title: Agrobacterium tumefaciens mediated transformation of Orychophragmus violaceus cotyledon and regeneration of transgenic plants.
abstract: Excised cotyledons of Orychophragmus violaceus were used as explants for tissue culture. They were cultured on the MS medium supplemented with BA (3 mg/L) and NAA (0.2 mg/L). When the regenerated buds were 2 cm long, they were excised and transferred onto 1/2 MS medium with IBA (0.03 mg/L), then the whole plants were regenerated. The frequency of plant regeneration was 100%. Subsequently, the genetic transformation of O. violaceus was studied. After 2-3 days of cocultivation with Agrobacterium tumefaciens strain A208se (pTiT37, pROA93), the cotyledons were transferred onto the selection medium containing 25 mg/L Km and 250 mg/L Ap. After shoots emerged, they were excised and transferred onto the rooting medium containing 25 mg/L Km and 100 mg/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 beta-glucuronidase and neomycine phosphotransferase II. Southern blot analysis confirmed that NPTII gene had been stably integrated into the chromosomal genome of O. violaceus. The transformation frequency was 5.6%. The first transgenic plant of O. violaceus is being reported.
authors.affiliation: National Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology, Chinese Academy of Sciences, Shangha, China.
authors.forename: J
authors.initials: J
authors.lastname: Zhou
authors.forename: Z
authors.initials: Z
authors.lastname: Wei
authors.forename: Z
authors.initials: Z
authors.lastname: Xu
authors.forename: S
authors.initials: S
authors.lastname: Liu
authors.forename: P
authors.initials: P
authors.lastname: Luo
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D018548:Cotyledon / Q000502:physiology; D046508:Culture Techniques / Q000379:methods; D005966:Glucuronidase / Q000235:genetics; D019868:Kanamycin Kinase; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D010945:Plants, Edible / Q000235:genetics*; D030821:Plants, Genetically Modified; D012038:Regeneration; D014170:Transformation, Genetic
chemical_list: D017853:Phosphotransferases (Alcohol Group Acceptor); D019868:Kanamycin Kinase; D005966:Glucuronidase