Isolation and molecular characterization of atrazine tolerant barley mutants.

Abstract

Two atrazine-tolerant barley mutants were isolated from atrazine-selection experiments performed on barley chloroplast-mutator plants. Genetic analysis demonstrated that atrazine tolerance was maternally inherited. Molecular characterization of the mutants was performed by PCR amplification of an internal fragment of the chloroplast gene psbA. The BstXI restriction patterns of the amplified fragments showed two bands in both tolerant barley mutants and only one in the atrazine-sensitive control. The 277-bp amplified fragments from the parental line and both atrazine-tolerant mutants were cloned and sequenced. Sequence analysis showed a single nucleotide substitution in both barley atrazine-tolerant mutants, i.e. A to G at the +790 position of the psbA gene-coding sequence. This point mutation corresponds to an amino-acid change of serine- to -glycine and creates a BstXI restriction site. Our results confirmed the conservative variability involved in atrazine tolerance which was previously reported for several other species. To our knowledge this is the first report on the obtention of atrazine-tolerant barley. This finding provides support to the hypothesis that, in addition to a wide variety of chlorophyll deficiencies, the barley chloroplast mutator genotype induces variability in other traits, which could include agronomically valuable mutants.