Development of a simple and efficient method for transformation of buckwheat plants (Fagopyrum esculentum) using Agrobacterium tumefaciens

Apical meristems of seedlings of buckwheat (Fagopyrum esculentum var. Shina no No. 1) were pricked with a needle and inoculated with Agrobacterium tume faciens (LBA4404, pB1121). The inoculated seedlings were grown to maturatio n and allowed to pollinate randomly to set the seeds (T1 plants). The trans formation efficiency of the T1 plants was estimated by germination in the p resence of geneticin (20 mu g/ml) and by detection of beta-glucuronidase (G US) gene with PCR, indicating that 36% and 70% of the T1 plants were transf ormed, respectively. Four plants taking on a mutated morphology were select ed from T1 plants which were transformed with the method using A. tumefacie ns harboring a modified pBI121 for plasmid rescue. Southern blot analysis o f plasmids rescued from the 4 T1 plants demonstrated that each plasmid cont ained a different flanking DNA of the buckwheat genome, an evidence that T- DNA was integrated in different sites of the genomic DNA among the 4 T1 pla nts.