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用RT-PCR从水稻品种日本晴的cDNA中扩增出苯达松和磺酰脲类除草剂抗性基因Bel,得到基因ORF全长1542 bp。将Bel片段克隆到pGEM-T Easy载体。将此基因插入pCAMBIA3301的Ca MV35 S启动子后取代原载体中的GUS基因构成表达载体pCAMB IA3301-Bel,通过电击转化法导入根癌农杆菌(Agrobacterium tumefaciens)GV3101,并转化拟南芥(Columbia生态型)获得转基因植株。对转化植株进行的基因组PCR和RT-PCR分析,结果表明Bel基因已成功地整合到拟南芥的基因组中并可以表达。研究结果为利用水稻苯达松和磺酰脲类除草剂抗性基因Bel开展油菜、棉花等双子叶作物的抗除草剂育种提供了实验依据。
The bentazon and the sulfonylurea herbicide resistance gene Bel were amplified by RT-PCR from the rice variety Nipponbare, and the full-length ORF of the gene ORF was 1542 bp. The Bel fragment was cloned into the pGEM-T Easy vector. This gene was inserted into the CaMV35S promoter of pCAMBIA3301 to replace the GUS gene in the original vector to construct the expression vector pCAMB IA3301-Bel. The Agrobacterium tumefaciens GV3101 was transformed into the Arabidopsis thaliana Type) obtained transgenic plants. Genome and RT-PCR analyzes of the transformed plants showed that the Bel gene has been successfully integrated into the Arabidopsis genome and expressed. The results provided an experimental basis for the herbicide-resistant breeding of dicotyledonous crops such as rapeseed and cotton by using Bendazon and sulfonylurea herbicide resistance genes Bel.