Agrobacterium Rhizogenes-Mediated High Frequency Hairy Root Induction and Genetic Transformation in Tea Plant

An efficient protocol for the establishment of transgenic high hariy root tea plant(Camellia sinensis) system induction infected by Agrobacterium rhizogenes is reported using two wild type strains of A.rhizogenes,three different co-cultivation media and six different explants.The highest induction frequency was obtained from stem segments of aseptic seedlings infected by A.rhizogenes and co-cultivated on YMB solid media with 100 mmol/L AS for 2 d.The hairy roots grew rapidly on LG0 medium without phytohormone and produced a plenty of branches and root hairs.PCR confirmed that rolA,rolB and rolC have inserted into the genomic DNA of hairy root induced by wild type A.rhizogenes.Hairy roots were also induced from explants of tea plant infected by A.rhizogenes 15834 carrying pCAMBIA2301 binary vector harboring Bt gene,transgenic root were confirmed by PCR and GUS histochemical staining that the Bt exogenous gene was inserted and expressed.