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以高油大豆中豆32开花后30 d的种子为材料,根据已报道的拟南芥脂肪酸合成相关转录因子LEC1序列设计简并引物,采用同源序列法从大豆种子中分离了大小为850 bp的cDNA片段,测序结果表明,该片段与拟南芥中已克隆的脂肪酸合成基因高度同源,包含完整的读码框,采用酶切连接和gateway技术构建了该基因的超量表达和RNAi植物表达载体。为借助农杆菌介导法将LEC1基因转化到大豆再生植株中,对分离的LEC1基因进行功能验证,培育高油大豆新品种奠定了基础。
Based on the reported LEC1 transcription factor related to fatty acid synthesis in Arabidopsis thaliana, the degenerated primers were designed and the size of 850 bp was isolated from soybean seeds by homologous sequence method. The results of sequencing showed that the fragment was highly homologous to the cloned fatty acid synthase gene in Arabidopsis thaliana, and contained the complete reading frame. The overexpression of this gene and the RNAi plant Expression vector. In order to transform the LEC1 gene into soybean regenerated plants by Agrobacterium-mediated method, the function of LEC1 gene was verified and the new breeds of high-oil soybean were established.