Agrobacterium-mediated sorghum transformation frequency has been enhanced significantly via medium optimization using immature embryos from sorghum variety TX430 as the target tissue. The new transformation protocol includes the addition of elevated copper sulfate and 6-benzylaminopurine in the resting and selection media. Using Agrobacterium strain LBA4404, the transformation frequency reached over 10% using either of two different selection marker genes, moPAT or PMI, and any of three different vectors in large-scale transformation experiments. With Agrobacterium strain AGL1, the transformation frequencies were as high as 33%. Using quantitative PCR analyses of 1,182 T0 transgenic plants representing 675 independent transgenic events, data was collected for T-DNA copy number, intact or truncated T-DNA integration, and vector backbone integration into the sorghum genome. A comparison of the transformation frequencies and molecular data characterizing T-DNA integration patterns in the transgenic plants derived from LBA4404 versus AGL1 transformation revealed that twice as many transgenic high-quality events were generated when AGL1 was used compared to LBA4404. This is the first report providing molecular data for T-DNA integration patterns in a large number of independent transgenic plants in sorghum.

中文翻译：

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优化的农杆菌介导的高粱转化方案和转基因高粱植物的分子数据。

通过使用来自高粱品种TX430的未成熟胚作为目标组织的培养基优化，农杆菌介导的高粱转化频率已显着提高。新的转化方案包括在静置和选择培养基中添加硫酸铜和6-苄基氨基嘌呤。使用农杆菌菌株LBA4404，在大规模转化实验中，使用两种不同的选择标记基因moPAT或PMI以及三种不同的载体中的任一种，转化频率均达到了10％以上。用农杆菌菌株AGL1，转化频率高达33％。通过对代表675个独立转基因事件的1,182个T0转基因植物进行定量PCR分析，收集了有关T-DNA拷贝数，完整或截短的T-DNA整合的数据，并将载体骨架整合到高粱基因组中。比较源自LBA4404与AGL1转化的转基因植物中转化频率和表征T-DNA整合模式的分子数据，发现与LBA4404相比，使用AGL1产生的转基因高质量事件多两倍。这是第一份报告，提供了高粱中大量独立转基因植物中T-DNA整合模式的分子数据。