Germinal and somatic activity of the maize element Activator (Ac) in Arabidopsis.
We have investigated the germinal and somatic activity of the maize Activator (Ac) element in Arabidopsis with the objective of developing an efficient transposon-based system for gene isolation in that plant. Transposition activity was assayed with a chimeric marker that consists of the cauliflower mosaic virus 35S promoter and a bacterial streptomycin phosphotransferase gene (SPT). Somatic activity was detected in seedlings germinated on plates containing streptomycin as green-resistant sectors against a background of white-sensitive cells. Germinal excisions resulted in fully green seedlings. The transposition frequency was extremely low when a single copy of the transposon was present, but appeared to increase with an increase in Ac copy number. Plants that were selected as variegated produced an increased number of green progeny. The methylation state of the Ac elements in lines with either low or high levels of excision was assessed by restriction analysis. No difference was found between these lines, indicating that the degree of methylation did not contribute to the level of Ac activity. Germinal excision events were analyzed molecularly and shown to carry reinserted transposons in about 50% of the cases. In several instances, streptomycin-resistant siblings carried the same transposed Ac element, indicating that excision had occurred prior to meiosis in the parent. We discuss parameters that need to be considered to optimize the use of Ac as a transposon tag in Arabidopsis.
Keller
J
J
DNA Plant Technology Corporation, Oakland, California 94608.
Lim
E
E
James
D W
DW
Dooner
H K
HK
D004247:DNA / Q000032:analysis; D004251:DNA Transposable Elements; D015870:Gene Expression; D005819:Genetic Markers; D008745:Methylation; D010858:Pigmentation / Q000235:genetics; D010944:Plants / Q000235:genetics*; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014170:Transformation, Genetic; D003313:Zea mays / Q000235:genetics*
D004251:DNA Transposable Elements; D005819:Genetic Markers; D011993:Recombinant Fusion Proteins; D004247:DNA