• 关键词：铜绿微囊藻  细胞活性  氯化三苯基四氮唑(TTC)  脱氢酶

• 基金项目：国家自然科学基金项目(No.50678024);国家高技术研究发展计划项目(No.2007AA06Z301)

• 梁文艳
• 北京林业大学环境科学与工程学院, 省部共建森林培育与保护教育部重点实验室, 北京 100083

• 王珂
• 北京林业大学环境科学与工程学院, 省部共建森林培育与保护教育部重点实验室, 北京 100083

• 阮清鸳
• 北京林业大学环境科学与工程学院, 省部共建森林培育与保护教育部重点实验室, 北京 100083

• 王金丽
• 北京林业大学环境科学与工程学院, 省部共建森林培育与保护教育部重点实验室, 北京 100083

• 摘要：利用氯化三苯基四氮唑(TTC)-脱氢酶还原法定量地测定了铜绿微囊藻细胞的活性,并对影响活性测定的因素进行了分析与研究.结果显示,TTC浓度大于0.6%时会抑制酶活性,最适TTC浓度为0.2%;脱氢酶还原反应在Tris-HCl缓冲液中能正常进行,而在磷酸盐缓冲液中受到抑制,最适pH值为7.5～8.0;培养时间和培养温度对TTC脱氢酶还原反应的影响较大,最适培养时间为8～16h,最适培养温度为30～35℃;反应不需加入非离子去垢剂,吐温20和曲拉通X-100对脱氢酶反应会产生较强抑制作用.TTC还原所生成的三苯基甲臢(TPF)可采用50%乙醇提取.研究显示TTC-脱氢酶还原法可以很好地进行藻细胞活性定量化测定.

• Abstract：In this paper,the cell viability of Microcystis aeruginosa was quantitatively determined by the 2,3,5-triphenyl tetrazolium chloride(TTC)-dehydrogenase assay and some major factors influencing the determination were investigated.Results showed that the enzyme viability could be inhibited when the TTC concentration was higher than 0.6%,and the optimum TTC concentration was 0.2%.The dehydrogenase reduction reaction was inhibited in the phosphate buffer solution,while the Tris-HCl buffer solution with pH of 7.5～8.0 was most appropriate for this reaction.Incubation time and temperature affected the dehydrogenase reaction significantly,and their optimum values were 8～16 h and 30～35℃,respectively.On the other hand,the addition of non-ionic detergents in the incubation period,such as Tween 20 or Triton X-100,was not necessary due to their inhibition effects on the dehydrogenase reduction reaction.The formazan reduction product could be efficiently extracted by 50% ethanol efficiently.This study demonstrated that the TTC-dehydrogenase assay might be successfully applied to the determination of Microcystis aeruginosa viability under the optimumal conditions.

• Key words：Microcystis aeruginosa  algae viability  2,3,5-tripheyl tetrazolium Chloride(TTC)  dehydrogenase

• 
  摘要点击次数： 2463 全文下载次数： 2714