Abstract
We isolated the dihydroflavonol 4-reductase (DFR) gene from tomato (Lycopersicon esculentum) using a previously characterized cDNA as probe. Earlier studies had indicated that the DFR gene is present in tomato as a single gene located on chromosome 2 near the locus anthocyanin without (aw). Mutant alleles of the aw locus result in the complete absence of anthocyanin pigmentation throughout all stages of plant development. When the genomic DFR clone was introduced by Agrobacterium-mediated transformation into plants bearing the aw mutation, primary transgenic seedlings accumulated anthocyanins that could be observed while the plants were still in tissue culture and which continued to be observed as the plants matured. Progeny of self pollinated and backcrossed transgenic plants segregated for anthocyanin pigmentation, and Southern hybridization analyses indicated the presence of the DFR transgene exclusively in those plants with pigmentation. These data indicate that the aw locus likely corresponds to the structural gene for DFR and that DFR can be used as a visual, nondestructive, plant-derived marker gene for tomato.
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- Beld M., Martin C., Huits H., Stuitje A. R., Gerats A. G. Flavonoid synthesis in Petunia hybrida: partial characterization of dihydroflavonol-4-reductase genes. Plant Mol Biol. 1989 Nov;13(5):491–502. doi: 10.1007/BF00027309. [DOI] [PubMed] [Google Scholar]
- Bevan M., Barnes W. M., Chilton M. D. Structure and transcription of the nopaline synthase gene region of T-DNA. Nucleic Acids Res. 1983 Jan 25;11(2):369–385. doi: 10.1093/nar/11.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. doi: 10.1093/nar/12.22.8711. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirsch P. R., Wang C. L., Woodward M. J. Construction of a Tn5 derivative determining resistance to gentamicin and spectinomycin using a fragment cloned from R1033. Gene. 1986;48(2-3):203–209. doi: 10.1016/0378-1119(86)90078-8. [DOI] [PubMed] [Google Scholar]
- Jefferson R. A., Kavanagh T. A., Bevan M. W. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 1987 Dec 20;6(13):3901–3907. doi: 10.1002/j.1460-2075.1987.tb02730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jones J. D., Shlumukov L., Carland F., English J., Scofield S. R., Bishop G. J., Harrison K. Effective vectors for transformation, expression of heterologous genes, and assaying transposon excision in transgenic plants. Transgenic Res. 1992 Nov;1(6):285–297. doi: 10.1007/BF02525170. [DOI] [PubMed] [Google Scholar]
- Meyer P., Heidmann I., Forkmann G., Saedler H. A new petunia flower colour generated by transformation of a mutant with a maize gene. Nature. 1987 Dec 17;330(6149):677–678. doi: 10.1038/330677a0. [DOI] [PubMed] [Google Scholar]
- Murphy G., Kavanagh T. Speeding-up the sequencing of double-stranded DNA. Nucleic Acids Res. 1988 Jun 10;16(11):5198–5198. doi: 10.1093/nar/16.11.5198. [DOI] [PMC free article] [PubMed] [Google Scholar]
- O'Reilly C., Shepherd N. S., Pereira A., Schwarz-Sommer Z., Bertram I., Robertson D. S., Peterson P. A., Saedler H. Molecular cloning of the a1 locus of Zea mays using the transposable elements En and Mu1. EMBO J. 1985 Apr;4(4):877–882. doi: 10.1002/j.1460-2075.1985.tb03713.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ow D. W., DE Wet J. R., Helinski D. R., Howell S. H., Wood K. V., Deluca M. Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Science. 1986 Nov 14;234(4778):856–859. doi: 10.1126/science.234.4778.856. [DOI] [PubMed] [Google Scholar]
- Thomas C. M., Meyer R., Helinski D. R. Regions of broad-host-range plasmid RK2 which are essential for replication and maintenance. J Bacteriol. 1980 Jan;141(1):213–222. doi: 10.1128/jb.141.1.213-222.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yenofsky R. L., Fine M., Pellow J. W. A mutant neomycin phosphotransferase II gene reduces the resistance of transformants to antibiotic selection pressure. Proc Natl Acad Sci U S A. 1990 May;87(9):3435–3439. doi: 10.1073/pnas.87.9.3435. [DOI] [PMC free article] [PubMed] [Google Scholar]
