Agrobacterium-mediated transformation of Orychophragmus Violaceus Cotyledon and Regeneration of Transgenic Plants
根癌土壤杆菌介导转化诸葛菜子叶获得转基因植株

Excised cotyledons of Orychophragmus violaceus, were used as explants for tissue culture. They were cultured on MS medium supplemented with BA 3 mg/L and NAA 0. 2mg/L. When the regenerated buds were 2cm long, they were excised and transferred onto 1/2MS medium with IBA 0. 03mg/L, then the whole planticts were regenerated . The frequency of plant regenration was 100%. Then we began to study the genetic transformation of O. violaceus, After 2-3 days of cocultivation with Agrobacteri-um tumefaciens strain A208se (pTiT37 pROA93), the cotyledons were transferred onto the selection medium containing 25mg/L Km and 250mg/L Ap. After shoots emerged, they were excised and transferred onto the root medium containing 25mg/L Km and 100mg/L Cef, the roots were formed within 4-5 weeks. The whole plants were transplanted into pots and grew well. The frequency of plant regeneration was about 51%. The regenerated plants showed high enzymatic activities of B-glucuronidase and neomycine phosphotransferase I . Southern blot analysis confirmed that NPT I gene had been stably integrated into the chomosomal genome of O. violaceus. The transformation frequency was 5. 6%. In this paper, the first transgenic plant of O. violaceus is being reported.