目的 克隆红花花瓣中的黄酮醇合成酶(flavonol synthase,FLS)基因并研究其在不同开花时期的表达量.方法 根据红花花瓣转录组测序结果挑选FLS基因的设计引物,以红花花瓣总 RNA为模板,采用RT-PCR的方法扩增FLS基因片段并连接到PEASY-T1载体上,阳性克隆经PCR检测后进行测序.结果 获得了224 bp的序列,将获得的序列在NCBI上进行Blast比对,该基因与其他物种的FLS基因具有较高的同源性.结论 克隆了红花FLS基因中间片段,根据FLS基因片段设计引物,对红花不同品种不同开花时期进行荧光定量PCR分析,红花FLS基因在吉红油姊妹系的盛花期表达量最高.

Objective To clone the flavonol synthase (FLS) gene from Carthamus tinctorius (safflower) pigments and study its expression in different blossom periods. Methods Primers were designed according to FLS which was selected from transcriptome sequencing results of safflower petal. Taking total RNA of safflower petal as template, FLS genes were amplified by RT-PCR and connected to PEASY-T1 carrier, and positive cloning was detected by PCR and then sequenced. Results Sequencing results showed that 224 bp sequence was acquired, to which the Blast comparison was carried out on NCBI. The gene had the higher homology compared with the FLS from other species. Conclusion The fragment of FLS gene is cloned from safflower, and PCR primers of safflower are designed based on FLS gene for real-time PCR in different blossom of periods different varieties in safflower. The results show that the expression of safflower FLS genes in the full bloom of auspicious red oil sisters line is the highest.

国家高技术研究发展计划(863)(2011AA100606);国家自然科学基金资助项目(31101172,31201237);吉林省科技厅中青年科技领军人才及优秀创新团队项目(20111815);教育部博士点基金(20122223120002)