喷雾：一种简便的拟南芥转化方法

Spraying:A Simplified Method for Transformation of Arabidopsis

目的：建立一种改良的且操作更为简便可行的拟南芥转化方法。方法：用含有OD600等于0．8的农杆菌,5%蔗糖和0. 2 ml/L表面活性剂Silwet L-77的MS液体培养基喷洒正在发育的花器官即可实现基因转化。植物生长到高约4cm时进行转化,此时植物具有大量的花和极少数的果荚,可以得到比较高的转化效率。喷洒溶液后将植物用保鲜膜等材料包裹起来以保持湿度,可有利于转化。结果：得到了365棵转化植株,转化效率和花器官浸泡法的基本一致。结论：这种方法可适用于多种生态型的拟南芥植物转化,并且方便大规模的拟南芥植物的转化,便于人们快速获得带有T-DNA标记的植株或基因遗传互补工作的顺利开展。

Objective：To establish a modified and simplified transformation method for Arabidopsis thaliana. Methods： In this method, an MS liquid culture medium containing Agrobacterium whose OD600 value was 0.8, 0.5%sucrose and 0.2ml/L suifactant Silwet L- 77 was used to spray developing flowers. Plants inoculated when numerous immature floral buds and few siliques were present produced transformed progeny at a relatively higher rate. Covering plants for two to three days to retain humidity after inoculation was conducive to transformation. Results： With this method 365 transformants were obtained. The transformation rate was almost the same as that of floral dip method. Conclusion： Multiple ecotypes were transformable by this method. The modified method should facihtate high - throughput transformation of Arabidopsis for efforts such as T - DNA gene taggine or attempts at targeted gene replacement.