气相色谱检测芦荟中海藻糖方法的建立

用1-甲基咪唑、氯化羟胺、乙酸酐将海藻糖衍生化，以气相色谱法作为定量分析手段，建立了植物组织中微量海藻糖定量检测方法.用该方法对同一种糖类衍生物进行多次分析，其特征峰保留时间误差在3 s内；能将D-葡萄糖、乳糖、蔗糖和海藻糖进行分离.海藻糖在（3.697～28.661）×10^-9 g检测量范围内其相关系数为0.998 6.利用本实验建立的植物组织中微量海藻糖定量检测的方法，分别对3年生、5年生库拉索芦荟和半年生海藻糖合成酶基因转化芦荟凝胶中海藻糖的含量进行测定，结果表明：5年生库拉索芦荟凝胶中海藻糖含量是3年生的1.59倍，每10 g凝胶匀浆中分别为1.103×10^-5 g和6.905×10^-6 g；每10 g半年生海藻糖合成酶基因转化芦荟凝胶匀浆中海藻糖含量为1.614×10^-5 g，是3年生的2.33倍.证明目的基因成功转入芦荟并已经表达.

Establishment of GC to Detect Trehalose in Aloe

To determine the content of trehalose in transgenic Aloe by gas chromatography (GC),1-Methyl-imidazol,Hydroxylammonium chloride and Acetic anhydride were used to derivative the trehalose.The difference of the retention time among the same carbohydrate derivatives is 3s by this method.And it can separate the glucose,milk sugar,sucrose and trehalose clearly.The results showed a good linear relationship within a range from 3.697×10^-9　g to 28.661×10-9　g,and the liner correlation coefficient of the trehalose is 0.998　6.The content of trehalose in the three-year,five-year aloe (Aloe vera.L) and aloe transformed by ostA were determined by GC.The results show that the trehalose of the five year aloe (Aloe vera.L) is 1.59 times as great as the three-year.The content of trehalose in five-year aloe is 1.103×10^-5　g per 10g,and 6.905×10^-6　g per 10g in three-year aloe.The content of trehalose is 1.614×10^-5　g per 10g in aloe transformed by ostA which is 2.33 times as great as the three-year aloe (Aloe vera.L).The results prove that the ostA gene from E.coli is successfully transformed to aloe and expressed.