| 采用Extracellular Flux Analyzer方法测定植物线粒体能量代谢 |
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| 引用本文： | 纪翔宇,刘翔宇,张志翔.采用Extracellular Flux Analyzer方法测定植物线粒体能量代谢[J].中国农学通报,2015,31(21):268-272. |
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| 作者姓名： | 纪翔宇 刘翔宇 张志翔 |
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| 作者单位： | 北京林业大学自然保护区学院,北京林业大学自然保护区学院,北京林业大学自然保护区学院 |
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| 基金项目： | 中央高校基本科研业务费专项资金 “利用非损伤活体成像技术研究线粒体能量代谢对玉兰开花生热的调控机制”(YX2014-14)；国家自然基金 “木兰科植物开花生热效应表型及调控机制研究”(31100450)。 |
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| 摘 要： | 线粒体是植物进行呼吸和能量代谢的主要场所,但目前关于线粒体能量代谢的测定方法较为繁琐,亟需一种快速有效测定线粒体能量代谢的手段。本研究采用Extracellular Flux Analyzer(Seahorse Bioscience,America)方法对玉兰(Magnolia denudata)花部器官线粒体代谢情况进行测定,并对测试技术进行改进。通过直接测定氧气消耗速率,进而检测组织细胞内的线粒体呼吸速率和能量代谢。采用该方法对同一组织样品进行多次测量,测量结果稳定,表明该方法是一种可靠的测定线粒体呼吸和能量代谢的方法。并且,通过加入不同的呼吸抑制剂来鉴定细胞的呼吸途径,可快速获取有效数据。此外,还可直接对组织内线粒体的呼吸代谢进行测定,简便性、实时性均优于其他线粒体代谢测定方法,是一种快速有效,且可以进行实时动态观测的试验方法。
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| 关 键 词： | 线粒体 玉兰 呼吸 花器官 能量代谢 实时观测 |
| 收稿时间： | 4/8/2015 12:00:00 AM |
| 修稿时间： | 7/1/2015 12:00:00 AM |
Measuring Mitochondrial Energy Metabolism Using Extracellular Flux Analyzer |
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| Abstract: | Mitochondria are the major sites of respiration and energy metabolism in plants. However, previous methods for measuring respiration metabolism of mitochondria are complex and time-consuming. Thus, accurant and efficient methods are in urgent need for measurements of mitochondrial respiration. In the current study, we developed a simple and efficient method to test respiration metabolism of mitochondria in living cells and tissues by using Extracellular Flux Analyzer. Oxygen consumption rates were identified quickly in floral cells of Magnolia denudata, which is a perennial woody plant. In the measurements, stable repeated data of samples were obtained, indicating the reliability of this method in measuring mitochondrial respiration and energy metabolism. Furthermore, were identified in the cells via using various respiratory inhibitors. In addition, the method allows dynamic and real-time monitoring of mitochondrial respiration in plant tissues and individual plant cells, which outperforms existing methods. Above all, this method shows a broad application potential in measuring mitochondrial energy metabolism of plant cells in the future. |
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| Keywords: | mitochondria Magonolia denudata respiration floral organ energy metabolism real-time observation |
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