山葡萄CBF基因表达载体构建及对拟南芥根生长的影响

The Construction of Expression Vector of Gene CBF from Vitis amurensis and the Effects of It on the Root Growth of Arabidopsis

为了研究山葡萄CBF基因调节植物对盐胁迫的应答机理,分别构建了山葡萄Va CBF1、Va CBF2和Va CBF3的植物过表达载体。经酶切及琼脂糖电泳检测证实3个基因均插入到p BASTA中,表明表达载体构建成功。然后,分别将3个植物过表达载体转入农杆菌EHA105中,并通过浸花法浸染拟南芥。利用除草剂筛选获得3个基因的拟南芥过表达株系。最后,对野生型拟南芥与转基因拟南芥进行盐胁迫处理,发现OE-CBF2转基因植株的主根伸长长度显著长于其它植株,3个转基因株系的侧根长度也明显长于野生型植株。上述结果表明山葡萄CBF基因可能在植物盐胁迫中对根部生长发育起到非常重要的调控作用。

In order to investigate the response mechanism of CBF gene in the regulation of plant to the salt stress, the plant overexpression vectors of Va CBF1, Va CBF2 and Va CBF3 from Vitis amurensis were constructed. The results by restriction enzyme digestion and agarose gel electrophoresis showed that 3 genes were correctly inserted into p BASTA, indicating that the expression vectors were successfully constructed. Then, 3 over-expressed vectors were transformed into Agrobacterimu tumefaciens EHA105, and Arabidopsis thaliana was leached using floral dip method. The transgenic plants of A. thaliana with over-expression of 3 genes were screened by herbicide Basta. Furthermore, wild and transgenic plants were treated with salt stress. The results revealed that the elongation length of primary root of OE-CBF2 transgenic plant was longer than others, and the lateral roots in 3 genes transgenic plants were longer than wild ones. These results indicate that the CBF genes of V. amurensis play the critical regulation role in the root development during salt stress.