Abstract
Over 5000 transgenic families of Arabidopsis thaliana produced following seed transformation with Agrobacterium tumefaciens were screened for embryonic lethals, defectives, and pattern mutants. One hundred and seventy-eight mutants with a wide range of developmental abnormalities were identified. Forty-one mutants appear from genetic studies to be tagged (36% of the 115 mutants examined in detail). Mapping with visible markers demonstrated that mutant genes were randomly distributed throughout the genome. Seven mutant families appeared to contain chromosomal translocations because the mutant genes exhibited linkage to visible markers on two different chromosomes. Chromosomal rearrangements may therefore be widespread following seed transformation. DNA gel blot hybridizations with 34 tagged mutants and three T-DNA probes revealed a wide range of insertion patterns. Models of T-DNA structure at each mutant locus were constructed to facilitate gene isolation. The value of such models was demonstrated by using plasmid rescue to clone flanking plant DNA from four tagged mutants. Further analysis of genes isolated from these insertional mutants should help to elucidate the relationship between gene function and plant embryogenesis.
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Bauwens S, Van Oostveldt P, Engler G, Van Montagu M (1991) Distribution of the rDNA and three classes of highly repetitive DNA in the chromatin of interphase nuclei of Arabidopsis thaliana. Chromosoma 101:41–48
Burnham CR (1962) Discussions in cytogenetics. Burgess Publishing Co., Minneapolis
Castle LA, Meinke DW (1993) Embryo-defective mutants as tools to study essential functions and regulatory processes in plant embryo development. Semin Dev Biol 4:31–39
Deng X-W, Matsui M, Wei N, Wagner D, Chu AM, Feldmann KA, Quail PH (1992) COP1, an Arabidopsis regulatory gene, encodes a protein with both a zinc-binding motif and a Gβ homologous domain. Cell 71:791–801
Deroles SC, Gardner RC (1988) Analysis of the T-DNA structure in a large number of transgenic petunias generated by Agrobacterium-mediated transformation. Plant Mol Biol 11:365–377
Errampalli D, Patton D, Castle L, Mickelson L, Hansen K, Schnall J, Feldmann K, Meinke D (1991) Embryonic lethals and TDNA insertional mutagenesis in Arabidopsis. Plant Cell 3:149–157
Feldmann KA (1991) T-DNA insertion mutagenesis in Arabidopsis: mutational spectrum. Plant J 1:71–82
Feldmann KA, Marks MD (1987) Agrobacterium-mediated transformation of germinating seeds of Arabidopsis thaliana: a nontissue culture approach. Mol Gen Genet 208:1–9
Gheysen G, Villarroel R, Van Montagu M (1991) Illegitimate recombination in plants: a model for T-DNA integration. Genes Dev 5:287–297
Graham MW, Doherty JP, Woodcock DM (1990) Efficient construction of plant genomic libraries requires the use of mcr − host strains and packaging mixes. Plant Mol Biol Rep 8:18–27
Heath JD, Weldon R, Monnot C, Meinke DW (1986) Analysis of storage proteins in normal and aborted seeds from embryolethal mutants of Arabidopsis thaliana. Planta 169:304–312
Herman PL, Marks MD (1989) Trichome development in Arabidopsis thaliana. II. Isolation and complementation of the GLABROUS1 gene. Plant Cell 1:1051–1055
Hobbs SLA, Kpodar P, DeLong CMO (1990) The effect of T-DNA copy number, position and methylation on reporter gene expression in tobacco transformants. Plant Mol Biol 15:851–864
Jones JDG, Gilbert DE, Grady KL, Jorgensen RA (1987) T-DNA structure and gene expression in petunia plants transformed by Agrobacterium tumefaciens C58 derivatives. Mol Gen Genet 207:478–485
Jorgensen R, Snyder C, Jones JDG (1987) T-DNA is organized predominantly in inverted repeat structures in plants transformed with Agrobacterium tumefaciens C58 derivatives. Mol Gen Genet 207:471–477
