Zou Qing-Jun, Wang Tao, Guo Qiao-Sheng, Xiao You-Mei, Wu Li-Wei
Institute of Chinese Medicinal Materials, Nanjing Agricultural University, Nanjing 210095, China.
Zhongguo Zhong Yao Za Zhi. 2018 Jan;43(1):52-57. doi: 10.19540/j.cnki.cjcmm.20171030.008.
To investigate the effects of the expression of flavonoid 3' hydroxylase gene ( and active ingredients in under flooding stress, we cloned F3'H from Hangju (temporarily named ) and conducted bioinformatics analysis. During the flower bud differentiation stage, we flooded the and then used the Real-time PCR to detect the relative expression of ; Finally, active ingredients of the inflorescence were measured by HPLC.The sequencing results showed that 1 562 bp sequence was acquired with the largest open reading frame of 1 527 bp, which encoded 508 amino acids. The phylogenetic tree found that was highly homologous to other species of Compositae. Real-time PCR results showed that had a significant response to flooding stress and had the highest expression level after flooding for 24 h, which was about 9 times as that of the control group. The results of HPLC showed that luteolin and luteoloside, the downstream products catalyzed by the F3'H, were significantly higher than those in the control group. It was also found that the contents of chlorogenic acid and 3,5- acid were also significantly higher than those of the control group. Therefore, regulates the synthesis of downstream products by regulating the expression of in the flavonoid synthesis pathway under flooding stress, thereby responding to flooding stress. The flooding stress during flower bud differentiation can significantly enhance the accumulation of active ingredients.
为研究黄酮类3'羟化酶基因（F3'H）的表达及杭菊活性成分在淹水胁迫下的影响，我们从杭菊中克隆了F3'H（暂命名）并进行了生物信息学分析。在花芽分化期对杭菊进行淹水处理，然后用实时荧光定量PCR检测F3'H的相对表达量；最后，通过高效液相色谱法测定花序的活性成分。测序结果显示获得了一条1562 bp的序列，最大开放阅读框为1527 bp，编码508个氨基酸。系统发育树分析发现，该序列与菊科其他物种高度同源。实时荧光定量PCR结果表明，F3'H对淹水胁迫有显著响应，淹水24 h后表达量最高，约为对照组的9倍。高效液相色谱法结果显示，F3'H催化的下游产物木犀草素和木犀草苷显著高于对照组。还发现绿原酸和3,5-二咖啡酰奎宁酸的含量也显著高于对照组。因此，F3'H在淹水胁迫下通过调控黄酮合成途径中F3'H的表达来调节下游产物的合成，从而响应淹水胁迫。花芽分化期的淹水胁迫可显著提高活性成分的积累。
