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PMID: 10639185 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

An Arabidopsis histone H2A mutant is deficient in Agrobacterium T-DNA integration.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 97 ·No. 2 ·2000-01-18 ·Pages 948-53

Mysore KS, Nam J, Gelvin SB

Abstract

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.

MeSH Terms
Agrobacterium tumefaciens/genetics Amino Acid Sequence Arabidopsis/genetics DNA, Bacterial/chemistry,genetics DNA, Plant/chemistry,genetics Genetic Complementation Test Glucuronidase/genetics Heterozygote Histones/genetics Molecular Sequence Data Mutation Plant Roots/genetics,microbiology Plants, Genetically Modified/genetics Recombinant Fusion Proteins/genetics Sequence Analysis, DNA Transformation, Genetic
Chemicals
DNA, Bacterial DNA, Plant Histones Recombinant Fusion Proteins T-DNA Glucuronidase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Mysore K S
Department of Biological Sciences, Purdue University, West Lafayette, IN 47907-1392, USA.
Nam J
Gelvin S B
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.97.2.948
Published
2000-01-18
Pages
948-53
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC15436
Subset
IM
Databases
GENBANK
AF204967, AF204968
External Links
PubMed source DOI: 10.1073/pnas.97.2.948 PMC Full Text View journal details
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