Purdue Research Foundation

Other

Pioneer Hi-bred International, Inc.

Iowa State University Research Foundation Inc

Biotechnology Research and Development Corporation

Lubrizol Genetics, Inc.

Stanton B Gelvin

Kirankumar S Mysore

William James Gordon-Kamm

Keith S Lowe

Yumin Tao

Matthew A Bailey

Kan Wang

Saikat Bhattacharjee

Nagesh Sardesai

Min Ni

Lan-Ying Lee

Heiko Oltmanns

Randal Hauptmann

Lan Ying Lee

Yaling Wang

Bronwyn Frame

Decai Cui

Bronwyn R Frame

K Veluthambi

The present invention provides methods and compositions for enhancing the efficiency of-mediated transformation of host cells, such as plant cells, wherein the host cells overexpress light-dependent short hypocotyls 10 (LSH10) protein. The methods of the invention comprise-mediated transfer of T-DNA to a plant cell, wherein the T-DNA contains a DNA of interest to be inserted into a plant genome at a homologous position.

Gene targeting using mutantstrains

A genetic screen formutants displaying a hyper-susceptible totransformation (hat) phenotype was performed. The gene disrupted in the hat3 mutant encodes a putative myb-family transcription factor (MTF) that negatively regulates susceptibility to-mediated transformation. Over-expression of an integrin-like protein results in plants that are hyper-susceptible to transformation. Manipulation of MTF, members of this protein family, and members of the integrin domain-like protein family for example At14a allows improved control oftransformation, including in crops.

Methods and compositions to regulate plant transformation susceptibility

Methods and compositions reduce copy number of transgenes and minimizing vector backbone sequences in plant transformation.strains with T-DNA integrated into the chromosomal DNA of thereduce the copy number and vector backbone sequences. Chromosomal integration vectors to integrate T-DNA into a specific locus ofchromosome are disclosed.

Backbone-free low transgene copy transgenic plants

The invention relates to the use of a modifiedto deliver proteins directly to plant cells. Proteins of interest are delivered to the plant host in the form of a fusion protein with thevirulence protein VirF. Nucleotide sequences encoding such fusion proteins of VirF and a protein of interest are provided. Also provided are bacteria modified to comprise such fusion proteins of VirF and a protein of interest. Methods of introducing such fusion proteins into a plant host are provided. The invention finds use in facilitating plant transformation and particularly in the bio-engineering of desirable traits into crop plants.

Method for delivery of proteins to plant cells

Methods and compositions to increasetransformation efficiency (frequencies) in both dicot and monocot host plants include adding histones to the host plant at most transiently, and using histones and L-cysteine at certain stages in monocot transformation.

Methods and compositions for enhanced plant cell transformation

Adding at least one gene involved in plant host cell T-DNA integration enhances transformation by. The histone H2A gene encoded by theRAT5 gene increases transformation frequencies of plants, most likely by causing overexpression of a product needed for T-DNA integration.genetically transforms plant cells by transferring a portion of the bacterial Ti-plasmid, designated the T-DNA, to the plant, and integrating the T-DNA into the plant genome. However, not all plants are transformable byand transformation frequencies may be too low to be useful. Little is known about the T-DNA integration process, and no plant genes involved in integration have been identified prior to the present invention.

Enhanced plant cell transformation by addition of host genes involved in T-DNA integration

Adding at least one gene involved in plant host cell T-DNA integration enhances transformation by Agrobacterium. The histone H2A gene encoded by the Arabidopsis RAT5 gene increases transformation frequencies of plants, most likely by causing overexpression of a product needed for T-DNA integration. genetically transforms plant cells by transferring a portion of the bacterial Ti-plasmid, designated the T-DNA, to the plant, and integrating the T-DNA into the plant genome. However, not all plants are transformable by Agrobacterium and transformation frequencies may be too low to be useful. Little is known about the T-DNA integration process, and no plant genes involved in integration have been identified prior to the present invention.

Chimeric regulatory regions and gene cassettes for expression of genes

Transcription of the vir genes of the Ti-plasmid in Agrobacterium species, typically induced by phenolic compounds secreted by wounded plant cells, is enhanced when induction is carried out in the presence of effective amounts of an opine from the Octopine Family of Nopaline Family. Production of T-strands in Agrobacterium species can be potentiated by induction preferably in the presence of octopine, nopaline, succinamopine, or leucinopine. Agrobacterium species induced in the presence of an opine compound of the Nopaline or Octopine Family effect highly efficient transformation of even recalcitrant plant species.

Opine enhancement of vir gene induction

A promoter region that drives expression of a 1450 base T.sub.R transcript in octopine-type crown gall tumors can also promote expression of a foreign structural gene in bacteria. Use of this dul-purpose promoter region to drive expression of a single copy of a foreign structural gene in both plants and bacteria is taught. The construction of a selectable marker functional in eukaryotes and prokaryotes is exemplified, as are vectors useful in efforts to transform plants.

Growing community of inventors

West Lafayette, IN, United States of America

Stanton B Gelvin