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Zhou J, Wei Z, Xu Z, Liu S, Luo P: Agrobacterium tumefaciens mediated transformation of Orychophragmus violaceus cotyledon and regeneration of transgenic plants. Chin J Biotechnol. 1996;12(1):39-45.
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. |
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