Effects of heterologous expression of Populus euphratica brassinosteroids biosynthetic enzyme genes CPD (PeCPD) and DWF4 (PeDWF4) on tissue dedifferentiation and growth of Arabidopsis thaliana seedlings

To understand the functions of Populus euphratica CPD (PeCPD) and DWF4 (PeDWF4), the responses to exogenous phytohormone in Arabidopsis-PeCPD and -PeDWF4 transgenic lines (PeCPD-TL and PeDWF4-TL) and corresponding wild type (WT) seedlings were investigated. Results showed that all of PeCPD-TL, PeDWF4-TL and WT seedlings cultured on the mediums containing 2,4-dichlorophenoxyacetic acid (2,4-D) + 6-benzylaminopurin (6-BA) or 2,4-D + 6-BA + brassinolide (BL) could be dedifferentiated to callus with 100% frequency, but they displayed strong differences in callus formation sites, callus growth rates (CGRs) and tissue dedifferentiation degrees. On the medium containing 2,4-D alone, the seedlings of all the plants could formed callus, but callus formation times (CFTs) were delayed, and callus formation rates (CFRs) were differentially decreased. After adding lower concentrations of BL, their CFRs were all restored to 100%, but tissue dedifferentiation degrees were obviously lower than these on the mediums with 2,4-D + 6-BA or 2,4-D + 6-BA + BL. On the mediums containing 6-BA or 6-BA + BL, the seedlings of all the plants failed to produce callus. Semi-quantitative RT-PCR analysis also showed that the transcription levels of PeCPD, PeDWF4, AtCPD and AtDWF4 in PeCPD-TL, PeDWF4-TL and WT were evidently different. These results suggest that PeCPD and PeDWF4 play similar but not exactly the same roles in the regulation of callus morphogenesis of Arabidopsis seedlings, and that BL can partially replace the role of cytokinin to induce callus formation through interacting with auxin.