甘蓝型油菜BnBRI1基因过量表达载体和RNA干扰载体的构建及遗传转化

Construction of the over-expression vector and the RNAi vector of BnBRI1 gene from Brassica napus L.and its transformation

构建了含甘蓝型油菜BnBRI1基因过量表达载体pFGC5941-BnBRI1和RNA干扰载体pFGC5941-BnBRI1-Ri,以根癌农杆菌介导的方法转化甘蓝型油菜"W679"下胚轴外植体,经PCR鉴定,获得18株转基因阳性植株,其中1株为BnBRI1基因过量表达转基因植株,17株为BnBRI1基因RNA干扰转基因植株.经RT-PCR鉴定显示,在过表达转基因植株中BnBRI1基因表达量比野生型甘蓝型油菜高,而在RNA干扰转基因植株中表达量比野生型低,RNA干扰BnBRI1基因的表达导致了植株的矮化,矮化植株将可用于油菜株型的遗传改良,以培育适于机械化生产的中矮秆油菜新品种.

In order to investigate the effects of BRI1 gene in Brassica napus（BnBRI1）,the over-expression vector pFGC5941-BnBRI1 and the BnBRI1 targeted RNA interference（RNAi）expression vector pFGC5941-BnBRI1-Ri were constructed and transferred into Brassicanapusby Agrobacterium tumefaciens.One over-expression transgenic plant and seventeen RNAi transgenic plants were obtained by PCR.RT-PCR analysis showed that,compared with the wild-type＂W679＂,the expression level of BnBRI1 in over-expression transgenic plant had increased,while the expression level of BnBRI1 in RNAi transgenic plants were down-regulated.RNA interference the expression of BnBRI1 leading to dwarf plants.Dwarf plants will be used for the genetic improvement of rapeseed,and it conducive to mechanized production.