Abstract
A two-component transposon system based on the Ac element of maize was used as a mutagen in Arabidopsis thaliana. Transposition of a Ds element marked with a hygromycin-resistance gene was activated from four different locations in the Arabidopsis genome. The progeny of 201 plants carrying independent transposition events were screened for mutants with severe, visible phenotypes. Seven mutants were identified and four of them were analyzed genetically. Three mutations were shown to be very closely linked to a transposed copy of the element. Moreover, a mutation (alb3) causing an albino phenotype was conclusively shown to be caused by insertion of the Ds element: somatic and germinal reversion of the mutation occurred in the presence of the transposase gene but not in its absence, and in three revertants the Ds had excised from its position in the mutant line. The DNA adjacent to Ds in the mutant was isolated and it was demonstrated that revertants retained part of the 8-bp duplication caused by insertion of Ds. These experiments indicate that the Ac/Ds system can be used as an insertional mutagen in the heterologous host Arabidopsis, which will permit the isolation of genes from this species by transposon tagging.
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Selected References
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- Baker B., Schell J., Lörz H., Fedoroff N. Transposition of the maize controlling element "Activator" in tobacco. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4844–4848. doi: 10.1073/pnas.83.13.4844. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bancroft I., Bhatt A. M., Sjodin C., Scofield S., Jones J. D., Dean C. Development of an efficient two-element transposon tagging system in Arabidopsis thaliana. Mol Gen Genet. 1992 Jun;233(3):449–461. doi: 10.1007/BF00265443. [DOI] [PubMed] [Google Scholar]
- Bingham P. M., Levis R., Rubin G. M. Cloning of DNA sequences from the white locus of D. melanogaster by a novel and general method. Cell. 1981 Sep;25(3):693–704. doi: 10.1016/0092-8674(81)90176-8. [DOI] [PubMed] [Google Scholar]
- Chuck G., Robbins T., Nijjar C., Ralston E., Courtney-Gutterson N., Dooner H. K. Tagging and Cloning of a Petunia Flower Color Gene with the Maize Transposable Element Activator. Plant Cell. 1993 Apr;5(4):371–378. doi: 10.1105/tpc.5.4.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fedoroff N. V., Furtek D. B., Nelson O. E. Cloning of the bronze locus in maize by a simple and generalizable procedure using the transposable controlling element Activator (Ac). Proc Natl Acad Sci U S A. 1984 Jun;81(12):3825–3829. doi: 10.1073/pnas.81.12.3825. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feldmann K. A., Marks M. D., Christianson M. L., Quatrano R. S. A Dwarf Mutant of Arabidopsis Generated by T-DNA Insertion Mutagenesis. Science. 1989 Mar 10;243(4896):1351–1354. doi: 10.1126/science.243.4896.1351. [DOI] [PubMed] [Google Scholar]
- Haring M. A., Rommens C. M., Nijkamp H. J., Hille J. The use of transgenic plants to understand transposition mechanisms and to develop transposon tagging strategies. Plant Mol Biol. 1991 Mar;16(3):449–461. doi: 10.1007/BF00023995. [DOI] [PubMed] [Google Scholar]
- Koncz C., Martini N., Mayerhofer R., Koncz-Kalman Z., Körber H., Redei G. P., Schell J. High-frequency T-DNA-mediated gene tagging in plants. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8467–8471. doi: 10.1073/pnas.86.21.8467. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koncz C., Mayerhofer R., Koncz-Kalman Z., Nawrath C., Reiss B., Redei G. P., Schell J. Isolation of a gene encoding a novel chloroplast protein by T-DNA tagging in Arabidopsis thaliana. EMBO J. 1990 May;9(5):1337–1346. doi: 10.1002/j.1460-2075.1990.tb08248.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Martin C., Carpenter R., Sommer H., Saedler H., Coen E. S. Molecular analysis of instability in flower pigmentation of Antirrhinum majus, following isolation of the pallida locus by transposon tagging. EMBO J. 1985 Jul;4(7):1625–1630. doi: 10.1002/j.1460-2075.1985.tb03829.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meisler M. H. Insertional mutation of 'classical' and novel genes in transgenic mice. Trends Genet. 1992 Oct;8(10):341–344. doi: 10.1016/0168-9525(92)90278-c. [DOI] [PubMed] [Google Scholar]
- Meyerowitz E. M. Arabidopsis, a useful weed. Cell. 1989 Jan 27;56(2):263–269. doi: 10.1016/0092-8674(89)90900-8. [DOI] [PubMed] [Google Scholar]
- Pereira A., Saedler H. Transpositional behavior of the maize En/Spm element in transgenic tobacco. EMBO J. 1989 May;8(5):1315–1321. doi: 10.1002/j.1460-2075.1989.tb03511.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sutton W. D., Gerlach W. L., Peacock W. J., Schwartz D. Molecular analysis of ds controlling element mutations at the adh1 locus of maize. Science. 1984 Mar 23;223(4642):1265–1268. doi: 10.1126/science.223.4642.1265. [DOI] [PubMed] [Google Scholar]
- Swinburne J., Balcells L., Scofield S. R., Jones J. D., Coupland G. Elevated levels of Activator transposase mRNA are associated with high frequencies of Dissociation excision in Arabidopsis. Plant Cell. 1992 May;4(5):583–595. doi: 10.1105/tpc.4.5.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Valvekens D., Van Montagu M., Van Lijsebettens M. Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5536–5540. doi: 10.1073/pnas.85.15.5536. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van Sluys M. A., Tempé J., Fedoroff N. Studies on the introduction and mobility of the maize Activator element in Arabidopsis thaliana and Daucus carota. EMBO J. 1987 Dec 20;6(13):3881–3889. doi: 10.1002/j.1460-2075.1987.tb02728.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weck E., Courage U., Döring H. P., Fedoroff N., Starlinger P. Analysis of sh-m6233, a mutation induced by the transposable element Ds in the sucrose synthase gene of Zea mays. EMBO J. 1984 Aug;3(8):1713–1716. doi: 10.1002/j.1460-2075.1984.tb02036.x. [DOI] [PMC free article] [PubMed] [Google Scholar]