以携带有潮霉素标记基因的RNAi转基因大麦和常规大麦为材料，采用叶片涂抹法进行了潮霉素筛选浓度确定及转基因大麦T2代和回交后代的筛选和PCR检测。结果表明，3 000 mg/L潮霉素连续涂抹3次，5 d后转基因大麦涂抹区域无受害症状，生长受害率显著低于无涂抹处理。潮霉素涂抹检测结果与其PCR检验结果的符合度为96.2%，目的基因筛选准确率可达61.8%。表明所建立的活体幼苗潮霉素叶片涂抹法筛选转基因大麦快速，直观、准确，具有一定可行性。

Using RNAi transgenic barley and conventional barley carrying hygromycin marker gene as experimental materials， the concentration of hygromycin screening by leaf smearing method was determined， and the screening and PCR detection of T2 generation and backcross progenies of transgenic barley were carried out. The results showed that 3 000 mg/L hygromycin was applied for three times continuously. After 5 days， no symptoms were found in the smearing area of GM barley， and the rate of growth damage was significantly lower than that of non-smearing treatment. The coincidence between the results of hygromycin smear test and those of PCR test was 96.2%， and the accuracy of target gene screening was up to 61.8%. Therefore， the hygromycin leaf smear method for screening transgenic barley was rapid， intuitive and accurate， and had certain feasibility.

S512.3

国家自然科学基金(31660391、31460350)。