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。
