The dsdA gene from Escherichia coli provides a novel selectable marker for plant transformation.
Plants are sensitive to D-serine, but functional expression of the dsdA gene, encoding D-serine ammonia lyase, from Escherichia coli can alleviate this toxicity. Plants, in contrast to many other organisms, lack the common pathway for oxidative deamination of D-amino acids. This difference in metabolism has major consequences for plant responses to D-amino acids, since several D-amino acids are toxic to plants even at relatively low concentrations. Therefore, introducing an enzyme specific for a phytotoxic D-amino acid should generate a selectable characteristic that can be screened. Here we present the use of the dsdA gene as a selectable marker for transformation of Arabidopsis. D-serine ammonia lyase catalyses the deamination of D-serine into pyruvate, water and ammonium. dsdA transgenic seedlings can be clearly distinguished from wild type, having an unambiguous phenotype immediately following germination when selected on D-serine containing medium. The dsdA marker allows flexibility in application of the selective agent: it can be applied in sterile plates, in foliar sprays or in liquid culture. Selection with D-serine resistance was compared with selection based on kanamycin resistance, and was found to generate similar transformation frequencies but also to be more unambiguous, more rapid and more versatile with respect to the way the selective agent can be supplied.
Erikson
Oskar
O
Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, Umeå.
Hertzberg
Magnus
M
Näsholm
Torgny
T
D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D004351:Drug Resistance; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D005819:Genetic Markers / Q000235:genetics*; D012695:L-Serine Dehydratase / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012694:Serine / Q000378:metabolism / Q000494:pharmacology; D014170:Transformation, Genetic / Q000235:genetics*
D029968:Escherichia coli Proteins; D005819:Genetic Markers; D012333:RNA, Messenger; D012694:Serine; D012695:L-Serine Dehydratase