Bialaphos-resistant cells of dallisgrass (Paspalum dilatatum Poir.) through particle bombardment with a simple self-built inflow gun

摘要

A simple and inexpensive, self-built particle acceleration apparatus is described, and the special features of the device are emphasized. We have employed this easy-to-use gene gun for efficient direct delivery of DNA to cultured cells of an important monocotyledonous forage crop, dallisgrass (Passpalum dilatatum Poir.). High levels of transient expression of the beta-glucuronidase gene were obtained following bombardment of suspension cells. Furthermore, stable transformed cells of this grass havebeen obtained after intrusion of the bar (bialaphos resistance) gene at optimized delivery conditions. Fifteen calli were selected on solidified medium. After transfer to liquid selection medium, one of these calli provided a stable suspension culture tolerant at 1 mg~(-1) bialaphos. Integration of the transgene in the suspended cells was confirmed by PCR amplification analysis.