napus flowers with the gene Pis 63/Gus fusion, the Pis 63 promoter was shown to direct GUS expression to the stigma. This promoter will be useful for the temporal and spatial control of gene expression in plants.

Brassica sp. gene promoter highly expressed during stigma development

The initial steps in photosynthesis are the conversion of light energy into chemical energy. This conversion is performed by the multisubunit protein-pigment complexes of the thylakoid membranes. Oxygen-evolving photosystems contain photosystem I (PSI) and photosystem II (PSII), which act in tandem. In PSII, the majority of light-adsorbing chlorophylls are attached to LHCII, the light harvesting complex associated with this photosystem. LHCII is the most abundant member of the family of chlorophyll a/b binding (CAB) proteins. A gene encoding a type I chlorophyll a/b binding protein of LHCII (ICABPSII) has been cloned from Brassica napus L. An anti-sense RNA of this gene has been used to reduce the amount of chlorophyll a/b binding protein and thus reduce the amount of chlorophyll in the resulting transgenic plants. By using 'site' specific promoters the reduction of chlorophyll can be targeted to specific organelles in the transgenic plant and thus can be used to reduce the green color at these sites. Thus it is possible to use the anti-sense RNA of a chlorophyll a/b binding protein as a means for degreening, for example fruits, seeds or floral parts in transgenic plants.

Anti-sense RNA for CAB transcript to reduce chlorophyll content in plants

A Brassica sp. genomic clone containing a polygalacturonase gene promoter was isolated. This promoter directs high levels of transcription in the pollen. When the promoter was fused with the GUS gene and introduced into B. napus by Agrobacterium mediated transformation, this promoter controlled GUS expression in microspores and pollen in transgenic B. napus plants. Expression occurred between the late uninucleate microspore stage and the mature pollen grain stage. GUS activity was also identified in tapetal tissue at the binucleate microspore stage. There was no GUS expression in other tissues such as root, stem, leaf, sepal, petal or pistil. This promoter will be useful for the temporal and spatial control of endogenous gene expression in plants.

Growing community of inventors

Gatineau, Canada

Laurian Sylvio Robert