农杆菌介导法向小麦茎尖导入DREB1A基因的研究初报

Preliminary Study on Agrobacterium-mediated Transformation of DREB1A Gene into Apical Point of Wheat(Triticum aestivum L.)

为了研究以小麦茎尖为受体进行农杆菌转化的可行性,选用小麦品种H6756,以茎尖为受体进行了农杆菌转化。构建了含有拟南芥逆境诱导转录因子DREB1A及除草剂bar基因的表达载体pC3300IS-DREB1A,酶切鉴定此载体,结果证明其连接完全正确,具有转录和表达的功能。农杆菌介导法向小麦茎尖导入DREB1A基因,共获得247株转化苗。转化苗经除草剂筛选,获得66株抗性苗,对抗性苗进一步进行PCR检测,有30株抗性苗扩增出特异性片段,转化率达到12.1%,初步说明本试验中以小麦茎尖为受体的转化系统用于基因转化是可行、高效的。

The stress-induced DREBIA gene was transferred into the shoot apical meristem of wheat （Triticurn aestivum L. ） cultivar H6756 by Agrobacterium-mediated transformation to study the feasi- bility of this receptor in Agrobacterium-mediated transformation. The expression vector pC3300IS-DREB1A, containing transcription factor DREB1A gene and herbicide bar gene, was constructed and then cut by enzyme to identify the correct ligation, the function of transcription and expression. The DREB1A gene in the plasmid pC3300IS-DREB1A was transferred into the shoot apical meristem of wheat cuhivar H6756 by Agrobacteriurn-mediated transformation, and 247 seedlings were obtained. 66 plants survived after selecting with 100 mg/L Basta. Detecting the target gene by PCR amplification, 30 plants were screened with the integrated DREBIA gene, the frequency of transformation reached 12.1%. This study showed that the shoot apical meristem of wheat could be feasible and high efficiency as gene transformation receptor system .