We have established an efficient particle-bombardment transformation protocol for the diploid non-apomictic genotype of the warm season forage crop Paspalum notatum (bahiagrass). A vector containing a herbicide resistance gene (bar) together with the GUS reporter gene was used in transformation experiments. The bar gene confers resistance to the herbicide bialaphos. An improved culture system, highly regenerative callus, dense in compact polyembryogenic clusters, was produced on medium with a high CuSO4 content at elevated temperature. Target tissue (360 calli) produced under these conditions yielded 52 rooted plants on herbicide-containing medium, and 22 of these plants were PCR-positive. DNA gel blot analysis revealed a copy number of 1-5 for the GUS gene in different independent transformants. There was no correlation between copy number and GUS activity. While conventional cultures yielded exclusively albino plants on herbicide-containing medium, improved culture conditions for the target tissue resulted in the recovery of 100% green transgenic plants. All green herbicide-resistant regenerants were morphological normal and fertile.

中文翻译：

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通过粒子流入枪介导的基因转移到二倍体巴伊亚草（Paspalum notatum）的绿色抗除草剂植物

我们已经为暖季饲料作物雀稗 (bahiagrass) 的二倍体非无融合生殖基因型建立了有效的粒子轰击转化方案。在转化实验中使用含有除草剂抗性基因 (bar) 和 GUS 报告基因的载体。bar 基因赋予对除草剂双丙氨磷的抗性。在升高的温度下，在高 CuSO4 含量的培养基上产生了一种改进的培养系统，即高度再生的愈伤组织，密集在致密的多胚簇中。在这些条件下产生的靶组织（360 个愈伤组织）在含有除草剂的培养基上产生了 52 株生根植物，其中 22 株为 PCR 阳性。DNA凝胶印迹分析揭示了不同独立转化体中GUS基因的拷贝数为1-5。拷贝数和 GUS 活性之间没有相关性。虽然常规培养在含除草剂的培养基上只产生白化植物，但改善目标组织的培养条件导致 100% 绿色转基因植物的恢复。所有绿色抗除草剂再生株形态正常且能育。