红花黄酮醇合成酶基因片段的克隆及表达分析

Cloning and expression analysis of flavonols synthetase gene from Carthamus tinctorius

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 expre ssion 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 co nnected to PEASY-T1 carrier,and positive cloning was detected by PCR and then sequenced.Results Sequencing results showed th at 224 bp sequence was acquired,to which the Blast comparison was carried out on NCBI.The gene had the higher homology co mpared with the FLS from other species.Conclusion The fragment of FLS gene is cloned from safflower,and PCR primers of s afflower are designed based on FLS gene for real-time PCR in different blossom of periods different varieties in safflower.The res ults show that the expression of safflower FLS genes in the full bloom of auspicious red oil sisters line is the highest.