Abstract
Transgenic forage-type Italian ryegrass (Lolium multiflorum Lam.) plants have been obtained by microprojectile bombardment of embryogenic suspension cells using a chimeric hygromycin phosphotransferase (hph) gene construct driven by riceActl 5′ regulatory sequences. Parameters for the bombardment of embryogenic suspension cultures with the particle inflow gun were partially optimized using transient expression assays of a chimericβ-glucuronidase (gusA) gene driven by the maizeUbi1 promoter. Stably transformed clones were recovered with a selection scheme using hygromycin in liquid medium followed by a plate selection. Plants were regenerated from 33% of the hygromycin-resistant calli. The transgenic nature of the regenerated plants was demonstrated by Southern hybridization analysis. Expression of the transgene in transformed adult Italian ryegrass plants was confirmed by northern analysis and a hygromycin phosphotransferase enzyme assay.
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Abbreviations
- 2,4-D 2,4:
-
Dichlorophenoxyacetic acid
- GUS :
-
Glucuronidase
- Hm :
-
Hygromycin
- HPH :
-
Hygromycin phosphotransferase
- MS :
-
medium Murashige and Skoog medium
- PCR :
-
Polymerase chain reaction
- X-Gluc :
-
5-Bromo-4-chloro-β-indolyl-β-D-glucuronic acid
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Communicated by N. Amrhein
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Ye, X., Wang, Z.Y., Wu, X. et al. Transgenic Italian ryegrass (Lolium multiflorum) plants from microprojectile bombardment of embryogenic suspension cells. Plant Cell Reports 16, 379–384 (1997). https://doi.org/10.1007/BF01146777
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DOI: https://doi.org/10.1007/BF01146777