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为建立小麦遗传转化中高效的潮霉素筛选体系，以小麦品种郑麦9023的幼胚愈伤组织为材料，探讨了潮霉素浓度、处理时间、恢复培养与否对愈伤组织分化和再生的影响，并对基因枪转化的郑麦9023、邯6172、龙麦30、龙麦31的愈伤组织进行了筛选。结果表明，对小麦幼胚愈伤组织基因枪遗传转化后的筛选以50 mg·L^-1潮霉素处理30 d为宜；恢复培养不能提高筛选阶段愈伤组织的存活率和分化率，但能提高转化后期的成苗率。经过筛选发现郑麦9023抗性愈伤和T₀代抗性植株根系及花器官中均有GUS基因的表达；4个小麦品种抗性植株中潮霉素磷酸转移酶基因(hpt)和候选基因PCR阳性率分别为78.95%和73.68%以上；龙麦30、龙麦31的T₁代植株的候选基因PCR阳性率分别在33.33%和73.33%以上，T₂代阳性率分别在71.43%和75.00%以上。

英文摘要:

Effective selection system is essential to high efficiency genetic transformation. In this study,the effects of different hygromycin concentrations,screening time and convalescence or not on differentiation and regeneration of immature embryo callus of Zhengmai 9023 were investigated，and callus of Zhengmai 9023，Han 6172，Longmai 30，Longmai 31 transformed by particle bombardment were selected using hygromycin. The results showed that 50 mg·L^-1 of hygromycin applied for 30 d was suitable for screening after transformation; convalescence could not improve the survival rate and differentiation rate of callus.However,it could promote plantlet establishment for transformational callus. GUS expressions were found in Zhengmai 9023 resistant tissues,roots and flower of T₀ generation. In addition,the expressions of hygromycin phosphotransferase gene (hpt) and candidate gene were confirmed by PCR in the T₀ resistant plants of four cultivars,positive ratio were up to 78.95% and 73.68%, respectively. For Longmai 30 and Longmai 31,the positive ratio of candidate gene in T₁ generation were higher than 33.33%and 73.33%,respectively; T₂ generation were over 71.43% and 75.00%,respectively. Results above proved that this established hygromycin selection system for immature embryo callus of wheat was efficient and feasible.

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