河南农业大学学报2017，Vol.51Issue(6)：781-785,5.
农杆菌介导小麦遗传转化方法的优化与应用
Improvement and application of agrobacterium mediated genetic transformation of wheat
摘要
Abstract
In order to optimize the method of wheat agrobacterium mediated genetic transformation,the embryonic calli derived from the immature embryos of a wheat cultivar"Zhengmai 9023"were used as materials,to study the effects of gamma irradiation on agrobacterium-mediated genetic transformation. The results indicated that 4 Gy is the optimal dose for agrobacterium-mediated genetic transformation of Zhengmai 9023,using GUS transient expression rates as the indicator. Based on the survival rate and GUS transient expression rate,further observation found that 24 hours reculture after gamma irradiation is the most appropriate for agrobacterium infection. Another negative pressure treatment results showed that pulling 10 times in enclosed injector was the optimal manner for transformation. Bar gene PCR as-say results of positive regeneration plantlets showed that transformation efficiency was enhanced from 2.2% to 3.5% by using the improved method (4 Gy irradiation+24 h reculture+10 times negative pressure treatment) described above. The optimied agrobacterium-mediated transformation method can be used in the genetic transformation of wheat.关键词
小麦/再生体系/农杆菌/遗传转化/辐照/负压处理/GUS瞬时表达Key words
wheat/regeneration system/agrobacterium/genetic transformation/gamma irradiation/negative pressure treatment/GUS transient expression分类
农业科技引用本文复制引用
张磊,李国领,张建周,张德奇..农杆菌介导小麦遗传转化方法的优化与应用[J].河南农业大学学报,2017,51(6):781-785,5.基金项目
国家自然科学基金面上项目(31371707) （31371707）
