Co-transformed, diploid Lolium perenne (perennial ryegrass), Lolium multiflorum (Italian ryegrass) and Lolium temulentum (Darnel) plants produced by microprojectile bombardment

Susan Dalton, Andy J. E. Bettany, Emma Timms-Taravella, Phillip Morris

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Abstract

A total of 37 plants (30 Lolium multiflorum Lam., 6 L. perenne L., 1 L. temulentum L.) were regenerated from cell suspension colonies bombarded with plasmid DNAs encoding a hygromycin resistance gene (HYG) expressed under a CaMV35S promoter and a β-glucuronidase (GUS) gene expressed under a truncated rice actin1 promoter and first intron, or a maize ubiquitin promoter and first intron. Resistant plants were regenerated under hygromycin selection and transferred to soil. PCR analysis showed that the co-transformation frequency of the GUS gene varied from 33% to 78% of transformants, while histochemical staining of leaf tissue from soil-grown plants showed that the co-expression frequency varied from 37% to 50%. The transgenic nature of the plants was demonstrated by Southern hybridisation analysis, which also showed that the non-selected (GUS) gene was generally present at a higher copy number than the selected (HYG) gene.
Original languageEnglish
Pages (from-to)721-726
Number of pages6
JournalPlant Cell Reports
Volume18
Issue number9
DOIs
Publication statusPublished - May 1999

Keywords

  • Co-expression
  • Co-transformation
  • Lolium
  • Microprojectile bombardment
  • Transgenic plants

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