PLANT REGENERATION AND GENETIC TRANSFORMATION AND DEVELOPMENT OF LATE FLOWERING CULTIVARS OF RADISH(RAPHANUS SATIVUS L.)

摘要

PURPOSE: Provided are plant regeneration and genetic transformation and development of late flowering cultivars of Radish(Raphanus sativus L.) by Agrobacterium tumefaciens. CONSTITUTION: The transformation method is characteristically comprises the steps of: inserting Radish-CONSTANS(CO) gene (RsCO) into Agrobacterium tumefaciens by using electroporation of a vector pCAMBIA 3301 containing the RsCO gene and bar gene as an antibiotic resistant gene to prepare Agrobacterium tumefaciens AGL1; co-cultivating Agrobacterium tumefaciens AGL1 and radish tissue to induce a callus and a shoot for 1 to 3 hours; drying the co-cultivated medium using filter paper for 1 to 3 hours; and redifferentiating the callus and shoot and transplanting them to a pot, in which the liquid medium containing BAP and NAA is used for pretreatment of the Agrobacterium tumefaciens AGL1 and radish before their co-cultivation; and the Agrobacterium tumefaciens AGL1 is pretreated by centrifuging its cultivated medium to recover the pellet, and diluting the pellet in the pretreatment liquid medium before co-cultivation with the radish tissue.