Jürgens G, Mayer U, Torres Ruiz RA, Berleth T, Miséra S (1991) Genetic analysis of pattern formation in the Arabidopsis embryo. In: Roberts K, Coen E, Dean C, Jones J, Chater K, Flavell R, Wilkins A, Holder N (eds) Molecular and cellular basis of pattern formation. Development Suppl 1, pp 27–38
Kieber JJ, Rothenberg M, Roman G, Feldmann KA, Ecker JR (1993) CTRL, a negative regulator of the ethylene response pathway in Arabidopsis, encodes a member of the Raf family of protein kinases. Cell 72:427–441
Kilby NJ, Leyser HMO, Furner IJ (1992) Promoter methylation and progressive transgene inactivation in Arabidopsis. Plant Mol Biol 20:103–112
Koornneef M, Stam P (1988) Procedures for mapping by using F2 and F3 populations. Arabidopsis Inf Serv 25:35–40
Koornneef M, Dresselhuys HC, Sree Ramulu K (1982) The genetic identification of translocations in Arabidopsis. Arabidopsis Inf Serv 19:93–99
Lee M (1988) The chromosomal basis of somaclonal variation. Annu Rev Plant Physiol Plant Mol Biol 39:413–437
Maluszynska J, Heslop-Harrison JS (1991) Localization of tandemly repeated DNA sequences in Arabidopsis thaliana. Plant J 1:159–166
Mayer U, Torres Ruiz RA, Berleth T, Miséra S, Jürgens G (1991) Mutations affecting body organization in the Arabidopsis embryo. Nature 353:402–407
Mayerhofer R, Koncz-Kalman Z, Nawrath C, Bakkeren G, Crameri A, Angelis K, Rédei GP, Schell J, Hohn B, Koncz C (1991) T-DNA integration: a mode of illegitimate recombination in plants. EMBO J 10:697–704
Meinke DW (1985) Embryo-lethal mutants of Arabidopsis thaliana: analysis of mutants with a wide range of lethal phases. Theor Appl Genet 69:543–552
Meinke DW (1991a) Perspectives on genetic analysis of plant embryogenesis. Plant Cell 3:857–866
Meinke DW (1991b) Embryonic mutants of Arabidopsis thaliana. Dev Genet 12:382–392
Meinke DW (1992) A homoeotic mutant of Arabidopsis thaliana with leafy cotyledons. Science 258:1647–1650
Müller AJ (1963) Embryonentest zum Nachweis rezessiver Letalfaktoren bei Arabidopsis thaliana. Biol Zentralbl 82:133–163
Patton DA, Franzmann LH, Meinke DW (1991) Mapping genes essential for embryo development in Arabidopsis thaliana. Mol Gen Genet 227:337–347
Peterhans A, Schlüpmann H, Basse C, Paszkowski J (1990) Intrachromosomal recombination in plants. EMBO J 9:3437–3445
Spielmann A, Simpson RB (1986) T-DNA structure in transgenic tobacco plants with multiple independent integration sites. Mol Gen Genet 205:34–41
Tovar J, Lichtenstein C (1992) Somatic and meiotic chromosomal recombination between inverted duplications in transgenic tobacco plants. Plant Cell 4:319–332
Van Lijsebettens M, den Boer B, Hernalsteens J-P, Van Montagu M (1991) Insertional mutagenesis in Arabidopsis thaliana. Plant Sci 80:27–37
Velten J, Schell J (1985) Selection-expression plasmid vectors for use in genetic transformation of higher plants. Nucleic Acids Res 13:6981–6998
Walden R, Hayashi H, Schell J (1991) T-DNA as a gene tag. Plant J 1:281–288
Ward JE, Akiyoshi DE, Regier D, Datta A, Gordon MP, Nester EW (1988) Characterization of the virB operon from an Agrobacterium tumefaciens Ti plasmid. J Biol Chem 263:5804–5814
Weigel D, Alvarez J, Smyth DR, Yanofsky MF, Meyerowitz EM (1992) LEAFY controls floral meristem identity in Arabidopsis. Cell 69:843–859
Yanofsky MF, Ma H, Bowman JL, Drews GN, Feldmann KA, Meyerowitz EM (1990) The protein encoded by the Arabidopsis homeotic gene agamous resembles transcription factors. Nature 346:35–39
Yeung EC, Meinke DW (1993) Embryogenesis in angiosperms: development of the suspensor. Plant Cell 5:1371–1381
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Communicated by E. Meyerowitz
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Castle, L.A., Errampalli, D., Atherton, T.L. et al. Genetic and molecular characterization of embryonic mutants identified following seed transformation in Arabidopsis . Molec. Gen. Genet. 241, 504–514 (1993). https://doi.org/10.1007/BF00279892
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DOI: https://doi.org/10.1007/BF00279892