Laboratory of Phytoimmunology, Siberian Institute of Plant Physiology and Biochemistry, Siberian Branch of the Russian Academy of Sciences, Irkutsk, Russia.
Plant Cell Rep. 2011 Jan;30(1):125-32. doi: 10.1007/s00299-010-0950-5. Epub 2010 Nov 13.
Presently, there is no doubt about the functioning of the adenylate cyclase signaling system in plants, but the role of this system in various physiological-biochemical processes has been investigated insufficiently. Cyclic adenosine monophosphate (cAMP), the key component produced by adenylate cyclase, whose concentrations in plant cells vary rather widely, is the indicator of functional activity for this signaling way. In the latter case, in the process of determination of concentrations of this messenger, one encounters difficulties related to insufficient sensitivity of the methods most frequently applied. In this connection, the proposed mechanism is a modification of the method of the enzyme immunoassay (EIA), which is based on immediate measurement of cAMP concentrations in the sample with the use of antibodies. This modification allows us to determine the concentrations of cAMP with the precision of 5 pM, which exceeds the sensitivity of other methods by approximately 10 times. The specificity of the assay has been confirmed by other two independent tests--the capillary electrophoresis and the nuclear magnetic resonance (NMR). It has also been compared to the data obtained with the use of the commercial kit from Sigma-Aldrich. The modification has been tested on such plant objects as in vitro potato plants, and suspension cells of potato and Arabidopsis.
目前，植物中的腺苷酸环化酶信号系统的功能无疑已得到证实，但该系统在各种生理生化过程中的作用还没有得到充分的研究。环腺苷酸（cAMP）是由腺苷酸环化酶产生的关键成分，其在植物细胞中的浓度变化很大，是该信号通路功能活性的指示剂。在后一种情况下，在确定这种信使的浓度的过程中，人们遇到了与最常应用的方法的灵敏度不足有关的困难。有鉴于此，提出的机制是对酶免疫测定（EIA）方法的改进，该方法基于使用抗体立即测量样品中的 cAMP 浓度。这种改进使得我们能够以 5 pM 的精度来确定 cAMP 的浓度，其灵敏度比其他方法高约 10 倍。该测定的特异性已通过另外两项独立测试——毛细管电泳和核磁共振（NMR）得到证实。它还与使用 Sigma-Aldrich 的商业试剂盒获得的数据进行了比较。该改进已在体外马铃薯植株和马铃薯及拟南芥悬浮细胞等植物对象上进行了测试。
