Isolation of transgenic progeny of maize by embryo rescue under selective conditions

Kris Weymann, Katharina Urban, Daniel M. Ellis, Roseann Novitzky, Erik Dunder, Susan Jayne, Gary Pace

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Fertile transgenic maize plants (T 0) and progeny (T 1) were obtained using microprojectile bombardment and callus selection on hygromycin B. To quickly identify progeny expressing the transgene, embryos from T3 generation kernels were excised 20 days after pollination and exposed to different concentrations of hygromycin B. Surviving and non-surviving embryos were assayed for the presence of the hygromycin phosphotransferase (aphIV) gene using polymerase chain reaction. Embryos that germinated and survived on 25, 50, or 100 mg/liter hygromycin possessed the aphIV gene. Embryos that did not germinate lacked the gene. Progeny surviving selection were transferred to the greenhouse and tested for expression of the gene using a leaf disc assay. The results demonstrated that the gene construct was expressed in both embryo and leaf tissue and that selection during germination successfully eliminated progeny lacking the gene of interest. This method is also useful for rapid-cycling of maize generations.

Original languageEnglish (US)
Pages (from-to)33-37
Number of pages5
JournalIn Vitro Cellular & Developmental Biology - Plant
Volume29
Issue number1
DOIs
StatePublished - Jan 1 1993

Keywords

  • embryo rescue
  • hygromycin
  • selection
  • transgenic maize

ASJC Scopus subject areas

  • Biotechnology
  • Plant Science

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