Ge Yaxin, Wang Zeng-Yu
Forage Improvement Division, The Samuel Roberts Noble Foundation, Ardmore, OK, USA.
Methods Mol Biol. 2006;344:47-54. doi: 10.1385/1-59745-131-2:47.
Bermudagrass is an important warm-season forage and turf species widely grown in the southern United States. This chapter describes a rapid and efficient protocol that allows for the generation of a large number of transgenic bermudagrass plants, bypassing the callus formation phase. Stolon nodes are infected and co-cultivated with Agrobacterium tumefaciens harboring pCAMBIA binary vectors. Hygromycin phosphotransferase gene (hph) is used as the selectable marker and hygromycin is used as the selection agent. Green shoots are directly produced from infected stolon nodes 4 to 5 wk after hygromycin selection. Without callus formation and with minimum tissue culture, this procedure allowed us to obtain well-rooted transgenic plantlets in only 7 wk and greenhouse-grown plants in only 9 wk.
狗牙根是一种重要的暖季型牧草和草坪草种,在美国南部广泛种植。本章描述了一种快速高效的方法,该方法无需愈伤组织形成阶段,就能产生大量转基因狗牙根植株。匍匐茎节与携带pCAMBIA双元载体的根癌农杆菌进行侵染和共培养。潮霉素磷酸转移酶基因(hph)用作选择标记,潮霉素用作选择剂。在潮霉素选择4至5周后,受感染的匍匐茎节直接产生绿色芽。该方法无需愈伤组织形成且组织培养最少,使我们仅在7周内就能获得根系良好的转基因幼苗,仅在9周内就能获得温室种植的植株。