A highly sensitive quantification method for the accumulation of alarmone ppGpp in Arabidopsis thaliana using UPLC-ESI-qMS/MS
A highly sensitive quantification method for the accumulation of alarmone ppGpp in Arabidopsis thaliana using UPLC-ESI-qMS/MS
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DOI：
10.1007/s10265-015-0711-1
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发表时间：
2015-05-01
影响因子：
2.8
通讯作者：
Masuda, Shinji
中科院分区：
文献类型：
--
作者：
Ihara, Yuta;Ohta, Hiroyuki;Masuda, Shinji
最近，已经在叶绿体中检测到细菌第二信使，鸟苷5 '-二磷酸3'-二磷酸（ppGpp）。然而，由于植物中的ppGpp浓度远低于细菌中的浓度，因此尚未对植物中的ppGpp进行详细分析。一个高度敏感的定量方法是需要进一步的特点ppGpp功能的叶绿体。在这里，我们报告了一种新的方法，允许高灵敏度和选择性的高通量定量ppGpp的超高效液相色谱法（UPLC）与串联四极杆质谱仪（qMS/MS）配备了电喷雾接口（ESI）。该方法仅需要近似100 mg植物组织用于ppGpp定量。我们使用这种方法来测量拟南芥在不同光照条件下的ppGpp水平。A.拟南芥在黑暗期积累ppGpp。该方法将有助于进一步表征高等植物的严紧反应。
Recently, a bacterial second messenger, guanosine 5'-diphosphate 3'-diphosphate (ppGpp), has been detected in chloroplasts. However, because ppGpp concentration in plants is much lower than that in bacteria, detailed analysis of ppGpp in plants has not been performed. A highly sensitive quantification method is required for further characterization of ppGpp function in chloroplasts. Here, we report a new method that allows for the highly sensitive and selective high-throughput quantification of ppGpp by ultra-performance liquid chromatography (UPLC) coupled with a tandem quadrupole mass spectrometer (qMS/MS) equipped with an electrospray interface (ESI). This method requires only similar to 100 mg of plant tissue for ppGpp quantification. We used this method to measure ppGpp levels in Arabidopsis thaliana under different light conditions. A. thaliana accumulated ppGpp during dark periods. This method will be helpful to further characterize the stringent response in higher plants.