Abstract: Agrobacterium strains with mutations in the VirD2 gene and protein and expression constructs and genome-editing systems for leveraging the same. Methods are also provided for Agrobacterium-mediated incorporation of exogenous expressible nucleic acids into a host plant material using the Agrobacterium strains, expression constructs, and/or genome-editing systems.

Abstract: The present disclosure provides transgenic plants and/or plant cells comprising overexpressed VirE2 gene or VirE2 protein in plant cytoplasm that upregulates or downregulates certain plant gene and/or proteins to facilitate transformation. The present disclosure further provides transgenic plants and/or plant cells comprising overexpressed plant gene or protein that upregulated by VirE2 gene or VirE2 protein for facilitating transformation. The transgenic plants and/or plant cells comprising downexpressed or knockout plant gene or protein that downregulated by VirE2 gene or VirE2 protein for facilitating transformation are also provided. Methods of making and using the transgenic plants and/or plants cells are also provided.

Abstract: The present disclosure provides a series of mutant Agrobacterium strains generated by random mutagenesis of a wide-type or ? mutant VirD2 gene or VirD2 protein. The mutant Agrobacterium strains of the present disclosure transiently express T-DNA-encoded transgenes in a target plant but do not stably integrate these genes into the plant genome.

Abstract: The present invention provides methods and compositions for enhancing the efficiency of Agrobacterium-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 Agrobacterium-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.

Abstract: Mutation of the rice ortholog of the Arabidopsis MTF1 gene results in increased rice transformation. CRISPR was used to generate several different mutations in the rice MTF1 gene. Agrobacterium-mediated transformation of these homozygous mutants indicated 5- to 10-fold increased transient and stable transformation, using GUS and luciferase assays.

Abstract: A genetic screen for Arabidopsis mutants displaying a hyper-susceptible to Agrobacterium transformation (hat) phenotype was performed. The gene disrupted in the hat3 mutant encodes a putative myb-family transcription factor (MTF) that negatively regulates susceptibility to Agrobacterium-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 of Agrobacterium transformation, including in crops.

Abstract: The present invention provides methods and compositions for enhancing the efficiency of Agrobacterium-mediated transformation of host cells, such as plant cells, wherein the host cells overexpress LSH10 protein. The methods of the invention comprise Agrobacterium-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.

Abstract: Peptide aptamers and the methods to produce cassettes including the aptamers and manipulating them, are described. The peptide aptamer cassettes are useful to, e.g., inhibit protein function such as proteins necessary for the transformation of plants, or to replicate cells.

Abstract: A genetic screen for Arabidopsis mutants displaying a hyper-susceptible to Agrobacterium transformation (hat) phenotype was performed. The gene disrupted in the hat3 mutant encodes a putative myb-family transcription factor (MTF) that negatively regulates susceptibility to Agrobacterium-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 of Agrobacterium transformation, including in crops.

Abstract: A genetic screen for Arabidopsis mutants displaying a hyper-susceptible to Agrobacterium transformation (hat) phenotype was performed. The gene disrupted in the hat3 mutant encodes a putative myb-family transcription factor (MTF) that negatively regulates susceptibility to Agrobacterium-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 of Agrobacterium transformation, including in crops.

Abstract: Peptide aptamers and the methods to produce cassettes including the aptamers and manipulating them, are described. The peptide aptamer cassettes are useful to, e.g., inhibit protein function such as proteins necessary for the transformation of plants, or to replicate cells.

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

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

Abstract: The invention relates to the use of a modified Agrobacterium to deliver proteins directly to plant cells. Proteins of interest are delivered to the plant host in the form of a fusion protein with the Agrobacterium virulence 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.

Abstract: Methods and compositions to increase Agrobacterium transformation 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.

Abstract: 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. Agrobacterium tumefaciens 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.

Abstract: 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. Agrobacterium tumefaciens 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.

Abstract: The invention relates to the use of a modified Agrobacterium to deliver proteins directly to plant cells. Proteins of interest are delivered to the plant host in the form of a fusion protein with the Agrobacterium virulence 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.

Abstract: Methods and compositions to increase Agrobacterium transformation 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.

Abstract: 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. Agrobacterium tumefaciens 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.