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利用RACE方法,从菊花(Chrysanthemum morifolium)‘神马’中分离得到细胞分裂素合成异戊烯基转移酶基因的全长cDNA序列,命名为DgIPT3,基因登录号为JQ711176。序列分析结果表明,DgIPT3的cDNA全长为1171bp,开放阅读框ORF编码331个氨基酸,具有IPT家族典型的ATP/GTP结合位点MGATGTGKS。系统进化分析显示,DgIPT3与毛果杨(Populus trichocarpa)的PtXP02321061亲缘关系最近。qRT-PCR分析表明,DgIPT3在菊花根、茎、叶中均有表达,其表达量为叶>茎>根。瞬时转化拟南芥(Arabidopsis thaliana)原生质体表明DgIPT3蛋白定位于细胞质中。过表达DgIPT3异源转化野生型拟南芥,莲座侧枝明显增多,表明DgIPT3可能是参与菊花侧枝形成的关键基因。
The full-length cDNA sequence of cytokinin-producing prenyltransferase gene was isolated from Chrysanthemum morifolium ’Shenma’ by RACE method and named as DgIPT3 with accession number JQ711176. Sequence analysis showed that the full-length cDNA of DgIPT3 was 1171bp, and the open reading frame ORF encoded 331 amino acids, with a typical ATP / GTP binding site MGATGTGKS of IPT family. Phylogenetic analysis showed that DgIPT3 was the closest to PtXP02321061 in Populus trichocarpa. qRT-PCR analysis showed that DgIPT3 was expressed in roots, stems and leaves of chrysanthemum, and its expression level was leaf> stem> root. Transient transformation of Arabidopsis thaliana protoplasts indicated that the DgIPT3 protein is localized in the cytoplasm. Overexpression of DgIPT3 heterologously transformed wild-type Arabidopsis, rosette axons significantly increased, indicating that DgIPT3 may be involved in chrysanthemum collateral formation of key genes.