Agrobacterium Tumefaciens Mediated Transformation In Lolium Perenne L.

Perennial ryegrass (Lolium perenne L.) (2n=2x=14), a graminaceous crop, is one of the most widely distributed grass in the temperate regions for forage and turf. Because of the highly self-incompatible, the conventional plant breeding is difficult and selection schemes are complex. Biotechnological approaches, especially genetic transformation for the direct introduction of agronomically useful genes, hold great promise.A method Agrobacterium tumefaciens transformation for perennial ryegrass was developed using the calli derived from mature embryos, and the transgenic plants were successfully obtained.In delivering report genes, nine ryegrass varieties were employed and their calli were inoculated with a disarmed Agrobacterium tumefaciens strain EHA105 harboring binary vector pCAMBIA2301 or p132. Transgenic plants were obtained from 4 varieties of perennial ryegrass ( Gold Medalist, Top Gun, Tove and Limes) by selection with paromomycin.The transgenic seedlings of Gold Medalist, Top Gun, and Tove were produced by using the plasmid pCAMBIA2301. The transgenic calli and plants were identified by PCR and GUS histochemical assay, which showed that the foreign genes were presented in host genome and expressed. Overall, 9 transgenic seedlings were obtained from Gold Medalist in one transformation event; 4 and 12 ones of Top Gun were gained from 2 events separatively; and 17, 18 and 15 transgenic seedlings were produced from Tove in 3 independent events, respectively. Dig-labelled Southern-bloting analysis for the 9 transgenic seedlings of Gold...