Pioneer Hi-bred International, Inc.

University of Missouri

Deping Xu

Dwight T Tomes

Haiyin Wang

Xiaomu Niu

Dilbag Singh Multani

Zuo-Yu Zhao

Sobhana Sivasankar

Timothy George Helentjaris

Myeong-Je Cho

David Charles Cerf

Xinli Emily Wu

Igor Cunha Oliveira

Anna Lyznik

Myoeng-Je Cho

Timothy J Helentjaris

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

A method for increasing efficiency of germplasm screening for transformability may include providing a plurality of lines of plant target tissue to be transformed, characterizing each of the lines to provide characterization data, the characterization data comprises DNA or nucleic acid delivery technique response data and tissue culture response data, eliminating one or more of the plurality of lines based on the characterization data without performing transformation of the plurality of lines, such that a subset of the plurality of lines remains, and performing transformation experiments on the subset of the plurality of lines. The method may also include selecting a DNA or nucleic acid delivery technique protocol and a tissue culture protocol prior to the characterization.

Method for increasing efficiency of germplasm screening in plant transformation

The present invention provides for rapid and large scale evaluation of expression of, or function of, nucleotide sequences in plants. The invention comprises three specific components which provide for fast and large scale evaluation of nucleotide sequences. The first component includes delivery in either a single event a library of different engineered vectors or a single engineered vector for a single target nucleotide sequence comprising sequences the function of which is desired to be known in plant cells. Surprisingly, applicants have discovered that, the introduction of multiple vectors to plant cells predominantly results in individual transgenic plants which contain only a single transformation event. The second feature of the invention involves a highly transformable, fast cycling and/or miniature size plant and the final step involves mass scale analysis of T0 plants for various phenotypes and plasmid rescue to identify the nucleotide sequence present in a particular phenotype.

Methods for large scale functional evaluation of nucleotide sequences in plants

The invention relates to the regulation of gene expression, particularly to methods and compositions for regulating gene expression in plants. Expression cassettes comprising a polynucleotide molecule of interest operably linked to the Brachytic2 (Br2) promoter, or a functional fragment or variant thereof, are provided. Methods of using such expression cassettes, as well as plant cells, plant tissues, plants, and seeds transformed with these expression cassettes, are additionally provided. The methods and compositions find use in regulating the expression of a polynucleotide molecule of interest in a plant.

Brachytic2 (Br2) promoter from maize and methods of use

The present invention provides compositions and methods for regulating expression of nucleotide sequences in a plant. Compositions comprise novel nucleic acid sequences encoding a transcription factor involved in modulating gene expression in response to abiotic stress such as cold or drought. Methods for expressing the nucleic acid sequence in a plant and improving cold and/or drought tolerance of plants are also provided.

Transcriptional activators involved in abiotic stress tolerance

Brachytic2 (BR2) promoter from maize and methods of use

Growing community of inventors

Johnston, IA, United States of America

Deping Xu