液质联用法测定干旱胁迫下红砂(Reaumuria soongorica)叶片F3H、DFR酶活性  

Determination of Reaumuria soongorica leaf F3H and DFR activities under drought stress by using RP-HPLC-MS

作　　者：刘美玲[1,2] 刘玉冰[1] 曹波[1,2] 

机构地区：[1]中国科学院寒区旱区环境与工程研究所,沙坡头沙漠试验研究站,植物逆境生理生态与生物技术实验室,兰州730000 [2]中国科学院研究生院,北京100049

出　　处：《生态学杂志》2012年第8期2158-2162,共5页Chinese Journal of Ecology

基　　金：国家自然科学基金项目(30800122;31070358)资助

摘　　要：黄烷酮3-羟化酶(flavanone 3-hydroxylase,F3H)和二氢黄酮醇4-还原酶(di-hydroflavonol 4-reductase,DFR)是类黄酮合成途径中的2个关键酶,研究其活性的变化对探讨类黄酮合成途径的调控有重要的意义。以荒漠植物红砂(Reaumuria soongorica)为材料,建立了类黄酮合成途径关键酶F3H、DFR活性的液质联用检测法,研究干旱胁迫对红砂叶片F3H、DFR酶活性的影响。结果表明,F3H酶作用的底物柚皮素和DFR酶作用的底物二氢槲皮素的检测含量线性范围分别为0.04～1.20μg(r=0.998)和0.08～2.40μg(r=0.998)。因此,本方法可用于测定F3H、DFR酶活性。根据以上建立的方法,对干旱胁迫处理下,不同处理时间的红砂植株叶片进行F3H、DFR酶活性的测定,结果表明,干旱胁迫下,F3H酶活性随着干旱胁迫处理时间的延长,先升高后降低至处理前水平。DFR酶活性随干旱胁迫时间的延长逐渐升高。表明干旱胁迫对红砂叶片F3H、DFR酶活性有诱导作用。Flavanone 3-hydroxylase （F3H） and dihydroflavonol 4-reductase （DFR） are the key enzymes in flavonoids biosynthesis pathway, and to study their activities is a crucial step to explore the regulation of this pathway. In this paper, a RP-HPLC-MS determination method of F3H and DFR activities was established to investigate the effects of drought stress on the activities of F3H and DFR in Reaumuria soongorica leaves. The calibration range of F3H substrate naringenin and of DFR substrate dihydroquertin was 0.04-1.20 μg （r=0.998） and 0.08-2.40 μg （r=0.998）, respectively, suggesting that this method was convenient and accurate to determine the activities of F3H and DFR. By using this method, the F3H and DFR activities in R. soongorica leaves were measured, and the results showed that under drought stress, the F3H activity increased first and then dropped to the same level as the control, while the DFR activity increased all along with increasing drought time, indicating that drought stress had induction effect on the F3H and DFR activities in R. soongorica leaves.

关 键 词：干旱胁迫 红砂 黄烷酮3-羟化酶 二氢黄酮醇4-还原酶 反相高效液相色谱-质谱法 

分 类 号：Q946[生物学—植物学]