Embryonic lethals and T-DNA insertional mutagenesis in Arabidopsis

Deena Errampalli, David Patton, Linda Castle, Leigh Mickelson, Karl Hansen, Jennifer Schnall, Kenneth A Feldmann, David Meinke

Research output: Contribution to journalArticle

110 Citations (Scopus)

Abstract

T-DNA insertional mutagenesis represents a promising approach to the molecular isolation of genes with essential functions during plant embryo development. We describe in this report the isolation and characterization of 18 mutants of Arabidopsis thaliana defective in embryo development following seed transformation with Agrobacterium tumefaciens. Random T-DNA insertion was expected to result in a high frequency of recessive embryonic lethals because many target genes are required for embryogenesis. The cointegrate Ti plasmid used in these experiments contained the nopaline synthase and neomycin phosphotransferase gene markers. Nopaline assays and resistance to kanamycin were used to estimate the number of functional inserts present in segregating families. Nine families appeared to contain a T-DNA insert either within or adjacent to the mutant gene. Eight families were clearly not tagged with a functional insert and appeared instead to contain mutations induced during the transformation process. DNA gel blot hybridization with internal and right border probes revealed a variety of rearrangements associated with T-DNA insertion. A general strategy is presented to simplify the identification of tagged embryonic mutants and facilitate the molecular isolation of genes required for plant embryogenesis.

Original languageEnglish (US)
Pages (from-to)149-157
Number of pages9
JournalPlant Cell
Volume3
Issue number2
StatePublished - Feb 1991
Externally publishedYes

Fingerprint

insertional mutagenesis
Mutagenesis
Insertional Mutagenesis
lethal genes
Arabidopsis
Embryonic Development
Genes
embryogenesis
nopaline
DNA
Seeds
Plant Tumor-Inducing Plasmids
mutants
Kanamycin Kinase
Kanamycin Resistance
Plant Genes
genes
Agrobacterium tumefaciens
Plant Development
Essential Genes

ASJC Scopus subject areas

  • Plant Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Errampalli, D., Patton, D., Castle, L., Mickelson, L., Hansen, K., Schnall, J., ... Meinke, D. (1991). Embryonic lethals and T-DNA insertional mutagenesis in Arabidopsis. Plant Cell, 3(2), 149-157.

Embryonic lethals and T-DNA insertional mutagenesis in Arabidopsis. / Errampalli, Deena; Patton, David; Castle, Linda; Mickelson, Leigh; Hansen, Karl; Schnall, Jennifer; Feldmann, Kenneth A; Meinke, David.

In: Plant Cell, Vol. 3, No. 2, 02.1991, p. 149-157.

Research output: Contribution to journalArticle

Errampalli, D, Patton, D, Castle, L, Mickelson, L, Hansen, K, Schnall, J, Feldmann, KA & Meinke, D 1991, 'Embryonic lethals and T-DNA insertional mutagenesis in Arabidopsis', Plant Cell, vol. 3, no. 2, pp. 149-157.
Errampalli D, Patton D, Castle L, Mickelson L, Hansen K, Schnall J et al. Embryonic lethals and T-DNA insertional mutagenesis in Arabidopsis. Plant Cell. 1991 Feb;3(2):149-157.
Errampalli, Deena ; Patton, David ; Castle, Linda ; Mickelson, Leigh ; Hansen, Karl ; Schnall, Jennifer ; Feldmann, Kenneth A ; Meinke, David. / Embryonic lethals and T-DNA insertional mutagenesis in Arabidopsis. In: Plant Cell. 1991 ; Vol. 3, No. 2. pp. 149-157.
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