农杆菌介导法将海藻糖合成酶基因转入芦荟的研究

Agrobacterium-mediated Transformation of Aloe with Trehalose Synthase Gene (otsA)

采用农杆菌介导法将海藻糖合成酶基因otsA导入芦荟,共培养后的芦荟外植体在含10-15 mg·L^-1G418筛选剂的MS培养基上经多次筛选和分化,获得了一定数量的抗性再生芽,对移栽成活的抗性再生植株进行PCR检测,表明otsA基因已经导入芦荟基因组中,转化率约为0.77%。Southern检测表明,目的基因在60%的阳性植株基因组中表现为单拷贝整合,进一步证明otsA基因已经整合到芦荟基因组中。气相色谱法测定转基因植株的海藻糖含量表明,转基因植株中的海藻糖含量为未转化植株的2.934倍,证明otsA基因在转基因芦荟植株中已得到表达。

Aloe has been deeply researched recently because of its attracting function on medical treatment, hairdressing, health protection and edibility. Trehalose can protect organism and biomolecule, and has been applied widely in foods, cosmetics and molecule biology researches. In this experiment, aloe stems were used as explants for otsA gene transformation mediated by Agrobacterium. PCR and Southern blotting analyses demonstrated that otsA gene was successfully transferred into aloe genome with a transformation efficiency of 0.77 %, and the single copy insert of the alien gene was found in about 60 % positive aloe plants. Gas Chromatogram analysis showed that the content of trehalose in transgenic aloe plants was 2. 934 times of that in untransformed aloe plants, revealing that otsA gene had been expressed in the transgenic aloe plants.