Characteristics of in vitro plant tissue culture of four widely applied elite Indica hybrid rice maintainer lines (Zhong9B, Jin23B,Ⅱ–32B,Chuanxiang29B)and two widely applied elite restorer lines(9311 and Minghui63) were investigated systematically in this study. The Indica rice cultivar-Chuanxiang29B was identified with excellent tissue culture response. It is shown that the callus induction rate of Chuanxiang29B from mature embryos was 96% and the plantlet regeneration rate of it was 82%, indicating higher rates of callus induction, proliferation, subculture tolerance and plantlet regeneration of Chuanxiang29B than most of other indica rice cultivars. An optimized and stable genetic transformation system mediated by Agrobacterium tumefacien for Chuanxiang29B was then established, in which the transformation efficiency of resistant plants reached up to 19.6% with the following conditions: the period of the calli inoculation with Agrobacterium tumefacien EHA105 (OD600 nm=0.1) in AAM with acetosyringone (200 μmol/L) for 30 min and cocultivation at 19 ℃ in dark for 72 h.