Abstract
A seedling lethal mutant of Nicotiana plumbaginifolia (sdl-1) was isolated by transposon tagging using a maize Dissociation (Ds) element. The insertion mutation was produced by direct co-transformation of protoplasts with two plasmids: one containing Ds and a second with an Ac transposase gene. sdl-1 seedlings exhibit several phenotypes: swollen organs, short hypocotyls in light and dark conditions, and enlarged and multinucleated cells, that altogether suggest cell growth defects. Mutant cells are able to proliferate under in vitro culture conditions. Genomic DNA sequences bordering the transposon were used to recover cDNA from the normal allele. Complementation of the mutant phenotype with the cDNA confirmed that the transposon had caused the mutation. The Dselement was inserted into the first exon of the open reading frame and the homozygous mutant lacked detectable transcript. Phenocopies of the mutant were obtained by an antisense approach. SDL-1 encodes a novel protein found in several plant genomes but apparently missing from animal and fungal genomes; the protein is highly conserved and has a potential plastid targeting motif.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aeschbacher, R.A., Hauser, M.T., Feldmann, K.A. and Benfey, P.N. 1995. The SABRE gene is required for normal cell expansion in Arabidopsis. Genes Dev. 9: 330–340.
An, G., Ebert, P.R., Mitra, A. and Ha, S.B. 1988. Binary vectors. In: S.B. Gelvin, R.A. Schilperoort and D.P.S. Verma (Eds) Plant Molecular Biology Manual, Kluwer Academic Publishers, Dordrecht, Netherlands, pp. A3: 1-19.
Arioli, T., Peng, L., Betzner, A.S., Burn, J., Wittke, W., Herth, W., Camilleri, C., Höfte, H., Plazinski, J., Birch, R., Cork, A., Glover, J., Redmond, J. and Williamson, R.E. 1998. Molecular analysis of cellulose biosynthesis in Arabidopsis. Science 279: 717–720.
Barfield, D.G., Robinson S.J. and Shields R. 1985. Plant regeneration from protoplasts of long term haploid suspension cultures of N. plumbaginifolia. Plant Cell Rep. 4: 104–107.
Baskin, T.I., Betzner, A.S., Hoggart, R., Cork, A. and Williamson, R.E. 1992. Root morphology mutants in Arabidopsis thaliana. Aust. J. Plant Physiol. 19: 427–437.
Bourgin, J.P., Chupeau, Y. and Missonier, C. 1979. Plant regeneration from mesophyll protoplasts of several Nicotiana species. Physiol. Plant. 45: 288–292.
Dellaporta, S.L., Wood, J. and Hicks, J.B. 1983. A plant DNA minipreparation: version II. Plant Mol. Biol. Rep. 1: 19–21.
Dower, W.J., Miller, J.F. and Ragsdale, C.W. 1988. High efficiency transformation of E. coli by high voltage electroporation. Nucl. Acids Res. 16: 6127–6145.
El Malki, F. and Jacobs, M. 2001. Molecular characterization and expression study of a histidine auxotrophic mutant (his1-) of Nicotiana plumbaginifolia. Plant Mol. Biol. 45: 191–199.
Fedoroff, N.V. and Smith, D.L. 1993. A versatile system for detecting transposition in Arabidopsis. Plant J. 3: 273–289.
Fraisier, V., Dorbe, M.F. and Daniel-Vedele, F. 2001. Identification and expression analyses of two genes encoding putative low-affinity nitrate transporters from Nicotiana plumbaginifolia. Plant Mol. Biol. 45: 181–190.
Gabard, J., Marion-Poll, A., Chérel, I., Meyer, C., Müller, A.J. and Caboche, M. 1987. Isolation and characterization of Nicotiana plumbaginifolia nitrate reductase-deficient mutants: genetic and biochemical analysis of the NIA complementation groups. Mol. Gen. Genet. 209: 596–606.
Gierl, A. and Saedler H. 1992. Plant transposable elements and gene tagging. Plant Mol. Biol. 19: 39–49.
Greco, R., Ouwerkerk P.B.F., Sallaud, C., Kohli, A., Colombo, L., Puigdomènech P., Guiderdoni, E., Christou, P., Hoge, J.H.C. and Pereira, A. 2001. Transposon insertional mutagenesis in rice. Plant Physiol. 125: 1175–1177.
Guerineau, F., Lucy, A. and Mullineaux, P. 1992. Effect of two consensus sequences preceding the translation initiator codon on gene expression in plant protoplasts. Plant Mol. Biol. 18: 815–818.
Hajdukiewicz, P., Svab, Z. and Maliga, P. 1994. The small, versatile pPZP family of Agrobacterium binary vectors for plant transformation. Plant Mol. Biol. 25: 989–994.
Hall, T.C., Ma, Y., Buchbinder, B.U., Pyne, J.W., Sun, S.M. and Bliss, F.A. 1978. Messenger RNA for G1 protein of French bean seeds: cell-free translation and product characterization. Proc. Natl. Acad. Sci. USA 75: 3196–3200.
Holding, D.R., McKenzie, R.J. and Coomber, S.A. 1994. Genetic and structural analysis of five Arabidopsis mutants with abnormal root morphology generated by the seed transformation method. Ann. Bot. 74: 193–204.
