Nontaswatsri Chalermsri, Fukai Seiichi
Department of Horticulture, Faculty of Agricultural Production, Maejo University, Chiangmai, Thailand.
Methods Mol Biol. 2010;589:87-96. doi: 10.1007/978-1-60327-114-1_9.
This chapter describes a rapid and efficient protocol for explant preparation and genetic transformation of carnation. Node explants from greenhouse-grown plants and leaf explants from in vitro plants are infected with Agrobacterium tumefaciens AGL0 harboring pKT3 plasmid, consisting of GUS and NPTII genes. Explant preparation is an important factor to obtain the transformed plants. The GUS-staining area was located only on the cut end of explants and only explants with a cut end close to the connecting area between node and leaf, produced transformed shoots. The cocultivation medium is also an important factor for the successful genetic transformation of carnation node and leaf explants. High genetic transformation efficiency of node and leaf explants cocultured with Agrobacterium tumefaciens was achieved when the explants were cocultivated on a filter paper soaked with water or water and acetosyringone mixture (AS).
本章介绍了一种用于康乃馨外植体制备和遗传转化的快速高效方案。来自温室种植植物的节段外植体和来自离体培养植物的叶片外植体,用携带pKT3质粒(由GUS和NPTII基因组成)的根癌农杆菌AGL0进行感染。外植体制备是获得转化植株的一个重要因素。GUS染色区域仅位于外植体的切口端,只有切口端靠近节与叶连接区域的外植体才能产生转化芽。共培养基也是康乃馨节段和叶片外植体成功进行遗传转化的一个重要因素。当外植体在用水或水与乙酰丁香酮混合物(AS)浸湿的滤纸上共培养时,节段和叶片外植体与根癌农杆菌共培养可实现较高的遗传转化效率。