The tomato gene for the chloroplastic Cu,Zn superoxide dismutase: regulation of expression imposed in transgenic tobacco plants by a short promoter.
The chloroplastic Cu,Zn superoxide dismutase (SOD) has an important role in the defense against damage by oxygen radicals in the chloroplasts. Here, for the first time, we report on the isolation of a genomic DNA clone from tomato that contains all the coding sequence for the chloroplastic Cu,Zn SOD as well as a 442 bp DNA fragment upstream of the translational initiation site. The latter upstream sequence has a putative TATA box and a 285 bp promoter region, 5' of the apparent transcriptional initiation and a 157 bp leader region. The coding sequence is composed of 8 exons that are interspaced by 7 introns; we termed this gene SODCp;Le:1. The 442 bp fragment was cloned into a pBI101 vector, upstream of the uidA (GUS) gene, via transcriptional fusion. Agrobacterium-mediated transformation resulted in transgenic tobacco plants. The progeny (after self-pollination) of 14 transformed plants, which expressed GUS above a threshold of 1 nmol/min per mg protein, were found to fall into two distinct groups. In the seedlings of 10 lines (group A) GUS expression was enhanced by exposure to light. In 4 lines of this group maintenance for 3 days in the dark eliminated GUS activity. The seedlings of group B expressed GUS regardless of the light/dark regime. In plants of group A, GUS expression was also developmentally regulated: high GUS activity in young leaves, low activity in mature leaves and no activity in the roots. The results suggest that this short chloroplastic Cu,Zn SOD promoter contains motifs for developmental (spatial) regulation as well as motifs responsive to light (or to oxygen radicals resulting from light-driven photosynthesis).
Kardish
N
N
Department of Plant Genetics, Weizmann Institute of Science, Rehovot, Israel.
Magal
N
N
Aviv
D
D
Galun
E
E
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002736:Chloroplasts / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic / Q000528:radiation effects; D017930:Genes, Reporter; D005822:Genetic Vectors; D015698:Genomic Library; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics; D008027:Light; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified / Q000528:radiation effects; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012639:Seeds / Q000254:growth & development / Q000528:radiation effects; D017422:Sequence Analysis, DNA; D013482:Superoxide Dismutase / Q000096:biosynthesis / Q000235:genetics*; D014026:Nicotiana / Q000235:genetics / Q000502:physiology / Q000528:radiation effects; D014675:Vegetables / Q000201:enzymology / Q000235:genetics*
D011993:Recombinant Fusion Proteins; D013482:Superoxide Dismutase; D005966:Glucuronidase