University of California

Pioneer Hi-bred International, Inc.

Other

Myeong-Je Cho

Peggy G Lemaux

Bob B Buchanan

Zuo-Yu Zhao

Joshua H Wong

Gregorio Del Val

Maxime Caillau

David Charles Cerf

Xinli Emily Wu

Deping Xu

Shibo Zhang

Corina Marx

Phillip Bregitzer

Robert B Buchanan

The present invention provides assays and methods for efficiently testing a polynucleotide of interest for a phenotype in a root. In some embodiments, the assays and methods include regenerating green tissue that is transgenic for at least one polynucleotide of interest into one or more transgenic plantlets that have at least one transgenic root. Further provided are methods of making a root assay by contacting green tissue with a first rooting medium to produce a plantlet and a plurality of roots. Additionally provided are methods of assaying for insecticidal activity on a live root. Accordingly provided herein is a substantially contamination-free, root bioassay. Further provided are methods of identifying a promoter having activity in a root.

In vitro plant bioassay methods

The invention provides improved plant transformation methods. In particular the method provides increased transformation frequency, especially in recalcitrant plants. The method includes various transformation protocols for monocots, such as maize and sorghum, using a combination of media and light conditions to achieve increased efficiency of monocot transformation and increased callus initiation frequencies.

Methods for improving monocot transformation

The present invention is directed to a transgenic plant wherein at least a part of said plant includes a recombinant nucleic acid with a promoter active in the part operably linked to a nucleic acid encoding a thioredoxin polypeptide wherein the promoter is a seed or grain maturation-specific promoter and the thioredoxin polypeptide includes the amino acid sequence WCGPC. The present invention is further directed to transgenic plants that overexpress thioredoxin in seed wherein the overexpression of thioredoxin h effects a significant increase in the reduction of proteins (&#x2014;SH as compared to S&#x2014;S) of the albumin fraction of the seed.

Transgenic plants with elevated thioredoxin levels

Transgenic monocot plants and seeds comprising monocot seed-specific promoters from seed-storage protein genes and monocot seed-specific targets sequences of expression of heterologous proteins in subcellular compartments to protect the protein are disclosed. The transgenic monocot plants include maize, wheat, rice and barley. Also disclosed are methods of producing transgenic monocot seed from the transgenic plants.

Production of proteins in plant seeds

Improved compositions and methods for transformation and regeneration of plants from embryogenic callus are disclosed that include, for example: use of an intermediate-incubation medium after callus induction to increase the competence of the transformed cells for regeneration; dim light conditions during early phases of selection; use of green callus tissue as a target for microprojectile bombardment; and media with optimized levels of phytohormones and copper concentrations.

Compositions and methods for plant transformation and regeneration

The present invention provides barley thioredoxin h nucleic acids and NADP-thioredoxin reductase nucleic acids, the respective encoded proteins and methods of use.

Barley gene for thioredoxin and NADP-thioredoxin reductase

Compositions and methods of use are provided herein to make and use transgenic plants with value-added traits.

Value-added traits in grain and seed transformed with thioredoxin

Methods for producing proteins in plant seeds are disclosed. Expression of the protein is driven by a seed-specific promoter and the protein is preferably expressed as a fusion polypeptide that includes a signal peptide that causes the protein to accumulate in a subcellular compartment to protect the protein.

A method of preparing green regenerative tissue of barley suitable for transformation is presented. The method includes incubating barley tissue on a callus induction medium under dim light. The dim light conditions are sufficient to produce green regenerative tissue. The callus induction media includes auxin and copper.

Method for preparing barley green regenerative tissue

Methods for transforming plants, particularly commercial genotypes of cereals, are provided. The methods involve transformation of meristematic organogenic tissue, and include the use of defined plant growth media. The methods disclosed provide more stable transgenic plants, and permit the transformation of varieties of cereals that are not amenable to transformation by conventional approaches.

Methods and compositions for transformation of cereals using cultured shoot meristematic tissue

The present invention provides barley thioredoxin h nucleic acids and proteins. The barley thioredoxin h nucleic acid may be isolated or it may be an expression vector. The expression vector may be operably linked to a transcriptional regulatory sequence. The invention also provides for transgenic plants comprising recombinant barley thioredoxin h. The invention also provides methods of expressing and purifying barley thioredoxin h.

Growing community of inventors

Alameda, CA, United States of America

Myeong-Je Cho