论文部分内容阅读
目的:灯盏花的有效成分灯盏乙素是一种具有显著药理活性的黄酮类化合物,目前仍缺乏对其合成途径的相关认识。查尔酮合成酶(CHS)是黄酮类生物合成的一个关键酶,该研究旨在通过研究CHS表达水平与灯盏花中各组织灯盏乙素含量的变化规律,阐明该基因的表达模式与灯盏乙素含量之间的关系。方法:通过RT-PCR及RACE方法从灯盏花中克隆CHS基因全长,利用荧光定量PCR方法检测该基因在灯盏花各组织中的表达量,采用HPLC分析各组织中灯盏乙素的含量。结果:序列全长1 270 bp,编码405氨基酸,该基因DNA序列与菊科植物的同源基因相似性在80%左右,荧光定量显示CHS在叶中表达量最高,远高于根、茎和花;HPLC发现灯盏乙素在叶中含量最高,其次是花和茎,而在根中未检测到。结论:相关性分析表明CHS相对表达量与灯盏花不同部位灯盏乙素含量间呈正相关关系(r=0.761,P<0.05),说明灯盏乙素的生成与CHS基因的表达密切相关。
Objective: Erigeron is the active ingredient of Erigeron is a pharmacologically active flavonoids, there is still a lack of understanding of its synthetic pathway. Chalcone synthase (CHS) is a key enzyme in the biosynthesis of flavonoids. The aim of this study was to investigate the expression pattern of CHS and the variation of scutellarin content in each tissue of Breviscapus, The relationship between the content of the elements. Methods: The full length of CHS gene was cloned from Erigeron breviscapus by RT-PCR and RACE. Fluorescent quantitative PCR was used to detect the expression of CHS gene in Erigeron breviscapus tissues. The content of scutellarin in each tissue was analyzed by HPLC. Results: The full length cDNA was 1 270 bp encoding a protein of 405 amino acids. The homology of this gene with that of Compositae was about 80%. Fluorescence quantitative analysis showed that the expression of CHS in leaves was the highest, much higher than that of roots and stems The results showed that the content of scutellarin was highest in leaves, followed by flowers and stems, but not detected in roots. Conclusion: The correlation analysis showed that there was a positive correlation between CHS expression and scutellarin content (r = 0.761, P <0.05), indicating that scutellarin production is closely related to the expression of CHS gene.