B, D, E transgenic clones with rolABC genes and wildtype plants were used as materials to construct binary standard curves of rol genes and β-actin gene with SYBR Green I fluorescent dye. After normalizing the influence of amplification efficiency with different primers in qRT-PCR, comparative Delta-delta Ct method was applied to develop suitable qRT-PCR method for analysising the expression of rolA, rolB, rolC genes in 5 tested tissues of B, D, E clones. The primary result showed that rolC expressed highest among 3 rol genes in tender stem, tender leaf, functional leaf and bark, rolA gene’s expression in the middle level and mainly in tender stem, and rolB expressed lowest and mainly in root.