铁皮石斛原球茎对潮霉素及卡那霉素的敏感性研究

Effect Sensitivity of Hygromycin and Kanamycin on Tissue Culture Protocorms of Dendrobium officinale

为了探索农杆菌介导的遗传转化中铁皮石斛对抗生素的耐性,本试验以铁皮石斛原球茎为原材料,在培养基中分别添加不同浓度潮霉素和卡那霉素进行培养,筛选出最适合铁皮石斛遗传转化的抗生素种类和浓度。研究表明:潮霉素浓度为40 mg/L时为临界浓度,接种一周后,植株出现黄化现象,2周后,出现死亡植株,培养17周后,铁皮石斛完全死亡。潮霉素浓度越大,铁皮石斛褐化和黄化速率增加,死亡速率越快,浓度低于40 mg/L,铁皮石斛则拥有一定的存活率,为15%左右。当卡那霉素浓度为600 mg/L,铁皮石斛的褐化和黄化现象出现在培养后第3周,培养17周后,浓度为200 mg/L、400 mg/L、600 mg/L卡那霉素,铁皮石斛原球茎死亡率依次为72%、68%、78%,之后仍保持一定的存活量。实验结果表明,铁皮石斛对潮霉素敏感性远大于卡那霉素,由此说明,铁皮石斛遗传转化中应选用潮霉素作为抗生素,而且使用浓度应在40 mg/L以上,卡那霉素不能用于铁皮石斛遗传转化。

For exploring the resistance of Dendrobium officinale to antibiotics in the process of agrobacte-rium-mediated genetic transformation, in this experiment, Protocorms of D. officinale, used as the raw material, were cultured on the medium which Hygroscopious and Kanamycin with different concentrations were added respectively to the basic medium for selecting the best antibiotics and its concentrations in genetic transformation of D. officinale. The results showed that Hygroscopious concentrations of 400 mg/L are as critical concentration in which plant became yellow after inoculated one week;a number of them died after two weeks and died totally after 17 weeks. The higher the concentration of Hygromycin is, the higher the rate of browning and yellowing is, and finally it can speed the higher mortality of D. officinale. When the Hygroscopious concentrations are below 400 mg/L, there is still small amount survival rate, 15%. When Kanamycin’s concentrations were 600 mg/L, the yellow and brown phenomenon presented after inoculated three weeks. When the concentration of Kygromycin was 200 mg/L, 400 mg/L, and 600 mg/L, the mortality was 72%, 68% and 78% after 17 weeks, respectively. After that, no death phenomenon appeared again. The experimental results show that D. officinale is more sensitive to hygromycin than to kanamycin. It is noted that Hygroscopious is subjected to be selected as antibiotics and its concentration should be above 40 mg/L, and Kanamycins cannot be used as antibiotics in genetic transformation of D. officinale.