转bar基因小麦和非转基因小麦抗除草剂鉴定方法比较

方便、快捷、准确地对转基因小麦中的bar基因进行检测,对于筛选纯合稳定转基因植株、获得无筛选标记转基因植株、鉴定常规小麦品种和商品小麦中的bar基因成分等具有一定价值。本试验对叶片涂抹、植株喷洒、培养基添加除草剂3种方法鉴定转bar基因小麦植株的效果进行了比较,表明3种方法都能很好鉴定转基因小麦中的bar基因,叶片涂抹200mg/LLiberty鉴别的准确性高于PCR检测,植株喷洒Basta的适宜浓度为100mg/L,喷洒Liberty的适宜浓度为150mg/L,培养基添加Bialaphos的适宜浓度为5～8mg/L。叶片涂抹和植株喷洒除草剂方法受环境条件影响较大,区别转基因植株和非转基因植株的标准不够明确。相比之下,成熟胚离体培养除草剂筛选不受外界环境条件的影响,具有鉴定效果直观明了、操作简单、试验周期短等优点,在检测小麦转入或飘入的bar基因方面具有潜在应用前景。

Comparison of Different Methods for Bar Gene Detection in Trangenic or Non-transgenic Wheat Using Herbcide as Tool

It is very useful to detect bar gene in transgenic and non-transgenic wheat by a method with convenience, swiftness, and reliability for the screening of stable transgenic lines, the development of marker-free transgenic plants, inspection of commercial wheat varieties and products. Three identification methods of leaf painting, plant spraying, and mature embryos in vitro culture with the herbicides corresponding to bar gene were compared and evaluated in this study. Results indicated that transgenic or non-transgenic wheat plants and seeds with or without bar gene could be distinguished well by the three methods mentioned above. Application of leaf painting with 0.1% Liberty for the confirmation of the herbicide transgenic plants was better than PCR analysis in accuracy. The optimal concentrations for plant spraying were 100ppm for Basta and 150ppm for Liberty, and the available amount of bialaphos in medium for the intro mature embryos is 5-8 mg L-1. However, the first two methods were influenced in some extent by environmental conditions such as temperature, light, and nutrition in practice, which often led to an unclear observation in symptom for detecting the bar plants. In comparison, mature embryo in vitro culture on the medium with herbicide was less dependant on environment, and showed some advantages including clear observation, easy manipulation, and short time expense. Therefore, mature embryo in vitro culture identification was strongly suggested to be used in the detection and inspection of bar gene in transgenic and commercial wheat.