TRANSGENIC ALL FESCUE (FESTUCA-ARUNDINACEA) AND RED FESCUE (F-RUBRA) PLANTS FROM MICROPROJECTILE BOMBARDMENT OF EMBRYOGENIC SUSPENSION CELLS

Transgenic forage- and turf-type tall fescue (Festuca arundinacea Schreb.) and red fescue (F. rubra L.) plants have been obtained by microprojectile bombardment of embryogenic suspension cells using chimeric hygromycin phosphotransferase (hph) gene constructs driven by CaMV 35S promoter or rice actin 1 5' regulatory sequences. Bombardment parameters of embryogenic suspension cultures with the particle inflow gun were partially optimized using transient expression assays of a chimeric beta-glucuronidase (gusA) gene construct. For the recovery of stably transformed clones, hygromycin selection using liquid and solidified media was tested. Initial selection in liquid culture medium allowed for a twofold - compared with continuous plate selection using solid medium - recovery efficiency of transformed hygromycin resistant clones. Plants were regenerated from 35 % and 85 % of the hygromycin resistant calli obtained in tall and red fescue, respectively. The transgenic nature of the regenerated plants was demonstrated by Southern hybridization analysis. Expression of the transgene in transformed mature tall and red fescue plants was confirmed by northern analysis and hygromycin phosphotransferase enzyme assay.