An Arabidopsis histone H2A mutant is deficient in Agrobacterium T-DNA integration.
Agrobacterium tumefaciens genetically transforms plant cells by transferring a portion of the bacterial Ti-plasmid, the T-DNA, to the plant and integrating the T-DNA into the plant genome. Little is known about the T-DNA integration process, and no plant genes involved in integration have yet been identified. We characterized an Arabidopsis mutant generated by T-DNA insertional mutagenesis, rat5, that is resistant to Agrobacterium root transformation. rat5 contains two copies of T-DNA integrated as a tandem direct repeat into the 3' untranslated region of a histone H2A gene, upstream of the polyadenylation signal sequence. Transient and stable beta-glucuronidase expression data and assessment of the amount of T-DNA integrated into the genomes of wild-type and rat5 Arabidopsis plants indicated that the rat5 mutant is deficient in T-DNA integration. We complemented the rat5 mutation by expressing the RAT5 histone H2A gene in the mutant plant. Overexpression of RAT5 in wild-type plants increased Agrobacterium transformation efficiency. Furthermore, transient expression of a RAT5 gene from the incoming T-DNA was sufficient to complement the rat5 mutant and to increase the transformation efficiency of wild-type Arabidopsis plants.
Mysore
K S
KS
Department of Biological Sciences, Purdue University, West Lafayette, IN 47907-1392, USA.
Nam
J
J
Gelvin
S B
SB
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D005816:Genetic Complementation Test; D005966:Glucuronidase / Q000235:genetics; D006579:Heterozygote; D006657:Histones / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D018517:Plant Roots / Q000235:genetics / Q000382:microbiology; D030821:Plants, Genetically Modified / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic
D004269:DNA, Bacterial; D018744:DNA, Plant; D006657:Histones; D011993:Recombinant Fusion Proteins; C036483:T-DNA; D005966:Glucuronidase