滇水金凤黄烷酮3-羟化酶基因(IuF3H)的克隆及表达分析

为探究黄烷酮3-羟化酶(flavanone 3-hydroxylase, F3H)基因对滇水金凤(Impatiens uliginosa)花色的调控机制。本研究以红色滇水金凤花器官为试验材料,通过RT-PCR及RACE技术分离克隆F3H基因,发现其c DNA序列全长1 104 bp,编码367 aa,故命名为Iu F3H;同时通过氨基酸比对发现其蛋白与其他植物F3H蛋白同源性较高,约为78.91%,说明F3H基因具有较高的保守性;通过进一步克隆其g DNA序列,发现其全长为1 221 bp,其中包含3个外显子和2个内含子。q RT-PCR结果显示,Iu F3H基因在4种不同花色滇水金凤及其不同发育阶段均有不同程度的表达,其中以深红色的始花期表达量最高,粉红色的盛花期表达量最低。本研究结果表明,Iu F3H基因参与了相关色素生物合成,同时可能在滇水金凤花色素合成中具有关键作用,本研究为探讨Iu F3H基因在4种不同颜色滇水金凤中的分子机制提供数据参考。

Cloning and Expression Analysis of IuF3H Gens in Impatiens uliginosa

In order to explore the regulation mechanism of flavanone 3-hydroxylase(F3 H)gene on flower color of Impatiens uliginosa.In this study,the F3 H gene was isolated and cloned by RT-PCR and RACE technology,which c DNA sequence was 1104 bp,encoding 367 aa,and named Iu F3 H.Meanwhile,through amino acid comparison,it was found that its protein had a higher homology with F3 H protein of other plants,about 78.91%,indicating that F3 H gene had a higher conservation.By further cloning its g DNA sequence,it was found that its full length was 1221 bp and contained 3 exons and 2 introns.The results of q RT-PCR showed that Iu F3 H gene implied that there existed some certain differences in its expression levels,not only between different flower colors of I.uliginosa,but also between different flower development stages,among which the expression level was highest in the beginning flowering stage of deep red and lowest in the blooming stage of pink.In this study,the above-mentioned results showed Iu F3 H gene not only took part in the biosynthesis of related pigments,but also was considered as one of the key gene responsible for flower color of I.uliginosa.To explore the molecular mechanism of Iu F3 H gene in four different colors of I.uliginosa.