Ri质粒介导的DNA转移及诱发甘草毛根再生植株

本文报道了通过三亲交配将PB1121双元中间载体从大肠杆菌C600转移到发根农杆菌R1000(PRiA4b)中,在含Km、Sm的MinA选择平板上获得了转移子,上述转移子菌液用注射法感染甘草胚轴、茎等外植体,在无激素含1mg/mlcb,0.1mg/mlkm的MS培养基上培养,两周后诱发出毛根,切下毛根在同上培养基上6周长出不定芽,继续培养长成再生植株,经检测转化株毛根中存在甘露碱和GUS基因产物,具有Km抗性和激素自主性生长特性,证明Ri质粒不仅可介导GUS基因转移还可诱发毛根再生植株,提供了简化、高效的植物遗传转化系统。

DNA Transference by Introduction of Ri Plasmid and regeneration plant from Glycyrhiza Hairy Roots

The present paper reports the transFerence of bi-nary vector pBI121 from E. coli to Agrobac-terium rhizogenes R1000pRIA4b by triparental mating, transferants were obtained on selective plates of MinA medium containing Km and Sm. Embryo axis and stem explants of Glycyrhiza were infected by these transfants with injection,and cultured on hormone-free MS medium with 1g/ml Cb and 0.1g/ml Km,hairy roots formed after two weeks,The hairy roots were cut down and cultured on the same medium for six weeks,adventitious bud formed,continued to be cultured and regenerated plants. Manopine and the products of Gus gene were detected in hairy roots. The transformed plants showed Km resistance and the character of self sufficient hormone. This indicates Ri plasmid not only introduct the transference of Gus gene but also induce hairy roots to regenerate plants. It supplies a simple and high-effective system for genetic transformation of plants.