rol genes of Agrobacterium rhizogenes cucumopine strain: sequence, effects and pattern of expression.
By sequencing the central region of the cucumopine-type T-DNA of Agrobacterium rhizogenes strain 2659, we identified three open reading frames homologous, to different extents, to ORFs 10, 11 and 12 (rolA, B and C) of the agropine-type (1855) T-DNA. Recombinant Agrobacterium strains encompassing the ORFs of 2659 T-DNA--which we refer to as rol alpha, beta and gamma--were utilized to infect carrot discs and to obtain transgenic tobacco plants, in order to compare the morphogenetic capabilities to those of the 1855 rol genes. Moreover, a long segment of the 5' non-coding region of rol alpha and rol beta was fused to the GUS reporter gene and the pattern of expression and the responsiveness to auxin of the constructs was analysed in transgenic tobacco. Differences in the auxin requirement for root induction between the 2659 rol genes and their respective 1855 counterparts were pinpointed. These differences are not due to gene regulation and presumably reflect functional differences in the proteins encoded. Differences were also observed in the pattern of expression of rol beta in roots of transgenic plants, as compared to rolB. In addition, the pattern of expression of rol alpha-GUS construct in roots was found to be analogous to that observed for a construct driven by two of the five regulatory domains of the rolB promoter.
Serino
G
G
Centro per lo Studio degli Acidi Nucleici, CNR, Roma, Italy.
Clerot
D
D
Brevet
J
J
Costantino
P
P
Cardarelli
M
M
D001426:Bacterial Proteins / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D018552:Daucus carota / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D007093:Imidazoles; D007210:Indoleacetic Acids / Q000494:pharmacology; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D018517:Plant Roots / Q000187:drug effects / Q000254:growth & development; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic / Q000235:genetics; D011523:Protoplasts; D011725:Pyridines; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012231:Rhizobium / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014026:Nicotiana / Q000235:genetics
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D007093:Imidazoles; D007210:Indoleacetic Acids; D011725:Pyridines; D011993:Recombinant Fusion Proteins; C036483:T-DNA; C082688:cucumopine; C075969:rol protein, Bacteria; D005966:Glucuronidase