Transgenic tea [ Camellia sinensis (L.) O. Kuntze cv. Kangra Jat] plants obtained by Agrobacterium -mediated transformation of somatic embryos

摘要: A protocol for the production of transgenic tea [Camellia sinensis (L.) O. Kuntze cv. Kangra Jat] was developed via Agrobacterium-mediated genetic transformation somatic embryos. Two disarmed Agrobacterium tumefaciens strains, EHA 105 and LBA 4404, both carrying binary plasmid p35SGUSINT with nptII gene gus-intron were evaluated as vector systems. number parameters tested respect to maximizing efficiency. While pre-culture, wounding acetosyringone treatment inhibitory, bacterial growth phase (optical density; OD600 = 0.6), cell density (109/ml), co-cultivation period (5 days) pH medium (5.6) had positive effects on transformation. Following co-cultivation, globular embryos placed multiplication stressed kanamycin (50 µg/ml). Further selection occurred in maturation germination at an elevated level (75 An average 40% transient expression evident based GUS histochemical assay. Kanamycin-resistant, GUS-positive germinated, resulting microshoots multiplied vitro. Integration transgenes into nuclear genome confirmed by PCR analysis using nptII- gus-specific primers Southern hybridization nptII-specific probe. The shoots micrografted onto seed-grown rootstocks Jat eventually hardened a walk-in polyhouse. This is first report tea.