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生态学杂志 ›› 2012, Vol. 31 ›› Issue (08): 2158-2162.

• 方法与技术 • 上一篇    

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

刘美玲1,2,刘玉冰1**,曹波1,2   

  1. (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)
  • 出版日期:2012-08-10 发布日期:2012-08-10

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

LIU Mei-ling1,2, LIU Yu-bing1**, CAO Bo1,2   

  1. (1Laboratory of Plant Stress Ecophysiology and Biotechnology, Shapotou Desert Research & Experiment Station, Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China; 2 Graduate University of Chinese Academy of Sciences, Beijing 100049, China)
  • Online:2012-08-10 Published:2012-08-10

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摘要： 黄烷酮3-羟化酶(flavanone 3-hydroxylase, F3H)和二氢黄酮醇4-还原酶(dihydroflavonol 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0.04～1.20 μg (r=0.998)和0.08～2.40 μg (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。

关键词: 马铃薯, 大豆, 套作, 光合特性, 产量, 补偿效应

Abstract: 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.

Key words: potato, soybean, intercropping, photosynthetic characteristics, yield, compensation effect.

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