A routine system for generation of transgenic rice (Oryza sativa L.) plants by microprojectile bombardment of embryogenic cell clusters.

摘要

A routine transformation system, based on microprojectile bombardment of embryogenic cell clusters, was developed for the recovery of fertile transgenic rice (Oryza sativa) plants. Embryogenic cell clusters were established within 3-4 months from calluses derived from mature seeds of 9 diverse cultivars. Cell clusters were bombarded with plasmids carrying genes encoding hygromycin phosphotransferase (hph, conferring resistance to hygromycin B) and beta-glucuronidase (gusA [uidA]), both driven by the cauliflower mosaic virus CaMV 35S promoter. Resistant calluses were readily recovered from hygromycin-containing medium. On average, 4.6 resistant calluses were recovered from each bombarded plate. Transgenic plants were routinely regenerated from suchresistant calluses (a regeneration frequency of 69%). Data from Southern blot analysis of T0 and T1 plants confirmed that the transgene was stably integrated into the host genome and inherited in a Mendelian fashion.