To develop a transformation protocol of Rosa hybrida Samantha via Agrobacterium tumefaciens, the authors examined the effect of different factors on T-DNA transfer by measuring transient expression levels of an intron...To develop a transformation protocol of Rosa hybrida Samantha via Agrobacterium tumefaciens, the authors examined the effect of different factors on T-DNA transfer by measuring transient expression levels of an intron-containing -glucuronidase gene. The results indicate that explant, light condition, salt concentration and acetosyringone (AS) concentration in co-culture medium are the most important factors, and factors like co-culture temperature, co-culture period and bacteria density have a strong effect on the growth of bacteria and then T-DNA transfer. Optimized co-cultivation was performed by inoculation of embryogenic callus with bacteria at a density of OD600= 0.50.8 for 20 min and co-culture in darkness under 23 C on medium with 1/2 MS salts and 300 mol稬1 AS for 3 d.展开更多
基金Supported by the National Natural Science Foundation of China (Grant No.30170666)
文摘To develop a transformation protocol of Rosa hybrida Samantha via Agrobacterium tumefaciens, the authors examined the effect of different factors on T-DNA transfer by measuring transient expression levels of an intron-containing -glucuronidase gene. The results indicate that explant, light condition, salt concentration and acetosyringone (AS) concentration in co-culture medium are the most important factors, and factors like co-culture temperature, co-culture period and bacteria density have a strong effect on the growth of bacteria and then T-DNA transfer. Optimized co-cultivation was performed by inoculation of embryogenic callus with bacteria at a density of OD600= 0.50.8 for 20 min and co-culture in darkness under 23 C on medium with 1/2 MS salts and 300 mol稬1 AS for 3 d.
基金Foundation project: This paper was supported by Zhejiang Provincial Science and Technology Plan of China (Grant No. 2003C30053) and Zhejiang Provincial Natural Science Foundation of China (Grant No.Y504076).
文摘高教鞭(Festuca arundinacea Schreb ) 是在高尔夫球场在航路上使用的一棵凉爽季节的草地草。这研究的目标是开发更多有效的,可靠,;在用 Agrobacterium (EHA105 ) 转变草重复有能力的途径,在β - glucuronidase 基因(uidA ) 被用作一个记者的地方;hygromycin phosphotransferase 基因(hyg ) 作为一个可选的标记。transgene 的一个有效表达式在转变调用 i 从成熟种子导出的 2-month-old 被观察(cv。宾果游戏) 在与 9 mg · L 补充的 MS 媒介上有教养[1 ] 2, 4-D。有增加 hygromycin 集中的一种二拍子的圆舞固体媒介选择(从 30 ~ 50 mg · L [1 ]) 被用来获得抵抗电话 i。转基因的植物从许多独立转变电话 i 被生产了。功能的β - glucuronidase 基因(uidA ) 的存在在 hygromycin 抵抗的电话 i 被检测。转基因的植物被改革;PCR;南部的污点在高教鞭染色体证实了 transgene 集成。