Houba-Hérin, N., Domin, M. and Pédron, J. 1994a. Transposition of a Ds element from a plasmid into the plant genome in Nicotiana plumbaginifolia protoplast-derived cells. Plant J. 6: 55–66.
Houba-Hérin, N., Domin, M. and Leprince, A.S. 1994b. Some features about transposition of the maize element Dissociation in Nicotiana plumbaginifolia. Genetica 93: 41–48.
Jasinski, M., Stukkens, Y., Degand, H., Purnelle, B., Marchand-Brynaert, J. and Boutry, M. 2001. A plant plasma membrane ATP binding cassette-type transporter is involved in antifungal terpenoid secretion. Plant Cell 13: 1095–1107.
Lane, D.R., Wiedemeier, A., Peng, L., Höfte, H., Vernhettes, S., Desprez, T., Hocart, C.H., Birch, R.J., Baskin, T.I., Burn, J.E., Arioli, T., Betzner, A.S. and Williamson, R.E. 2001. Temperature-sensitive alleles of RSW2 link the KORRIGAN endo-1,4-?-glucanase to cellulose synthesis and cytokinesis in Arabidopsis. Plant Physiol. 126: 278–288.
Langeron, M. 1949. Précis de Microscopie. Techniques-Expérimentation-Diagnostic. Collection de Précis Médicaux. Masson, Paris.
Leprince, A.S., Grandbastien M.A. and Meyer, C. 2001. Retrotransposons of the Tnt1B family are mobile in Nicotiana plumbaginifolia and can induce alternative splicing of the host gene upon insertion. Plant Mol. Biol. 47: 533–541.
McElroy, D., Louwerse, J.D., McElroy, S.M. and Lemaux, P.G. 1997. Development of a simple transient assay for Ac/Ds activity in cells of intact barley tissue. Plant J. 11: 157–165.
Marchuk, D., Drumm, M., Saulino, A. and Collins, F.S. 1991. Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products. Nucl. Acids Res. 19: 1154.
Marin, E., Nussaume, L., Quesada, A., Gonneau, M., Sotta, B., Hugueney, P., Frey, A. and Marion-Poll, A. 1996. Molecular identification of zeaxanthin epoxidase of Nicotiana plumbaginifolia, a gene involved in abscisic acid biosynthesis and corresponding to the ABA locus of Arabidopsis thaliana. EMBO J. 15: 2331–2342.
Marion-Poll, A., Marin, E., Bonnefoy, N. and Pautot, V. 1993. Transposition of the maize autonomous element Activator in transgenic Nicotiana plumbaginifolia plants. Mol. Gen. Genet. 238: 209–217.
Meyer, C., Pouteau, S., Rouzé, P. and Caboche, M. 1994. Isolation and molecular characterization of dTnp1, a mobile and defective transposable element of Nicotiana plumbaginifolia. Mol. Gen. Genet. 242: 194–200.
Müller, J.F. and Caboche, M. 1983. Low density growth of cells derived from Nicotiana and Petunia protoplasts: influence of the source of protoplasts and comparison of the growth promoting activity of various auxins. Physiol. Plant. 57: 35–41.
Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bio-assays with tobacco tissue culture. Physiol. Plant. 15: 473–497.
Negrutiu, I. 1990. In vitro mutagenesis. In: P.J. Dix (Ed.) Plant Cell Line Selection: Procedures and Applications, VCH, Weinheim/New York/Basel/Cambridge, pp. 19–38.
Parinov, S. and Sundaresan, V. 2000. Functional genomics in Arabidopsis: large-scale insertional mutagenesis complements the genome sequencing project. Curr. Opin. Biotech. 11: 157–161.
Sugimoto, K., Otsuki, Y., Saji, S. and Hirochika, H. 1994. Transposition of the maize Ds element from a viral vector to the rice genome. Plant J. 5: 863–871.
van Engelen, F.A., Molthoff, J.W., Conner, A.J., Nap, J.P., Pereira, A. and Stiekema, W.J. 1995. pBINPLUS: an improved plant transformation vector based on pBIN19. Transgenic Res. 4: 288–290.
Vaucheret, H., Marion-Poll, A., Meyer, C., Faure, J.D., Marin, E. and Caboche, M. 1992. Interest in and limits to the utilization of reporter genes for the analysis of transcriptional regulation of nitrate reductase. Mol. Gen. Genet. 235: 259–268.
Walbot, V. 1992. Strategies for mutagenesis and gene cloning using transposon tagging and T-DNA insertional mutagenesis. Annu. Rev. Plant Physiol. Plant Mol. Biol. 43: 49–82.
Walbot, V. 2000. Saturation mutagenesis using maize transposons. Curr. Opin. Plant Biol. 3: 103–107.
Zuo, J., Niu, Q.-W., Nihizawa, N., Wu, Y., Kost, B. and Chua, N.-H. 2000. KORRIGAN, an Arabidopsis endo-1,4-?-glucanase, localizes to the cell plate by polarized targeting and is essential for cytokinesis. Plant Cell. 12: 1137–1152.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Majira, A., Domin, M., Grandjean, O. et al. Seedling lethality in Nicotiana plumbaginifolia conferred by Ds transposable element insertion into a plant-specific gene. Plant Mol Biol 50, 551–562 (2002). https://doi.org/10.1023/A:1019851913083
Issue Date:
DOI: https://doi.org/10.1023/A:1019851913083