摘要： 在生物或非生物胁迫的条件下,小麦的产量和质量常会受到一定的影响.基因工程技术是小麦品质改良的有效手段,但目前小麦的转化效率还比较低.通过克隆大肠杆菌otsA和otsB 基因,将目的基因和p2300-GFP相连,成功构建p2300-otsAB表达载体.以普通小麦品种为材料,在建立较完善再生体系的基础上,利用根癌农杆菌介导法进行转化,将otsAB导入到受体细胞,进而获得耐盐的转基因小麦新品种.遗传转化条件的优化结果表明,当农杆菌的OD₆₀₀=0.5,侵染成熟胚或愈伤组织的时间为30 min,共培养时间为3 d,小麦的转化效率最高.结果可以为今后通过基因工程技术提高小麦的抗逆能力提供参考.

关键词: 小麦, otsA, otsB, 成熟胚, 愈伤组织, 农杆菌

Abstract: The yield and quality of wheat,a kind of important food crops,are often influenced under the condition of biotic or abiotic stresses.Genetic engineering technology is an effective method for wheat quality improvement,however,the transformation rate of wheat is still relatively low by far.In this experiment, E.coli genomic DNA was as template to clone otsA and otsB genes by the mean of PCR method.Then the target genes were connected with p2300-GFP,which would lead to the construction of p2300-otsAB vector.On the basis of establishing a perfect regeneration system,the common wheat was as the material.The p2300-otsAB expression vector was introduced into the cells of mature embryo and callus of wheat by Agrobacterium-mediated method.And then the new variety of salt-tolerant transgenic wheat was obtained.The genetic transformation results showed that when the concentration of Agrobacterium was OD₆₀₀=0.5,the time of infection of mature embryo or callus was 30 min,and the co-culture time was 3 d,the transformation efficiency of wheat was the highest.The experimental results can provide the reference to improve the stress resistance of plants through genetic engineering in the future.

Key words: Wheat, otsA gene, otsB gene, Mature embryo, Callus, Agrobacterium

中图分类号: 

• S432.4

• Q78