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棉花叶片硝酸还原酶活性的测定方法
引用本文:刘洁,王省芬,张桂寅,吴立强,李志坤,马峙英.棉花叶片硝酸还原酶活性的测定方法[J].河北农业大学学报,2010,33(4).
作者姓名:刘洁  王省芬  张桂寅  吴立强  李志坤  马峙英
作者单位:华北作物种质资源研究与利用省部共建实验室,河北省作物种质资源重点实验室,河北农业大学,河北,保定,071001
基金项目:河北省科技支撑计划重大项目,河北省自然科学基金重大项目 
摘    要:本研究应用磺胺比色法测定棉花叶片硝酸还原酶活性(NRA),探讨了体内法测定棉花叶片NRA的最适条件。通过分析比较硝酸盐诱导、2,4-二硝基苯酚、三氯乙酸及煮沸等因素对NRA测定结果的影响,建立了体内法测定棉花叶片NRA的最佳条件,即用50 mmol/L的KNO3溶液诱导棉花叶片48 h、抽气前后分别加入2,4-二硝基苯酚和1,6-二磷酸果糖、暗保温35 min后煮沸10 min可以使NRA的活性达到最高。棉花叶片NRA测定方法的建立为进一步研究棉花的氮素代谢奠定了基础。

关 键 词:棉花  硝酸还原酶  酶活性测定

Determination method of nitrate reductase activity in cotton leaves
LIU Jie,WANG Xing-fen,ZHANG Gui-yin,WU Li-qiang,LI Zhi-kun,MA Zhi-ying.Determination method of nitrate reductase activity in cotton leaves[J].Journal of Agricultural University of Hebei,2010,33(4).
Authors:LIU Jie  WANG Xing-fen  ZHANG Gui-yin  WU Li-qiang  LI Zhi-kun  MA Zhi-ying
Abstract:The method of sulfonamides colorimetry was used to determine the nitrate reductase activity(NRA) in cotton leaves.The optimum factors for determination of NRA in cotton leaves by in vivo method were discussed.Through comparative analysis of several main factors of in vivo method such as nitrate induction,2,4-dinitrophenol,3-chloroactic acid and boiling etc.,the optimum condition was identified for determination method of NRA in cotton leaves.The induction of 50 mmol/L KNO3 solution for 48 h,appendment of 2,4-dinitrophenol before elicitting air and of 1,6-diphosphate fructose after elicitting air,boiling 10 min after incubated for 35 min at 30 ℃ in the dark could obtain the highest NR activity.The results showed that in vivo method was reliable and easy to determine the NR activity of cotton leaves,and this method lays the foundation for further study on nitrogen metabolism of cotton.
Keywords:cotton  nitrate reductase  determination of enzyme activity
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