基因枪法介导转ZmalⅠ基因小麦植株的获得和鉴定

Acquisition and identification of wheat with ZmalⅠ gene by biolistic particle and molecular identification

【目的】获得转玉米ZmalⅠ基因的安全转基因小麦植株,为研究玉米ZmalⅠ基因在增加小麦籽粒大小和粒质量方面的作用,以及利用基因工程提高小麦产量奠定基础。【方法】以陕农138为供试材料,绿色荧光蛋白基因(GFP)为安全标记基因,利用基因枪法,将由谷蛋白胚乳特异表达启动子启动的ZmalⅠ+GFP融合蛋白基因的过表达载体pGlu-exbessa::ZmalⅠ导入小麦中,并对转基因小麦进行PCR检测。【结果】采用基因枪法共轰击3 000个幼胚,获得再生植株59株,移栽至培养钵后成活52株,成活率为88.1%;利用载体上安全标记基因GFP的特异引物对成活的转化植株进行PCR检测,获得含有GFP基因的阳性安全植株16株,转化率为0.53%。【结论】将ZmalⅠ基因成功地整合到了小麦基因组中。

【Objective】 The study was done to obtain safe transgenic wheat with the gene of ZmalⅠ,pave the way for studying the function of the gene ZmalⅠ in enlarging seed volume and grain weight and use the genetic engineering to improve the wheat yield.【Method】 With the wheat variety Shaannong 138 as the test material and application of GFP as a reporter gene,the over expressed vector pGlu-exbessa：：ZmalⅠ,which contained the fusion protein gene ZmalⅠ and GFP that were promoted by the special glutelin albumen promoter,was introduced into wheat by biolistic particle,and the transgenic plants were tested by PCR.【Result】 Then 3 000 embryos of Shaannong 138 were bombarded by biolistic particle,and 59 regenerated plants were obtained.Then 52 plants survived after transplanted to train bowls,the living ratio being 88.1%;16 safe transgenic plants with GFP gene were identified by PCR assay in the T0 generation with the transformation frequency of 0.53%.【Conclusion】 The ZmalⅠ gene has successfully integrated into the wheat genome.