人参细胞TTC-脱氢酶活力测定条件的优化  

The Optimization of TTC- dehydrogenase Method for Determination of Panax ginseng Cell Activity

作　　者：王晗[1] 雷秀娟[1] 宋娟[1] 尹红新[1] 姚丽娜[1] 梁韶 侯志芳[1] 王英平[1] 

机构地区：[1]中国农业科学院特产研究所,长春130112

出　　处：《特产研究》2016年第1期13-20,共8页Special Wild Economic Animal and Plant Research

基　　金：吉林省青年科研基金项目(20140520165JH)

摘　　要：以人参愈伤组织为试材,采用单因子试验和L16(45)正交试验,对2,3,5-氯化三苯基四氮唑(2,3,5-Triphenyltetrazolium Chloride,TTC)-脱氢酶还原法测定人参细胞活力的条件进行优化。选取TTC浓度、p H值、反应温度、反应时间作为试验中的4个因子,以吸光光度值和酶活力值为考察指标,探讨TTC-脱氢酶还原法测定人参细胞活力的最佳条件。结果表明,反应温度对人参细胞活力影响达到极显著水平(P<0.01),反应时间对人参细胞活力影响达到显著水平(P<0.1),得出人参细胞活力测定的最佳条件为TTC溶液浓度0.4%、p H 7.5、反应温度25℃、反应时间18h。This article focused on the optimization of test conditions of Panax ginseng cells activity through setting L16（ 4^5） orthogonal test and single factor test by using 2,3,5- Triphenyltetrazolim Chloride dehydrogenase assay. Taking TTC concentration,p H,reaction temperature and time as four factors of test,this paper explore the best test condition of Panax ginseng cells activity by using TTC dehydrogenase assay based on OD and U value as reviewindicators. The results showed reaction temperature and time had significantly influence on ginseng cells activity（ P〈 0. 1）,especially the reaction temperature（ P 〈0. 01）,We conclude that the optimum condition in determination of gensing cells vitality as： 0. 4% TTC,p H 7. 5,reaction temperature 25℃,reaction time 18 h.

关 键 词：人参愈伤组织 2 3 5-氯化三苯四氮唑-脱氢酶还原法 单因子试验设计 正交设计 细胞活力 

分 类 号：S567.51[农业科学—中草药栽培]