Pioneer Hi-Bred International, Inc.

E.i. Dupont De Nemours and Company

Xinli Emily Wu

William James Gordon-Kamm

Keith S Lowe

Kevin E McBride

Christopher Jay Scelonge

Zuo-Yu Zhao

Theodore Mitchell Klein

Yue Yun

Ning Wang

George J Hoerster

Shiv Bahadur Tiwari

Alexandre Da Silva Conceição

Craig E Hastings

Justin Schares

Randal Arnold

Matthew Paul Cope

Roberto Barreiro

Carlos M La Rota

Ajith Anand

Clifford Paul Hunter

Todd J Jones

Maren L Arling

Dennis L Bidney

Michael D Miller

Mark E Williams

Myeong-Je Cho

Deping Xu

Sheila E Maddock

Joanne E Barton

Tanveer Hussain

Bing-Bing Wang

Evan Dale Elsing

Myoeng-Je Cho

Justin Andrew Schares

Evan D Elsing

The disclosure pertains to methods and compositions for the rapid and efficient transformation of plants. The disclosure further provides methods for producing a transgenic plant, comprising (a) transforming a cell of an explant with an expression construct comprising (i) a nucleotide sequence encoding a WUS/WOX homeobox polypeptide; (ii) a nucleotide sequence encoding a polypeptide comprising two AP2-DNA binding domains; or (iii) a combination of (i) and (ii); and (b) allowing expression of the polypeptide of (a) in each transformed cell to form a regenerable plant structure in the absence of exogenous cytokinin, wherein no callus is formed; and (c) germinating the regenerable plant structure to form the transgenic plant. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present disclosure.

Methods and compositions for rapid plant transformation

Methods for preserving viability of plant tissues such as plant embryos are provided herein. Also included are methods for storing genomic DNA and/or molecular marker assay materials in an oil bilayer as part of a high-throughput molecular characterization system. Moreover, plant embryos may be treated while in an oil matrix. The treatment may include chromosome doubling,-mediated transformation, or herbicide selection as part of an embryo rescue process.

Methods and devices for creating doubled haploid embryos using oil matrices

Methods for creating doubled haploid maize embryos using oil matrices

Methods for producing homozygous plants, seeds, and plant cells are provided. The methods comprise transforming a somatic cell of a maize haploid embryo with a polynucleotide of interest and treating the transformed cell with a chromosome doubling agent. One or more growth stimulation proteins, such as, for example, RepA or Lec1, may also be provided.

Methods of transforming somatic cells of maize haploid embryos

Methods for producing doubled haploid corn () plants, seeds, and plant cells are provided.

Doubling of chromosomes in haploid embryos

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

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

Growing community of inventors

Johnston, IA, United States of America

Xinli Emily Wu