Efficient transformation of wheat by using a mutated rice acetolactate synthase gene as a selectable marker.
Acetolactate synthase (ALS) is a target enzyme for many herbicides, including sulfonylurea and imidazolinone. We investigated the usefulness of a mutated ALS gene of rice, which had double point mutations and encoded an herbicide-resistant form of the enzyme, as a selectable marker for wheat transformation. After the genomic DNA fragment from rice containing the mutated ALS gene was introduced into immature embryos by means of particle bombardment, transgenic plants were efficiently selected with the herbicide bispyribac sodium (BS). Southern blot analysis confirmed that transgenic plants had one to more than ten copies of the transgene in their chromosomes. Adjustment of the BS concentration combined with repeated selection effectively prevented nontransgenic plants from escaping herbicide selection. Measurement of ALS activity indicated that transgenic plants produced an herbicide-resistant form of ALS and therefore had acquired the resistance to BS. This report is the first to describe a selection system for wheat transformation that uses a selectable marker gene of plant origin.
Ogawa
Taiichi
T
Plant Genome Research Unit, National Institute of Agrobiological Sciences, Kan-non-dai 2-1-2, Tsukuba, Ibaraki, 305-8602, Japan. togawa@affrc.go.jp
Kawahigashi
Hiroyuki
H
Toki
Seiichi
S
Handa
Hirokazu
H
D000095:Acetolactate Synthase / Q000235:genetics*; D015139:Blotting, Southern; D053209:Herbicide Resistance / Q000235:genetics; D006540:Herbicides / Q000494:pharmacology; D009154:Mutation; D012275:Oryza / Q000201:enzymology / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics; D016133:Polymerase Chain Reaction; D014170:Transformation, Genetic; D014908:Triticum / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*
D006540:Herbicides; D010940:Plant Proteins; D000095:Acetolactate Synthase