An Efficient Tissue Culture System And Microprojectile-mediated Transformation Of Kentucky Bluegrass(Poa Pratensis L.)

Six breeding lines of DP-37-61, DP192-10, 82-37, DP-LF-300, 2W42-117, PRO-4287 were cultured using meristem and mature seed as explants to establish the tissue culture system of Kentucky bluegrass (Poapratensis L.) . The embryogenic calli of breeding line DP-37-61 and PRO-4287 which had higher embryogenic-calli induction rate and regeneration rate were co-transformed with plasmid pAct1IHPT-4 containing hpt gene and plasmid M14 containing transcription factor DREB. Bombarded calli were selected on the selection medium supplemented with 100 mg/L hygromycin. Eighty-four resistant calli of breeding line DP-37-61 survived after 12-20 weeks selection on the selected medium, of which 59 resistant calli regenerated successfully with the very high transformation rate of 13%. Eight resistant calli of breeding line PRO-4287 survived after 10-16 weeks selection on the selected medium, of which 6 resistant calli regenerated successfully with the high transformation rate of 3%. The independent nature of transformed plants was verified by PCR analysis. An efficient and reproducible transformation system for Kentucky bluegrass (Poa pratensis L.) was set out with a microprojectile bombardment delivery system.